I. Delphinium alkaloids. II. Halogenation of l-menthone

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NORTHWESTERN UNIVERSITY

I. II*

DELPHINIUM ALKALOIDS

HALOGENATION OP 1-MENTHONE

A DISSERTATION SUBMITTED TO THE GRADUATE SCHOOL IN PARTIAL FULFILMENT OF THE REQUIREMENTS for the degree DOCTOR OF PHILOSOPHY

DEPARTMENT OF CHEMISTRY

BY ROBERT CLARENCE K0DER

EVANSTON, ILLINOIS JUNE,

1942

P ro Q u e s t N u m b e r: 10101624

All rights reserved INFORMATION TO ALL USERS The q uality o f this rep ro d u c tio n is d e p e n d e n t u p o n th e q uality o f th e c o p y sub m itted . In th e unlikely e v e n t th a t th e author did n o t send a c o m p le te m anuscript a n d th e re are missing p a g e s , th e s e will b e n o te d . Also, if m a te ria l h a d to b e re m o v e d , a n o te will in d ic a te th e d e le tio n .

uest ProQ uest 10101624 Published by ProQ uest LLC (2016). C opyright o f th e Dissertation is held by th e Author. All rights reserved. This work is p ro te c te d a g a in s t unauthorized co p yin g u n d er Title 17, United States C o d e M icroform Edition © ProQ uest LLC. ProQuest LLC. 789 East Eisenhower Parkw ay P.O. Box 1346 Ann Arbor, Ml 48106 - 1346

ACKNOWLEDGMENT The author wishes to express grat­ itude to Dr, C, M, Suter, under whose helpful direction this research was carried out. Acknowledgment is also made of financial assistance provided In the form of Research Assistantships by the Graduate School of Northwestern Uni~ versity and a Research Fellowship by the Allied Chemical and Dye Corporation

TABLE OF CONTENTS Part One DELPHINIUM ALKALOIDS I. INTRODUCTION..................................... . i II. TAXONOMY OP THE DELPHINIUM GENUS 2 III. HISTORICAL BACKGROUND. . .......... 24 A. Chemical and Physical Properties of lphinium. Alkaloids . 24 B* Pharmacology of Delphinium Species and 'their Alkaloids.......................... .i 54 1. General physiological action. . ...... 54 2. Delphinosls. ...... 60 5. Therapeutic value................,......., 65 4. Insecticidal action*...................... 70 71 IV. DISCUSSION OP RESULTS........... A. The Alkaloids of D, Ajacis L................. 71 B. The Alkaloids of Annual Larksput*1i .••*••.••••• 75 1. Extraction of the a l k a l o i d s , ..........; 75 2. Properties of a ^ a c i n i n e . 75 5. Properties of delpholine.;................. 81 4. Pharmacology of ajaclnine, delphinoline, and delpholine ............. 85 C. The Alkaloids of Perennial Larkspur........... 86 V. EXPERIMENTAL DETAILS.777777777......... 88 A* D. Ajacis L. ...... 88 B# Annual Larkspur. ..... •.105 G . Perennial Larkspur. 142 VI. SUMMARY ............................... ......146 BIBLIOGRAPHY. ...148

Part Two HALOGEN AT ION OF 1-MEN THONE I. II.

HISTORICAL INTRODUCTION...........................154 DISCUSSION OF RESULTS..................... ........159 A, Bromination of 1-Menthone ...... 159 B. Reactions of 4-Bromomenthone . ....... .165 C * Chlorination of 1-Menthone. ........ 165 D. Reactions of 4-Chloromenthone and the Dicbloromenthones ..... 167 III. EXPERIMENTAL DETAILS..............................172 IV. SUMMARY. ......................................... 189 BIBLIOGRAPHY.......................................190 VITA.

191

PART ONE

DELPHINIUM ALKALOIDS

I*

INTRODUCTION

Although many Delphinium species have been recognized during the past several decades as a major cause of cattle poisoning in the western United States and medicinal prep­ arations from certain Delphinium species have been in com­ mon use for centuries, the chemistry of the alkaloids of this genus Is very little known.

It has been recognized

that a close relationship exists between the delphinium alkaloids and the highly toxic aconitum alkaloids, but the chemistry of the latter is equally unknown,

This investi­

gation has for Its purpose the furthering of the knowledge concerning the delphinium alkaloids* As starting material was available seed of commercial origin.

In order to indicate the botanical identity of the

seed there will first be a discussion of the taxonomlcal relationships of the more important Delphinium species and varieties*

Following this will be a history of the previous

work on the delphinium alkaloids, special note being made of the physical properties and elementary analyses, for it will be necessary to consider these details in correlating the experimental results with the earlier data*

2.

II,

TAXONOMY OP THE DELPHINIUM GENUS

The Delphinium genus Is a member of the family Rairnnoulaceae (order Ranales or Ranunculales), falling In a group of genera commonly known as the Hellebores*

Davis (15) has

classified North American genera of Ranunculaceae according to the following scheme: I* II, III,

CROSSOSOMEAE: FAEONIAE:

Cross os oma,

Paeonla,

HELLEBOREAE:

Hydrastis#, Caltha, Trolllus. Cam-

marom, Helleborus*. NigelTa#, tsoisyrum#, 6optis#, Xanthorrhiza#, Anemonopsis, Clmlcifuga, Actaea."“Aqullegla. Delpfaln ium» . Xconitum*, IV,

CLEMATIDEAE: Clematis,

V, ANEMONEAE: Anemone. Hepatica,Syndesmon*, Myosurus, Trautvetterla, Batrachlunu Ranunculus, Kumlienla, fflcaria. Cyrtorhyncha, Arcteranthus, Pay graphis, Thailetrum, Adonis, 3foose genera marked with an asterisk have been reported to contain alkaloids (54): it will be noted that with one exception all the alkaloid-containing genera fall In the Hellebore group.

The structures of the alkaloids are known

for only five of the genera; of these five, four (Hydrastis, Copt Is, Xanthorrhiza , and Syndesmon) contain alkaloids of the Isoquinoline group, while the other (Nlgella) contains a simple amine, methyl 2-methylamlno-3-methoxybenzoate (20), The structural types of the alkaloids of the remaining four genera are unknown but their empirical formulas and pharma-

cologlcal properties are closely Interrelated The genus Delphinium ( T o u m . ) L* consists of about 250 species of perennial (and occasionally annual) erect, branching herbs of world-wide distribution.

The leaves

are alternate, palmately veined, and palmately lobed, cleft, or divided*

The showy flowers are terminally racemose or

paniculate; there are five irregular petal-like sepals, the upper one prolonged Into a spur at the base; there are usu­ ally four (sometimes two, united Into one) Irregular petals, the upper pair continuing backward into long spurs which are enclosed within the calyx spur, the lower pair having a pair of short claws; the stamens are numerous and there are from one to five pistils*

The carpels form many-seeded

follicles in the fruit,(64, 66) The genus acquired its name from the Greek word delphis, since the shape of the flower suggests the classical figure of the dolphin* (29)

Also suggestive of the flower form

are many of the trivial names which have been attached from time to time to the more common species:

Dolphin Flower,

Dauphinelle, Knight's Spur, HIttersporn, Calcatrlppe, Lark's Heel, Lark's Toes, Lark's Claw, Lerchen Klaue, Larkspur. Other names applied to the Delphiniums are Horn Kummel, Pied d'Allouette, Herbe Sainte-Athalie, Fleur d'Amour, Sperone, King's Consound, Stavesacre, Lousewort, Staggerw^ed, Poison Weed, Cow Poison, and Peco* (59, 66. 85)

A convenient botanical classification of the natural Delphinium species is both morphological and histological* The following outline shows the classification of some of the North American species according to a combination of Bailey*s morphological arrangement (2) and Marsh, Clawson, and Marsh*s arrangement based on the histology of the cross sections of the stems (59)• NATURAL DELPHINIUMS I*

II,

Annuals (the European consollda group)--only two petals, united; one follicle-**D. A.jacis L., D. consollda L* Perennials— four petals; three to five follicles* A,

The Low Larkspurs 1# D* Andersonll. D. bicolor. D. decorum* ~^D* depaupe ra turn* B. Mangle's 11* D. nudicaule. D. t r l ^ r n e * “ 2. D. blochmannae. f>* cardinale* 5* B* Carolinianum*"*D* recurvatum. D. aim~plex Dougl*. D* varlegatum. D* varlegatum aplculatum* — ' 4* D. (Sever!. D* scaposum. D, virescens,

B*

The Tall Larkspurs D* Barbeyl, D, Calif ornlcum. D. cucul~"latum* D, geranlif ollum. B. glaucum* B . occ i5entale * P. robus turn. D* sapelTonTs* £>* scopulorum* b. trolTifbiium.

The annual species, D. A.jacis and D* consollda. are of European origin but were introduced to the United States as garden varieties; having escaped somewhat from the gar­ dens to uncultivated habitat they may be considered as nat­ uralized species of the east and east central states. two species are often confused.

These

D. A.jacis Is usually described

5*

as being about 18 Inches in height with a few spreading branches; the stem leaves are sessile and deeply cut into fine linear segments; the root leaves are similar but shortpetioled; the flowers, in a splcate raceme, are showy, blue or violet varying to white, and more numerous than in D* consollda; the ealyx-spur about equals the rest of the flower; the follicle is pubescent,

to 1^ inches long; the seeds

have wrinkled, broken ridges*(15)

Likewise, D* consollda

is an erect, hairy annual, 12 to 18 inches high; the blue, violet, or white flowers are few and loosely panlcled, with the pedicels shorter than the bracts; the follicle is glab­ rous; the seeds have broken, transverse ridges* (2) In differentiating between the two species, Marsh, Clawson, and Marsh (59) make use of the stem cross sections* In £♦ A.jacis the stem is circular and has a relatively small lacuna; there are about 46 bundles of two sizes, large and small arranged alternately; there is a distinct row of endodermal cells*

All the cell walls are much thickened and

the shape of the fibrovascular bundles Is quite character­ istic*

The wedge-shaped bast is eemposdd of cells with

walls thickened so that the lumen is reduced almost to a point; the small phloem is completely enclosed by the bast and xylem; the elongated xylem mass is larger than the bast and includes a large amount of xylem parenchyma*

D* consol—

Ida is similar to D* Alacls but has fewer bundles, thicker cell walls in the pericycle, and part of the cells of the cortical parenchyma with thickened walls*

Other annual larkspurs are D* cardlopetalum. D* cinereum. D* dlvarlcatum* and D # plctum (D* Recrulenl). all natives of Asia Minor or southern Europe* The low larkspurs attain a height of about one or two feet, blossom early in the spring, and die down soon after flowering; they are found in a variety of habitats from sealevel to foothills and sometimes as high as 10,000 feet* The tall larkspurs, on the other hand, may reach a height of five feet or more, blossom in mid-summer or in autumn, and grow until frost; they are found chiefly in forest ranges usually at altitudes of 8,000 to 11,000 feet* (17) The classification of the cultivated Delphinium species falls into three main groups:

the so-called "botanical*

larkspurs, the annual larkspurs, and the true horticultural "Delphiniums11*

Tfcie "botanical" larkspurs are perennials

grown as wild flowers or as border flowers and have not yet been developed into modified or highly-bred horticultural forms*

Among this group are the species D* Andersonll

(Anderson Larkspur), D* bicolor (Purple Larkspur), D* Bul­ lsvanum* D* CarolInlanum (Carolina Larkspur), D* decorum* D* exaltatum (Tall Larkspur), D. Menzlesil (Menzies Lark­ spur), D. scapo sum, D* trie orne (Dwarf Larkspur), D* varlegatum (Royal Larkspur), and D* virlscens (Prairie Lark­ spur)* (38) The cultivated annual larkspurs are of three types, derived from the three species D* Ajacls. D. consollda. and

.grandIf1 orum*

D* A.jacis. presumably the "Flower of

Rjax", Is said to be named from the marks at the base of the united petals suggesting the letters AIAI; It Is often called the Rocket Larkspur because of the narrow spike-like erect racemes* (3)

This species originated in the Swiss

Alps and was one of the first foreign larkspurs to be intro­ duced to medieval England (during Elizabethan times, 1573). There are now recognized three subdivisions of this type. (68) (1)

J£* A.lac is major, the Tall Rocket Larkspur, grows

-to a height of three to four feet and produces both single and double flowers of many colors*

The outstanding varieties

are coelestinum. exceptionally bright Cambridge blue flow­ ers, double; coeruleum. Indigo blue; and roseum. pure bright pink, double* £• Ajacls minus, the Dwarf Rocket Larkspur, reaches a height of

to 2 feet and produces double flowers, of

which the main varieties are:

album, white; azureum. sky

blue; coeruleum. deep gentian; and roseum. deep rose pink* (3) D* A.jacls hvaclnthlflorum. the Hyacinth-flowered Larkspur, possesses a dwarf habit and produces spikes of a tapering hyacinth-like appearance* consollda. the Branching Larkspur of English corn­ fields, is also known as the Forking Larkspur, the Field Larkspur, and the Knight*s Spur; it grows to about three feet In height.

Consollda is the ante-Linnaean substantive

name, referring to the consolidated petals. (3)

The two

main subdivisions are the Double Stock Flowered Larkspurs, including such varieties as Dark Blue, Empress,Rose, Lilac Supreme, Lustrous Carmine (Newport Fink), Purple, Rose Queen, Salmon Rose, Sky-Blue, and or Double Giant

White; and the Emperor

Imperial Larkspurs (D. consollda var.

imperial is). among which are Blue Bell,

Carmine King, Coral

King, Exquisite Pink Improved, Lilac Queen, Sweet Lavender, and White Spire. (84) D. grandlflorum. the Great-flowered Larkspur or the Siberian Larkspur, gives rise to a type of Delphiniums known as the Bouquet Larkspurs.

These larkspurs are descendants

of a short-lived perennial species but are usually treated as blennuals or even annuals in regions of long growing seasons or severe winters.

They attain a height of up to

four feet, but the most common form, the annual D. grandiflorum var. chlnense or D. chinense. the Chinese Larkspur, usually reaches a stature of only li- to 2 feet*

Among the

Chinese Larkspurs are the varieties Album (white), Azure Fairy (bright Cambridge blue), Blue Butterfly (bright sal­ via blue), Blue Gem (deep gentian), and Blue Mirror (navy blue). (68, 84) The third group of cultivated Delphiniums is the highly developed perennial group, of which three types are recog­ nized:

the Candle Larkspurs, the Garland Larkspurs, and

the Red Larkspurs*

The first type is usually considered as

descending from D* elatum of Europe, and the second type from D* chellanthum of Siberia and China; these two types are so confused by long cultivation and interbreeding, how­ ever, that they are usually listed under the one general heading D* cultorura: probably other species, such as D* formosum of the Caucasus region, have also entered Into the ancestry of the hybrid perennial Delphiniums*

There are

several races within the cultorum group, differing in stat­ ure, flower characteristics, foliage, etc*; these races have more or less recognized garden names but botanic ally they cannot be clearly defined as to species* £• elatum* the Candle Larkspur, Bee Larkspur, or Blue Larkspur, originated in the Swiss Alps and Is the chief ancestor of the taller garden types*

It grows to a height

of two to six feet; It is glabrous but with the leaves some­ what pubescent, five- to seven-parted; parts rather narrow, cut-lobed; the upper leaves are three- to five-parted; petioles are not dilated at the base*

the

The raceme is much

like that of D. eacaltatum (with which D* elatum is often confused) or more spike-like; the flowers are blue with dark violet petals; the sepals are ovate, glabrous, and nearly equaling spurs*

There are three follicles; the seeds arc

transversely wrinkled, not scaly* (2) D* che llanthum * the Garland Larkspur, has its origin in Siberia and is of smaller stature than D* elatum*

It

has a simple or branched stem two to three feet in height;

the leaves are glabrous or slightly pubescent, five-parted, with the lobes pointed, sub-trifid, and somewhat toothed* The flowers are dark blue, the upper petals sometimes pale yellow, the lower ones Inflexed, ovate, and entire; the spur Is rather long and straight or somewhat curved.

The follicles

are three in number, either glabrous or pubescent; the seeds are three-cornered, three-winged, and not scaly. (2)

This

species Is supposedly the parent of the races belladonna* bellamosum* coelestinum* formosum* and others.

