BIOLOGICAL ASSAY

MIXTURES OF ANTIBIOTICS
THEY ARE PRODUCED DURING FERMENTATION Penicillins, not less than 95% of total penicillins and not less that 90% of bencil penicillin. Tyrothricin ( Gramicidin and Tyrocidin) and Natural Cephalosporins ( C, N, and P) THEY ARE COMPONENT OF A FORMULATION Related compounds: Streptomycin and Dihidrostreptomycin or diferents salts of the same active substance Unrelated compounds: Streptomycin and Penicillin

IN WICH CASES, THE ACTIVITY MUST BE DETERMINED SEPARATELY OR TOTALY

BIOLOGICAL ASSAY
MIXTURES OF ANTIBIOTICS
SELECTIVE DETERMINATION SEPARATION METHODS Solvents In situ removal with ionic exchange resins Bioautograph SELECTIVE DESTRUCTION Test Organism entirely selective Consider interferences of mixed compounds DIFFERENTIAL ASSAYS Differing responses by different test organism to the individual components in a mixture of antibiotic substance

BIOLOGICAL ASSAY
MIXTURES OF ANTIBIOTICS
Comparative autobiograph of Antibiotic E129 to determine proportion of componenets G and A test organism:. M. luteus Sample: provisional test standart Response: Inhibition zone areas mesures as weigh of paper cut from methylene blue print DOSE 800 600 400 300 200 Comp A 219 196 169 132 96 Comp G 112 86 52 26 -250 200 ACTIVITY 150 100 50 0 2.3000 2.5000 2.7000 2.9000 3.1000 LOG DOSE

For responce of 104 the interceps are: Line A: Log dose = 2.333 dose = 215 UI Line G: Log dose = 2.865 dose = 733 UI The % of G activity is: (100*215)/(733+215) %G = 22.7%

BIOLOGICAL ASSAY
MIXTURES OF ANTIBIOTICS
STIMATING THE PROPORCION WITH DIFERENTIAL RESPONSES The Potency of compounds would remain constant The posible contaminans in pure compounds have negligible influence The activities of tow compounds are purely additive. Neomycin B sulfate S. Aureus: 850U/mg

M. Luteus: 950U/mg Neomycin C sulfate 410U/mg S. Aureus: M. Luteus: 15U/mg Potency in mix is: 850b +410c vs S. Aureus 410b + 15c vs. M. luteus
%B = 100b/(b+c) %C = 100c/(b+c)