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HPLC法检测万古霉素血药浓度
引用本文:林忠,周鹏,孙渊,杜有功,杨建苗,林建群,林赛赛. HPLC法检测万古霉素血药浓度[J]. 中国临床药学杂志, 2014, 0(2): 92-96
作者姓名:林忠  周鹏  孙渊  杜有功  杨建苗  林建群  林赛赛
作者单位:浙江省台州医院临床药学实验中心,台州317000
基金项目:恩泽科研基金(编号12EZD34)
摘    要:目的利用单因素实验法建立和优化临床万古霉素HPLC分析方法,为临床监测万古霉素血药浓度、调整万古霉素临床给药剂量和个体化差异给药提供定量分析方法。方法比较国内外已报道的万古霉素HPLC测定方法,利用本院WA—TERS液相系统进行方法验证,以40%的硫酸锌沉淀处理血液标本,最终选择Waters0DS2(250mm×4.6mm,5ttm)色谱柱为分析柱。实验设计通过梯度改变柱温、pH、流动相的组成比例,验证每种条件对实验结果的容量因子(K),拖尾因子(T),分离度(R)的影响,最终得到最佳色谱条件为流动相0.05mol·L-1 KH2P04缓冲溶液(用磷酸调pH至2.85)-乙腈-甲醇(91.5:8:0.5),紫外检测波长为200nm,流速为1mL·min-1,柱温30℃,进样量为20止。结果万古霉素的最低检测质量浓度为O.05mg·L-1,在0.1~64.0mg·L。时线性较好(r=0.9992),相对回收率为97.5%~100.1%,日内、日问RSD均〈2%,R为2.1。结论该方法灵敏,快速,简便,特异性且稳定可靠,适用于临床万古霉素血药浓度的监测及临床药学的研究。

关 键 词:HPLC  万古霉素  血药浓度

HPLC determination of vancomycin in human plasma
LIN Zhong,ZHOU Peng,SUN Yuan,DU Yougong,YANG Jianmiao,LIN Jianqun,LIN Saisai. HPLC determination of vancomycin in human plasma[J]. Chinese Journal of Clinical Pharmacy, 2014, 0(2): 92-96
Authors:LIN Zhong  ZHOU Peng  SUN Yuan  DU Yougong  YANG Jianmiao  LIN Jianqun  LIN Saisai
Affiliation:( Clinical Pharmacy Experimental Center of Taizhou Hospital in Zhejiang Province, Taizhou 317000, China )
Abstract:AIM To establish and optimize HPLC ultraviolet detection analytical method of plasma drug concen- tration of vancomycin for monitoring clinic plasma drug concentration, adjusting dosage and individual differences in drug dosage by single factor experiment design. METHODS We reviewed papers about the HPLC detection method of van- comycin at home and abroad and carried out method validation based on the HPLC system of our hospital in order to choose the compatible method for our system. Specimens were disposed by zinc sulfate solution(40% ). Chromatographic separation was carried out on a Waters ODS2 column(250 mm×4.6 mm, 5μm). HPLC detecting method was optimized via single factor experiment design. We changed column temperature, pH, the proportion of KH2PO4 buffer solution and verified the impact of all factors on experimental results of corresponding capacity factor, tailing factor and resolution. Fi- nally, the optimized chromatographic conditions were as follows: a mixture of KH2PO4 buffer solution (adjusted pH to 2.85 using phosphoric acid) -acetonitrile- methanol (91.5:8:0.5) as mobile phase at the wavelength of 200 nm, the flow rate was 1.0 mL·min-l with the column temperature at 30℃ and 20 μL injection volume. RESULTS The mini- mum detectable concentration was 0.05 mg· L-1, the linear equation was fine( r = 0.999 2) in the range of 0.1 - 64.0 mg·L- 1, and the relative recovery was 97.5 % - 100.1%, the RSD of intra-day and inter-day assays were less than 2 %, and the resolution was 2.1. CONCLUSION The result of validation study demonstrates that the method is simple, sensitive, precise and selective for monitoring clinic plasma drug concentration.
Keywords:HPLC  vancomycin  plasma drug concentration
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