DESCRIPTIONS OF ANTIBIOTICS 



287 



Positive Sakaguchi, Elson-Morgan, Alolisch, and 

 Benedict tests (I). Negative biuret, ninhydrin, 

 Fehling, and maltol tests (2). Rf = 0.09 to 0.11 

 (n-butanol, 2 per cent piperidine, 2 per cent p- 

 toluene sulfonic acid monohydrate) (1). About 

 50 per cent inactivation at 100°C for 5 to 10 min- 

 utes (1). Trihydrochlonde: White substance (4). 

 [a Id = —95° (c = 1 per cent in water) (4). C = 

 35.6%; H = 5.95%; N = 13.9%; CI = 14.8%. 

 CaiHsgNyOnrSHCl. Helianthate: Reddish brown 

 crystals. Darken at about 220°C and char without 

 melting. X-ray diffraction pattern similar to 

 streptomycin helianthate (4). Yields a dihydro 

 derivative on catalytic hydrogenation (3, 8). 

 Alkaline hydrolysis products include pyromeconic 

 and isokojic acids, but no maltol (2, 10). Isokojic 

 acid (2-hydroxymethyl-3-hydro.\y-l,4 pyrone) : 

 m.p. 154-157°C. Ultraviolet absorption spectrum 

 maximum at 274 m^ (£"1™ 690) in 0.1 N HCl. Posi- 

 tive FeCls test (2, 3). 



Biological activity: Similar to that of strepto- 

 mycin (1, 2). Inactivated by cysteine and reacti- 

 vated by iodine (1). 



Toxicity: LDjo (mice) 154 mg per kg intrave- 

 nously, 865 to 948 mg per kg subcutaneously (2, 7). 

 Acute and chronic toxicity said to be essentially 

 the same as for streptomycin. 



Utilization: None. Presents no known advantage 

 over streptomycin. 



References : 



1. Hosoya, S. et al. Japan. J. Exptl. Med. 



20: 327-337, 1949. 



2. Grundy, W. E. et al. Arch. Biochem. 28: 



150-152, 1950. 



3. Benedict, R. G. et al. Science 112: 77-78, 



1950. 



4. Stodola, F. H.e/aL J. Am. Chem. Soc. 73: 



2290-2293, 1951. 



5. Benedict, R. G. et al . J. Bacteriol. 62: 



487-497, 1951. 



6. Grundy, W. E. et al. Antibiotics & Chemo- 



therapy 1: 309-317, 1951. 



7. Ambrose, A. M. Proc. Soc. Exptl. Biol. 



Med. 76: 466, 1951. 



8. Benedict, R. G. and Stodola, F. H. U. S. 



Patent 2,617,755, November 11, 1952. 



9. Hosoya, S. et al. J. Antibiotics (Japan) 



5:525-527, 1952. 



10. Hosoya, S. et al. J. Antibiotics (Japan) 



6A: 102, 1953 (abstr. of 6B: 61-66, 1953). 



11. Hosoya, S. Quoted in Benedict, R. G. 



Botan. Rev. 19: 229-320, 1953. 



12. Nakazawa, K. et al. Ann. Rept. Takeda 



Research Lab. 13: 67-77, 1954. 



Hygroiiiyciii 



Produced by: Streptoniyces hygroscopicu.'i (1) and 

 S. noboritoensis. The latter organism also produces 

 blastmycin and an antibiotic active on gram- 

 positive bacteria (7). 



Synonym: Homomycin (7). 



Method of extraction: I. Broth-filtrate satu- 

 rated with (NH4)2S04 and extracted with n- 

 butanol. Extract concentrated in vacuo, filtered, 

 and petroleum ether added to precipitate hygro- 

 mycin. Chromatographed on carbon from a 0.001 N 

 sulfuric acid solution and developed with an aciue- 

 ous solution containing 10 per cent n-butanol and 

 30 per cent acetone. Active fractions concentrated 

 in vacuo with addition of n-butanol to remove 

 water. Antibiotic precipitated on addition of 

 petroleum ether. Further purified by counter- 

 current distribution between n-butanol or n-amyl 

 alcohol and water-glacial acetic acid (2). II. Cul- 

 ture media acidified, stirred with acidic clay, and 

 filtered. Filtrate neutralized and stirred with 

 active carbon. Eluted from the carbon with 80 

 per cent aqueous acetone. Eluates concentrated 

 in vacuo and acetone added to the concentrate to 

 precipitate impurities. Concentrated in vacuo, 

 then lyophilized. Chromatographed on alumina 

 from methanol-ethanol (1:1) and developed with 

 acetone containing 20 per cent 0.5 N HCl. Further 

 purification by chromatography on silica gel with 

 water-saturated n-butanol as solvent and devel- 

 oper. Active fractions treated with carbon, con- 

 centrated in vacuo, and precipitated by adding 

 ether or petroleum ether. Final purification by 

 countercurrent distribution, first in an n-butanol- 

 0.25 M phosphate buffer (pH 4.6), then in n-bu- 

 tanol-ethyl acetate-water (1.2:0.5:1.9) (7). 



Chemical and physical properties: Weakly acidic 

 substance (2). White powder. Gradually melts at 

 80-90°C. Colors above 155°C (7). Very soluble in 

 water and ethanol; essentially insoluble in less 

 polar solvents. Infrared absorption spectrum 

 given in reference 2. Ultraviolet absorption spec- 

 trum maxima in dilute acid at 214 m/u (Eum 416) 

 and 272 m^ (ElL 306) (2) ; in dilute alkali at 254 

 m,x (£'}L 350), 286 m^ (£"5^ 194), and 323 m,x 

 (E'lcm 116) (6); in water at 270 to 272 m,u (E'i'L 

 291) (3). [a]f = -126° (c = 1 per cent in water) 

 (2). Positive Folin-Ciocalteau, Fehling, Benedict, 

 diazo, iodoform, Nessler, ToUen, indole, and 

 carbazole tests. Negative FeCla , Ehrlich, nin- 

 hydrin, biuret, phloroglucinol, Seliwanoff, gluco- 

 samine, anthrone, Molisch (doubtful), and maltol 

 tests (2, 3, 7, 8). Acetylation product is biologically 

 inactive (2, 3). pKa' = 8.9 (water) (6). Rf = 0.63 

 in water-saturated n-butanol. Cannot be hvdro- 



