June 20, 1954
NOTES
quired 24 hours a t O0 followed by 48 i 12 hours at room temperature for complete reaction. With isoamyl bromide the reaction was run at 0" for 12 hours and then for 4 days at room temperature; only then was a negative Beilstein test obtained. Isobutyl bromide after 24 hours a t 0" and 96 hours at room temperature gave only 37570 of the theoretical amouut of silver bromide and ca. d 40Yc recovery of isobutyl bromide (which was, however, not quite pure). The reaction time can be shortened considerably by running the reaction a t 0" for 3 hours, then at room temperature for 5 hours and, finally, refluxing until a negative Beilstein test is obtained (ca. 4 hours of refluxing for iodides and 18 hours for straight chain bromides). However, the product is then a more complex mixture and the sulfuric acid treatmcnt' is iiecessary to ensure t h a t pure nitroparaffin is obtained; the yields are generally about 15-207c lower. Spectrophotometric Determination of Nitrite Esters.-Since the lower molccular weight nitrites cannot be separated quantitatively from the reaction solvent i t was necessary to devise a method for their determination in ether yolution. The absorption spectrum of nitrite esters extends further into the visible than t h a t of any other component of the reaction mixture; a t 410 mg the nitroparaffins, alkyl nitrates, alkyl halides, alcohols and carbonyl compounds do not interfere and, therefore, photometric readings were taken a t this wave length on the ether solutions obtained by filtering out the silver salts. The amounts of each nitrite were determined from a calibration curve set up by using known concentrations of t h a t particular alkyl nitrite. The accuracy of the method as checked by means of known mixtures of nitrite and nitroparaffin appears to be good t o within 2%. Furthermore, the spectrophotometric technique was applied in the n-octyl iodide case Here the amount of
nitrite ester found spectrophotometrically was 14% while, upon rectification, 11% of pure 1-octyl nitrite was isolated. The primary nitrite esters needed as reference compounds were synthesized by the excellent procedure of Chretien and Longilg in which a 40Yc aqueous solution of Alz(S04)3.18Hz0is added t o a cold mixture of saturated aqueous sodium nitrite and the alcohol. After a few hours yields ranging from 7 0 4 0 % were obtained in all cases (cf. Table W). TABLE IV ALKYL l T ~ ~
3211
1,it. value
h-itrite
R.p. ' C . Rfm.
li2OD
13,s. ' C . him.
llpu0
Kef.
n-Butyl 78 1.3762 7 5 1,3760 6 %-Hexyl 64 76 1.3990 50 48 1.3977/23' c n-Heptyl 41 5 1.4063 58 20 1.4060 d n-Octyl 56 9 1.4127 55 8 1.4129 e Isobutyl 66" 1.3702" 67 1.3715/22' f Isoamyl 51 115 1.3870 99 1.3871 g Isobutyl nitrite prepared from isobutyl alcohol and nitrosyl chloride has identical b.p. and n%. b IV. A . Noyes, THISJOURSAL, 55,3883 (1933). L. M . Soffer, E. IV. Parrotta and J . D . Domenico, i b i d . , 74, 5301 (1952). d R e f . 10. e T. C. Krantz. U. S. Patent 2.101.358: C..4.. 33. J . 330° (uncor.) from 95% ethanol. ExperirnentaF.6 Anal. Calcd. for C12H12Ns02: N, 12.96. Found: N, 13.11. Ethyl 3a-Hydroxy-A11-cholenate (11) .-3a-Hydroxy-A"XI1 was berizoylated under conditions similar to those cholenic acid7.8 was esterified with absolute ethanol essentially according to the method used by Kendall and his associused by de Arce, Greene and Cappss for the benzoylation of ~5-arnino-8-bromo-6-methylquinoline; yield 42%, m.p. > ates for the preparation of the methyl ester.9 However, whereas esterification with methanol is complete in less 315". Anal. Calcd. for C17H14Nn02: N, 10.07. Found: N, than an hour, with ethanol 27% of unreacted acid was recovered even after 22 hours. The ethyl ester did not 10.00. crystallize from aqueous ethanol, but separated satisfac8-Methyl-5-quinoline~usonic Acid (XV) , 2-Chloro-8- torily from purified Skellysolve F as colorless needles melting methyl-5-quinolinearsonic Acid (XVIII) and 2-Hydroxy-8- a t 81-82', [ a I z 8 D 30' (c 2.01, chf.). Anal. Calcd. for methyl-5-quinolinearsonic Acid (XXI) .-The hydrochlorides C2&203: C,77.56; H , 10.52. Found: C,77.3; H, 10.6. of X (12.0 g.), X I (9.0 g.) and XI1 (10.0 g.) were diazotized Ethyl 3P-Chl0ro-A~~-cholenate (I).lO-To a stirred solution and converted into arsonic acids according tcr t h e p r m d r e of 500 mg. of I1 in 28 ml. of chloroform, in a flask equipped reported by Capps and Hamilton" for changing certain 2- with a drying tube and immersed in an ice-bath maintained chloroaminoquinolines into khloroquinolinearsonic acids. a t O', 800 mg. of powdered calcium carbonate and, in two XV, XVIII and X X I resulted in yields of 12.8, 11.9 and portions with a 20-minute interval, 1.2 g. of phosphorus 14 S'%, respectively. X V melted a t 224-226' while XVIII pentachloride were added. Stirring was continued for 100 dud X X I melted above 315" (uncor.). minutes