盐酸阿那格雷胶囊人体药代动力学研究
发布时间：2019-07-09 07:56
【摘要】：研究目的： 为测出十二名健康志愿者单次口服盐酸阿那格雷胶囊和多次口服给药后，不同时刻血浆样品中阿那格雷的浓度。本次研究建立了一种灵敏度高、分析时间短、重现性好、样品处理简便，适于样品高通量分析的LC-MS/MS（液相色谱-质谱法）定量方法。通过试验数据对口服盐酸阿那格雷胶囊后的体内药动学进行分析，为阿那格雷后期临床安全性、合理用药提供了可靠的依据。 研究方法： 使用乙醚-二氯甲烷（2:1，v/v）作为提取试剂，对阿那格雷血浆样品采用液-液萃取法进行前处理，柱温调节到35C，流速选择1.0mL/mi（n分流体积比1:1），吸清液层30μL进行LC-MS/MS检测分析。阿那格雷与内标格列吡嗪经AscetnisC18柱（4.6×150mm I.D.，5μm粒径），在流动相为甲醇-0.1%甲酸水（80:20，v/v）的色谱系统中予以分离之后，再通过高效液相质谱进行分析检测。选用MRM模式，即多重反应监测技术，选择ESI (电喷雾离子化源)作为离子源，正离子模式扫描阿那格雷和内标格列吡嗪，分别以m/z256.3m/z199.0和m/z446.1m/z321.0为定量分析的离子反应。 为了确保建立的阿那格雷定量分析方法有效可靠，试验对如下指标进行了全面的方法学确证：包括其特异性、线性范围、最低定量下限、准确度、精密度、基质效应、提取回收率和稳定性等。 对招募的十二名健康志愿者，口服给予盐酸阿那格雷胶囊各1mg以及多次口服1mg剂量后，为测出不同时刻采集的血浆中阿那格雷的浓度，使用已经建立好的HPLC-MS/MS定量方法来进行分析。为了了解阿那格雷在人体内的药代动力学所具备的特点，我们使用DAS3.0和SPSS17.0软件算出相应的药代动力学参变量并用统计学方法阐明其药代动力学参变量的特征。 研究结果： 为了测出人体血浆中阿那格雷的浓度，此次实验创建了快速灵敏、准确可靠、重现性极好且稳定的HPLC-MS/MS测定方法。结果表明：在待测物和内标格列吡嗪出峰时间处均没有外源性和内源性物质影响的情况下，阿那格雷血浆样品的定量线性范围是0.1-100ng/mL，LLOQ是0.1ng/mL，本方法线性良好(r=0.9971)，且定量范围较宽。日内精密度(日内RSD)5.92，日间精密度(日间RSD)10.4，准确度为3.11%-4.04%，以上值均±15%。阿那格雷低、中、高三个浓度回收率分别是96.1±3.6%、99.5±1.2%、104±0.91%。内标格列吡嗪回收率也稳定，结果精密可重现。阿那格雷低、中、高三浓度基质效应分别是95.4±3.9%、97.1±5.0%、91.1±5.6%以及格列吡嗪基质效应是95.6±4.0%，即基质效应干扰不了阿那格雷和格列吡嗪的测定，能够满足测定的基本要求。经过反复3次-20C冻融，血浆样品置于室温4h后，提取处理后室温放置4h后，-20C冰冻放置80d后，储备液室温放置6h和4C放置7d后都很稳定。测定阿那格雷血浆样本的分析方法可用于阿那格雷人体药代动力学试验。 单次服药和多次服药后，得到的药代动力学参变量分别是：Cmax：8.47±2.05ng/mL和8.85±2.30ng/mL；t1/2：1.68±0.71h和1.82±0.70h；Tmax：0.60±0.13h和0.77±0.13h；AUC0-t：16.49±4.30ng/mL*h和13.46±3.60ng/mL*h；AUC0-∞：16.49±4.30ng/mL*h和13.46±3.60ng/mL*h；MRT：2.25±0.40h和1.97±0.27h；Vz/F：151.31±62.20L和155.15±61.29L；CLz/F：64.78±17.99L/h和65.75±15.80L/h。 经过统计分析，，在给药剂量为1mg时，阿那格雷单次口服与连续给药的t1/2、Cmax、CLz/F、Vz/F（P0.05）等主要药代动力学参变量没有明显性差别。即药物的连续口服不会改变药物在体内的药代动力学特征。阿那格雷在多次给药后，AUC下降，MRT变小，而阿那格雷的t1/2小，CLz/F较大，表明多次给药后阿那格雷快速地被代谢并排出到体外，它与组织亲和力小，不会导致药物浓度偏高而在体内产生蓄积。全程试验周期内，自愿者均没有不适反应现象出现，说明阿那格雷具备非常好的安全性能。
[Abstract]:The purpose of the study: in ord to measure that concentration of anaggrel in plasma samples at different time points after a single oral administration of the aggrel capsule and multiple oral administration of 12 healthy volunteers The method has the advantages of high sensitivity, short analysis time, good reproducibility and simple sample treatment, and is suitable for quantitative prescription of high-throughput analysis of the sample by LC-MS/ MS (liquid chromatography-mass spectrometry). Methods: The pharmacokinetics of the oral hydrochloride capsules were analyzed by the test data, which provided a reliable basis for the clinical safety and reasonable medication in the later stage. According to. Research The method comprises the following steps of: taking diethyl ether-dichloromethane (2:1, v/ v) as an extraction reagent, performing pretreatment on the sample of the aggrel plasma by a liquid-liquid extraction method, adjusting the temperature of the column to 35C, selecting a flow rate of 1.0 mL/ mi (n-split volume ratio of 1:1), and performing LC-MS/ M on the liquid-absorbing liquid layer 300.mu. L S. Analysis of S. Agaggrel and the inner standard column were separated by an Ascension C18 column (4.6-150 mm I. D.,5. mu.m particle size), after being separated in a chromatography system with a mobile phase of methanol-0.1% formic acid (80:20, v/ v), and then by high-performance liquid-phase mass spectrometry. The MRM mode, i.e. the multi-reaction monitoring technique, was selected to select the ESI (electrospray ionization source) as the ion source. The positive ion mode was used as the ion source. The positive ion mode was used as the positive ion mode. In order to ensure the effective and reliable method for the quantitative analysis of the agrela, a comprehensive methodology for the determination of the following indexes is carried out: including its specificity, linear range, minimum limit of quantification, accuracy, precision, matrix effect, and extraction The yield and stability of the 12 healthy volunteers were given orally with 1 mg of the aggrel hydrochloride capsule and once a dose of 1 mg for several times, the concentration of the aggrel in the plasma