The Bella­

donna Delphiniums are of considerable interest and impor­ tance, including such varieties as Blue Grotto (deep indigo), Cliveden Beauty (Cambridge blue), Kelway*s Azure (coerulean blue), Lamartine (deep purplish blue), Lohengrin (deep blue), Moerhelml (pure white), Theodora (pale to mid-blue, slightly shaded rosy purple), Wendy (very bright deep cobalt), and many others. (68) The third type of perennial Delphiniums, the Red Lark­ spurs, are sometimes grouped under the name D* Rays 11* These larkspurs, hybrids with D, nudicaule apparently one of the ancestors, have been developed fairly recently, chiefly in California* A summary of the classification of cultivated Delphini­ ums is given below.

CULTIVATED DELPHINIUMS I* II*

III.

The "botanical" larkspurs The annual larkspurs A#

The Alacls type 1* Tall Socket Larkspurs 2* Dwarf Rocket Larkspurs 3 . Hy ac Inth- f1 ower ed Larks purs

B.

The consollda type 1* Double Stock Flowered Larkspurs 2. Emperor or Double Giant Imperial Larkspurs

C*

The grandif1orum type Bouquet Larkspurs, Chinese Larkspurs

The perennial Delphiniums A*

The elatum type Candle Larkspurs

B.

The che llanthum type Garland larkspurs; Belladonna type

C*

The nudicaule or Buys11 type Red Larkspurs

The following reference list includes the most impor­ tant species and varieties of the Delphinium genus, arranged alphabetically*

Under each entry Is given the botanical

name, the common or trivial name, the height of the fulLgrown plant, the color of the flower, and the habitat or country of origin*

The list has been compiled from many

sources (12, 3, jt, J3, Ji5, 24* 2 5 * 2 8 * 2 9 * 3 7 * 38* 59, 60* 6 4 * 66* 68* 70* 71* 72* 81) * as no one list hitherto pub­ lished is complete*

12*

DELPHINIUM SPECIES D* abletinum TIdest* A. Nels.

Possibly the same as D* cucullatum

£♦ A lac is L* (D* addendum McNab; D. consollda Sibth* $ Sm.; D* ornatum Bouche; D* pubes cens Sriseb,; Ceratosanthus A.jaols Sehur.); Rocket Larks pur, Annual Xarkspur; l-'ii ft*; originally blue, now blue, white, violet, or pink; Swiss Alps, now naturalized in U.S. and escaped from gardens; fields, roadsides, wet places, and around dwell­ ings, Vt* to N. Car* west to Tex*, Mo*, and Mont. D* albescens Rydb.: same as D* virescens Nutt* and D, Penardi “Buth. ~ “ £• ftlb^stre Rydb.; 4-9 in.; medium to dark blue with yellow tinges; rocky heights, Colo., N. Mex. D.

alplnum Waldst* & Kit.; same asD. elatnm

L.

£•

Altissimum Walllchs 3 ft.; blue to purple;Himalayas.

D* amoenum; 2 ft*; blue; Siberia. D. Anders onil Gray (D. Leonardil Rydb.); Anderson Larkspur, "" fibw Larkspur; l-'S ft*; deep blue; semidesert, Sierras and other mts*, Utah, Nev*, Calif*, Ore* D*

atropurpureum; same as D* elatum L*

D. ""

attenuatum (M, E. Jones) Rydb*; dark blue upper petals; along brooks, Utah*

with white

D* agureum MIehx. (once considered a variety of D. Carolinianum IFalt.) ; Larkspur; 1-2 ft*; pale blue, slcy blue, or whitish; Va. to Ga* west to Tex., Ark.,Mo.$WIs. to Man* west to Sask. and the Dakotas and south; northern Rocky Mts*; Calif., Ore* D* azureum Mlchx* var* album: white form of above; northern "" Rocky Mts • D. Barbeyl Huth* (D. scopul orum Gray var. sub alplnum Gray; D* sub alplnum 7Lm He Is.); Tall Larkspur; 1-7 ft *7 dark o p violet blue, upper petals yellow with blue tinge; open woods and grassy mt. parks, occasionally partially shaded; 8,000-11,000 ft., Mont. to Wyo., Colo., Utah and south. D. Barlowli Hort*; the plant now grown under this name Is of the P* che llanthum group but the original Barlowli (1837) is a very double form regarded as a hybrid of D* elatum and D. grand If 1 orum.

13

£• Belladonna Hort.; a race of* D. ^heilanthum formosum: light blue. £♦ Bel lamp sum Hort.; a race of D, cheilanthum formosum: dark blue • "" D. bieolor Nutt.; Purple Larkspur, Low Larkspur; 6-20 in.; spur and sepals rich blue or dark purple, upper petals pale or brownish yellow or white, lower petals blue; dry woods, 4,000-10,000 ft*, H. & S. Dak. south to Colo., west to Ore., and north to Alaska. D. blochmannae. D. brachycentr on:

2 ft.; blue; Siberia.

D* Breckli Hort*; double blue form of D* grandlflorum L. var. Chinense Fischer. £• Bi*ownii Rydb.; 5 ft.; dark blue or purple; Mont., Sask., Alta., to Alaska. D. Brunon 1anum Royle; Musk Larkspur (so-called because of "" its musk scent); 6-20 in.; light blue with purple mar­ gins, center black; Himalayas, China. D. Bulleyanum; 2-4% ft.; deep blue; China. D. Burkei Greene; blue and white; arid hills, Ida. D. CalIf ornlcum T. & G. (D. exaltatum Hook. & A m . ); Coast Larkspur; 7 ft.; white, whitish, or pale blue, sometimes purplish inside; low hills and peaks of coast ranges, Calif. D. camp orum Greene; same as D. Penardi Huth. D* candelabium; a form of D. Ajacls L. D. candldum Hemsi.; dwarf; pure white with purple anthers; tfganda, mts. of tropical E. Afr.; recently Introduced to N. Am. D. canmorense Rydb.; 6 dm. or more; brownish or dark blue; Rocky Mts., Canmore, Alta. £• cardinale Hook.; Scarlet Larkspur; 2-6 ft.; bright red or scarlet with petal limbs yellow; Calif. D. cardiopetalum. DC. (D. halter aturn Sibth# & Sm. var. cardio­ petalum lluth.); 10^20 in.; blue to purple; S. France and Mediterranean region; also cultivated.

14.

£•

Carollnianum Walt# (B. azureum Michx.; D. virescens Nutt.); Carolina Larkspur; ft.; medium or azure blue vary­ ing to whitish or white, sepals often with a brown spot; II. Gar. to Ga. west to 111., Iowa,Colo.

B.

Carollnlanum Walt. var.album Hort. 2-3 ft.; creamy white.

£*

Carollnlanum Walt. var. Nortlanum (B, Nortlanum): taller than the parent type; deep bluish purple; Ozarks, Mo.

(var. albidum Hort.): —

B. C ar olinlanum Walt. var. vlmlneum Gray (B. vlmlneum; perhapa B. azureum Mlchx.); leaves broade"? than In parent type, seeds also different; 2-4 ft.; blue-, violet, white, or whitish; prairies, Texas. B. carporum: 1 ft.; white and pink; western Rocky Mts. B.

cashmirianum

Hort.; one of the B. eultorum Voss group.

B.

cashmirianum Royle; 10-20 in.; pale to deep azure blue, upper petals almost black, lateral ones greenish; high Himalayas, Cashmlr.

B. cashmirianum Royle var. Walker! 1 Hook.; 1 f t.; pale or light blue sepals striped dark blue, petals yellow, wye yellow; Cashmlr. B. caucaslcum Hort.; one of the B. eultorum Voss group. B. caucasloum C. A. Mey (B. speciosum var. caucaslcum Huth. ); 4 in.; azure blue backed purple; Caucasus. B. che llanthum Fischer (B. magnlflcum): Garland Larkspur; — 2-3 ft.; dark blue varying to whitish, upper petals some­ times pale yellow; Siberia, China. B. che llanthum formosum: contains the horticultural races Belladonna and &ellamosum; rich blue. B, che llanthum Moerheimil; varying to white. B. chinense; same as B. grandif1 orum L. B. clnereum; apparently a sport from B. grand if1 orum L.; blue; Asia Minor. B. coelestinum; 4 ft.; bright azure blue; eastern Szechuan. B. coelestinum Rydb.; 4-5 dm.; light blue; arid regions, ~ Ariz., southern Utah.

15.

£♦ coeleatlat a horticultural name, probably belonging to I>» che llanthum jTormosum. D* coerulescens Freyn.; Asia. £♦ ^oeraleum; 1 ft*; Cambridge blue; Tibet, Sikkim* D* Columblanum Greene; same as D* Huttallll Gray* J)# consollda L*(often confused with D. A.jacis L*); Field Larkspur, Forking Larkspur, Branching larkspur, Knights Spur, Ritterspom; 1-3 ft*; white, blue, or violet; north­ ern Europe; naturalized InU*S.; roadsides, old grain fields, Va. to N. J. and Penna* north and west to Iowa* J2» consolida L* var. Imperial is Hort. (D. imperial is fl. pi. Hort*); Emperor Larkspur; flowers d'ouble; from English gardens* D. consollda L* var. ornatum Candelabrum; a horticultural variety. D* corymbosums 2 ft. or more; pale violet and greenish on "" back; mts., Turkestan. D# crassifolium; l|r ft.; light blue; Turkestan, Siberia, China* £• cucullatum A, Nels. (apparently a form of L* occidentale "" "^S* Wats.T; Tall Larkspur; 6-8 ft*; yellowish white to grayish blue; wet meadows, Mont., Ida., Utah, Colo* £• eultorum Voss; the horticultural or perennial garden "" ^larkspurs, hybrids of the elatum and che llanthum groups; 3-5 ft.; light or dark blue, violet to white. £• cyan ore ios Piper (D. simplex A* Gray); 3-6 dm*f blue; ~ damp prairies and~wet rocky places, B. C. to Ida., Ore. D. dasyanthum Kar. et Kir.; 1% ft.; light or deep brilliant "" blue; central Asia, Turkestan, W. India, Siberia. £• Davldii Franch; 3 ft.; clear light blue; eastern Tibet, China. D* decorum Flsch* & Meyer (the name decorum has also been applied to a horticultural form of the elatum group); 6-24 In.; sepals bright or violet blue turning to violet purple, upper petals at least tinged with yellow; Calif. D. decorum F. & M. var. patens Gray; Smooth Larkspur; mts., Calif.

16#

£• Delavayl (the plant cultivated under this name Is prob­ ably one of the elatum group); 9-12 in.; bright blue; Yunnan, West China# £• degauperaturn Nutt. (probably more closely allied to B. columblanum Greene than to B. Nuttalllanum Pritzel.T; 12-18 in.; deep or dark blue to purple, upper petals yellowish or whitish; poor soil, Ore., Ida., Nev., Calif, B# dietyocarpum; 2 ft# or more; sky blue; Caucasus, Siberia. £• discolor: one of the elatum forms. £* distlchum Geyer (very similar to D. simplex Dougl.); 3-6 am#; blue; meadows, Wash, and Ore# to Mont. B. dlvaricatum Ledeb. (allied to B. consolida L. but taller, more branched, with smaller more abundant flowers); 14^*2 ft#; purplish blue; s.w. Asia, Caspian and Caucasus regions; also cultivated. B* diversicolor Rydb.; 4 dm.; blue and white; bogs, Mont. B* Bqhmbergll; 2-6 ft.; blue to white; Russia, Turkestan# B# dumetoram Greene (a relative of B# Nelsonll Greene); 4-6 dm.; light blue; dry hills, s.wT Colo. B# elatum L# (B. alplnum Waldst. & Kit.; B. pyramidale Royle); Candle LarTcspur, ^ee Larkspur, Blue L'arkspur; 2-6 ft.; heavenly blue, petals dark violet; Swiss Alps. B# elegans BC#; introduced to the U.S. B# elongaturn Ifydb#; Tall Larkspur; 1-5 ft.; pale to dark blue; hills and mts., along streams and moist places up to 9,000 ft., Colo, to Mont. and Alta. B.

Bmlllae; 12-18 in.; sky blue; Calif.

B#

exaltatum Ait. (B. urceolatum Jacq.); TallLarkspur; 1-5 ft#; bright blue or bluish triolet, sometimes white, yel­ low on upper petals; borders of woods, rich soil, Ala. and S. Car. north to Ky., Ohio, Pa., and Canada, west to Minn., Iowa, and Nebr#

B# exaltatum Nuttallil; 2 ft.; deep blue; wooded banks of the Columbia River. £* exaltatum Hook. & A m . ; same as B. Calif o m l c u m T. & G. £• barges11; 1 ft.; dazzling blue; Szechuan#

It)

17.

It)

• j£lssum; 3 ft.; blue; eastern hills of Transylvania.

It)

• formosum Boiss. & Hult.; 2-3 ft.; blue with indigo mar­ gin, spur violet; Asia Minor, Caucasus.

It)

• formosum Hort.; one of the D. eultorum Voss group.

It)

• Froynll« l ft.; bright blue; Caucasus.

It)

. geranilfollum Rydb*; 3-4 dm.; dark blue and yellowish; dry hills, Ariz. to Colo.

It)

• Geyerl Grreene; Geyers Larkspur, Plains Larkspur, Poison Weed; 2-7 dm.; azure or deep blue and yellow; high plains, dry mesas, rocky hills, and foothills, 4,000-8,000 ft., central Rocky Mts., Colo., Wyo., Mont., Utah.

It)

• glabellums a name without botanical definition.

It)

• glabratum (a supposed variety of D. caucaslcum C. A. Mey); 3 ft.; deep azure blue; Himalaya's.

It)

. glaciale; 9 in.; medium blue; alps of Sikkim.

It)

. grandlflorum L. (D. Slnense Fischer); Great-flowered Larkspur, Bouquet Larkspur, Siberian Larkspur; 1-4 ft.; rich blue or violet, varying to white, spur and lower petals often violet, upper petals often yellow; Russia, Siberia, China.

It)

• glancum Wats.; Tall Larkspur, Tall Mountain Larkspur; possibly the same as L. Barbeyi Huth.; Rocky Mt. region, Wyo., Ore., Calif.

It)

• glances cens Rydb.; 3-5 dm.; dark blue with yellow tinges; among rocks, canyons and mt. sides, 6,000-9,000 ft., Wyo., Mont., Ida., and possibly adjacent states.

It)

• gpandiflorum k. var. albo-pleno Hort.; flowers double and pure white. • grand if 1 orum L. var. album Hort.; pure white.

It)

. grandiflorum L. var. Chinense Fischer; Chinese Larkspur; 1-2 ft.; China.

D. grandiflorum L. var. chinense Huth. (D. chinense Fisch. ); ~~ China. D. grandlflorum L. var. cineraria coeruleum; 6-9 in.; rich "" Cambridge blue; Russia, Siberia"^ China.

is.

£• grandlflorum L. var* flore-pleno Hort* (var. hybrldum Hort*); flowers double, blue* £♦ grandlflorum L. var* variegatum Hook. & A m . ; same as t>m variegatum* £• bait era turn Sibth* & Sm* var. cardlopetalum Hutb.; same as cardlopetalum DC* D* hamatum; 6 In*; pale blue; limestone rocks of Yunnan* £* Hansenls pinkish mauve; Calif. D* He H e r 1 Rydb* (originally named as a form of D* Huttalll~ asra** Prltzel.); 3 dm.; dark blue; plains, Ida* D* besperium Cray; Western Larkspur; from sea level to lower mts., 6alif. £• besperium Cray var. recurvaturn Jepson; white to pink and lavender* rarely blue; dalif.

which on contact with water or steam immediately became crystalline again. Analysis of crystalline lycoctonine. Pound: N, 2.63, 2.75, 2.68, 2.94* Hg°, 4.02, 3.92, 4.09, 3.96 MeO, 24.45, 25.26, 25.02 MeR, 2.63 Equlv. Wt., 472.6, 474.1, 473.5 (titration) Mol. Wt., 444.5, 442.8, 449.8 (naphthalene) Calc, for CggH3 gN07.H2 0 : N, 2.90; T1q09 3.73; 4 MeO, 25.66; MeN, 3.18; Mol. Wt., 483.6.

