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  • 许小建,仰先蜜,王学彬,王卓.高效液相色谱法测定左氧氟沙星微透析探针的回收率[J].第二军医大学学报,2020,41(1):110-114    [点击复制]
  • XU Xiao-jian,YANG Xian-mi,WANG Xue-bin,WANG Zhuo.Determination of recovery rate of levofloxacin microdialysis probe by high-performance liquid chromatography[J].Acad J Sec Mil Med Univ,2020,41(1):110-114   [点击复制]
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高效液相色谱法测定左氧氟沙星微透析探针的回收率
许小建1,2,仰先蜜1,2,王学彬2,王卓1,2*
0
(1. 安徽中医药大学药学院, 合肥 230013;
2. 海军军医大学(第二军医大学)长海医院药学部, 上海 200433
*通信作者)
摘要:
目的 研究左氧氟沙星微透析探针的体内外相对回收率。方法 建立高效液相色谱法(HPLC)检测左氧氟沙星的浓度,采用Kromasil-C18色谱柱(150 mm×4.6 mm,5 μm),流动相为乙腈-10 mmol/L磷酸二氢钾溶液(15:85),柱温30℃,紫外检测波长为294 nm,进样量10 μL。利用透析法、反透析法测定血管同心圆微透析探针的体外相对回收率,分析药物浓度、流速对相对回收率的影响。利用反透析法测定比格犬颈静脉、前列腺中探针的体内相对回收率,考察体内12 h的回收率稳定性。结果 在0.1~50.0 μg/mL浓度范围内左氧氟沙星的HPLC分析方法线性关系良好(r=0.998 7),且专属性良好,高浓度质控样品、中浓度质控样品、低浓度质控样品、定量下限样品的精密度和准确度等均符合分析要求。体外实验中,血管同心圆微透析探针的正向回收率和反向回收率差异无统计学意义(P>0.05),药物浓度不影响体外回收率,但回收率随灌流液流速的增大而减小。体内实验中,颈静脉中探针的相对回收率为(48.46±1.94)%,前列腺中探针的相对回收率为(13.23±1.44)%,且相对回收率在12 h内稳定。结论 建立的HPLC可用于左氧氟沙星微透析探针的测定,反透析法可用于测定左氧氟沙星微透析探针的体内相对回收率。
关键词:  高效液相色谱法  微透析  相对回收率  颈静脉  前列腺
DOI:10.16781/j.0258-879x.2020.01.0110
投稿时间:2019-08-07修订日期:2019-11-04
基金项目:国家自然科学基金(81870520),上海市卫生计生系统重要薄弱学科建设计划(2016ZB0303 01),国家临床重点专科军队建设项目(临床药学).
Determination of recovery rate of levofloxacin microdialysis probe by high-performance liquid chromatography
XU Xiao-jian1,2,YANG Xian-mi1,2,WANG Xue-bin2,WANG Zhuo1,2*
(1. School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230013, Anhui, China;
2. Department of Pharmacy, Changhai Hospital, Naval Medical University(Second Military Medical University), Shanghai 200433, China
*Corresponding author)
Abstract:
Objective To investigate the in vitro and in vivo relative recovery rates of levofloxacin from microdialysis probes. Methods High-performance liquid chromatography (HPLC) was established for determination of levofloxacin concentrations. Column was Kromasil C18 (150 mm×4.6 mm, 5 μm), the mobile phase consisted of acetonitrile and 10 mmol/L of potassium dihydrogen phosphate solution (15:85), with column temperature being 30℃, detection wavelength being 294 nm, and injection volume being 10 μL. The relative recovery rates of vascular microdialysis probes in vitro were measured by dialysis and reverse dialysis. The effects of drug concentration and flow rate on probe deliveries were investigated. The in vivo recovery rates of the probe in the jugular vein and prostate were measured by reverse dialysis, and the stability of 12 h recovery rate in vivo was investigated. Results When the concentration of levofloxacin was 0.1-50.0 μg/mL, the linear relationship was good in the HPLC analysis (r=0.998 7), and its specificity was good. The precision and accuracy of high-, medium-, low-concentration quality control samples, and lower limit of quantitation all met the analysis requirements. In the in vitro relative recovery experiments, there was no statistical difference between the recovery rate and reverse recovery rate of the vascular microdialysis probes (P>0.05). Drug concentrations did not affect in vitro recovery rate. The recovery rate decreased with the increase of flow rate. In vivo, the relative recovery rate of the probe was (48.46±1.94)% in the jugular vein and (13.23±1.44)% in the prostate. The relative recovery rates were stable during 12 h. Conclusion The established HPLC can be used for the determination of levofloxacin microdialysis probe. Reverse dialysis can be used to detect the relative recovery rate of levofloxacin microdialysis probe in vivo.
Key words:  high-performance liquid chromatography  microdialysis  relative recovery  jugular vein  prostate