Frita Yanuar Angriani and Achmad Toto Poernomo and Isnaeni (2015) Perbandingan Metode Penentuan Kadar Oksitetrasiklin Dalam Udang Dengan KLT-Densitometri dan KLT-Bioautografi. Berkala Ilmiah Kimia Farmasi, 4 (2). pp. 25-34. ISSN 2302-8270

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Abstract

A simple Thin Layer Chromatography (TLC) Densitometry and TLC-Bioautography has been used for identification and quantification of oxytetracycline. Validation both of the methods has been developed for determination of oxytetracycline in shrimp, included parameter linearity, accuracy (recovery), precision (CV), limit of detection, and limit of quantification, as a purpose to verifying that the parameters is appropriate with literature. The TLC of oxytetracycline has been carried out by using silica gel GF254 impregnated by EDTA 1% was used as a stationary phase, while choloroform: methanol : sodium EDTA 1% (55:25:10, v/v) was used as mobile phase. Retardation factor (Rf) of Oxytetracycline obtained by this system was 0,44. Staphylococcus aureus was used as a bacterial test on the bioautography method. The analyte spots were quantified by using densitometry method at 365 nm. The response was found to be linear at the amount of oxytetracycline between 100 ppm - 500 ppm. Oxyetracycline’s regression line equation was Y = 9992,1091X – 1861,706, r value = 0,9916, and Vxo = 7,9332%for densitometry and Y = 0,8573X – 7,5901.10-3, r value = 0,9942, and Vxo = 1,6743% for bioautography. Recovery and CV were 61,7450% ± 2,4737 and 6,8402% for densitometry, and 59,7287% ± 2,9133 and 4,8775% for the bioautography. Limit of detection value was 12,10 ppm and 20,04 ppm for densitometry and bioautography respectively. Limit of quantitation value was 40,35 for densitometry, and 50,10 ppm for bioautography. Conclusion, all the validation parameters were appropriate with standard reference, except accuracy and precision, but the methods can be applied for determining oxytetracycline in shrimp, and there’s no meaning difference between both of them for determining oxytetracycline in shrimp for accuration parameter. The precision of TLC-Densitometry was better than TLC-Bioautography.

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