DESCRIPTIONS OF ANTIBIOTICS 



329 



the same ultraviolet absorjjtion spectrum, with 

 maxima at 225 and 232 ± 0.5 m^u (in absolute 

 ethanol). Molar extinction values; A, 20,200 and 

 18,200; B, 18,800 and 17,000; and C, 23,200 and 

 21,600, respectively. Inflections at 220, 240 ± 0.5, 

 and 285 m^ (A and C) or 295 m^ (B). Infrared 

 spectrum and crystallographic data given in 

 reference 7. All give a yellow to cherry-red color 

 with concentrated H-2S04 , the color varying with 

 the concentration. Positive 2,4-dinitrophenyl- 

 hydrazine, Fehling, ammoniacal silver nitrate, 

 bromine, and KMn04 tests. Negative tests for 

 phenol, steroid, coumarin, tlavone and flavone- 

 related compounds (1, 7). Thermostability at 

 95''C: C > A > B (6). 



Hiolngiral activity: Active on certain fungi, 

 including Tilletia caries (2, 10), but not on bacteria 

 (1,7). In general the activity of the components is 

 quantitatively A > B > C, and qualitatively the 

 same, although A is active on Rhodotonda glutinis 

 and B and C are not. The diacelate is inactive on 

 Glomerella cingulata (only organism tested) (7). 

 Active on the soil nematode Rhabditis briggsae (1). 

 Effective on post-harvest decay {Hotrytis and 

 Rhizopus) of strawberries (9), downy mildew 

 {Phytuphthora phaseoli) and stem anthracnose 

 {Colletotrichion truncatum) of lima beans, rust 

 (Uromyce.s phaseoli var. typica) and anthracnose 

 (('. liN(U)iiiithianiinii) of beans (4), tomato early 

 l)light (Alternaria solani) (3), and brown rot 

 {Sclerotiniafructicola) of peach (5). Blocks a reac- 

 tion involved in phosphate transfer, l)ut not elec- 

 tron transport per sc. inhibiting respiration and 

 phosphate uptake with certain substrates and 

 inhibiting mitochondrial adenosine trii)hosphate 

 (8). 



Toxicity: LD.^i (mice) 1.5 mg per kg (A), 2.9 mg 

 per kg (B), and 8.3 mg per kg (C) intraperitoneally 



(7). 

 References: 



1. Smith, R. M. et al. Antibiotics & Chemo- 



therapy 4: 962-970, 1954. 



2. Newburgh, R. W. and Cheldelin, V. H. 



Plant Disease Reptr. 39: 684. 1955. 



3. Leben, C. et al. Phytopathology 46: 333- 



335, 1956. 



4. Zaumeyer, W. J. and Wester, R. E. Phyto- 



pathology 46: 478, 1956. 



5. Szkolnik, M. and Hamilton, J. M. Plant 



Disease Reptr. 41: 289^292, 1957. 



6. Sehgel, J. M. et al. Abstr. 132nd Meeting 



Am. Chem. Soc. 46C, 1957. 



7. Masamvme, S. et al. J. Am. Chem. Soc. 



80:6092-6095, 1958. 



8. Lardy, H. A. et al. Arch. Biochem. Bio- 



phys. 78:587-597,1958. 



9. Becker, R. F. ct al. Plant Disease Reptr. 

 42: 1066-1068, 1958. 



10. Marty, E. W., Jr. and McCoy, E. Antibio- 



tics et Chemotherapy 9: 286-293, 1959. 



11. Larson, M. H. and Peterson, W. H. Appi. 



Microbiol. 8: 182-189, 1960. 



Oxy tetracycline 



Produced by: Streptomyces rimosus strains (15, 

 27, 69); S. pla'.ensis, closely related to <S. hy- 

 groscopicus (56, 84); S. anuillatus (65) (this may 

 be the same culture as that described in reference 

 83 as Streptomyces sp.); S. vendargensis (86) (this 

 culture produces vengicide, and five or six other 

 antibiotics) ; S. gilvus: Streptomyces sp. resembling 

 S. griseohis; and Sireptomi/ces sp. One of these 

 cvdtures produces a heptaene and a basic anti- 

 biotic active on gram-positive bacteria (66, 85). 



Synonym: Terramycin. 



Method of extraction: lA. Whole broth adjusted 

 to pH 2.0 and filtered. Filtrate adjusted to: (a) pH 

 3.5 and extracted with i)henyl Cellosolve; or (b) 

 pH 9.0 and extracted with n-butanol, benzyl al- 

 cohol, amyl alcohol, or phenyl Cellosolve. i-^xtract 

 concentrated in vacuo at 50°C and extracted with 

 0.1 A' HCl. Extract neutralized to precipitate 

 oxy tetracycline. Purified by chromatography on 

 Florisil from a dilute HCl solution, and developed 

 with water, followed by acetone. Acetone fractions 

 diluted with water, acidified to pH 2.0, concentrated 

 under reduced pressure to remove the solvent, 

 then freeze dried. Powder dissolved in water, ad- 

 justed to pH 7.5, and extracted with n-butanol. 

 Re-extracted into 0.05 .V HCl. Extract concen- 

 trated in vacuo to i)recipitation (2, 15, S3). IB. 

 After acidification and filtration of the whole 

 l)roth, a sequestering agent, such as citric acid, 

 sodium tetraphosphate, or ethylenediamine- 

 tetraacetic acid, is added to the filtrate to l)ind the 

 multivalent metallic ions with which oxytetra- 

 cycline forms complexes (.see II). The whole 

 adjusted to pH 10.1 and extracted with butanol. 

 Butanol adjvisted to 5.3 to precipitate oxytetra- 

 cycline (54). IIA. Precipitated as the mixed 

 barium-magnesium salt from culture-filtrate at 

 1)H 8.5. Precipitate suspended in water, and salt 

 complex cleaved by acidification to pH 1.5 with 

 sulfuric acid. Adjusted to pH 3.0. seeded, then 

 adjusted successively to pH 5.0 and 7.0 to precipi- 

 tate oxytetracycline. Purified by salt conversion 

 or as in IV (18). IIB: Precipitated from cullurt> 

 filtrate at pH 9.0 in the presence of multivalent 

 metallic ions (as a metallo-organic salt) with or- 

 ganic amine bases such as octylamine, or quater- 

 nary ammonium halide salts such as alkyl and 

 alkenvl trimethvlaininonium chlorides with eight 