Analysis of anhydrous lycoctonine. Pound: C, 64.24, 64.14, 63.90, 64.16, 64.17 H, 9.17, 8.42, 8.61, 8.63, 8.72 N, 3.05; MeO, 26.24; MeN, 2.57. Calc, for C2 5 H 3 9 N07 : C, 64.46; H, 8 .4 4 ; N, 3.01; 4 MeO, 26.64; MeN, 3.23. Schulze and Bierling found that lycoctonine was a strong tertiary base containing one methylimlde, four methoxyl, and at least two hydroxyl groups; it could be titrated sharply in the presence of either iodoeosin or methyl red; ammonia precipitated lycoctonine incompletely in crystalline fonn from its salt solutions only after a long time.

The hydro­

chloride, CggH^gNO^.HCl.HgO, melted at about 75° (foaming); thick, transparent prisms from alcohol and ether, becoming cloudy on exposure to air; very soluble in water.

The hydro-

bromide, thick prisms with two molecules of water of crystal­ lization, melted at 88-9° (air-dried).

The perchlorate,

thick prisms with 1.5 molecules of water, melted at 68-9° (foaming) and was readily soluble in water.

The methiodido,

CggHgQNC^.CHgl, yollowish needles from alcohol and ether, m.p. 178° (foaming), contained varying amounts of water of crystallization which was expelled at 100°. ide was a syrup.

The methochlor-

The chloroaurate, small thick yellow prisms,

was rather insoluble in water.

The platinum salt was easily

soluble in water. The lycoctonic acid, light brown needles or leaves, m.p* 179-80° on slow heating (decomposing above the melting point), was dibasic and was shown to be succlnylanthranlllc

acid by acid hydrolysis, which yielded succinic and anthranilic acids. Anthranoyllycoctonine,

formed light brown

glistening leaves from alcohol, m.p, 154-5°,

It was soluble

in water, alcohol, and ether, solutions in the latter two solvents showing a deep blue-violet fluorescence. wise was shown to groups.

The salts

It like­

contain one methylimlde and four methoxyl showed little tendency to crystallize;

the perchlorate, C^gH^NgOg.SHClO^, formed long acicular needles from alcohol and ether and began to decompose at 185°. Alkaline hydrolysis of anthranoyllycoctonine gave lycoctonine and anthranilic acid. The alkaloid myoctonlne was found to be a dimer, (^3 5 2 4 5 ^ 2 0 ^0 ), of lycaconitine.

It was an almost colorless

amorphous powder, soluble in cold water to the amount of about 1%; (Oi)p^ +44,79° (alcohol).

The alcoholic solution

showed a faint violet-blue fluorescence while the aqueous solution did not.Crystalline salts were It was a weak base

not obtainable.

and on hydrolysis gave the same products

as lycaconitine. The third base from A. lycoctonum gavo an insoluble thiocyanate; on alkaline hydrolysis it also yielded lycocto­ nine and lycoctonic acid.

33

£• chlnenae The presence of alkaloids In I>* chlnense was reported by Keller (4 7 ) but no further Information was given*

D* consolida L* Keller (46) obtained from D, consolida three alkaloids: Base A, crystalline, extraotable with ether; Base B, amor­ phous , almost insoluble in ether; and Base C, amorphous, eas­ ily soluble in ether*

Base A, six-sided plates, melted at

195-7° after five recrystallizations from alcohol; there was no effect on drying at 105-10°*

It was easily soluble in

alcohol; chloroform; acetone; and methanol; difficultly soluble in ether and ethyl acetate; and very little soluble in water*

The average of fifteen analyses gave C, 62*67;

H, 8*69; and N, 3*68$; there was no internal agreement among the analytical results, however, so that Keller proposed no formula; the average of three determinations gave MeO, 19*49#. Using the averages of the fifteen analyses, Markwood (55) later calculated the formula C2 0 H 3 3 KO 0 , for which the theo­ retical values are C, 62.63; 16*18;

H,

8

*6 6 ;

N, 3*65;

2 MeO,

3 MeO, 24.27#.

Markwood (5 5 ) found in D. eons olida three crystalline alkaloids, one of which could be extracted from oxalic acid solution by chloroform and yet was precipitated by ammonia; it melted between 153 and 160° and was biaxial negative* was insoluble in water, slightly soluble in ether, more so

It

in alcohol, and readily soluble in chloroform*

To obtain

the other two alkaloids, Markwood made basic with sodium hydroxide the oxalic acid solution from which the first alkaloid had been removed and then extracted the alkaline solution first with ether and then with chloroform*

Prom

the ether extract was obtained the alkaloid delsoline and from the chloroform extract the alkaloid delcoslne* Delsoline, CggH^-jNOg, brilliant crystals resembling octahedra but probably monoclinic, was biaxial positive and melted at 207-9°*

It was very soluble in alcohol and

chloroform and was fairly soluble in water.

The analytical

results were: Found: C, 62.07; H, 8.49; N, 2*88. Calc, for C 2 5 H 4 1 H 0 8 ; C, 62.07; H, 8.55; Deleosine, Cg^H^NOg,

N, 2.90*

198-9°, crystallized In the

orthorhombic system In six-sided plates or prisms which wcro biaxial negative.

The optical activity was not reported.

It was very soluble in chloroform and alcohol, appreciably soluble In water, and less so in ether.

Markwood regarded

his delcoslne as identical with Kellerfs Base A, although the carbon analysis differed by different formula.

1

#, necessitating a slightly

The analysis of delcoslne was:

Pound: C, 63.73; H, 8*55; N, 3.58. Calc, for C2 1 H 3 5 M06 : C, 63.77; H, 8.41;

H, 3.54.

Cionga and Iliescu (12) recently Investigated the alka­ loids of D* consolida and, like Markwood, also found delso­ line and delcoslne, hut again the analytical data vary from the previous investigators#

Petroleum ether removed from

the seeds 32# oil; the alkaloids were then extracted with benzene, giving on evaporation of the dried extracts 65 g# of a brown-red viscous residue from 1670 g# of seeds#

This

residue was taken up in 5# sulfuric acid and the alkaloids precipitated with ammonia and extracted with ether.

Prom

the ether extracts two crystalline fractions were obtained: crude delsoline, 0*75 g.; and crude delcoslne, 0*57 g.; total, 1*32 g., or 0*08#* The delsoline of Cionga and Iliescu melted at 207-9°; rhombohedrons from ether.

It was little soluble In water

and ether, easily soluble In benzene, and very soluble In 24 alcohol and chloroform* (od)c «48.12° (chloroform)* The chloroform solution of the alkaloid took up bromine very easily, giving a dibromo derivative, yellow, glassy, and transparent, rapidly becoming turbid.

The formula CggP^QNO^

with three methoxyls and three active hydrogens was pro­ posed; no molecular weight determinations were reported. Pound:

C, 64.57, 64.44; H, 8.93, 8.82; N, 3.11; MeO, 20*02; active H, 0.67 (OH, 11.45). Calc, for C2 5 H4 0 N07 : C, 64.33; H, 8.64; N, 3.00; 3 MeO, 19*94; 3 active H, 0*64 (3 OH, 10*09).

The delcosine of Cionga and Iliescu consisted of color­ less crystals, m.p. 198-9°.

It was very little soluble in

56

water and ether, soluble In benzene, and very soluble in alcohol and chloroform*

( o O § 4 -54.41° (chloroform).

A

dlbromide was obtained in the form of a transparent varnish, soon becoming dull.

The analytical data indicated that

delcosine was isomeric with delsoline; they were not identi­ cal, however, since the mixed melting point was 196-7°. Pounds

G, 63.96, 63.98; H, 8.82, 8.62; N, 3.17; MeO, 19.96| 20.35; active H, 0.75 (OH, 12.7). Calc, for C 2 5 H4 0 HO7 : C, 64.33; H, 8.64; H, 3.00; 3 MeO, 19.94; 3 active H, 0.64 (3 OH, 10.09). Only oily products were obtained from delcosine with benzoyl chloride and pyridine or with acetic anhydride and sodium acetate. It should be noted that the formula C2 5 H 4 QHO7 proposed by Cionga and Iliescu for both delsoline and delcosine is impossible; there must be an odd number of hydrogen atoms. The plausible formula Cg 5 H 4 1 N07 fits their data Just as w d l , as shown in the following tables, which summarize all the analytical data on the two alkaloids.

]?eund:

C H N MeO

Calc. for C H N 3 MeO

Delsoline Markwood 62.07 8.49 2,88

Cionga & Iliescu 64.57, 64.44 8.93, 8.82 3.11 20.02

C25H41N08 62.07 8.55 2.90 19,26

C25H40N07 64.33 8.64 3.00 19.94

C25H41N07 64.20 8.84 3.00 19.90

.

Pound :

C E N MeO

Calc# for C H N 3 MeO

Delcosine Markwood 6t>M 8*55 3.58 *•#*—

Keller 62*67 8#69 3*68 19.49 G20H33N06 62 #63 8*66

3#65 24.27

C21H33N06 63*77 8.41 3 *54 23.53

Cionga 8c Iliescu 63*96, 63.98 8.62 8 .82 , 3.17 19.96 C 2 5 H4 0 N 0 7 64.33 8*64 3.00 19.94

c 25H41N07

64.20 8.84 3.00 19.90

The presence of mannitol in D* eons olida was reported by Jaretzky and Schaub (42). who found this alcohol in some twenty species of Delphinium and concluded that since all species of this genus probably contain mannitol the latter should be regarded as a genetic characteristic#

D # cueullatum A # Nels* D, cueullatum was found by Beath and Nelson (8 ) to con­ tain 1*14-1#86$ alkaloids# isolated: ammonia; m*p# 162°;

Pour separate alkaloids were

(A), crystalline, m#p* 1*78°, precipitated by (B), crystalline, m*p# 225-7°;

(C), crystalline,

(D), crystalline, soluble in water, slightly

soluble in alcohol, and insoluble in ether or chloroform#

D. dasyanthum Kar. et Kir# A recent investigation (50) indicated the presence of alkaloids in D# dasyanthum#

* fe^nton L.

22

Prom an acidic solution of the alkaloids from one kg. of D. elatum seeds Keller (4 7 ) obtained, bjr neutralizing with ammonia and extracting with ether,

10

g. of a yellow

viscous mass from which 3.5 g. was obtained as a crystalline alkaloid, CggHg^NOg; methanol.

m.p. 218° from dilute alcohol or

The analytical results were:

Pound:

C, 66.59, 66.50, 66.59. 66.78, 66.71, 66.84, 66.79 (ave., 66*69). H, 8.87, 8.91, 8.67, 8.45, 8.70, 8.58, 8.69 (Ave., 8.69). N, 2.35. Calc, for C3 3 H 5 1 F08 : C, 67.20; H, 8.72; N, 2.38. Prom the mother liquors were obtained two amorphous compounds:

(1 ) soluble in ether, ethyl acetate, alcohol,

and chloroform; (2 ) soluble in alcohol and chloroform but Insoluble in ether.

D. form osuni Alkaloids are present in D. formosum according to Keller (47).

D. Geyeri Greene The presence of alkaloids in the leaves and roots of 22* Qeyerl was reported in 1913; mannitol was also found* (35) Several years later a more thorough Investigation was made by Beath (5); in the flowers of D* Cteyerl he found

0*245$ alkaloid, 5*02$ sucrose, some mannitol, and a mixture probably of arabinose and galactose.

Prom the leaves and

stems were obtained three alkaloids, one amorphous and two crystalline. The amorphous alkaloid corresponded in Its physical and chemical properties to the principal amorphous alkaloid of D. Barbeyl (p. 27) but was somewhat less toxic; Its melt­ ing point was indefinite (decomposition below 160°),

It was

soluble in chloroform, alcohol, and acetone and was precipi­ tated by ammonia.

Alkaline hydrolysis gave a crystalline

acid and a crystalline base, m.p. 197-8°, optically inactive. One of the crystalline alkaloids formed monoclinle pris­ matic needles, m.p. 207-8° from absolute alcohol; It was optically inactive.

The other crystalline alkaloid, extreme­

ly soluble in water, formed pseudotetragonal orthorhomblc plates, m.p. 214-5°.

Analyses were not given.

D. glaucescens Rydb. The seeds of D. glaucescens were found by Beath (5) to have an alkaloldal content of 0.56$.

Prom the leaves and

stems he obtained two alkaloids, one crystalline and one amorphous.

The crystalline alkaloid was orthorhomblc,

pyramidal, and melted at 208.5-9.5°; it was insoluble in water but soluble In chloroform and alcohol; It was strongly dextrorotatory (specific rotation or analysis not reported).

The amorphous alkaloid had an Indefinite melting point; it was insoluble in water, was not precipitated by ammonia, and was decomposed by alcoholic potassium hydroxide into non-alkaloidal substances.

D. glaucum Wats. Alkaloids were found in the flowers, leaves, and roots of D. glaucum: mannitol was also reported present. (55)

£• hybridum Keller (47) reported the presence of alkaloids in D. hybridum.

D. laxlflorum The alkaloidal content of D. laxlflorum was found to be 0.19-0.21&.

(90)

D. Menzlesii DC. Delpjiowxrarine (see p. 27) was obtained by Heyl (56) in 0.35$ yield from the roots of D. Menzlesii.

22* Nelsonii Greene G. Heyl (56) reported a 0.72$ yield of delphocurarine (see p. 27) from the roots of D, Nelsonll .

P. W. Heyl,

Hepner, and hoy (35) found alkaloids (along with mannitol)

41*

in the flowers, pods, seeds, leaves, and roots of this species* Beath and Nelson (8 ) isolated in 0*16# yield an amorphous alkaloid of indefinite melting point*

D. occidentale S. Wats. Couch (13) obtained 0$98# total alkaloid from the dried plant of D. occidentale*

By extracting with ether the sodium

hydroxide-precipitated alkaloid from 19*22 kg. of plant he obtained 109 g* of the crude alkaloid deltaline; recrysta.1 lized from 25# alcohol it formed long silky needles, m*p* 180-1°, which did not contain water of crystallization* formula proposed was C2 1 H 3 3 N 0 6 ;

The

it contained two methoxyls

and three hydroxyls which could be acetylated*

Deltaline

was soluble in methanol, alcohol, chloroform, ether, and 24 acetone; 5 -27*86°* No crystalline salts were obtained. Analytical results were: ?ound:

63.67, 63.71, 63;56; H, 8.45, 8.41, 8.41; N, 3.62, 3.64; MeO, 15.68; Calc, for CgjBggNOg: C, 63.76; H, 8.40; N, 3,54; 2 MeO, 15.70*

D, rhinante According to Keller (47) alkaloids are contained in D. rhinante.

D* scopulorum Gray Delphocurarine (see p, 27) was reported by Heyl (36) to occur to an extent of 1.30# In the roots and 1.18# in the seeds of D, scopulorum var. stachyd.

D» semibarbatum Bienert. Alkaloids were recently reported present in D. semi­ barbatum. (50)

staphisagrla k* More work has been done on the alkaloids of this species than on those of any other Delphinium species and consequent­ ly more confusion exists in the literature than for any other species.

Some half-dozen different alkaloids have been re­

ported over a period of about eighty years but not until the last several years have any results of lasting value appeared. The named alkaloids present in D. staphisagria are staphisagrine, staphisagroine, staphisagroidine, delphinoidine, del­ phisine, delphinine, and staphisine. Staphisagrine was obtained as an amorphous compound (m.p. over 90°) by Marquis (57) in 1877.

It was very slight­

ly soluble in ether, slightly soluble in water, and very sol­ uble In alcohol and chloroform. optically inactive*

In alcohol solution it was

Marquis gave it the formula as rhombic aclcn&ar crystals, m.p. 189.2°; it was easily soluble In benzene, chloroform, ether, absolute ether, and 90# alcohol; slightly soluble in petroleum ether and abso­ lute alcohol; very slightly soluble in water.

Delphinine was reported in 1864 a compound easily soluble in alcohol

by Erdmann(18)

to be

and etherbutalmost

insoluble in water; he assigned to it the formula C ^ ^ ^ K O g , based on the following analysis: /

, Q ) found a lethal dose of 19 mg.^cg. (rab­ bit) and a toxic dose of IS mg./kg. for the amorphous alka­ loid, precipitated by ammonia, from immature B. Severi plants; for the alkaloid obtained from the mature plants the corre­ sponding doses were 21 and 16 mg./kg*

For the base, m.p#

197-8°, obtained on hydrolysis of the amorphous alkaloid, the lethal dose was 35 mg./kg.; for the crystalline alkaloid, m.p. 207-8°, it was 50 mg./kg.; and for the crystalline alkaloid, m.p. 214-5°, it was 60 mg./kg.

