Direct determination of four fluoroquinolones, enoxacin, norfloxacin, ofloxacin, and ciprofloxacin, in pharmaceuticals and blood serum by HPLC
Direct determination of four fluoroquinolones, enoxacin, norfloxacin, ofloxacin, and ciprofloxacin, in pharmaceuticals and blood serum by HPLC
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DOI：
10.1007/s00216-003-1749-9
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发表时间：
2003-03-01
影响因子：
4.3
通讯作者：
Papadoyannis, IN
中科院分区：
文献类型：
--
作者：
Samanidou, VF;Demetriou, CE;Papadoyannis, IN
建立了一种快速、准确、灵敏的定量测定四种氟喹诺酮类抗菌药依诺沙星、诺氟沙星、氧氟沙星和环丙沙星的方法，该方法对多种革兰氏阴性和革兰氏阳性菌具有较高的体外活性。色谱柱为Kromasil 100 C-8 250 mm×4 mm，5微米，流动相为乙腈-甲醇-柠檬酸0.4mol L-1(7：15：78%，v/v)。检测波长为275 nm，最低检测限为：依诺沙星0.02 ng/20Mul，氧氟沙星、诺氟沙星和环丙沙星0.01 ng。以氢氯噻嗪(HCT)为内标，浓度为2 ng/L。依诺沙星、氧氟沙星、诺氟沙星和环丙沙星的线性关系分别为2、12、3和5 ng。在10min内实现分离。对该方法进行了日内(n=8)和日间(n=8)精密度测定，结果令人满意，具有较高的准确度和精密度。方法用于人血清中4种氟喹诺酮类药物的直接测定。样品前处理只需用乙腈沉淀蛋白质。在0.1-0.5 ng/L范围内，加标样品中分析物的回收率为97+/-6%。
A rapid, accurate and sensitive method has been developed for the quantitative determination of four fluoroquinolone antimicrobial agents, enoxacin, norfloxacin, ofloxacin and ciprofloxacin, with high in-vitro activity against a wide range of Gram-negative and Gram-positive organisms. A Kromasil 100 C-8 250 mmx4 mm, 5 mum analytical column was used with an eluting system consisting of a mixture of CH3CN-CH3OH-citric acid 0.4 mol L-1 (7:15:78 %, v/v). Detection was performed with a variable wavelength UV-visible detector at 275 nm resulting in limits of detection: 0.02 ng per 20 muL injection for enoxacin and 0.01 ng for ofloxacin, norfloxacin and ciprofloxacin. Hydrochlorothiazide (HCT) was used as internal standard at a concentration of 2 ng muL(-1). A rectilinear relationship was observed up to 2 ng muL(-1) for enoxacin, 12 ng muL(-1) for ofloxacin, 3 ng muL(-1) for norfloxacin, and 5 ng muL(-1) for ciprofloxacin. Separation was achieved within 10min. The statistical evaluation of the method was examined by performing intra-day (n=8) and inter-day precision assays (n=8) and was found to be satisfactory with high accuracy and precision. The method was applied to the direct determination of the four fluoroquinolones in human blood serum. Sample pretreatment involved only protein precipitation with acetonitrile. Recovery of analytes in spiked samples was 97 +/- 6% over the range 0.1-0.5 ng muL(-1).