LC-MS/MS法同时测定人血浆中谷氨酸和精氨酸  

Simultaneous determination of glutamate and arginine in human plasma by LC-MS/MS

作　　者：张丽娜[1] 刘曼[1] 张丹[1] 王晓琳[1] 韩静[1] 刘会臣[1] 

机构地区：[1]航天中心医院临床药理室,北京100049

出　　处：《药物分析杂志》2015年第7期1172-1178,共7页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的：建立测定人血浆中谷氨酸和精氨酸的方法,以应用于人体药代动力学研究。方法：血浆样本经与丹酰氯衍生化反应后,选用Synergi 4μHydro-RP 80A柱（150 mm×2.0 mm,4μm）,以甲醇-10 mmol·L-1乙酸铵（含0.1%甲酸）（50∶50）为流动相,流速0.3 m L·min-1,采用API3200型三重四极杆串联质谱仪的多重反应监测（MRM）扫描方式进行监测,电喷雾离子化源,正离子方式,选择监测离子反应分别为m/z 381.0→170.2（谷氨酸丹酰氯衍生化产物）、m/z 408.0→170.3（精氨酸丹酰氯衍生化产物）和m/z 353.0→157.1（内标D-高丝氨酸丹酰氯衍生化产物）。结果：谷氨酸、精氨酸和内标D-高丝氨酸的丹酰氯衍生化产物的保留时间分别为4.62、3.21和4.39 min;血浆中谷氨酸和精氨酸的线性范围均为3.00-300μg·m L-1（r〉0.99）,定量下限（LLOQ）均为3.00μg·m L-1;批内、批间精密度（RSD）均小于15%;准确度（RE）均在±15%的范围以内;谷氨酸和精氨酸平均提取回收率分别为（85.6±3.3）%和（79.9±3.9）%;平均基质效应因子分别为（116.3±4.9）%和（102.4±7.3）%;稳定性试验中,在各种贮存条件下血浆中谷氨酸和精氨酸均较稳定。结论：该方法适用于谷氨酸精氨酸注射液人体药代动力学研究。Objective：To develop an LC-MS/MS method for determination of glutamate and arginine in human plasma for human pharmacokinetic studies. Methods： After precolumn derivatization with dansyl chloride,the ana- lytes and internal standard（ I. S. ）were separated on a Synergi 4 μm Hydro-RP 80A analytical column（ 150 mm × 2. 0 mm,4μm）using the mobile phase of methanol and 10 mmol· L-1 ammonium acetate containing 0. 1% formic acid（50： 50）at a flow rate of 0. 3 mL· min-1. Detection was carried out by electrospray positive ionization mass spectrometry in multiple reaction monitoring（MRM） mode. The MRM transitions of m/z 381.0 →170. 2, m/z 408.0 →170. 3 and m/z 353.0→157. 1 were applied to quantify the derivatized products of glutamate, arginine and D- homoserine （I. S. ）, respectively. Results： The derivatized products of glutamate, arginine and I. S. were eluted at 4. 62 min,3.21 min and 4. 39 min,respectively. The calibration curves were linear over the concentration range of 3.00-300 μg· mL - 1 （ r 〉 0. 99 ） with the lower limit of quantitation （LLOQ） 3.00 μg·mL -1. Both Intra-and inter -batch relative standard deviations（RSD）were below 15% and the relative errors（RE） were in the range of ＋ 15%. The mean extraction recovery of glutamate and arginine was（ 85.6 ± 3.3 ）% and （79.9 ± 3.9 ）%, and the mean matrix effect factor was（ 116.3 ± 4.9 ） % and （ 102.4 ± 7.3 ） %. In the stability studies, glutamate and argi- nine were found to be stable in plasma under various storage conditions. Conclusion： The established method can be used for the clinical pharmacokinetie study of glutamate and arginine injection.

关 键 词：谷氨酸 精氨酸 LC-MS/MS法 药代动力学 人血浆 

分 类 号：R917[医药卫生—药物分析学]