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For the screening studies of new antibiotics, separation and identification using paperi,2,3) anc| thin-layer4>5>6) chromatography have been reported by many researchers. In general, thin-layer chromatography is effective in rapidly separating antibiotics soluble in organic solvents, and paper chromatography is useful for separating watersoluble antibiotics, although it takes much more time and mobility of the antibiotic may be influenced by the purity of the sample applied, especially by inorganic salts. We have found that high-voltage paper electrophoresis for 15 minutes gives a good separation of antibiotics, especially watersoluble basic or amphoteric antibiotics, even with crude preparations. The commercial apparatus, Model HV 5000-3 of Savant Instruments, Inc., was used in these experiments. The sample solution was spotted on strip of Toyo No. 51 filter paper (60cmx15cm) on a transverse line 16 cm from one end and the strip uniformly sprayed with an electrolyte solution (pH 1.8) of formic acid-acetic acid-water (22: 75: 900 by volume), similar to the technique reported by At field and Morris7) for the separation of amino acids. A constant current at 3,300V (about 40 mA/15cm) was then applied to the strip for 15 minutes. After electrophoresis, the strip was dried with flowing warm dry air to remove electrolyte solution. Antibiotics on the paper were detected by bioautography using test organisms such as Staphylococcus aureus, Micrococcus flavus, Bacillus subilis or Escherichia colt. In some cases, ninhydrin and Sakaguchi reactions and ultraviolet light were used for the detection of the antibiotics. Under standard conditions, alanine moved toward the cathode ca. 10cm. The mobilities of antibiotics relative to alanine were calculated as Rmvalues. The relative mobilities (Rm) of 92 antibiotics are listed in two groups according to their extraction properties in Tables 1 and 2. Most antibiotics assigned to group II are adsorbed on weak cation-exchange resins such as Amberlite IRC-50 or strong cationexchange resins such as Amberlite IR-120. Interestingly, antibiotics having an Rmvalue Table 1. Relative mobilities of antibiotics extracted by solvent process. (Group I)

APPLICATION OF HIGH-VOLTAGE PAPER ELECTROPHORESIS FOR SEPARATION AND IDENTIFICATION OF ANTIBIOTICS