SPE-HPLC法检测病毒灭活血浆中的残留亚甲蓝

目的建立SPE-HPLC法检测病毒灭活血浆中亚甲蓝浓度的方法并对病毒灭活血浆中的残留亚甲蓝进行质控检测。方法血浆样品经Waters Oasis HLB固相萃取柱提取,采用迪马DiamonsialC18色谱柱(250mm×4.6mm,5μm),以甲醇-0.1%乙酸(45∶55,V/V)为流动相,流速为1.0mL·min-1,柱温30℃,检测波长651nm。结果亚甲蓝在0.05～5μmol.L-1范围内线性良好(r=0.9998),最低检出浓度为0.05μmol·L-1。日内、日间RSD均小于8%。结论SPE-HPLC法具有灵敏度高、特异性强的特点,用此法检测的病毒灭活血浆样品中的残留亚甲蓝均达到要求。

Determination of methylene blue in human plasma by SPE-HPLC

OBJECTIVE To establish a SPE-HPLC method for the determination of the methylene blue concentration in human plasma that virus were deactivated.And then perform quality control of the human plasma deactivated.METHODS The methylene blue in plasma was processed by Waters Oasis HLB solid phase extractor and determined by SPE-HPLC method using a C18 column(Diamonsial,250mm×4.6mm,5μm)with a mobile phase of methanol-0.1% acetic acid(45∶55,V/V)at a flow rate of 1.0mL·min-1,the column temperature was 30℃ and the eluate was monitored at 651nm.RESULTS The linear range of the calibration curve was 0.05~5μmol·L-1 with r=0.9998,and the determination limit was 0.05μmol·L-1.Both of the intra-day and inter-day RSD were less than 8%.CONCLUSION The established SPF-HPLC method was suitable for the determination of methylene blue in human plasma and it was sensitive and specific.