Khuroo A H, Monif T, Verma P R P, Gurule S
Department CPP, Ranbaxy Research Laboratories, Plot-20, Sec-18, Udyog Vihar Industrial Area, Gurgaon, Haryana, India.
J Chromatogr Sci. 2008 Nov-Dec;46(10):854-61. doi: 10.1093/chromsci/46.10.854.
A simple, economical, and reproducible high-performance liquid chromatography mass spectrometric (MS) method is developed and validated for the determination of amoxicillin in human plasma. The present method has been successfully used to determine bioequivalence between a test and innovator formulation of amoxicillin 500 mg capsules. The method is validated in terms of selectivity, precision/accuracy, recovery, dilution integrity, matrix effect, effect of anti-coagulant, and stability studies. Sample preparation is carried out by solid-phase extraction (HLB Oasis cartridges). The processed sample is chromatographed on Hypersil Gold (4.6 x 50 mm); 3 microm C18 column, using 10mM ammonium formate buffer (pH 5.0) and acetonitrile, (10:90, v/v) as mobile phase. Amoxicillin is detected by MS-MS detection with turbo-ion spray in positive ion mode. The weighed (1/X2) calibration curves were linear over the range of 0.17 to 17.0 microg/mL. The intra day precision is from 1.3% to 8.8% and intra day accuracy is 94.1% to 108.5%. The inter day precision is from 1.8% to 6.2% and inter-day accuracy is 95.1% to 105.9%. Mean recovery of 66.3% is observed for amoxicillin and 71.6% for internal standard (ampicillin). The stability of amoxicillin is studied at -15 degrees C and -50 degrees C using human plasma with different anti-coagulants (citrate, monobasic sodium phosphate, dextrose, and adenine-citrate, monobasic sodium phosphate, dextrose, and adenine and ethylene diamine tetraacetic acid-ethylene diamine tetraacetic acid). No significant degradation is observed for 60 days.
开发并验证了一种简单、经济且可重复的高效液相色谱-质谱（MS）方法，用于测定人血浆中的阿莫西林。本方法已成功用于测定阿莫西林500mg胶囊的受试制剂与创新制剂之间的生物等效性。该方法在选择性、精密度/准确度、回收率、稀释完整性、基质效应、抗凝剂的影响以及稳定性研究方面进行了验证。样品制备采用固相萃取（HLB Oasis柱）。处理后的样品在Hypersil Gold（4.6×50mm）；3μm C18柱上进行色谱分析，使用10mM甲酸铵缓冲液（pH 5.0）和乙腈（10:90，v/v）作为流动相。通过MS-MS检测，采用涡轮离子喷雾正离子模式检测阿莫西林。加权（1/X2）校准曲线在0.17至17.0μg/mL范围内呈线性。日内精密度为1.3%至8.8%，日内准确度为94.1%至108.5%。日间精密度为1.8%至6.2%，日间准确度为95.1%至105.9%。阿莫西林的平均回收率为66.3%，内标（氨苄西林）的平均回收率为71.6%。使用含有不同抗凝剂（柠檬酸盐、磷酸二氢钠、葡萄糖、腺嘌呤-柠檬酸盐、磷酸二氢钠、葡萄糖、腺嘌呤和乙二胺四乙酸-乙二胺四乙酸）的人血浆，在-15℃和-50℃下研究了阿莫西林的稳定性。60天内未观察到明显降解。