collected at different times was measured, and the established HPLC-MS/ MS was used. In order to understand the characteristics of the pharmacokinetics of agana in human, we use the DAS3.0 and the SPSS17.0 software to calculate the corresponding pharmacokinetic parameters and clarify their pharmacokinetics by a statistical method dynamic parametric variation The results of the study are as follows: In order to measure the concentration of agana in human plasma, this experiment has created a fast, sensitive, accurate, reproducible and stable H The results of the method of PLC-MS/ MS show that the quantitative linear range of the samples in the AAGGREY plasma is 0.1 -100 ng/ mL, the LLOQ is 0.1 ng/ mL, and the linearity of the method is good (r = 0). 9971) with a wide range of quantitation. Intra-day precision (intra-day RSD) 5.92, day-to-day precision (day-day RSD) 10.4, accuracy of 3.11% -4 The recoveries were 96.1% 3.6% and 99.5%1% respectively. .2%,104% 0.91%. The yield is stable and the results are accurate and reproducible. The matrix effect of the medium and high three-concentration matrix is 95.4%, 3.9%, 97.1%, 5.0%, 91.1% and 5.6%, respectively, and the matrix effect of the gliclazide is 95.6% to 4.0%, that is, the matrix effect can not interfere with the test of Agaggrel and gliclazide. The method can meet the basic requirements of the measurement. After repeated 3 times to 20 C freeze-thaw, the plasma sample is placed at room temperature for 4 h, and after the extraction treatment, after being placed at room temperature for 4 h, the storage solution is placed at room temperature for 80d, and the stock solution is placed at room temperature for 6 days. Both h and 4C were stable after 7 d. The analytical method for the determination of the plasma samples of the Agaggrel plasma may be used The pharmacokinetic parameters obtained after single and multiple doses were: Cmax: 8.47, 2.05 ng/ mL and 8.85-2.30 ng/ mL; t1/2: 1.68, 0.71 h and 1.82-0.70h; Tmax: 0.60-0.13 h and 0.77-0.13 h; AUC0-t: 16.49-4.30 ng/ mL * h and 13.46-3.60 ng/ mL * h; AUC0-1: 16.49-4.30 ng/ mL * h and 13.46-3.60 ng/ mL * h; MRT: 2.25-0.40 h and 1.97-0.27 h; Vz/ F: 151.31-62.20 L and 155.15-61.29 L; CLz/ F: 64.78-17.99 L/ h And 65.75 to 15.80 L/ h. After a statistical analysis, the time t1/2, Cmax, CLz/ F, Vz/ F (P0.05), etc., of the single oral and continuous administration of anaggrel at the administration dose of 1 mg, etc. There was no significant difference in the main pharmacokinetic parameters, i.e., continuous oral administration of the drug The pharmacokinetic profile of the drug in the body was not changed. It can lead to high drug concentration and accumulation in the body. During the whole test period, there is no discomfort to the volunteers.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R969.1
本文编号：2511994
[Abstract]:The purpose of the study: in ord to measure that concentration of anaggrel in plasma samples at different time points after a single oral administration of the aggrel capsule and multiple oral administration of 12 healthy volunteers The method has the advantages of high sensitivity, short analysis time, good reproducibility and simple sample treatment, and is suitable for quantitative prescription of high-throughput analysis of the sample by LC-MS/ MS (liquid chromatography-mass spectrometry). Methods: The pharmacokinetics of the oral hydrochloride capsules were analyzed by the test data, which provided a reliable basis for the clinical safety and reasonable medication in the later stage. According to. Research The method comprises the following steps of: taking diethyl ether-dichloromethane (2:1, v/ v) as an extraction reagent, performing pretreatment on the sample of the aggrel plasma by a liquid-liquid extraction method, adjusting the temperature of the column to 35C, selecting a flow rate of 1.0 mL/ mi (n-split volume ratio of 1:1), and performing LC-MS/ M on the liquid-absorbing liquid layer 300.mu. L S. Analysis of S. Agaggrel and the inner standard column were separated by an