UPLC-ESI-MS/MS测定大鼠血浆降血压肽的方法学研究  

Methodological Study of the Determination of Antihypertensive Peptidein Rat Plasma by UPLC-ESI-MS/MS

作　　者：何利思[1] 王金林[1] 崔淑芬[1] HE Lisi;WANG Jinlin;CUI Shufen(School of Food and Drug,Shenzhen Polytechnic,Shenzhen,Guangdong 518055,China)

机构地区：[1]深圳职业技术学院食品药品学院,广东深圳518055

出　　处：《深圳职业技术学院学报》2022年第3期55-59,共5页Journal of Shenzhen Polytechnic

基　　金：深圳市科技计划资助项目(GJHZ20180928161212140).

摘　　要：建立了超高效液相色谱-电喷雾串联质谱(UPLC-ESI MS/MS)法测定大鼠血浆中的重组降血压肽VLPVPR的方法.大鼠血浆样品经甲醇沉淀蛋白处理后,经Thermo Hypersil GOLD C18柱(100 mm×2.1 mm,1.9μm)分离,流动相为10 mmol·L乙酸铵水溶液-乙腈.梯度洗脱流程为:0~4 min乙腈(vol%)10%~90%;4~4.5 min乙腈90%~10%,4.5~6.5 min乙腈保持10%,流速:0.2 mL·min^(-1).质谱条件采用ESI离子源,检测方式为正离子电离,选择性离子检测(MRM).VLPVPR和内标VLPVPR-R位C,N双标的选择性检测离子分别为m/z 680.4→255.2和m/z 690.6→264.2.VLPVPR浓度在0.1~100 ng·mL^(-1)范围内线性良好;定量下限为0.1 ng·mL^(-1);日内、日间RSD均小于15%;低、中、高3个浓度下的提取回收率分别为(101±7.4)%;(99±4.2)%;(100±3.3)%.该方法专属性强,灵敏度高,血浆中内源物对测定无干扰,适用于复杂生物基质中VLPVPR的检测,可用于VLPVPR药代动力学方面的研究.To establish an UPLC-ESI-MS/MS method for the determination of recombinant antihypertensive peptide VLPVPR in rat plasma,plasma samples were precipitated by methanol,and the analyte was then separated on a Thermo Hypersil GOLD C18(100 mm×2.1 mm,1.9μm)with the mobile phase of acetonitrile and 10 mmol•L^(-1) ammonium acetate at a flow rate of 0.2 mL•min^(-1).Gradient elution program:acetonitrile 10%~90%,0~4 min;acetonitrile 90%~10%,4~4.5 min;acetonitrile 10%,continue 2 min.Detection was carried out by electrospray positive ionization mass spectrometry in the multiple reaction monitoring(MRM)mode of the triple quadrupole rod tandem mass spectrometer.Monitoring the transitions m/z 680.4→255.2 for VLPVPR and m/z 690.6→264.2 for VLPVPR-R C,N double labeled.Good linearity was obtained over the range of 0.1~100 ng•mL^(-1),and the LOQ was 0.1 ng•mL^(-1).The intra-and inter-day previsions were all below 15%(RSD).The three different concentrations(low,medium,high)recovery of VLPVPR was(101±7.4)%,(99±4.2)%and(100±3.3)%respectively.The UPLC-ESI MS/MS method for VLPVPR determination is specificity and sensitivity,and could be applied to the pharmacokinetic study.

关 键 词：超高效液相色谱-电喷雾串联质谱 重组降血压肽 内标法 大鼠血浆 

分 类 号：R917[医药卫生—药物分析学]