B, glaucescans Rydb. The crystalline alkaloid, m.p. 208.5-9.5°, from the leaves and stems of B # glaucescens was found to be lethal in a dose of 35 mg./kg. (rabbit) and the amorphous alkaloid was lethal to the same degree. (£5)

Later it was reported

(8) that for the alkaloid, precipitated by ammonia, from the mature plant the lethal dose was 26 mg./kg. and the toxic dose 21 mg./kg.

B. Menzlesii BO. For the properties of the delphocurarine obtained from this species, see under B. bicolor Nutt. (p. 54).

B* Nelsonii Greene

For the amorphous alkaloid, precipitated by ammonia, from the mature plants of B. Nelsonli the lethal dose was 10 mg./kg. (rabbit) and the toxic dose 6 mg./kg. (8).

See

also delphocurarine under B. bicolor Nutt. (p. 54).

B* sconulorum Gray var. stachyd. See delphocurarine under B. bicolor Nutt.

(p.

54).

B# staphisagrla L. Since delphinine and other B. staphisagria alkaloids resemble the aconite alkaloids both in chemical and pharma­ cological properties, it will be well to describe briefly the physiological action of aconitine and lycaeonitine. Aconitine is a highly toxic alkaloid, lethal on sub­ cutaneous injection in the following doses:

0.05-0.14 mg./

kg. for frogs, 0.012 mg./kg. for guinea pigs, and 0.014mg./ kg. for rabbits; for cats or dogs the lethal dose is 0.020.1 mg./kg.; 3 mg. was found lethal to a horse.

Aconitine

produces respiratory paralysis and acts directly on the heart, often causing ventricular fibrillation. (34, 4 4 ) In general, lycaconitine Is similar to aconitine in action but much weaker.

Lycaconitine In frogs increases

the indirect sensitiveness of the muscles (due to action on the nerve endings), but there is no action on the nerve

trunks or spinal cord; the lethal dose for frogs is 0.S-0.4 S.As-

In warm-blooded animals lycaconitine increases the 40

reflex excitability; It causes mydriasis, convulsions, general paralysis, and finally death by stoppage of the heart and respiration; the lethal dose for cats is 0,012 g./kg, (9) Delphinine has also been compared to veratrine in its physiological action.

The veratrine action consists of

stimulation of the sensory nerve endings, followed by a sensation of numbness and cold; intensification and pro­ longation of the contraction of striated muscle; prelimi­ nary excitation and then depression of the central nervous system; and death due to respiratory paralysis; there Is a circulatory action similar to that of aconitine but much weaker* (5 4 ) The action of delphinine itself has been the object of considerable study.

In frogs the alkaloid causes fibril­

lary convulsions beginning in the abdominal muscles; opening of the mouth; broken movement, and progressive motor paraly­ sis (paralysis of the heart vagus nerve, Ate.).

The lethal

dose for Rana temp or aria is 0.1 mg. (9) In warm-blooded animals (cats and dogs) delphinine causes diarrhea, nausea, vomiting, and excessive salivation. An uneasiness and staggering gait are followed by stronger convulsions and spasms of the muscles of the legs and lower jaws.

Retardation of breathing, dyspnea (shortness of breath)

accompanied by long expiratory pauses, and coma are followed by death due to respiratory paralysis. 1.5 mg./kg.

The lethal dose is

Autopsy reveals inflamed or congested mucous

membranes, the heart and great veins gorged with dark-colored 4 4 blood, and the lungs covered with ecchymotic spots. (9, 44. 59) Delphinine has no influence on the eyes, ears, or muscular system.

It acts on the spinal cord, resulting In

depression and causing the loss of its exclto-motor power. The chief difference between the action of delphinine and that of aconitine and veratrine Is the direct depressing action of delphinine on the vasomotor and respiratory cen­ ters of the cord, medulla oblongata, and afferent vagus fib­ ers, resulting in death by asphyxiation (death may be delayed by artificial respiration) rather than by paralysis of the heart muscles as In the case of aconitine and veratrine poisoning; because of the circulatory effect of delphinine, however, the heart action is weak and stops about as soon as respiration ceases.

(59)

Injection of delphinine causes at first a decrease in both the pulse rate and the blood pressure, due to stimu­ lation of the vagus.

Then follows a rise in pulse rate and

blood pressure, caused by paralysis of the vagus through continued action of the poison.

Finally there is another

decrease and the heart eventually stops in diastole. (J39J

For a detailed account of the physiological action and therapeutic uses of delphinine, Waugh and Abbot1s text­ book may

be

consulted (87).

Kara-Stojanow (44) reported that the lethal dose of delphisine for cats and dogs was 0.7 mg./kg. and of delphino­ idine 5 mg./kg.

2.

Delphinosis

Delphinosis, the poisoning of cattle resulting from eating delphinium plants,

is a com m on cause of stock losses

on the ranges of the western United States.

It

Is recog­

nized that larkspurs cause m ore loss am ong cattle

0

other poisonous plant except locoweeds

than any

*

(49, J>9, .64)*

The

following species have been found to be toxic. Low Larkspurs

Tall Larkspurs

D. Andersonii Gray 15. agureum Michx.

£* D. 5# ■ D *

5* Uicoloy Nutt. D. Carolinlanum Walt. 5# decorum Fjsch; & Meyer

Barbeyl Huth. Californlcum T. & G. cucullatum A. Nels. dlstichum Geyer

5 # eloftgafrum Bydb. D. decorum F.& M, var, patens Gray D. exaltatum Ait. Seyerl Greene 15. glancum Wats. glaucescens Rydb.

5* hesperium (xray D. Menziesii DC. ^alsonii Greene 15. Darryl "Uray Fanardi Huth. soaposum Greene 5* tricorne Michx. D. venenosum A. Nels. 15. virescens Nutt.

15. occidentale Wats. robustum Rydb. sapellonls D. scopulorum Gray scopulorum Gray var. glancum Gray D. scopulorum Gray

var. subalpinum Gray D; simplex Dougl, D. trollifollum Gray £* A.jacis L. D, consolida L.

61.

The larkspurs are poisonous for horses and cattle, hut usually not for sheep; horses, however, rarely eat enough of the plants for the harmful effects to he noticed, especi­ ally on the open range.

The low larkspurs are poisonous

throughout their entire life and all parts of the plants are toxic.

The tall larkspurs, on the other hand, are the

most poisonous in their early stages of growth; the toxicity gradually diminishes after flowering and disappears as the plant dries up, except that the seeds remain very toxic. The leaves of larkspur are particularly poisonous when wet with dew, rain, or snow.

Most poisoning cases occur in

early spring when the cattle graze on the green plants, but where the snows remain late the poisoning may take place late in the summer or even in autumn. (17. 59. 64) A quantity of plant material of about two to three percent of the weight of the animal is necessary for serious poisoning or death.

Symptons of larkspur poisoning are

stiffness of the limbs, a straddling or staggering gait, and finally falling with the feet extended more or less rigidly; bloating (especially immediately after death), constipation, abdominal pain, nausea, vomiting, salivation, frequent swallowing, and loss of appetite; quivering of the muscles, general weakness (legs crumpling up under the animal), retardation of heart action, and paralysis of the respira­ tory centers (respiration is at first slow and then rapid). Death, due to respiratory failure, comes to the animal in convulsions. * rapid.

If recovery takes place it is usually veiy 22)

Fleming, Miller, and Vawter (21 ) found that animals killed toy delphinosis presented an external appearance of marked bloating with cyanosis of the visible mucous membranes and areas of the skin thinly covered by hair, particularly the Inguinal and perineal regions. The subcutis manifested a decided peripheral venous engorgement. Focal areas of submucous hemorrhage and congestion were found in both the small intestine and the caecum. Small scattered petechial hemorrhages were noted on the diaphragm and epicardium. Hie lungs exhibited well-marked passive con­ gestion. Other post-mortem Indications are congestion of the heart and central nervous system and Inflammation especially of *

*

the rumen, the oesophagus, and the pyloric end of the fourth stomach. ( 80 ) From the above description of the symptons of delph­ inosis it Is evident why several of the commonly occuring Delphinium species have acquired the names ncow poison1*, ‘•poisonweed1*, "staggerweed**, etc.

The general symptons

are apparently the same whatever the species causing the poisoning.

The most widely distributed of the tall lark­

spurs is D. Barbeyl and Is responsible for greater losses of cattle than any other species; the most common low lark­ spur is D. Menziesii and It Is probably the most destruct­ ive low larkspur because of its occurrence in such enormnous masses (58).

A brief summary of the distribution and

importance of larkspurs in several states where they have been recognized as injurious follows; this summary includes only those species which have been made the object of special

study in each state; the occurrence of* other species may he ascertained by consulting the list on page 60 and the reference list on pages 12-23. ALABAMA.

D # tricorne occurs In the Tennessee Valley

and lower hills. (11) CALIFORNIA.

Larkspurs are probably the most important

cause of cattle loss in California.

The poisonous tall

larkspurs are D. califomlcum. D. glaucum. D. scopulorum var. glaucum. and D. troillfollum: the poisonous low lark­ spurs are D. Andersonii. D. decorum. D. decorum var. patens, £• hesperlum. D. hesperium var. recurvatum. D. Menzlesii. and D. Parryl. COLORADO.

(31, 72) The larkspurs are the most important source

of cattle losses in the mountainous regions of Colorado and second only to the locoweed In the state as a whole. The four species growing in greatest abundance are, in order of decreasing importance:

D. Nelsonii. D. elongatum. D.

Geyerl, and D. Barbeyl. (2 7 )

Among the many other spedies

growing in Colorado, the following are known to be poison­ ous:

D. bicolor. D. Carollnianum. D. cucullatum. D. Penar-

dii, D. robustum. D. sapellonis. D. scaposum. and D. virescens. (17) IDAHO.

D. Menzlesii Is the most important poisonous

larkspur in this state. (23)

64.

INDIANA.

The occurrence of D. tricorne has been noted

in Indiana; It rarely poisons horses and apparently is harm­ less to sheep. (53) IOWA.

Poisonous larkspurs observed In Iowa are D.

Carolinianum. D. consolida. D. exaltatum Ait., and D. tri­ corne. (67) KANSAS.

Losses from larkspurs are Irregular In Kan­

sas and occur only in the spring when they are unusually abundant.

The chief species are D # trie o m e and D. vires-

cens. (26) MONTANA,

D # bicolor and

D,

cucullatum have been noted

to be causes of cattle poisoning, (79) ✓

NEVADA.

Feeding experiments showed that D. Andersonii.

which occurs in abundance in certain parts of Nevada, Is toxic to cattle. (81. 6 5 ) NEW MEXICO.

Johnson and Archer (££) state that four­

teen Delphinium species, several of them thought to be poisonous, occur in New Mexico. NORTH DAKOTA.

D. bicolor Is recognized as a cattle

poison In western North Dakota. (76) OREGON.

Delphinosis is responsible for more cattle

losses in Oregon than any other plant poisoning.

D. trolii-

folium causes most of the losses In the western part of the

65.

state while D. Menzlesii is the most important in eastern Oregon*

Other injurious species are D. bicolor. D. glaucum.

D # scopulorum. and D. simplex Dougl. (4 9 ) PENNSYLVANIA*

D. tricorne is found in the extreme

southwestern part of Pennsylvania but it is not abundant and is of minor importance. (50) WYOMING.

The most common of the Wyoming larkspurs Is

D, Geyerl: it Is responsible for more losses among cattle than are all the other poisonous plants of the state com­ bined.

D # Barbeyl and D* glaucum also cause heavy losses . ✓

Of lesser importance are the low larkspurs D. bicolor. j-J. *

*

'’

*

*

*

+

glancescens. D* Nelsonll. and D. venenosum. (.5, 6, 7, J35, 52)

5#

Therapeutic value D* A.jacis L.

The drug, Larkspur N.P. VI, consists of the dried ripe seeds of D* A.jacis»

Besides the alkaloldal content

the seeds also contain volatile and fixed oils, gums, res­ ins, and gallic and aconitic acids. (62)

The official prepa­

rations are Tinotura Delphlnli N.P. VI (Tincture of Lark­ spur) and Tinctura Delphlnli Ace tic a N.P. VI (Acetic Tinc­ ture of Larkspur).

The Tincture of Larkspur is made from

100 g. of the drug in moderately coarse powder form and sufficient alcohol to make 1000 cc.; It is used externally*

undiluted or diluted with an equal volume of water, as a parasiticide in pediculosis capitis and pubis.

The Acetic

Tincture of Larkspur (also known as Larkspur Lotion) con­ sists of 100 g. of the coarse powdered seeds, 50 cc. of aeetic acid, 100 cc. of alcohol, 50 cc* of glycerin, and water to make 1000 cc.; it also Is used (undiluted) externally as a parasiticide. (65)

Among physicians the acetic del­

phinium extract Is considered superior to the ordinary del­ phinium extract in removing vermin from the body, (78) Apparently some preparation of D. A 1acis was used In medieval England as a doubtful cure for the stings of scorpions. (68) The TJnground Larkspur Is described in the U. S. Dis­ pensatory (85) as Irregularly tetrahedral, acute at one end, obtuse or rounded at the other, about 2 mm. in length, and nearly as wide, surface black or blackish brown, occasionally grayish, with from 8 to 12 ridges transversely encirc­ ling the seed and forming'wavy, continuous, vertical walls or ruffles, occasionally inter­ secting, and with narrow channels between, seed coat crustaceans, endosperm whitish, fleshy and oily, embryo small, embedded in the fleshy endosperm. Odor very faint, taste bit­ ter, afterward biting and acrid.,.Epidermis of non-lignifled cells with thick, light yel­ lowish walls, some of the cells radially elon­ gated from the ruffle-like projections, a layer of cells containing a dark brown pigment, in­ ner seed coat composed of non-lignifled cells with thick, porous walls, occasionally cells near the micropyle containing a few starch grains, a large endosperm of thick-walled par­ enchyma, the cells filled with fixed oil and aleurone grains.

Likewise the Powdered Larkspur is described as Gray-brown, numerous fragments of endo­ sperm, 'aleurone grains up to 0.012 mm. in length, fragments composed of elongated epi­ dermal cells, the latter up to about 0.045 ram* in diameter and 0.300 mm. In length, groups of elongated cells from the Inner layer'of the seed coat up to 0.010 mm. in width, with characteristic beaded walls.

22* oo^solida L. Preparations from D. consol Ida were at one time used for their anthelmintic, diuretic, aperient, and. emmenagogic action; the flowers were used in the treatment of dysentery, dholera morbus, dropsy, gout, vesical affections, and vom­ iting of autumnal fevers; and the seeds for dropsy, spas­ modic asthma, and calculus. (61)

At the present time, how­

ever, the seeds and flowers are almost never employed as internal remedies, but are only used as a local application for

the destruction of lice in the hair. (83)

According

to White (88) a tincture of a mixture of D* consollda and Lobelia inflata (Indian tobacco) has enjoyed great popularity In the United States as a parasiticide.

D. scopulorum Gray The uses of the drug delphocurarine are the same as those of curare, which Is used da an antitetanic and a nerv­ ine because of its paralytic action on the ends of the motor nerves of the voluntary (striated) muscles.

Delphocurarine

does not act through the stomach and hence is used hypo­ dermically as an antidote for hydrophobia, tetanus, and strychnine. (61)

D. s_taphisagria L. The drug Staphisagria U.S.P. IX

or Staphisagrlae Sem­

ina B.P. consists of the ripe seed of D. staphisagria. the Stavesacre or Lousewort.

The seeds are also known as lice

grains or Stephans grains and contain, besides all the staph­ is agr la alkaloids, malic acid and fixed oil* (62)

Staves­

acre seeds have been used since the time of Pliny as a louse ✓

preventive. (41. 59)

Official preparations are:

Fluldextractum Staphisagrlae U.S.P. (Fluldextract of Staphisagria).

100 g. of the drug in No. 20 powder and

enought 95$ alcohol to produce 1000 cc. Lotlo Staphisagria B.P.C. Hair Lotion).

1 In 10.

(Stavesacre Lotion or Nursery

Employed as a lotion for children's

hair, being used to kill pediculi and their ova. Unguentum Staphisagria B.P.

(Stavesacre Ointment).