a t 0'. The reaction product was poured into 200 Anal. Ccilctl for CIOHIOASXO~: As, 28.05; N, 5.27. ml. of 5% sodium bicarbonate solution containing ice, and Found: As, 27 92; S , 5.09. Calcd. for CIOH~ASCINO~:ether was added. The resulting mixture was stirred until As, 24 84; S , 4 65. Found: As, 24.69; N, 4.75. Calcd. the ice had melted, transferred to a separatory funnel and for CIOHIOASNO~ H10: As, 24.87; N, 4.65. Found: As, shaken thoroughly. The organic layer, which still retained 24.79; N, 4.67. a small amount of an insoluble, colorless, inorganic solid, was washed with water, dried (Drierite), filtered and evapoD Capps and C. S. Hamilton, THIS JOURNAL, 60, 2105 (11) J rated (reduced pressure) to a colorless residual oil. This oil (1938). dissolved in the minimum amount of warm methanol, on refrigeration for 2 hoours yielded 380 mg. (73%) of colorless Ross CHEMICAL LABORATORY Two recrystallizat$ms from methacrystals m.p. 69-73 ALABAMA POLYTECHNIC INSTITUTE nol gave thin plates melting at 74-76 , [ C Y ] ~ ~ D25" AUBURN, ALABAMA (c 2.05, chf.). Anal. Calcd. for C&,102Cl: C, 74.16; H , 9.82; CL8.42. Found: C, 74.3; H, 9.8; C1,8.8. Catalytic Hydrogenation.-I in acetic acid solution and Seroflocculating Steroids. I. Ethyl 3p-Chlorothe presence of Adams catalyst absorbed 1.03 moles of A1l-cholenate hydrogen within 20 minutes. After removal of catalyst and solvent, two crystallizations of the product from BY FREDERICCAASGA N D DOUGLAS H. S P K U N T methanol gave colorless, feathery crystals, m.p. 59-60.5', R E C E I V EAPRIL D 2, 1954 which did not depress the melting point of ethyl 3p-chlorocholanate (111) prepared from ethyl lithocholatellJ* by the During the course of our study of the relationship same method as used for the unsaturated derivative, crysof steroids2 to ininiunological phenomena associ- tallizing in methanol as colorless needles, m.p. 59-61.5', ~ (c 1.27, chf.). Anal. Calcd. for C2sHtaO&1: ated with injury, we have had the occasion to [ a I z e +18.5" investigate the seroflocculating reagents described C, 73.81; H , 10.24; C1, 8.38. Found: C, 73.5; H, 10.5; c1, 8.2.
+
.
+
e.
by Penn and his associate^.^ These reagents4 (5) Microanalyses by the Microchemical Laboratory of New York cause flocculation in a high percentage of the sera University. of patients with cancer and other diseases. (6) Melting points were taken on an electrically heated micro Our investigation of the flocculating reaction hot-stage and are uncorrected. led us to prepare ethyl 3P-~hloro-A~~-cholenate (7) J. Press and T. Reichstein, Helu. Chim. Acla, 26, 878 (1912); (I) which in preliminary testing we have found to B. F. McKenzie, W. F. McGuckin and E. C. Kendall, J. Biol. Chcm., 555 (1946) be a very satisfactory flocculating reagent. The 162, (8) Generously supplied by Merck and Co. through the kindness of conipuund. is c r y s t a l h e a d stable -m&ror&my Dr: M;rx Tishier conditions. I r e will discuss in detail elsewhere (9) L. L. Engle, V. R. Mattox, B. F. McKenzie, W. F. McGuckin implications of general significance in this field sug- and E. C. Kendall, J. B i d . Chem., 162, 565 (1946). (10) Although a double bond shift is considered unlikely under the gested to us by this finding. conditions of this reaction. experiments are under way to confirm the 'Treatmext of ethyl 3a-hydroxy-A1Wiolenate position of unsaturation. ( I 1) with phosphorus pentachloride in chloroforni (11) F. Reindel and K. Niederlander, Bey., 68, 1969 (1935). (12) We are indebted t o Ciba Pharmaceutical Products, Inc.. and at 0" yielded ethyl 3~-chloro-A11-cholenate(I), which could be quantitatively hydrogenated to Dr. H. B. MacPhillamy for a supply of lithocholic acid. AND ethyl 3,b-chlorocholanate (111). The latter was DIVISIONOF CHEMISTRY, DiVi310N O F PATHOLOGY AND- BACTERIOLOGY found- to be identicar by meiting point comparison UNIVERSITY O F TENNESSEE with the product of reaction between ethyl litho- MEMPHIS,TENNESSEE cholate and phosphorus pentachloride. (1) Aided in part by a grant from the United States Public Ifealth Service. (2) D. 1%.Sprunt, A. D. Dulaney and R. P. Conger, Cancer Research, 2 , 282 (1951). (3) H. S.Penn. J . S n l l C Q J I C P Y ~12, I S ~1389 . , (1952); A. H. Dowdy, H S. Penn, C i . Hall and A. Brllamy, Proc. A m . Assoc. for Cancer R P s r i l r < h . 1, I2 (19.54).
( 4 ) We wish to thank 131s. D o w d y and Penn aud their group for their coijperation in making available t o us procedures for preparing and testing both the liver and desoxycholic acid-derived flocculating reagents which they designate as "antigens."
The Preparation of Sarcosine and Methyl aMethylamino-0- (3-indolyl)-propionate BY F. F. BLICKEA N D PAULE. NORRIS RECEIVED FEBRUARY 15, 1954
The preparation of methyl a-methylaniino-P-(3indoly1)-propionate was undertaken since a supply of this ester was required as an intermediate.