Ascension C18 column (4.6-150 mm I. D.,5. mu.m particle size), after being separated in a chromatography system with a mobile phase of methanol-0.1% formic acid (80:20, v/ v), and then by high-performance liquid-phase mass spectrometry. The MRM mode, i.e. the multi-reaction monitoring technique, was selected to select the ESI (electrospray ionization source) as the ion source. The positive ion mode was used as the ion source. The positive ion mode was used as the positive ion mode. In order to ensure the effective and reliable method for the quantitative analysis of the agrela, a comprehensive methodology for the determination of the following indexes is carried out: including its specificity, linear range, minimum limit of quantification, accuracy, precision, matrix effect, and extraction The yield and stability of the 12 healthy volunteers were given orally with 1 mg of the aggrel hydrochloride capsule and once a dose of 1 mg for several times, the concentration of the aggrel in the plasma collected at different times was measured, and the established HPLC-MS/ MS was used. In order to understand the characteristics of the pharmacokinetics of agana in human, we use the DAS3.0 and the SPSS17.0 software to calculate the corresponding pharmacokinetic parameters and clarify their pharmacokinetics by a statistical method dynamic parametric variation The results of the study are as follows: In order to measure the concentration of agana in human plasma, this experiment has created a fast, sensitive, accurate, reproducible and stable H The results of the method of PLC-MS/ MS show that the quantitative linear range of the samples in the AAGGREY plasma is 0.1 -100 ng/ mL, the LLOQ is 0.1 ng/ mL, and the linearity of the method is good (r = 0). 9971) with a wide range of quantitation. Intra-day precision (intra-day RSD) 5.92, day-to-day precision (day-day RSD) 10.4, accuracy of 3.11% -4 The recoveries were 96.1% 3.6% and 99.5%1% respectively. .2%,104% 0.91%. The yield is stable and the results are accurate and reproducible. The matrix effect of the medium and high three-concentration matrix is 95.4%, 3.9%, 97.1%, 5.0%, 91.1% and 5.6%, respectively, and the matrix effect of the gliclazide is 95.6% to 4.0%, that is, the matrix effect can not interfere with the test of Agaggrel and gliclazide. The method can meet the basic requirements of the measurement. After repeated 3 times to 20 C freeze-thaw, the plasma sample is placed at room temperature for 4 h, and after the extraction treatment, after being placed at room temperature for 4 h, the storage solution is placed at room temperature for 80d, and the stock solution is placed at room temperature for 6 days. Both h and 4C were stable after 7 d. The analytical method for the determination of the plasma samples of the Agaggrel plasma may be used The pharmacokinetic parameters obtained after single and multiple doses were: Cmax: 8.47, 2.05 ng/ mL and 8.85-2.30 ng/ mL; t1/2: 1.68, 0.71 h and 1.82-0.70h; Tmax: 0.60-0.13 h and 0.77-0.13 h; AUC0-t: 16.49-4.30 ng/ mL * h and 13.46-3.60 ng/ mL * h; AUC0-1: 16.49-4.30 ng/ mL * h and 13.46-3.60 ng/ mL * h; MRT: 2.25-0.40 h and 1.97-0.27 h; Vz/ F: 151.31-62.20 L and 155.15-61.29 L; CLz/ F: 64.78-17.99 L/ h And 65.75 to 15.80 L/ h. After a statistical analysis, the time t1/2, Cmax, CLz/ F, Vz/ F (P0.05), etc., of the single oral and continuous administration of anaggrel at the administration dose of 1 mg, etc. There was no significant difference in the main pharmacokinetic parameters, i.e., continuous oral administration of the drug The pharmacokinetic profile of the drug in the body was not changed. It can lead to high drug concentration and accumulation in the body. During the whole test period, there is no discomfort to the volunteers.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R969.1
【参考文献】
相关期刊论文 前3条
1 齐美玲;;液相色谱-质谱法在生物样品药物定量分析中的基质效应[J];药物分析杂志;2005年04期
2 钟大放;以加权最小二乘法建立生物分析标准曲线的若干问题[J];药物分析杂志;1996年05期
3 蒋祖军;阿那格雷简介[J];中国医院药学杂志;2004年06期
本文编号：2511994
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