超高效液相色谱-四极杆飞行时间质谱法快速定性、定量分析人全血中米酵菌酸

Rapid Qualitative and Quantitative Analysis of Bongkrekic Acid in Human Whole Blood by Ultra-High Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry

采集疑似米酵菌酸中毒患者全血样品200μL,加入已于-20℃冷冻的甲醇800μL,振荡5 min,离心5 min,上清液按照优化的仪器工作条件分析。在Infinity Lab Poroshell 120 EC-C_(18)色谱柱(150 mm×3.0 mm, 2.7μm)上以不同体积比的含10 mmol·L^(-1)甲酸铵的0.1%(体积分数,下同)甲酸溶液和乙腈的混合溶液梯度洗脱分离米酵菌酸,以双喷射流电喷雾离子源负离子模式电离,以全离子采集一级质谱(MS1-Allions)模式定性鉴别,以基质匹配工作曲线法定量。结果显示,米酵菌酸的质量浓度在45~450μg·L^(-1)内与其对应的峰面积呈线性关系,检出限(3S/N)为3.2μg·L^(-1)。对空白全血样品进行3个浓度水平的加标回收试验,回收率为82.6%~90.1%,测定值的相对标准偏差(n=6)为4.5%~5.6%。方法应用于疑似中毒患者全血样品的分析,检出了米酵菌酸(检出量为2 225μg·L^(-1))。

The whole blood sample from suspected bongkrekic acid poisoning patient was collected, and 800 μL of methanol precooled at-20 ℃ was added into 200 μL of sample. After shaking for 5 min, the mixture was centrifuged for 5 min, and the supernatant was analyzed by the optimal instrumental working conditions. The bongkrekic acid was separated on the Infinity Lab Poroshell 120 EC-C_(18)column(150 mm×3.0 mm, 2.7 μm) by gradient elution using mixed solutions composed of 0.1%(volumn fraction) formic acid solution containing 10 mmol·L^(-1)ammonium formate and acetonitrile at various volumn ratios, ionzed by double jet stream electric spray ion source with negative mode, identified by MS1-Allions mode, and quantitatively analyzed by matrix-matched working curve. It was shown that the linear relationship between values of mass concentration and peak area was kept in the range of 45-450 μg·L^(-1), with detection limit(3S/N) of 3.2 μg·L^(-1). Test for the spiked recovery was made on blank whole blood samples at three concentration levels, giving values of recovery in the range of 82.6%-90.1%, and RSDs(n=6) of the determined values ranged from 4.5% to 5.6%. The proposed method was applied to the analysis of the whole blood sample from a suspected poisoning patient, and the bongkrekic acid was detected with detection amount of 2 225 μg·L^(-1).