GC-MS快速测定鱼腥草注射剂致过敏介质释放的方法研究

Rapid GC-MS analysis for plasma histamine induced by Yuxingcao Injection

目的建立犬血浆中组胺GC-MS定量测定方法。方法 12只比格犬随机分4组,给药组注射含高、中、低3种浓度吐温-80的鱼腥草注射剂和生理盐水组,30 min后取犬血浆,加入氯甲酸乙酯/三氯甲烷进行衍生化。组胺作为过敏标志物用气质联用法进行测定,DB-WAX色谱柱(30 m×0.25 mm,0.25μm);柱温从150℃开始,再以40℃/min的速度升至240℃。质谱条件采用SIM选择离子模式,选择m/z166为组胺的检测离子,选择m/z192为内标9-methylanthracene的检测离子,内标以及组胺标的色谱保留时间分别为11和13.5 min。结果组胺在2～32 mg/L范围内具有良好线性关系,r2=0.999,平均回收率为100.6%(RSD=1.86%),最低检测限为0.7 mg/L(S/N=3)。结论 本实验方法操作简便、准确、回收率好,可快速检测中药注射剂引起的组胺过敏介质的释放,为临床、制药企业解决中药注射剂的安全性问题提供科学方法。

AIM To measure histamine concentration in plasma of beagle dogs induced by Yuxingcao Injection by GC-MS.METHODS Twelve beagle dogs were randomly divided into four groups,Yuxingcao Injection groups with high,medium,and low dosages of tween-80,and saline group.Blood samples taken from dogs were derived with ethyl chlorocarbonate/trichloromethane.Histamine was selected as the marker to evaluation of allergic reactions.A DB-WAX（30 m×0.25 mm,0.25 μm） column was used for the GC-MS analysis.The column temperature was programmed from 150 ℃ to 240 ℃ at a rate of 40 ℃/min.The ion m/z 166 and 192 were chosen as the qualifying ion for histamine and internal standard,with the retention time of 13.5 min and 11 min,respectively.RESULTS The linear relationship was excellent within the range of 2-32 mg/L（r2=0.999,n = 5）,and the limit of detection was 0.7 mg/L（S/N=3）.The average recovery of histamine was 100.6%（RSD was 1.86%）.CONCLUSION The method is accurate,sensitive and convenient for the measurement of histamine concentration in plasma.