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.
出处
《分析测试学报》
CAS
CSCD
北大核心
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)
