高效液相色谱串联质谱法测定中药注射剂对P815细胞脱颗粒后上清液中组胺含量  

Determination of Histamine in the Supernatant of Traditional Chinese Medicine Injection on Degranulation in P815 Cells by High-performance Liquid Chromatography-Mass Spectrometry

作　　者：谭晓梅[1] 傅钧庭[1] 罗佳波[1] 汤庆发[1] 蒙瑞波[1] 

机构地区：[1]南方医科大学中医药学院广东省中药制剂重点实验室,广东广州510515

出　　处：《分析测试学报》2013年第4期483-487,共5页Journal of Instrumental Analysis

基　　金：广东省产学研重大项目(2010A090200076)

摘　　要：建立了液相色谱-串联质谱(LC-MS/MS)检测P815细胞脱颗粒后上清液中组胺含量的方法。采用C18柱(4.6 mm×100 mm,3.5μm)进行分离,以体积比为60∶40的乙腈-0.5 mmol/L乙酸铵溶液(含0.3%甲酸)为流动相,流速0.4 mL/min;在电喷雾正离子化模式下,采用多离子反应检测方式(MRM)进行检测;检测离子对为m/z 112>95、112>68。在优化条件下,样品可在6 min内完成分析,细胞液中组胺在4.0～128.0μg/L范围内线性关系良好(r2=0.997 9)。定量下限(S/N>10)为1.2μg/L,检出限(S/N=3)为0.3μg/L。在加标水平为16、64、128μg/L时,细胞上清液中组胺的平均回收率为90%～94%,相对标准偏差(n=6)为4.1%～5.9%。该方法简便、灵敏、重复性好,可用于中药注射剂对P815细胞脱颗粒后上清液中组胺含量的测定。A high-performance liquid chromatography tandem mass spectrometric （LC -MS/MS ） method was developed for the determination of histamine in the supernatant after degranulation of P815 cells. After pretreatment procedure, the sample was separated on a C18（4. 6 mm ×100 mm, 3.5 μm） column using acetonitrile -0. 5 mmol/L ammonium acetate solution with 0. 3% formic acid （60 ： 40） as mobile phase at a flow rate of 0. 4 mL/min. The target compound was detected by posi- tive ion ionization mass spectrometry（ ESI MS） under multiple reaction monitoring（MRM） mode using m/z 112 〉95 and m/z 112 〉68 as detection ion. Under the optimal conditions, the analysis of sam- ple could be completed within 6 min. A good linear relationship was obtained for histamine in the range of 4. 0 - 128.0 μg/L with a correlation coefficient（ r^2 ） of 0. 997 9. The limit of quantitation （ S/N 〉 10） and limit of detection（ S/N = 3 ） were 1.2 μg/L and 0. 3 μg/L, respectively. The mean recoveries for histamine in the supernatant of cells at three spiked concentration levels of 16, 64, 128 μg,/L were in the range of 90% - 94% with relative standard deviations （ RSDs, n = 6 ） of 4. 1% -5.9% . The proposed method was simple, sensitive and reproducible, and was suitable for the determination of histamin in the supernatant of Traditional Chinese Medicine Injection（TCMI） on degranulation in P815 cells.

关 键 词：组胺 高效液相色谱串联质谱法 中药注射剂 P815细胞 

分 类 号：O657.72[理学—分析化学] R283.62[理学—化学]