高效液相色谱法测定人血清和脑脊液中莫西沙星浓度

 目的 建立测定人血清和脑脊液中莫西沙星药物浓度的高效液相色谱方法。方法 血清和脑脊液取样量均为10 μL；采用Shima-pack CLC-ODS色谱柱（6.0 mm×150 mm，5 μm）进行分离；以0.01 mol·L-1磷酸缓冲液（含硫酸四丁基铵5 mmol·L-1，pH 2.5）-乙腈（80∶20）为流动相；吡哌酸为内标，荧光检测器波长：Ex=295 nm、Em=495 nm。结果 内标和莫西沙星的保留时间分别为2.93和6.39 min；血清和脑脊液标准曲线在0.2~20 mg·L-1内线性良好（r>0.999 9，n=5），其高、中、低质控样本方法学回收率分别在98.08%~104.81%和92.08%~101.73%间，方法学误差批内RSD均小于5.5%，批间RSD均小于7.8%。扩大取样量至0.1 mL，标准曲线下限可达2 ng·mL-1并良好检出。结论 该方法取样量小、操作简便、灵敏度高；为进行血清和脑脊液中莫西沙星临床PK/PD研究提供了数据保障。

Determination of Moxifloxacin in Human Serum and Cerebrospinal Fluid by HPLC

OBJECTIVE To establish a RP-HPLC method for the determination of moxifloxacin(MOFX) in human serum and cerebrospinal fluid(CSF) by HPLC with fluorescence detection. METHODS The determination of MOFX in 0.01 mL of human serum and CSF was performed on shima-pack CLC-ODS column (6.0 mm×150 mm, 5 μm) using fluorescence detection with Ex=295 nm and Em=495 nm . The mobile phase was composed of acetonitrile and 0.01 mol·L-1 phosphate buffer (pH 2.5) containing 5 mmol·L-1 teterabutylammonium hydrogen sulfate (20∶80). RESULTS The retention time were 6.39 min for MOFX and 2.93 min for the internal standard (pipemidic acid), respectively. The calibration curves were linear in the range of 0.2-20 mg·L-1 (r>0.999 9, n=5), and the limit of detection was 2 ng·mL-1 (sample size of 0.1 mL). The method was fully validated and validation parameters for the low, middle and high concentrations of check samples were: recovery was 98.08%-104.81% for serum and 92.08%-101.73% for CSF, intra-run and inter-run RSDs were <5.5% (n=10) and <7.8% (n=6) in plasma and CSF, respectively. CONCLUSION The method can be used for determination of moxifloxacin in serum and CSF of patients in clinical PK and PD studies.