Stavesacre seeds, 20 parts; yellow beeswax, 10 parts; benzoated lard, 85 parts*

Used as a parasiticide to kill

pediculi capitis et pubis. (10) It has been found recently (41) that as antiparasitics delphinine and staphisagrine are markedly inferior to sabadilla (a drug containing veratrine alkaloids) In the form of tinctures,

Delphinine was formerly employed for its antlspasmodic and antlneuralgic action and was used for facial neuralgia, chronic rheumatism, otalgia (earache), convulsions, palpi­ tation of the heart, asthma, etc*

At present it is only

rarely used, being restricted to external application in pediculosis and occasionally in scabies*(81* 62)

A compar­

ison of the medicinal properties of ttdelphiniatt (extract from D* staphisagria) with those of "veratria" (veratrine alkaloids) when used as relief for tic douloureux and other neuralgic ailments is given by Turnbull* (82) The use of larkspur tinctures for destroying lice is not without undesirable effects; it is often accompanied by acute dermatitis resembling eczema, evidenced by redness and vesicles*

Delphinine itself is a mild vesicatory, pro­

ducing burning and inflammation when applied to the skin; ointments containing delphinine, when rubbed on the skin, causes burning, prickling, and a transitory redness, such as veratrine produces. (88)

It has been reported by Hal­

stead (52) that the herbage of larkspur causes a burning sensation*

4,

Insecticidal action D, A.jacis L,

The oil from D, AJacis seeds possesses insecticidal properties *

The alkaloid from the seeds also has slight

Insecticidal value; an extract containing 1% alkaloid and no oil was found to be one-tenth as active (against bedbugs) as samples containing a high content of oil, (89)

D» Brownli Rydb. The alkaloid of D. Brownli has been found to be prom­ ising as an insecticide for mosquitos and potato beetles. It Is not expected to excel nicotine sulfate as a contact insecticide but it may prove to be a superior stomach poison and feeding repellent, (77 )

£• consolida L, Larkspur oil (from D. eonsolida) in the form of soap emulsions is toxic to red spiders and aphids.

Delsoline

and delcosine are highly toxic to aphids and thrips, inef­ fective against red spider*, and of value as stomach poi­ sons against certain leaf-feeders, (14)

D. staphisagrla L. Stavesacre oil, as a soap emulsion, is toxic to red spiders and aphids.

Delphinine Is toxic to thrips but not

to aphids, red spiders, etc; it Is also effective as a stomach poison.

In general, delphinine Is much less toxic

than the consolida alkaloids. (14)

IV.

DISCUSSION OP RESULTS

The Alkaloids of D # Ajacis L, The plants used for the investigation of D. Ajacls alkaloids were collected In the vicinity of Athens, Gra,, by W. T. Sumerford (to whom appreciation is hereby expressed) in full bloom during the latter part of May^ 1941.

The

whole plants were dried in the shade for about three weeks and then ground to a meal.

Two methods were used to esti­

mate the alkaloidal content of the plants:

(l) by succes­

sive extraction with petroleum ether and chloroform, and (2) by extraction with methanol. Prom the petroleum ether extract of the plant material was obtained a 0,013# yield of an amorphous alkaloid, m.p, 85-95°, which Is precipitated from solution by ammonia. Prom the chloroform extract of the petroleum ether-extracted material were obtained two alkaloids:

(a) in 0,023# yield,

an amorphous alkaloid precipitated by ammonia or sodium hydroxide; and (b) in 0,11# yield, an ether-insoluble amor-

phous alkaloid which is not precipitated by sodium hydroxide but must be extracted from alkaline solution by chloroform. Thus the total yield of alkaloids by this procedxxre is 0,15#. The methanol extraction of the plant material was based on a procedure outlined by Manske (53) but suitably modified and extended for this particular case.

The total alkaloids

obtained by this method amounted to 0.22#, consisting of four fractions:

(a) in 0.003# yield, an amorphous alkaloid

(designated later as ECB), m.p. 90-100®, which is precipi­ tated by ammonia but can also be extracted from hydrochloric acid solution by chloroform (but not by ether); (b) in 0,042# yield, an amorphous alkaloid (EAB), m.p. 100-30°, which Is precipitated by ammonia but cannot be extracted from hydro­ chloric acid solution by chloroform;

(c) in 0.006^ yield,

an amorphous alkaloid (EPB), jirecipitated by potassium hydroxide but not by ammonia and not extracted from acid by chloroform; and (d) in 0.17# yield, a glassy dextrorota­ tory alkaloid (ESC), m.p. 50-90°, precipitated neither by ammonia nor by potassium hydroxide, but extractable from alkaline solution by chloroform.

In addition to the alka­

loids there was also found a considerable amount of mannitol, paralleling the experience of Manske (54) and others; the mannitol was Identified through its hexaacetate. There was no evidence of the presence of the ajacine or ajaconine reported by Keller and Volker (48, .§£)•

Alka­

line hydrolyses of the alkaloids obtained above gave only oily or gummy products,

B,

The Alkaloids of Annual Larkspur 1. Extraction of the alkaloids

Since Keller and Volker (4 8 . 8 5 ) had reported a rela­ tively high yield (1,8#) of alkaloids In the seeds of D # Ajacis and had obtained the simplest alkaloids yet reported present in Delphinium, species, It was thought that for the present investigation the commercially available D, Aiacis seeds would be excellent starting material for examination of Delphinium alkaloids. The Annual Larkspur seed used was supplied by Vaughan’s Seed Store of Chlcagv under the name D # Aiacis. but actually was composed of the Hyacinth Flowered, the Stock Flowered, and the Giant Imperial strains and therefore was a mixture of both D. Ajacis and D. cons olida varieties (see pp. 7-8). Some Annual Larkspur seed also was obtained from the FerryMorse Seed Co. of Detroit; no difference could be detected in the alkaloldal content of the seeds from the two differ­ ent sources.

Although the seeds had been sold as ’’inert”

or low-germinating, It was found by P. J. Bacon of Western Reserve University that on planting a 50# germination could be obtained; no report Is yet available concerning the plants, The method of obtaining the alkaloids from the seed consisted of three consecutive extractions using the solvents petroleum ether, chloroform, and methanol, in that order;

the essentials of the method were first devised in this Laboratory by A. W. Weston.

The petroleum ether extracted

about 25# of the total alkaloid, the chloroform about 50#, and the methanol about 25#,

Although the use of methanol

alone would extract most of the alkaloids, so much other material Is also extracted from the seeds by this solvent that the Isolation of the Individual alkaloids becomes very difficult; hence It is more feasible to confine most of the attention to the petroleum ether and chloroform extracts. These two solvents also extracted 30 to 33# yield of an oil from the seeds, which has been Investigated by Erick­ son (19),

The total amount of alkaloids obtained from the

seeds by the three solvents was about 1.2#. The alkaloids were removed from the petroleum ether and chloroform extracts with dilute sulfuric acid.

On addi­

tion of ammonium hydroxide to the acid extracts there was obtained an amorphous precipitate to which the name "ajacinine11 Is applied.

On extraction of the filtrate from the

ajacinine precipitation with chloroform and subsequent removal of the solvent from the chloroform extracts there was obtained as a glassy residue a mixture of alkaloids about half of which was soluble in acetone.

Hie acetone-

insoluble material was a new crystalline alkaloid, m,p. 197-9° (corr.), for which the name "delpholine” Is proposed. From the methanol extract of the seeds there was ob­ tained, on exchange of the solvent for dilute acid, by

ammonia precipitation an amorphous alkaloidal mixture re­ sembling ajacinine in appearance but differing from It some­ what on alkaline hydrolysis.

Also extracted from the seeds

by the methanol was a considerable amount of sucrose, which was identified through its octaacetate.

2,

Properties of Ajacinine

The ajacinine was a white amorphous powder which could not be obtained crystalline.

As first precipitated It usually

was contaminated with 5-15# of ammonium sulfate, which could be removed by prolonged washing with water.

Even with the

ammonium sulfate removed, the melting point was Indefinite, varying from 100-10° to 110-20° or higher.

The specific

rotation in chloroform varied from sample to sample between the limits +37° and about +50°.

Although ajaclnlne may be

considered soluble in the ordinary reagents such as acetone, methanol, ethanol, ether, chloroform, carbon tetrachloride, and benzene, there was usually a small resinous part of the material which was insoluble; ajacinine is only very slightly soluble in water and petroleum ether.

Because of the evi­

dently heterogeneous nature of ajacinine, It cannot be re­ garded as a definite compound, but only as a mixture of alka­ loids; that it is a mixture was further suggested by the ob­ taining of two bases on hydrolysis.

Attempts to separate

a3acinine into definite components, however, by fractional extraction and fractional solution were unsuccessful.

Although ajaelnine is a mixture,

its

elementary analy­

sis gave som e Indication of* the type of alkaloids present.

The analytical data corresponded to the empirical formula ^33^48®2^10or

^34^50^2^10*

Qualitative tests for phosphor­

us and sulfur were negative; no carhonyl groups were detected. Attempts to obtain crystalline salts of ajaclnine or of one of its components failed.

The following salts were ✓

obtained as amorphous powders, however: uene sulfonate, and trichloracetate.

hydrlodide, jo-tol-

The trichloroacetate \

was particularly Interesting; It consisted of a water-soluble component containing 7,87# Cl and 3.69# N, and a water-in­ soluble component containing 8,56# Cl and 4.03# N; each com­ ponent had practically the same melting point and specific rotation.

The atomic Cl:N ratio is also the same, 0.84, in

each case; this low ratio:

is at present unexplainable; the

ratio should be at least 1.5, assuming one molecule of tri­ chloroacetic to two at*ms of nitrogen, Ajacinine reacted with bromine and iodine either in aqueous or In carbon tetrachloride solution.

Bromine In

carbon tetrachloride, gave a substance which was probably a mixture of the hydrobromides of brominated and unbrominated ajacinlne. Ajacinine did not absorb hydrogen In the presence of a platinum catalyst; the alkaloid recovered from the attempted hydrogenation had all the properties of the original ajacininefc

Acetylation gave no clear-cut results.

No product

could be obtained with methyl sulfate.

Permanganate was

easily reduced by an acidic solution of ajacinine. The most Important reaction of ajacinine was its alka­ line hydrolysis.

Since analysis had indicated two nitro­

gen atoms per molecule, it was expected that one of the hydrolysis products might be anthrantlic acid as in the case of lycaconitine (p. 30) and the amorphous alkaloid from £• Brownli (p. 28).

As was expected, anthranilic acid was

found among the hydrolysis products; It was not isolated as such but was Identified by both the N-phenacyl- and the 3,5-dibromo- derivatives.

Although succinic or methylsuc-

cinic acid may also have been formed, as in the cases of A. lycoctonum and D. Brownii, no evidenceof the presence of either was obtained. Of the basic hydrolysis products there were two:

(a)

a white crystalline solid, m.p. 131-3°, (oC)^*^ +49,2° (abs. ale,); and (b) m 27 (ot)p +32.9

amorphous solid, m.p. 50-60°,

(abs. ale.).

Both products were fairly strong

bases; I.e., they were precipitated by sodium hydroxide but not by ammonia. The base melting at 131-3°, to which the tentative name ’’delphinoline" has been given, was shown to be containing three methoxyl groups; the mole­ cule of water was lost by heating at 100° in a vacuum, leav­ ing an amorphous mass tahich on contact with water again formed

*78*

th© crystalline delphinoline. be obtained*

Crystalline salts could not

The action of bromine gave a tr lbromo-compound,

either 0 2 8H32H 0 6BrS

or

C22H34N06BrS*

Delphinoline was found to be identical with the hydro­ lytic base (see pp. 28-9).

obtained by Manske (54) from D. Brownli The Identity was established by direct com­

parison of delphinoline with a sample of the D* Brownii com­ pound generously furnished by Dr, Manske.

When Manske 1 s

base was recrystallized from aqueous alcohol it formed needles melting at 131-5°; this melting point was not depressed on o admixture with delphinoline, m.p, 131-3 . The recrystallized Manske !s base had

+50,9° (abs. ale.) and gave the

same analysis for water as did the delphinoline from ajacinine. The resemblance between delphinoline and lycoctonine (p. 30) is striking; the melting point is the same, the optical activity practically the same, and the behavior on heating the same.

The chief difference is in the analytical

data, which require a larger molecule with four methoxyls for lycoctonine.

The analyses of the two compounds are

summarized in the following table.

' Sound, d H N MeO H20 Calc*, C H N MeO H 20

Delphinoline ^°^e ^3 *^2° leader Manske 61.61 62.12, 62.14 8* 56 8.62; 8,86 3*24' 3,30^ 3.22 22*64, 26*10, 29*26, 22*33 23.51, 25*23 5.5, 3*

Lycoctonine G21H27N03 ^ ^ e ^4*-®20 Schulze & Bierling 2.63,'2*75, 2.68, 2.94 24.45, 25,26, 25.02 4.02, 3.92, 4.09, 3.96

61.80 8*73 3*28 21*78 4.2

62.09 8*56 2.90 25;66 3*73

No simple alkaloid resembling ajacine was found in the extracts of Annual Larkspur seeds,

A critical analysis of

Keller and Volkerfs data for ajacine, however, throws doubt on the supposed simplicity of this compound#

Keller and

Volker found the molecular weights 491*8, 568#5, 591*2, 580*8, and 535.6 for ajacine in benzene and 341.5, 339.6, and 319.0 by titration. (85)

Evidently neglecting entirely

the high benzene values and not considering that the titt*ation values could be one-half of the molecular weight, they assigned to ajacine the formula C - ^ 0 4 *HgO, for which the calculated molecular weight is 297*3.

Yet for the salts

they obtained they found it necessary to ascribe formulas of the type (C15H 2 iN 0 4 )2 *HC1.2H20.

It seems evident that

the formula for ajacine should be approximately doubled# Recalculation of the analytical data for ajaclne suggests the formula ^ 3 2 ^ 5 0 ^ 2 ^1 0 *

indicated below.

80. A.jacine

C 5 B HgO Mol.Wt. Eq. Wt.

Pound (ave.) 60.99 8 *3 8

4.59 5.08 553.6 333.4

Calc, for C31H 46^2°8#8 H2° 60.96 8.25 4.59 5.90 610.7 305.4

Calc, for G15H21^°4*h2^ 60.60 7.80 4.71 6.06 297;5 297.3

The larger formula for ajacine would thus put it into the class of alkaloids represented by the amorphous alka­ loids lycaconitine, ajacinine, and the alkaloid from D. Brownii.

Thus, C31h 50N2°10

Ajacine

G34H 50^2°10

A^aclnine

^36^46^2^10

Iycaconitine

When ajacinine was treated with bromine in glacial acetic acid there was obtained a brominated product which on alkaline hydrolysis did not give delphinoline but did yield 3,5-dibromoanthranilic acid.

Hence at least part

of the reaction of ajacinine with bromine is due to bromination of the anthranoyl residue. The alkaline hydrolysis of the amorphous alkaloid re­ sembling ajacinine which was obtained from the methanol extract of the seeds gave, in addition to delphinoline and the other strong base which were obtained from ajacinine, a third compound, an amorphous weak base, m.p. 145-55°, apparently possessing the formula

Whether this

compound is actually a hydrolysis product or is one of the components of the starting material is at present undeter-

mined; it seems unlikely that a hydrolysis product would have such a high molecular weight.

3*

Properties of Delpholine

Delpholine, m.p. 197~9° (corr.), ( ° 0 p 3 + 5 4 .5 ° (chlor­ oform), is a strong, water-soluble base, requiring concen­ trated alkali for its precipitation. C 2 3 H 3 7 HO 7 groups.

It has the formula

(or possibly C2 3 H 3 5 HO 7 ), with three methoxyl Crystalline salts have not been obtained.

In con­

trast to ajacinine and delphinoline, in aqueous solution it did not give a precipitate with bromine water.

The action

of concentrated hydrobromlc acid on delpholine gave a com­ plex mixture of amorphous halogen-containing compounds, Delpholine was unaffected by the alkaline conditions which caused the hydrolysis of ajaeinlne. On treatment with acetic anhydride in the presence of sulfuric acid, delpholine gave two different amorphous acetates, apparently varying in the degree of acetylation. Acetate-1 was a weaker base and more non-polar than Act?t ate2, which was more nearly like the original delpholine.

The

solubility relationships of the alkaloid and the two ace­ tates are summarized In the following table.

