二维色谱法测定人血清中文拉法辛浓度

Determination of Venlafaxine in Human Serum by 2D-LC-UV

目的建立二维液相色谱法(2D-LC-UV)测定人血清中文拉法辛血药浓度,并用高效液相串联质谱法(LC-MS/MS)比较该方法的一致性和相关性。方法用二维液相色谱法与高效液相串联质谱法分别测定文拉法辛已知浓度和未知浓度的血清样本,已知浓度样本为低、中、高3个质量浓度(40、300、750 ng·mL^(-1))质控样本,未知浓度样本来自广州医科大学附属脑科医院52份服用文拉法辛患者,并用统计学分析两种方法测得结果的一致性和相关性。结果文拉法辛在两种方法中标准曲线浓度与峰面积均呈良好的线性关系(r>0.995),方法回收率均高于95%;批内、批间的精密度RSD均小于15%;稳定性试验偏差均小于15%;按照Bland-Altman法计算两种方法测出的患者血清样品浓度差值,有98%的样品浓度差值小于5%,说明两种方法的一致性较好,按照Pearson线性回归分析,相关系数为0.999,P<0.01,显示两种方法具有良好的相关性。结论两种方法的方法学参数均符合2020年版中国药典附录9012相关要求,临床实际样品测定一致性良好,均可用于临床血药浓度监测。

OBJECTIVE To determine the plasma concentration of venlafaxine by establishing automatic two-dimensional liquid chromatography(2 D-LC-UV)method and to compare the consistency and correlation of the method by high performance liquid chromatography tandem mass spectrometry(LC-MS/MS),METHODS LC-MS/MS and 2 D-LC-UV were used to determine the serum samples with known and unknown concentrations of venlafaxine.The known concentration samples were low,medium and high concentrations of venlafaxine(40,300,750 ng·mL^(-1)).The unknown concentration samples were collected from 52 patients who had taken venlafaxine in the Affiliated Brain Hospital of Guangzhou Medical University.And the consistency and correlation of the results measured by the two methods were analyzed statistically.RESULTS There was a good linear relationship in the standard curve concentration and peak area of venlafaxine between the two methods(r>0.995),and the recovery rate was higher than 95%.RSD of inter-day and intra-day precision were all less than 15%.Deviation of stability tests were all less than 15%.According to the Bland-Altman method,98%of the sample concentrations difference was less than 5%,indicating that the two methods had good consistency.According to Pearson linear regression analysis,the correlation coefficient was 0.999(P<0.01),suggesting that the two methods had good correlation.CONCLUSION The methodological parameters of the two methods are in line with the relevant requirements of Appendix 9012 of Chinese Pharmacopoeia(2020 edition),and the consistency of clinical samples determination are good.Both of the two methods can be used for clinical monitoring of blood drug concentration.