Wu Ting, Zhang Jun, Tan Heng-Shan, Ju Wen-Zheng, Xu Xiang-Yang
Zhongguo Zhong Yao Za Zhi. 2014 May;39(10):1928-32.
To establish a LC-MS/MS method for quantification of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid in rats plasma and study its pharmacokinetics after administration of Mailuoning injection at a single dose to rats. Plasma samples were acidified with hydrochloric acid and extracted with ethyl acetate. The analytes were determined by LC-MS-MS using a ZOBAX SB C18 column with a mobile phase of methanol-water (containing 2 mmol x L(-1) ammonium acetic) (60:40)at a flow rate of 0.5 mL x min(-1) and detected using ESI with negative ionization mode. Ions monitored in the multiple reaction monitoring (MRM) mode were m/z 353.1/191.0 [M-H]- for chlorogenic acid, m/z 178.9/134.9 [M-H]- for caffeic acid, m/z 515.2/353.0 [M-H]-for 3,4-DCQA, m/z 193.0/133.9 [M-H]-for ferulic acid, m/z 146.9/102.9 [M-H]- for cinnamic acid and m/z 246.0/125.8 [M-H]- for tinidazole (IS). After administration of Mailuoning injection at a single dose to eight Sprague-Dawley rats, the concentrations of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid in plasma were determined by LC-MS/MS method. The main pharmacokinetics parameters of measured data were caluculated by using DASver 1.0 software. The linear concentration ranges of the calibration curves for chlorogenic acid, caffeic acid, 3,4-DCQA and cinnamic acid were 2.006-1,027 microg x L(-1) (r = 0.999 6), 1.953-1,000 microg x L(-1) (r = 0.999 7), 28.51-1.459 x 10(4) microg x L(-1) (r = 0.998 9), 1.836-940.0, g x L(-1) (r = 0.997 7) and 4.780-2,447 microg x L(-1) (r = 0.998 6) respectively. The inner and inter-days relative standard deviations were both less than 5.0%, indicating legitimate precise and accuracy to the requirement of biological sample analysis. For chlorogenic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (49.78 +/- 12.81) min, (123.55 +/- 14.82) mg x min x L(-1) and (0.004 3 +/- 0.000 5) L x min(-1), respectively. For caffeic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (36.65 +/- 10.59) min, (91.67 +/- 11.77) mg x min L(-1) and (0.005 7 +/- 0.000 7) L x min(-1), respectively. For 3,4-DCQA, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (50.08 +/- 13.78) min, (278.34 +/- 31.82) mg x min x L-1 and (0.001 6 +/- 0.000 2) L x min(-1), respectively. For ferulic acid, the pharmacokinetic parameter t1/2, AUC0-t, and CL were (51.39 +/- 15.52) min, (34.72 +/- 4.67) mg x min x L(-1) and (0.000 4 +/- 0.0001) L x min(-1), respectively. For cinnamic acid, the pharmacokinetic parameter t1/2, AUCo-t, and CL were (74.42 +/- 18.32) min, (34.63 +/- 4.82) mg x min x L(-1) and (0.007 7 +/- 0.001 1) L x min-', respectively. The assay method is proved to be sensitive, accurate and convenient. It can be applied to the pharmacokinetic study of chlorogenic acid, caffeic acid, 3,4-DCQA, ferulic acid and cinnamic acid.
建立一种用于定量大鼠血浆中绿原酸、咖啡酸、3,4 - 二咖啡酰奎宁酸、阿魏酸和肉桂酸的液相色谱 - 串联质谱(LC - MS/MS)方法,并研究大鼠单次静脉注射脉络宁注射液后的药代动力学。血浆样品用盐酸酸化后,用乙酸乙酯萃取。采用LC - MS/MS法,以ZOBAX SB C18柱为分析柱,流动相为甲醇 - 水(含2 mmol·L⁻¹乙酸铵)(60:40),流速为0.5 mL·min⁻¹,采用电喷雾电离(ESI)负离子模式进行检测。多反应监测(MRM)模式下监测的离子为:绿原酸m/z 353.1/191.0 [M - H]⁻,咖啡酸m/z 178.9/134.9 [M - H]⁻,3,4 - 二咖啡酰奎宁酸m/z 515.2/353.0 [M - H]⁻,阿魏酸m/z 193.0/133.9 [M - H]⁻,肉桂酸m/z 146.9/102.9 [M - H]⁻,替硝唑(内标)m/z 246.0/125.8 [M - H]⁻。对8只SD大鼠单次静脉注射脉络宁注射液后,采用LC - MS/MS法测定血浆中绿原酸、咖啡酸、3,4 - 二咖啡酰奎宁酸、阿魏酸和肉桂酸的浓度。用DASver 1.0软件计算实测数据的主要药代动力学参数。绿原酸、咖啡酸、3,4 - 二咖啡酰奎宁酸和肉桂酸校准曲线的线性浓度范围分别为2.006 - 1027 μg·L⁻¹(r = 0.999 6)、1.953 - 1000 μg·L⁻¹(r = 0.999 7)、28.51 - 1.459×10⁴ μg·L⁻¹(r = 0.998 9)、1.836 - 940.0 μg·L⁻¹(r = 0.997 7)和4.780 - 2447 μg·L⁻¹(r = 0.998 6)。日内和日间相对标准偏差均小于5.0%,表明该方法对生物样品分析的精密度和准确度符合要求。绿原酸的药代动力学参数t1/2、AUC0 - t和CL分别为(49.78 ± 12.81)min、(123.55 ± 14.82)mg·min·L⁻¹和(0.004 3 ± 0.000 5)L·min⁻¹。咖啡酸的药代动力学参数t1/2、AUC0 - t和CL分别为(36.65 ± 10.59)min、(91.67 ± 11.77)mg·min·L⁻¹和(0.005 7 ± 0.000 7)L·min⁻¹。3,4 - 二咖啡酰奎宁酸的药代动力学参数t1/2、AUC0 - t和CL分别为(50.08 ± 13.78)min、(278.34 ± 31.82)mg·min·L⁻¹和(0.001 6 ± 0.000 2)L·min⁻¹。阿魏酸的药代动力学参数t1/2、AUC0 - t和CL分别为(51.39 ± 15.52)min、(34.72 ± 4.67)mg·min·L⁻¹和(0.000 4 ± 0.0001)L·min⁻¹。肉桂酸的药代动力学参数t1/2、AUCo - t和CL分别为(74.42 ± 18.32)min、(34.63 ± 4.82)mg·min·L⁻¹和(0.007 7 ± 0.001 1)L·min⁻¹。该测定方法灵敏、准确、简便,可用于绿原酸、咖啡酸、3,4 - 二咖啡酰奎宁酸、阿魏酸和肉桂酸的药代动力学研究。