Water Acetone Ether Alcohol Chloroform Carbon Tet* Benzene Pet* Ether

Delpholine ♦

Acetate-2 + +

m

m

+

+ + sl* + -

Hh

sl* sl*

Acetate-1 m

+ + + + + + m

Oxidation of delpholine in basic solution with potasslum permanganate gave a crystalline compound, m*p* 205-7° (corr*), still containing three methoxyls.

The

other products of the oxidation were not identified* Delpholine resembles in its solubility relationships and elementary analysis the alkaloid delcosine obtained from D. cons olida*

The formula

posed for delcosine, however*

agrees with none pro­ Furthermore delcosine is

levorotatory and biaxial negative while delpholine is dextro­ rotatory and biaxial positive# same compound* lowing table#

Hence the two cannot be the

Their properties are summarized in the fol­

8o .

-.-aDelcosine_______________ Delpholine KeYler(46) MTarkwoocL(55) t?Ionga(1 2 )________ ___ 193-7 198-9 198-9 197-9 Alcohol Chloroform Methanol Acetone Ether Water ( ^ ) d (c h c i 3 ) Crystal type Crystal shape Optical axes Found

C H N MeO

Formula Calc*

C H N 3 MeO M.W.

4#

sol. sol. sol. sol. sl. v.sl.

sol. sol.

sol. sol.

tm tm

•mm

sl. sol.

V.sl. v.sl.

f

mm

ortho rhombic 6-sided 6-sided plates plates or prisms biaxial neg.

mm

62.67 8.69 3.68 19.49

sol. sol. sol. v.sl. Ins ol* sol.

63.73 8.55 3.58

«54#41 mm+9 **'■*

+54.5 or th orhomb Ic prisms biaxial pos.

63 .96 ,63 .98 63.00,62.93 8.82, 8.62 8.22, 8.45 3.17 3.40, 3.24 18.45,19.00 19.96

C20H33N06 G21H33E06 62.63 63.77 8.66 8.41 3.66 3.54 24.27 25.53 383.5 395.5

C21H40N07 G23E37N07 64.20 62.83 8.84 8.49 3.00 3.19 19.90 21.18 467.6 439.5

Pharmacology of Ajacinine, Delphinoline, and Delpholine The following pharmacological properties were deter­

mined by C. C. Pfeiffer of Parke-Davls & Co# The intravenous minimum lethal dose of ajaclnine for rabbits was between 7#5 and 10 mg*/kg#

The alkaloid pro-

duced death by direct cardiac toxicity which resulted in ventricular fibrillation.

The animals continued to breath

for 30 to 60 seconds after the heart had stopped.

Delpho-

84.

line produced the same cardiac effect except that about twice the dose (17.5-20 mg./kg. ) was required to produce death. For delphinoline the intravenous minimum lethal dose was slightly above 40 mg./kg.

This compound appeared to

be more stimulating to the central nervous system than either ajacinlne or delpholine*

It had an accelerating effect on

the heart and was not depressant* None of the three compounds, in doses of

0*2

to 2 mg.

intravenously, had any effect on the blood pressure of the anesthetized dog.

These experiments, however, were carried

out before the toxicity studies were made, and hence the doses were probably all inadequate.

Since the action is

mainly cardiac It was not expected that further blood pres­ sure studies would elicit more than the expected effedt on blood pressure usually produced by changes in cardiac output. The Langendorff Isolated heart technique was employed to study the effect of the alkaloids on the mammalian heart, using veratrine as a control.

Veratrine produces In the

isolated heart a marked increase in rate and coronary flow and simultaneously the muscle contracts simulating a marked Increase In tone*

None of the three alkaloids, with the

possible exception of delpholine, produced a similar effect. Both ajacinlne and delphinoline caused a marked decrease

in

amplitude of the heart beat and a slowing of the heart rate.

Veratrine was successful In counteracting the toxic effect of ajaclnlne and delphinoline.

The possible veratrine-like

effect of delpholine was evident from the fact that an in­ creased rate and amplitude occurred*

The effect on coronary

blood flow Is summarized in the table below.

Alkaloid Ajacinine De lphinoline

Coronary Perfusion Before during 14 cc. 29 cc. 8

Delpholine »

16.0 3.6

Veratrine t! tl

11

8

7

After 1 1 cc.

10

2

16 3,5

19 10

19 18 17

12

3,5 12

It can be readily seen that all of the alkaloids and veratrine produce an Increase in coronary flow.

In the case

of ajaclnlne and delphinoline, however, the flow after the period of dilation is definitely less.

Delpholine is not

as potent as veratrine but Is less toxic.

C.

The Alkaloids of Perennial Larkspur

The Perennial Larkspur seed was obtained from Vaughan1s Seed Store of Chicago and was composed of the Elatum and Belladonna types.

Hence the seeds may be considered to be

in part, at least, descendant from D. elatum and D. cheil*>nthum. The extraction of the perennial seeds was carried out in the same manner as the extraction of the annual seeds* The acid extracts of the petroleum ether extract gave on neutralization with ammonia not an amorphous precipitate but an oil, which was extracted with ether.

Removal of the sol­

vent from the ether extract left a 0.27$ yield of an amor­ phous alkaloid, one-tenth of which was later obtained crystalline by slow evaporation from ether.

The crystalline

alkaloid, for which the name "elatine** is proposed, melts at 213-4° (corr.) and has the formula

(or possibly

G3 6 &S 5 8 Q9 )* containing four methoxyl groups in contrast to delpholine and delphinoline which contain only three methoxyls each.

Elatine differs also from the Annual Larkspur

alkaloids in that it Is levorotatory;

( °^)j^

-16.8° (chlor­

oform). Elatine resembles, and may be Identical with, tho crystalline alkaloid, m.p. 218°, from D. elatum described by Keller (47), to which he gave no name but ascribed the formula CggHgjNOg,

A comparison of the analytical results

is shown In the following table.

Keller*s Cmpd., m.p, 218° (see p. 58) ^ound(ave.) Calc. for C3 3 H 5 2 NO 8 c H B 4 MeO

66.69 8.69 2.35

67.20 8.72 2.38 21.04

Elatine, m.p. 213-4° (corr.) Found Calc, for Calc, for °35H53N09 C36H55N09 C 66.51 66.92 66.57 H 3.54 3.46 8.58 B 2.10 2.22 2.17 19.21 4 MeO 20.89 19.64

From the chloroform extract of the perennial seeds thero was obtained, by neutralizing the acid solution with ammonia, a 0*58$ yield of an amorphous alkaloid closely resembling o 23 ajacinine; the melting point was 1 1 0 - 2 0 and (o O p was +37° in chloroform. The other alkaloids of the Perennial Delphinium have not yet been investigated.

V*

EXPERIMENTAL DETAILS £• AlacIs L.

Petroleum Ether Extraction In an extractor of the Soxhlet type, patterned after the model described by Drake and Spies (16), was placed 155 g. of the ground whole plant (p. 71) and extraction with hot petroleum ether, b.p. 62-93°, continued until the fr^sh extracts were only slightly colored.

The petroleum ether

extract was treated with charcoal, filtered, and extracted with 5$ sulfuric acid.

The acid extract was freed of the

organic solvent by warming on the steam bath and then cooled and basified with concentrated ammonium hydroxide.

A cloudy

solution first formed, which on standing overnight yielded a precipitate settling to the bottom.

When this precipitate

was filtered off, washed with a little water, and dried in vacuo, it amounted to about

0*02

g. of a light tan amorphous

powder, m.p. about 85-95°.

The filtrate from the precipitate

was made strongly alkaline with sodium hydroxide and th^n extracted with chloroform.

The alkaline layer was discarded

and evaporation of the dried chloroform extract left a negli­ gible residue. The acid-extracted petroleum ether solution was extracted with

5

$ sodium hydroxide and the alkaline extract acidified

with hydrochloric acid and extracted with chloroform.

The

extracted acid layer was discarded and the sodium sulfate-

dried yellow-orange chloroform extract evaporated, leaving an amorphous residue of 0.05 g. The petroleum ether solution which had now "been extracted by both acid and base was decanted from some unfilterable gelatinous precipitate which had formed during the sodium hydroxide extraction, dried over anhydrous calcium chloride, and the solvent evaporated.

The greenish residue amounted

to about 1.5 g.

Chloroform Extraction The plant material which had been extracted with petrol­ eum ether was then thoroughly extracted with hot chloroform. The petroleum ether- and chloroform-extracted material on drying in air then weighed 150 g., a loss of 5 g. or 3$# The chloroform extract was treated with a little char­ coal, filtered, and extracted with 5$ sulfuric acid.

The

acid extract was freed of dissolved chloroform by warming on the steam bath and then cooled in ice-water and made basic with concentrated ammonium hydroxide.

A dark brown precipi­

tate formed, which was filtered off, washed with a very little water, and dried in vacuo.

The filtrate was made

more strongly alkaline with sodium hydroxide; on standing more of the dark brown precipitate slowly formed In this solution. 35

The combined precipitates when dry amounted to

mg. of a chocolate-brown amorphous powder (insoluble in

acetone or chloroform but soluble in water) which did not

melt, but only charred, when held on a spatula directly in a flame.

The alkaline filtrate was then extracted with

chloroform;

the aqueous layer was discarded and the chloro­

form was evaporated from the extract, leaving a brownish residue of 0.17 g*, which was practically insoluble In dry ether. The original chloroform extract from which the alkaloid had been removed by acid was then extracted with

5

$ sodium

hydroxide; the alkaline extract was acidified with hydro­ chloric acid

and extracted with chloroform.

The acid lay^r

was discarded; evaporation of the solvent from the dried chloroform extract left a negligible residue. The original chloroform extract which now had been ex­ tracted by both acid and base was dried and the solvent evaporated, leaving a black residue of

2.10

g.

Methanol Extraction Another sample of 476 g. of ground plant material was extracted with methanol, which removed 124 g. or 26.0$ of soluble material.

The cooled extract consisted of a darx

green methanolic solution (E) and some precipitated solid material (P) which was separated by decantation.

The

description of the investigation of the solution and precipi­ tate

which follows may be more clearly pictured by con­

sulting the accompanying schematic flowsheets, in which all the final alkaloidal fractions are underlined.

Investigation of the Methanol Solution (E§ The methanol solution (E), amounting to about one litor, was concentrated on the steambath, diluted with 500 cc. of hot water, and acidified with hydrochloric acid.

The rest

of the methanol was then removed by heating on the steam bath under slightly reduced pressure.

On leaving the acid

solution overnight In the Ice-box the Insoluble fatty sub­ stances (ER) settled to the bottom and the clear aqueous layer (ES) was poured off*

The residue (ER) was digested

in boiling 1% hydrochloric acid and cooled as before; the clear aqueous solution was added to (ES) and the black resi­ due (ERR) Investigated as described later. The acid solution (ES) was treated with a little char­ coal, filtered, and extracted with chloroform.

The chloro­

form extract (EC) was a greenish-yellow solution with a red­ dish fluorescence In more concentrated solution.

The acid

layer (EA) was decanted from a small amount of a black pre­ cipitate (EP), Insoluble in both layers, which formed during the chloroform extraction.

The Chloroform Extract (EC) The chloroform extract (EC) was concentrated on the steam bath, treated with charcoal, the solvent removed, and the residue boiled with 5$ hydrochloric acid.

The acid solu­

tion (ECS) was decanted from the Insoluble residue (ECR) extracted with ether. 5

and

The residue (ECR) was dissolved in

$ sodium hydroxide and the solution extracted with chloro-

Methanol Extract (E)f cone, on steambath, dil. with HgO, acidify, remove MeOH, let stand in cold

Residue (ER); digest In boil­ ing 1% HC1 Residue (ERR); see p. 9 9

Acid soln*: add to (ES)

CHCI3 extract tEcT? cone* on steam bath, treat with chare oa1 , remove solvent, boil with 5% Bpl

Free.' (BP); see p* 1 0 1

Acid soln.(ES); treat with char­ coal, extract with chci3

Acid layer(EA); basify, extract with CHClg

Prec*{EAP); wash with CHClg, add washings to (EAC)

Residue (ECR); dissolve in 5% NaOH, extract with CHCI3

Acidsoln, (EGS); see p* 94

CHOl* extract (ECCi); residue negligible

Alkaline layer (EAA); carbo­ hydrates

(Jhd!jz extrac *t (EAC); see p, 96

Alkaline layer (ECE)7 acidify with H61, no prec.

form*

the extract (ECC) gave a negligible residue on evapo­

ration; the alkaline layer dried; yield, 0.90 g.; m.p. 103-10°; +52.1° (CHC13 ).

(a£

114.

A solution of ajacinine in dry methanol or in absolute alcohol on treatment with sodium iodide in glacial acetic acid gave no precipitate, even on standing in the ice-box for six hours.

At the end of this time, dilution of the

mixture with dry ether caused the formation of a dark r3 (mol.,wt. 439.5): H, 8.49;

N, 3.19;

C, 62.83;

MeO, 21.18.

Chemical Properties of Delpholine An acid solution of delpholine is not precipitated by ammonium hydroxide and even a considerable excess of concen­ trated alkali must be used for its precipitation*

An aqueous

solution of the alkaloid gives no color reaction with ferric chloride solution*

Addition of Wagnerfs reagent to an acidic

or neutral solution of delpholine gives a voluminous daxk redbrown amorphous precipitate which Is easily soluble In thio­ sulfate or sulfite-sulfate solution. When a solution of trichloroacetic acid In carbon tetra­ chloride was added to a solution of delpholine In the same solvent a cloudy solution was first formed, but cleared im­ mediately, solvent.

The same result was obtained using benzene as Addition of trichloroacetic acid to an aqueous solu­

tion of delpholine gave a small yield of a heavy oil. The salt with jy- toluene sulfonic acid prepared in chlor­ oform appeared to be an oil.

Several attempts to prepare a

picrate were unsuccessful, using a.lcohol, acetone, and water as s olvents•

A solution of delpholine in carbon tetrachloride gave an immediate precipitate with bromine in carbon tetrachlor­ ide;

the washed and dried product was a light yellow amor­

phous powder with no definite melting point (beginning to decompose at about 120°);

it was almost completely soluble

In water and the aqueous solution gave a precipitate with silver nitrate solution. No precipitate was obtained by the addition of bromine water to an aoueous solution of delpholine or by the addition of bromine in glacial acdtlc acid to a solution of delpholine in glacial acetic acid in the presence of sodium acetate.

Attempted Hydrolys is of Delpholine A solution of 1.00 g. of delpholine and 2 g. of potas­ sium hydroxide in 25 cc. of methanol was refluxed for 12 hrs., the reaction mixture then diluted with 10 cc. of water, and the methanol evaporated.

During this evaporation a white crys­

talline solid separated out, which on filtration, very slight washing with water, and drying amounted to 0.88 g., identical in physical properties with the starting material.

Action of Hydrobromic Acid on Delpholine fL solution of 0,2 g. of delpholine in 5 cc. of concen­ trated hydrobromic acid was refluxed for 8 hours.

The dark

brown reaction mixture was then poured into 20 cc. of water, forming a dark greenish-brown precipitate which was separated

by centrifugation and washed with water by decantation,

The

precipitate when dry was a blue-black amorphous powder, 0.05 g.

It did not melt when held on a spatula directly in the

flame of a microburner,

It gave a strongly positive test

for bromine (Beilstein test).

It was fairly soluble in

water, giving a yellowish-brown solution.

The aoueous solu­

tion gave no precipitate with bromine water or silver nitrate, but did give a precipitate with Wagnerfs reagent.

The com­

pound was also soluble ih methanol and slightly soluble In absolute alcohol, but insoluble in acetone, ether, chloro­ form, carbon tetrachloride, petroleum ether, benzene, dioxan, carbon disulfide, trichloroethylene, methyl acetate, and ethyl acetate. The filtrate (dark brown) and washings from the above precipitate were thoroughly extracted with chloroform (very little of the color was removed from the aoueous layer). The yellow extracts were dried over anhydrous sodium sulfate and the solvent evaporated, leaving a non-crystalline residue of 0,01 g, The chloroform-extracted acid solution was freed of chloroform by warming on the steam-bath and then (after cool­ ing) made basic with ammonia, giving an unfilterable colloidal precipitate.

On extracting with chloroform the precipitate

coagulated and was filtered off.

