HPLC-MS/MS法检测猪肉中苯并菲啶类生物碱的研究

Study of determination of quaternary benzo[c]phenanthridine alkaloids in pork using HPLC-MS/MS

关键词:  血根碱  白屈菜红碱  QBAs, HPLC-MS/MS  残留

Key words:  sanguinarine  chelerythrine  QBAs  HPLC-MS/MS  residues

摘要:

利用高效液相色谱串联三重四极杆质谱技术建立猪肉中痕量苯并菲啶类生物碱定量检测方法。将猪肉样品匀浆经1%盐酸-甲醇（10:90，V/V）超声提取后离心分离取上清液经脱脂后氮气吹干浓缩，甲醇复溶。采用C18色谱柱（2.1mm×150mm,2.7μm）分离，流动相为乙腈与0.2%甲酸组成梯度洗脱，柱温35℃，流速0.3mL/min；采用液相色谱-电喷雾离子源质谱检测。血根碱和白屈菜红碱在0.5~50ng/mL浓度范围内与峰面积具有良好的线性关系，线性相关系数分别为R2=0.9996和R2=0.9995，平均加标回收率分别为91.50%和91.30%，重复性RSD为11.68%和9.90%，精密度RSD为2.58%和3.89%。经测定，饲喂美佑壮的猪的肌肉中血根碱和白屈菜红碱的残留量分别为4.76ng/g和2.09ng/g。该方法灵敏度高，重复性好，适合于猪肉中苯并菲啶类残留的定量检测。

Abstract:

Using high performance liquid chromatography tandem triple quadrpole mass spectrometry to establish a method for quantitative detection of trace amounts of quaternary benzo[c]phenanthridine alkaloids in pork. Porks were homogenized, centrifuged and separated after ultrasonic extraction with 1% hydrochloric acid-methanol (10:90, V/V). The supernatant was degreased, and dried with nitrogen and concentrated, then melted again with methanol. Separation was performed on a C18 column (2.1mm×150mm, 2.7μm). Acetonitrile and 0.2% formic acid(v/v) solution was used for the mobile phase. Gradient elution was performed at column temperature of 35℃ with a flow rate of 0.3 mL/min. Liquid chromatography-electrospray ion source Mass was used for the detection. The concentration of sanguinarine and chelerythrine in the range of 0.5~50ng/mL showed a good linear relationship, and the regression coefficients were R2=0.9996 and R2=0.9995 respectively. The average recoveries were 91.50% and 91.30%, and the RSD of the repeatability was 11.68% and 9.90%, the RSD of the precision was 2.58% and 3.89% respectively. The method with high sensitivity and good repeatability is suitable for quantitative determination of quaternary benzo[c]phenanthridine alkaloids in porks.