论文部分内容阅读
目的:建立能够灵敏、特异、准确、可靠的测定血清中氯贝胆碱浓度的分析方法。用于针刺穴位药物增效机理的研究。方法:优化氯贝胆碱及内标氘代乙酰胆碱离子化及断裂方式,确定相关液相色谱条件;开展方法学考察,以验证新建分析方法的准确性、精密度等,在此基础上将该方法用于分析大鼠给药后所得的实际血清样品,以验证新建分析方法的适用性。同时测定了大鼠穴位给药及静脉给药后血清样品中氯贝胆碱的浓度。结果:ESI(+)为氯贝胆碱提供最佳的离子化条件,采用MRM工作模式,用m/z161.0→101.9来检测氯贝胆碱,同时,以氘代乙酰胆碱-d9m/z155→87为内标化合物。氯贝胆碱及氘代乙酰胆碱-d9色谱保留时间分别为5.4min和5.0min。氯贝胆碱在50～300ng·mL-1范围内线性良好,回归方程为Y=0.002397X+0.0266,r=0.9944,检测限为0.0025ng·mL-1。高、中、低浓度的绝对回收率为96.4%～102.1%,方法回收率为100.6%～108.9%。日内精密度(n=6)为2.6%～5.7%,日间精密度(n=3)为4.1%～8.4%。结论:应用LC-MS/MS技术建立测定血清中氯贝胆碱的新方法灵敏可靠,为针药联合干预后血药浓度的比较研究以及针药联合干预后靶器官药物浓度的检测等研究工作打下了重要的分析方法学基础。
OBJECTIVE: To establish a sensitive, specific, accurate and reliable method for the determination of chlorphenesin in serum. Study on Synergistic Mechanism of Acupuncture Point Drugs. Methods: To optimize the ionization and cleavage of chloralcholine and internal standard deuteroacetylcholine, to determine the relevant liquid chromatographic conditions; to carry out methodological investigation to verify the accuracy and precision of the new analytical method, based on which the The method was used to analyze actual serum samples obtained after rat administration to verify the applicability of the new assay. Simultaneously, the concentrations of chlorphenesin in serum samples after acupoint administration and intravenous administration were determined. Results: ESI (+) provides the optimal ionization conditions for chloranil. MRM is used to detect chlorphe- choline with m / z161.0 → 101.9. Meanwhile, deuterated acetylcholine-d9m / z155 → 87 is the internal standard compound. Chloranil choline and deutero acetylcholine-d9 chromatographic retention time was 5.4min and 5.0min. The calibration curve was linear over the range of 50 ~ 300 ng · mL-1. The regression equation was Y = 0.002397X + 0.0266, r = 0.9944. The detection limit was 0.0025 ng · mL-1. The absolute recoveries of high, medium and low concentrations ranged from 96.4% to 102.1%, and the recoveries were 100.6% -108.9%. Intra-day precision (n = 6) was 2.6% to 5.7%, day-to-day precision (n = 3) was 4.1% to 8.4%. Conclusion: The new method of LC-MS / MS for the determination of clopidogrel in serum is sensitive and reliable. It is a comparative study of plasma concentration after the combination of acupuncture and medicine, as well as the detection of target organ concentration after the intervention of acupuncture and medicine Laid the important foundation of analytical methodology.