The filtrate was made more

alkaline with sodium hydroxide and the extraction continued. The extracts were dried and the solvent evaporated, leaving

a glassy reddish-brown residue of 0,03 g. The washed and dried coagulated precipitate amounted to 0.03 g, of a brown amorphous powder. flame on a spatula It charred. Beilstein test for bromine.

On holding in taie

It gave a strongly positive

It was somewhat soluble in water,

giving a precipitate with Wagnerfs reagent but none with sil­ ver nitrate or with bromine water.

It was slightly soluble

in chloroform and absolute alcohol; very slightly soluble In acetone; and insoluble in ether, benzene, and petroleum ether. The chloroform-extracted alkaline filtrate when acidified still gave a precipitate with Wagnerfs reagent.

Action of Hydrochloric Acid on Delpholine A solution of 0.2 g. of delpholine in 5 cc. of concen­ trated hydrochloric acid was refluxed for 8 hrs, and the reaction mixture (dark brown) then poured into 20 cc. of water.

No precipitate was obtained even after standing sev­

eral days.

In view

of the complicated results obtained from

the hydrobromic acid treatment, this experiment was not further continued.

Acetylation of Delpholine (a) A solution

of 0.3 g, of delpholine and 0.15 g. of

sodium acetate in 5 cc. of acetic anhydride was heated on the steam bath for 24 hours.

When the reaction mixture was

then poured into water, the anhydride allowed to hydrolyze,

and the solution, then, neutralized with ammonia, only gummy products were obtained. (b) A solution of 0.23 g. of delpholine in 2 cc. of acetic anhydride was refluxed for 90 min. and then poured into 15 cc. of water*

When the excess anhydride had hydro­

lyzed the solution was made alkaline with ammonia, giving an immediate amorphous, slightly gummy, precipitate.

On filter­

ing, washing with w^ter, and drying there resulted 0.12 g. of a yellowish amorphous powder, m.p. about 110-5° (with soft­ ening below 110°).

Addition of sodium hydroxide solution to

the filtrate gave about 0.02 g. more of the precipitate. The alkaline filtrate and washings were then thoroughly ex­ tracted with chloroform; evaporation of the extracts left a glassy residue of 0.05 g, (c) A solution of 0*50 g. of delpholine in 2,5 cc. of acetic anhydride containing a drop of concentrated sulfuric acid was refluxed for 30 min* and then heated on the steam bath for 18 hours.

On pouring the reaction mixture into

25 cc. of water it all went into solution immediately. Addition of a^idnia to the aqueous solution gave no precipi­ tate.

Concentrated sodium hydroxide was then added, giving

a precipitate which on fl.ltering, washing with water, and drying amounted to 0*16 g. of an amorphous powder, m.p* 105-15°•

The filtrate was thoroughly extracted with chlor­

oform and the extracts evaporated, leaving a glassy residue of 0.20 g.

On dissolving this residue in benzene and pour-

138

ing the solution into petroleum ether it was precipitated as an amorphous powder melting at 100-5° (dec. 137°) when

dry. The material obtained in (b) and (c) by precipitation with ammonia or sodium hydroxide will be referred to as Acetate-1, while that obtained from the chloroform extracts will be designated as Acetate-2, Properties of Acetate-1,

Very slightly soluble in cold

water (can be detected with Wagner's reagent); a little more soluble in hot water; insoluble in petroleum ether.

Soluble

in acetone, alcohol, benzene, ether, chloroform, and carbon tetrachloride.

It is not precipitated from acid solution

by ammonia except from concentrated solution.

It is precipi­

tated from acid solution by Wagner's reagent; an acid solu­ tion of the acetate gives no reaction with bromine water. A carbon tetrachloride solution of the acetate gives a volumi­ nous precipitate with bromine in carbon tetrachloride.

An

acid solution decolorized permanganate. Properties of Acetate-2.

Insoluble in ether and petroleum

ether; slightly soluble in carbon tetrachloride. water, acetone, benzene, chloroform, and alcohol.

Soluble in An aqueous

solution gives no reaction with bromine water but does give a precipitate with Wagner's reagent.

A ehloroform-carbon

tetrachloride solution of the acetate gives a voluminous amor­ phous precipitate (yellow) with bromine in carbon tetrachlor­ ide,

An acid or neutral so3.ution decolorizes permanganate.

Oxidation of Delpholine A solution of 0.5 g. of delpholine, 0.1 g. of potas­ sium carbonate, and 0.5 g. of potassium permanganate in 100 cc. of water was refluxed for one hour, cooled, and the manganese dioxide destroyed by addition of sodium bisulfite solution.

On standing a short time white crystals began to

deposit from the solution, which was now slightly acid to litmus.

After standing overnight the crystals were filtered

off; weight, 0.27 g.

These crystals were shown to be man-

ganous sulfite by the following tests: melt when held in the flame.

The material did not

It was very slightly soluble

in water, but easily soluble in dilute hydrochloric acid. The neutralization of -the acidic solution with sodium hydrox­ ide gave an amorphous white precipitate (Mn(OH)g) turning brown.

soon

An ammoniacal solution of the material gave

a pink amorphous precipitate (MnS) with sodium sulfide solu­ tion.

Addition of alkaline hydrogen peroxide solution to

an aqueous solution of the material gave a dark brown pre­ cipitate (MnOg) and a yellowish-brown solution. cation was identified as manganese.

Hence the

The only two insoluble

manganous salta possible under the conditions of the reaction were the carbonate and the sulfite.

The amount of potassium

carbonate (0.1 g.) originally added would not permit the formation of 0.27 g. of manganous carbonate; furthermore dilute acid caused no evolution of carbon dioxide from the material.

Concentrated acid, however, did cause the evo­

lution of a gas which formed a precipitate in a drop of

barium hydroxide solution*

The addition of a drop of barium

chloride solution to a solution of the material In dilute hydrochloric acid caused only a cloudiness; if to the acid solution, however, was first added several drops of potas­ sium permanganate solution (instantly decolorized) and then a drop of barium chloride, a definite white precipitate (BaSO^) was formed*

Hence the anion was Identified as sul­

fite* The filtrate from the manganous sulfite was evaporated on the steam bath to a volume of about 30 cc. and made alka­ line with potassium hydroxide.

The precipitated manganese

hydroxide, which had turned dark on standing, was filtered off and dried; that no alkaloid had been precipitated along with the hydroxide was shown by digesting the dried precipi­ tate with chloroform; evaporation of the filtered chloroform left no residue.

The reddish filtrate from the manganese ✓

hydroxide was extracted with three portions of chloroform, the colorless extracts dried over anhydrous sodium carbamate, and the chloroform evaporated; residue:

a clear, almost

colorless glass, 0.27 g., m.p. 160-75°,

A further extraction

of the alkaline solution with three more portions of chlor­ oform yielded no further product. The extracted alkaline solution (orange-yellow) was acidified with hydrochloric acid (no precipitate formed); the acidic solution still gave a slight precipitate with Wagner's reagent.

The acidic solution was extracted first

with three portions of ether and then with three portions of chloroform; hoth extracts were colorless and on evapo­ ration of the solvent neither left as much as 0,01 g. of residue*

The extracted acidic solution was then evaporated

to dryness and the residue (mostly Inorganic salts) ground to a powder.

The powder was extracted with acetone (Soxhlet

extractor) until an aqueous solution of the extracted resi­ due gave no precipitate with Wagner's reagent.

The solvent

was then evaporated from the yellow acetone extract, leaving a yellowish-brown oily residue not completely soluble in water or alkali* Properties of the Oxidation Product.

The glass, m.p,

160-75°, which was the main product of the oxidation, was soluble in water, acetone, alcohol, and chloroform; slightly soluble in benzene and carbon tetrachloride; and insoluble in ether, Analysis (by T. S. Ma): Found: C, 58.52; H, 7.45; N, 3.22; Calc, for C18H24H 05(0Me)3 : 0, 58.99; 3.28; MeO, 21.78.

MeO, 21.64.' H, 7.78; H,

The aqueous solution of the compound gave no color with ferric chloride solution, did not decolorize potassium permangante solution, and did not decolorize or give a precipi­ tate with bromine water.

The aqueous solution, however, did

give a copious precipitate with Wagner's reagent, and a chloroform, carbon tetrachloride, or benzene solution gave an amorphous precipitate with, bromine in carbon tetrachloride.

When the compound was dissolved in absolute alcohol, dry ether added, and the solution allowed to evaporate slow­ ly, crystals were obtained melting at 204-7° (corr.),

Crys­

tals could likewise be obtained from a mixture of dry meth­ anol and isopropyl ether; m.p. 205-7° (corr.), with soften­ ing several degrees below*

C*

Perennial Larkspur

Petroleum Ether Extraction of the Seeds The seeds, 3710 g., were finely ground and extracted with petroleum ether (Skelly Solve "B", hexanes, b.p* 6071°); the petroleum ether solution (about 6 1.) was then ex­ tracted with 5 % sulfuric acid until the acid extracts showed ohly a faint reaction with Wagner's reagent; five 400 cc, portions of acid were required.

In order to remove any non-

basic material the two liters of acid solution were then extracted first with three 100 cc. portions of ether and then with three 100 cc. portions of chloroform.

Evaporation

of the dried chloroform extracts left a yellowish residue of 0.1 g.; evaporation of the dried ether extracts left a negligible residue.

The extracted acid solution was then

freed of organic solvents by warming on the steambath and then cooled In ice water and neutralized with concentrated ammonium hydroxide*

This treatment caused no precipitate

but only a milky solution with an oil slowly settling to the

bottom*

The milfcy solution was extracted with three 100 cc.

portions of ether and then with three 75 cc. portions of chloroform.

Evaporation of the ether extracts (which showed

a violet fluorescence) at reduced pressure left a light yellow glassy residue of 10,1 g.; evaporation of the chlor­ oform extracts left an orange glassy residue of 0*8 g., insoluble in ether.

The extracted ammonia solution was

made more strongly basic with potassium hydroxide and ag&in extracted first with three 85 cc. portions of ether and then with three 90 cc. portions of chloroform*

Evaporation of

the dried ether extracts left a residue of 0.1 g. and evapo­ ration of the dried chloroform extracts also left a residue of 0.1 g.

The extracted alkaline solution when acidified

gave no precipitate with Wagner’s reagent, showing that all the alkaloid had been extracted* Examination of the Ether-extracted Alkaloid The 10,1 g. glassy residue obtained above when pul­ verized melted at about 80-100° and was soluble in acetone, benzene, and alcohol; only moderately soluble in ether; in­ soluble in water and petroleum ether.

On dissolving the

material in ether and allowing the solvent to evaporate slowly at room temperature there was obtained 1*0 g. of a white crystalline solid melting at 203-5°; repeated washing with ether raised the melting point to a constant value of 212-4° (corr.),9 with softening several degrees below.

Finally, *

recrystallization from aqueous alcohol gave pure elatine, m.p* 213-4° (corr.).

Properties and AnalysIs of Elatlne The elatlne, m.p. 213-40 (Corr.), needles or prisms, crystallizes in the orthorhomhic system and is "biaxial neg­ ative, with 2v large (crystallographie examination by A. L. Howland);

( o O ^3 „i6.8° (CHC13 ).

Analysis {by T. S # Ma): Pound: C, 66*51; H 8*54; N, 2.10; Calc* for ^31^41^05(Wfe)^: C, 66*57; 2.22; MeO, 19*64. Calc, for Cs2H43N05(0M©)4 : C, 66.92; 2.17; MeO, 19,21*

MeO, 20.89.' H, 8.46; N, H, 8.58;

N,

Chloroform Extraction of the Seeds The petroleum ether-extracted seeds were then extracted with chloroform and the chloroform extract (about 6 1.) ex­ tracted with six 400 cc. portions of

sulfuric acid (some

difficulty with emulsions encountered here); the last ex­ tract still gave some reaction with Wagnerrs reagent and a slight cloudiness with concentrated ammonium hydroxide.

The

combined acid extracts were light yellow in color; a test showed that ether did not extract the colored substance and hence the acid solution was not washed with ether or chloro­ form.

The acid extracts were then freed of chloroform by

warming on the steam bath, cooled in ice water, and made basic to litmus with concentrated ammonium hydroxide, giving a voluminous white precipitate which was filtered by suction washed three times with water, and dried over calcium chlorldG at 15 mm.

The dried precipitate, 21.5 g., was contami-

nated with about 4$ of ammonium sulfate as shown by the solu­ bility in chloroform; the melting point was indefinite, about 110-20°;

(c*)^3 -^370 (CHCI3),

A solution of 20.5 g. of the crude amorphous alkaloid in 600 cc. of dilute hydrochloric acid was made basic with ammonia and extracted with three 80 cc. portions of ether and then with one 60 cc. portion and six 40 cc. portions of chloroform.

The combined ether extracts (yellow, show­

ing a strong violet fluorescence) were dried over anhydrous sodium sulfate and the solvent removed under reduced pres­ sure, leaving a glassy residue of 16.9 g.

The dried chlor­

oform extracts left a residue of 1.5 g. on evaporation.

VI.

SUMMARY

The whole plant of D, A.jacis L.

was found to contain

0.22# alkaloids. Prom the seeds of Annual Larkspur, consisting of vari­ eties of the species D, A.lacis L. and D. consolida L.

was

obtained 1.2# total alkaloids, from which the amorphous alkaloidal mixture ajacinine and the crystalline alkaloid delpholine were isolated.

Alkaline hydrolysis of ajacinine

gave anthranilic acid and the crystalline compound delphinoline, CjgH2gR03(0MeJg.HgO, m.p. 131-3°, identical with a compound obtained by Manske from the hydrolysis of an amor­ phous alkaloid from D. Brownii and very similar to lycoctonine*

The alkaloid delpholine, CgQHggRO^(OMe)3 , m.p. 197-9°

(corr.), on alkaline permanganate oxidation gave a crystal­ line compound, C^gHg^NOgfOMe)g, m.p, 205-7° (corr.). From the seeds of Perennial Larkspur, probably descended from D, elatum L, and D. cheilanthum Fischer, was obtained a crystalline alkaloid, elatine, G^^H^^HOgfOMe)^, m.p. 213-4° (corr.). The alkaloids ajacinine and delpholine differ from delphinine in that their toxic action is due to ventricular fibrillation rather than asphyxiation.

Delpholine resembles

veratrine in its effect on coronary blood flow in the iso­ lated mammalian heart but is less potent and less toxic,

Delphinoline has an accelerating effect on the heart and is not depressant; it is more stimulating to the central nervous system and less toxic than either ajacinine or delpholine.

Ajacinine is more toxic than delpholine hut

less toxic than delphinine or aconitine.

BIBLIOG RAPHY

p.) (2 ) (£) (4t) (6) . Dispensatory. 22nd Ed*, pp. 389-90 (Philadelphia: tippincott, 1937). 84 Vaughan’s Seed Store Catalog, (1942). 85 VOLKER, 0*: Dissertation, Univ. of Marburg,(1913)» 86 WALZ, T.: Arch, Pharm. 260. 9-26 (1922). WAUGH, W. F. and ABBOTT, W. C.: A Text-Book of -Alkaloidal W Therapeutics. 3rd Ed., pp* 341-7 (Chicago: The Abbott Press, 191lT.

s

«•* *

(88) WHITE, J. C.: Dermatitis Venenata, on. 119-20 (Bos-hrm* Cupples & Hurd, 1887)* ‘ £§2) WILLIAMS, J • B, s Am, J, Pharm, 86, 414-6 (1914)* (90) YAVEL*BERG, G, I,; Sovet. Veterinarlya 1938. Ho, 3 75-6; Khim, Referat, Zhur, 2 , Ho, 5, 59-60 (1939) Chem, Ahs, 34, 1091 (1940)* (91) ZEMELEN, G, and PACSU, E . : Ber, 62, 1613-4 (1929)*

PART TWO

HALOGENATION OP 1-MENTHONE

154 #

I*

HISTORICAL INTRO DUCTIO N

bromination of 1—menthone was first studied in 1896 4t "by Beckmann and Mehrlander* One mole of menthone in chloro­ The

form solution reacted with two moles of bromine, giving a brown oil with the empirical formula CioH^BraO.

This compound

filmed in moist air and gradually decomposed with the evolution of hydrogen bromide*

Beckmann and Mehr lander believed that the

tribromoraenthone contained only one substituted bromine atom, and that the other two bromine atoms formed an addition compound:

C 1oHjL7BrO«Br2 .

