Bhogte C P, Patravale V B, Devarajan P V
Department of Chemical Technology, University of Mumbai, Matunga, India.
J Chromatogr B Biomed Sci Appl. 1997 Jul 4;694(2):443-7. doi: 10.1016/s0378-4347(97)00137-0.
High-performance thin-layer chromatographic (HPTLC) analysis of gentamicin by in situ fluorodensitometric evaluation of its 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) derivative is presented. The aminoglycoside components separated on silica gel plates using chloroform-methanol-20% ammonium hydroxide (2.4:2.2:1.5, v/v/v) as the mobile phase were reacted with NB-Cl to yield highly fluorescent derivatives. The calibration curves of gentamicin in water, plasma and urine were linear in the range 40-200 ng. The mean values of intercept, slope and correlation coefficient were 16.82 +/- 0.473, 6.83 +/- 0.015 and 0.9968 +/- 0.0017 for standard curves in water, 17.35 +/- 0.375, 6.85 +/- 0.018 and 0.9941 +/- 0.0012 for standard curves in plasma and 14.35 +/- 0.286, 6.86 +/- 0.002 and 0.9933 +/- 0.0011 for standard curves in urine respectively. The analytical technique was validated for within-day and day-to-day variation. The results indicate that HPTLC, coupled with in situ fluorodensitometry, is a reliable and valuable technique for quantitative analysis of the bulk drug gentamicin and gentamicin from urine and plasma.
本文介绍了通过对庆大霉素的4-氯-7-硝基苯并-2-恶唑-1,3-二氮杂茂（NBD-Cl）衍生物进行原位荧光密度测定评估，采用高效薄层色谱（HPTLC）法分析庆大霉素。以氯仿 - 甲醇 - 20%氢氧化铵（2.4:2.2:1.5，v/v/v）为流动相，在硅胶板上分离氨基糖苷类成分，使其与NBD-Cl反应生成高荧光衍生物。庆大霉素在水、血浆和尿液中的校准曲线在40 - 200 ng范围内呈线性。水标准曲线的截距、斜率和相关系数的平均值分别为16.82±0.473、6.83±0.015和0.9968±0.0017；血浆标准曲线的分别为17.35±0.375、6.85±0.018和0.9941±0.0012；尿液标准曲线的分别为14.35±0.286、6.86±0.002和0.9933±0.0011。该分析技术针对日内和日间变化进行了验证。结果表明，HPTLC结合原位荧光密度测定法是一种可靠且有价值的技术，可用于对原料药庆大霉素以及尿液和血浆中庆大霉素进行定量分析。
