NI WAYAN RIANTINI, 050810224 (2012) VALIDASI METODE KCKT DENGAN KOLOM RP-18 SILIKA KEMURNIAN TINGGI PADA PENETAPAN KADAR TETRASIKLIN, OKSITETRASIKLN, DAN KLORTETRASIKLIN DALAM PAKAN TERNAK. Skripsi thesis, UNIVERSITAS AIRLANGGA.

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Abstract

High Performance Liquid Chromatography method using RP-18 high purity silica column was optimized and validated for determination of oxytetracycline (OTC), tetracycline (TC) and chlortetracycline (CTC) in feed. Separation was on reversed phased column (Purospher® STAR RP-18 endcapped 12.5 cm x 4 mm ID, 5 μm) by isocratic elution using oxalic acid 0,01 M pH 3:methanol:acetonitrile (65:23:12, v/v) as mobile phase and flow rate was 0.4 mL/minute which provide a best chromatographic peak resolution. Monitoring was at 365 nm. Validity studies of the method revealed that all obtained calibration of OTC, TC and CTC curves showed good linearity (r > 0.999) over the range from 3 ppm to 25 ppm. Feed sampel was extrated with McIlvaine-EDTA buffer pH 2.5 and methanol (70:30, v/v). Average recovery of OTC, TC, and CTC were 94.33%, 85.97%, and 85.86% respectively, with all RSD below 4%. Method detection limits for OTC, TC, and CTC were 0.0405 ppm, 0.0666 ppm, and 0.0559 ppm. Furthermore the quantitation limits for OTC, TC, and CTC were 0.1349 ppm, 0.2221 ppm, and 0.1863 ppm respectively

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