Beckmann and Eickleberg

3

found that one mole of d- or 1-

menthone in chloroform solution quickly decolorized two moles of bromine, giving a red-brown oil which on standing in air gave off hydrogen bromide and soon began to crystallize5

the

pure crystals were colorless, stable in air, and melted at 79-80°5 the elementary composition corresponded to the formula CloH 16Br20; [oc]j) in carbon tetrachloride solution was +199*4°. When the dibromomenthone was dissolved in ether and gaseous hydrogen bromide introduced, the red-brown oil, CioHi-yBrsO, of Beckmann and Mehr lander was obtained, which on standing in air reverted to the dibromo-compound with the evolution of hydrogen bromide.

Semmler

13

assumed that two hydrogen atoms

in the ring had been replaced by bromine and that a molecule of hydrogen bromide was loosely associated with the oxygen atom, thus: C loHi6Brs0*HBr, rather than the C lOHi7Br0-Br2 formulation proposed by Beckmann and Mehrlander*

When only one mole of bromine was allowed to react with one mole of menthone, part of the menthone remained unchanged, and the only product obtained was the dibromo-compound. The dibromomenthone was not affected by zinc dust and alcohol, showing that the two bromine atoms were not on adjacent carbon atoms*

Menthone was regenerated by the action

of zinc dust and acetic acid on the dibromide.

When one mole

of dibromomenthone was boiled with six moles of quinoline, thymol was obtained*

From these data, Beckmann and Eickleberg

proposed the following structures and reactions:

o

+■

2. 8

»

o

h-

2 H Br

Dibromomenthone

O

+ 2 Hdr

Ketone form of thymol

OH

'Thymol The intermediate ketone form of thymol was not isolated. Wallach

17

found that the dibromomenthone could be obtained

better and in greater yield by adding bromine quickly to a well-cooled solution of mehthone in acetic acid.

In this v/ay

a 75$ yield of dibromomenthone crystals were obtained with

156. 12 9 M

D

~~

ether solution.

He also observed that thymol

could be obtained by merely refluxing the dibromomenthone 5 a violet coloration accompanied the vigorous evolution of hydrogen bromide. Treatment of dibromomenthone with potassium hydroxide solution gave three products;

diosphenol (I), an isomeric

unsaturated hydroxy-ketone (II), and an acid (III) probably formed from (II).

6." ic (I)

(II)

C 00H

(III)

2 ,4-Dibromomenthone

From the above results, Wallach concluded that in di­ bromomenthone the bromine atoms were in the 2,4-positions, rather than in the 1,4-positions proposed by Beckmann and Eickleberg.

Wallach considered that the obtaining of thymol

from dibromomenthone was no argument for the 1,4-positions, since thymol can be obtained also from 1,2-dibromomenthone and from 4,5-dibromomenthone*

According to Oddo,

10

when the

bromination of menthone is carried out in alcoholic solution, thymol is also produced in addition to the dibromide. Baeyer and Seuffert

2

found that eight moles of bromine

could be absorbed per mole of well-cooled menthone in the absence of any solvent.

The chief products were hexabromo-

thymol (IV) and smaller amounts of tetrabromo-m-cresol (V). With alkali, (IV) split off hydrogen bromide with the formation of a red-colored substance; even a slight trace of water in

157 an ether solution of (IV) caused decomposition.

The product

was pentabromodehydrothymol (VI), which in the presence of alkalis again split off hydrogen bromide, forming tetrabromodimethylcumarone (VII). Or

£

off

ott

(IV)

(V)

(VI)

K & t z and Steinhorst

(VII)

obtained a monobromomenthone by

bubbling bromine vapor in carbon dioxide gas through an equimolar amount of menthone in the presence of an aqueous sus­ pension of calcium carbonate.

In this way a 40$ yield of

4-bromomenthone boiling at 120-2-j

resulted.

The bromine

was shown to be in the 4-position since the reaction with sodium acetate in boiling acetic acid gave 4-menthenone. Cusmano

?

reported that a chloroform solution of 4-bromo-

menthone reacted with an equimolar amount of bromine, giving the 2,4-dibromomenthone, which, when shaken with 2.5$ potassium hydroxide solution, gave diosphenol and other products. KiJtz and Steinhorst (b.p.,

9

also obtained 4-chloromenthone

in 40$ yield by passing chlorine into

menthone in the presence of aqueous calcium carbonate.

By

heating 4-chloromenthone with potassium hydroxide solution, 4-hydroxymenthone was obtained, an oil boiling at 14 and giving an oxime melting at 131-3°. Wagner prepared 4—hydroxymenthone by the oxidation of menthene with potassium permanganate 5 he reported the boiling point as 104 *5-105# 5-^

158. and the melting point of the oxime as 132-3°. &

Recently Cornubert and Humean

investigated the chlorination

of menthone using the calcium carbonate method of K5tz and Steinhorst.

From 200 g. of menthone was obtained 54 g. of

4-chloromenthone boiling at 113-20^0^#fthe analysis showing 16.1$ chlorine instead of the calculated 18.30$.

Cornubert

and Humeau also obtained a dichloromenthone melting at 64.565°, very soluble in alcohol.

When pure, it did not change

at ordinary temperature, but heating at mild temperature in alcohol solution caused hydrogen chloride to be eliminated and the solution to turn violet.

When Cornubert and Humeau

treated their impure 4-chloromehthone with methyl magnesium iodide, neither a pure methylmenthone nor a pure chloromenthol was obtained.

They reported a ketone formed in poor yield

which boiled at lC^-K^inmu and contained 2.7$ chlorine.

159

II. A.

DISCUSSION OF RESULTS Elimination of 1-Menthono

The work of Beckmann and Eickleberg3 and of Wallach17 on dibromomenthone was verified by treating menthone in acetic acid with two molar proportions of bromine o

Pure crystals of

2 ,4-dibromomenthone were obtained melting at 79-9.5° (corr.), which after standing for several weeks in a stoppered testtube slowly decomposed with the formation of a violet colored mixture. Several attempts were made to obtain 4-bromomenthone in good yield by reacting equimolar proportions of menthone and bromine under varying conditions and then fractionally distilling the products at pressures of 20 to 30 mm. the results 5

Table I summarizes

the yields shown In the third column of this

table are calculated from the weights of the fractions boiling in the range corresponding to that reported by K6tz and Steinhorst9 for 4-bromomenthone.

Redistillation and further

investigation of these fractions, however, revealed that they consisted not at all of 4-bromomenthone, but of a liquid 20

having a specific gravity of 0.9632O and containing only 1.3$ bromine5 at least part of this liquid was shown to be thymol.

The following alternatives are then possible:

(1)

the original fractions contained very little 4-bromomenthone, or (2) 4-bromomenthone was actually collected originally but on standing and redistilling it decomposed, losing bromine in the form of hydrogen bromide*

160# Table I Solvent I. II. III. IV.

AcOH AcOH CC14 H 20 + CaC0s

Temp* Room temp 17° 0° 10 °

Bromomenthone range

Recovered menthone

8$ 15 8 12

37 62 69

4 -Bromomenthone was finally obtained in 33$ yield by slow bromination at 5-10° in the presence of water and calcium carbonate; slightly less than the molar equivalent of bromine was used and the products were first rapidly distilled at 5 mm* pressure and then more carefully fractionated at 4 mm.;

the

4-bromomenthone was obtained in 96$ purity (the bromine analysis was about 2$ low) in the fraction boiling at 90-2° at 4 mm.

This fraction had n^° 1.4945 and M d 5 + 92.7° in 3.13$

carbon tetrachloride solution. A quantitative investigation was then undertaken in an attempt to explain the low yields of 4-bromomenthone and to gain an insight into the mechanism of the bromination. If the bromine first adds to the menthone forming an addition compound, C loHi7Br0*Br2, as proposed by Beckmann and Mehrlander, the loosely bound bromine atoms should be available as an oxidizing agent.

This hypothesis was tested

by mixing small measured equimolar amounts of bromine and menthone in carbon tetrachloride and determining the bromine remaining available for oxidation by shaking the carbon tetrachloride solution with aqueous potassium iodide solution and measuring the liberated iodine with standard thiosulfate. Thus from 50 millimoles each of menthone and bromine, only

161. 0.25 millimole of iodine was found*

that is, only 0.5# of the

original bromine remained available as an oxidizing agent. During the reaction, the bromine was instantly decolorized, but only 2.1 millimole of hydrogen bromine was evolved, as determined by collecting it in water and titrating with standard base. Further enlightening experiments were performed using 50 millimoles of bromine and checking the amount of hydrogen bromide at various steps of the run.

The conditions of temp­

erature, concentration, etc., used in the attempt to obtain 4-bromomenthone by bromination in carbon tetrachloride solution were simulated as nearly as possible.

The hydrogen bromide

was measured at three different places: (1) that evolved during the actual addition of bromine*

(2) that remaining in the

carbon tetrachloride solution, but which could be extracted with cold water;

and (3) that evolved during the distillation

at atmospheric pressure of the dried, extracted carbon tetra­ chloride solution.

The results are summarized in Table II. Table II

Menthone used (mmoles) I. II. III.

50 50 25

Evolved

HBr found (mmoles) Extracted Distilled

■ — -----29--------2.7 21.5 9 26

60 46

Total 49 78 84 81

If the bromination were a normal substitution reaction, 50 millimoles of hydrogen bromide would be expected to be given off during the addition of bromine*

however in all

three cases approximately only one-half of this amount was formed9 and most of that remained dissolved in the carbon

162. tetrachloride, hut could he removed hy water extraction* Since all the hromine was decolorized, and yet practically none of it remained available for oxidation, the mechanism must he something similar to:

This equation accounts for the formation of the observed amount of hydrogen bromide and at the same time includes the compound CioHj.7Br30 which was isolated hy Beckmann and Mehr lander and the structure of which is the same as that proposed hy Semmlero The existence of the loosely-coupled hydrogen bromide addition compound is further evidenced hy the evolution of hydrogen bromide during distillation.

The amount of hydrogen

bromide obtained during distillation corresponds to the equation: CioHi7Br30

— >

2 HBr + C loH 15Br0

It seems improbable that any raonobromide formed would be decomposed to the menthenone*

since this reaction has been known to take place only in the presence of sodium acetate in acetic acid. reaction nor the following

Neither this

accounts for the evolution of enough hydrogen bromide.

The

main reaction then must be a partial conversion of the tri— bromo compound into thymol.

This reaction seems plausible

since about 80—85/^ of the bromine was recovered as hydrogen bromide and only small amounts of brominated ketone were ob­ tained in the larger runs.

Or O' HBr

B.

Reactions of 4 -Brornamenthone

Hydrolysis of the 4-bromomenthone was attempted by refluxing it in bQfo aqueous acetone, but after nine hours only 5% was hydrolyzed as detected by titration of the hydrobromic acid formed.

This slow rate of hydrolysis may be at least

partly due to the comparatively low reaction temperature. The action of concentrated potassium carbonate solution on 4-bromomenthone gave chiefly a liquid which on the basis of its physical properties was probably 4(5)-menthenone, but no oxime or semicarbazone of this latter compound could be obtained.

There was no evidence of the formation of 4-hydroxy

menthone although this hydroxyketone was obtained by KStz and Steinhorst9 on the alkaline hydrolysis of 4-chloromenthone.

164.

Although K 6 t z and Steinhorst9 reported that heating a 4-halomenthone with sodium acetate and acetic acid, followed by a 15-minute treatment with sodium hydroxide, gave 4(5)-menthenone, in the present case this procedure yielded from 4-bromomenthone only the acetate of 4-hydroxymenthone, which required several hours further treatment with sodium hydroxide for saponification to the hydroxyketone •

The ester has never

been reported before in the literature and the only proofs that can be offered for its identity are its method of prep­ aration, its reaction with sodium hydroxide, and its physical properties (high boiling point and low refractive index)•

The

saponification product was identified as 4-hydroxy-menthone chiefly by its physical properties;

no oxime could be obtained

but a semicarbazone was obtained melting at 84-5° (the semicarbazone of 4-hydroxymenthone has not been reported in the literature);

no urethane could be obtained with phenyliso-

cyanate, but there was obtained in this reaction sym-diphenylurea, which could result only by the expected dehydration of a tertiary alcohol, as the reaction was carried out under anhydrous conditions* A summary of the physical properties of the compounds under discussion is given below:

(continued on next page)

165. OBSERVED B.P. 2° QD

130-5°

101.0-3.5° 17mm
3) Beckm ann and Eickleberg, Ber., 29, 418-21 (1896). (4) Beckm ann and Mehrlander, Ann., 289. 367-91 (1896). (5) Cook and Cook, Ind.Eng.Chem.(Anal.Bd.) 5, 186-8 (1933); ,as D J°Sdn and Rosanoff,J.Am.Chem.Soc., 38, 713-6 (1916). /o> £ornubert and Hum eau, Bull.soc.chim.(4) 49, 1468-97 (1931). (7) Cusm ano, Atti R. Accad. dei Llncel 22 (5) I I , 569-75 1913); C hem .Zentr. 1914. 976-7. (8) Ktitz and Gatz, Ann. 358. 183-204 (1908). (9) Katz and Steinhorst, Ann., 379. 1-27 (1911). (10) Oddo, Gazz.chim.ital., 2£ I I , 112 (1897). (11) Perkin, J.Chem.Soc., 62, 1183 (1896). (12) Sandborn, Organic Syntheses 9, 52-3 (1929).

(13) Semmler, 2ie Atherischen 'Ole. Ill, 299 (Leipzig: Verlag von Veit and Company, 1906)• (14) Wagner, Ber., 2£, 1636-54 (1894). (15) Wallach,Ann., 305. 261-76 (1899). (16) Wallach,Ann., 222,211-6 (1913). (17) Wallach,Am., 4 i £ , 337-49 (1918). (18) Wallach and Meister, Ann., 362, 269-78 (1908).

191. VITA NAME: Robert Clarence Kuder BORN: December 31, 1918, at North Baltimore, Ohio* EDUCATION: ' * Washington Township Schools, Tontogany, 0 .^ 1924-31. Washington Township High School, Tontogany. 0 .. 1931-35. ■ * University of Toledo, Toledo, 0 *, 1935-57. The Ohio'State University, Columbus, 0 ., 1937-39; A. B,, June 1939. ' f ? Northwestern University, Evanston, 111*, 1939-42* POSITIONS HELD: Junior Chemist, Ethyl Gasoline Corporation Research Laboratory, Detroit, Mich*, summer of 1939* Research Assistant, Northwestern University, 1939-40, 1940-41* Assistant to Dr, Gustav Egloff, Chicago, 1 1 1 ., summer of 1940* Graduate Assistant, Northwestern University, summer of 1941, Allied Chemical and Dye Corporation Research Fellow, Northwestern University, 1941-42. PUBLICATIONS: "Homologous Series of Alkanes: Density and its Temperature Coefficlent.,f G. CALINGAERT, H. A* BEATTY, R* C. -KUDER, and G. W. THOMSON: Ind. Eng* Chem. 33, 103-6 (1941). "Molal Volume Relations among Aliphatic Hydrocarbons at their Boiling Points." G. EGLOFF and R. C. KUDER; J. Phys. Chem* 45, 836-45 (1941); J. Inst. Petroleum 27, 260-74 (1941). "The Molal Volumes of Aliphatic Hydrocarbons at their Melting Points." G; EGLOFF and R. C. KUDER: J. Phys. Chem, 46, 296-304 (1942). "Molecular Volumes of Liquid Alkanes at Correspond­ ing Temperatures,"' G. EGIDFF and R. C* KUDER: Ind, Eng. Chem, 34, 372-3 (1942), "Studies of the Physical Properties of Alicyclic Hydrocarbons. I. Molal Volumes of Monoalicyclic Hydrocarbons at 20° C," G, EGLOFF and R. C. KUDER: J, Phys. Chem, 46. £81-295 (1942). "Studies of the Physical Properties of Alicyclic Hydrocarbons. II. Boiling Points of Monocyclic Hydrocarbons," G. EGLOFF and R. Ci KUDER: pre­ sented at Memphis A. C. S. meeting, Apr* 1942. "Statistical Analysis of Physical-Chemical Data. G. EGLOFF and R. C. KUDER: presented at Atlantic City A. C. S. meeting, Sept. 1941, AFFILIATIONS: Phi Beta Kappa Sigma Xi Phi Lambda Upsilon American Association for the Advancement of Science American Chemical Society