【摘要】：研究目的： 為測(cè)出十二名健康志愿者單次口服鹽酸阿那格雷膠囊和多次口服給藥后，不同時(shí)刻血漿樣品中阿那格雷的濃度。本次研究建立了一種靈敏度高、分析時(shí)間短、重現(xiàn)性好、樣品處理簡(jiǎn)便，適于樣品高通量分析的LC-MS/MS（液相色譜-質(zhì)譜法）定量方法。通過(guò)試驗(yàn)數(shù)據(jù)對(duì)口服鹽酸阿那格雷膠囊后的體內(nèi)藥動(dòng)學(xué)進(jìn)行分析，為阿那格雷后期臨床安全性、合理用藥提供了可靠的依據(jù)。 研究方法： 使用乙醚-二氯甲烷（2:1，v/v）作為提取試劑，對(duì)阿那格雷血漿樣品采用液-液萃取法進(jìn)行前處理，柱溫調(diào)節(jié)到35C，流速選擇1.0mL/mi（n分流體積比1:1），吸清液層30μL進(jìn)行LC-MS/MS檢測(cè)分析。阿那格雷與內(nèi)標(biāo)格列吡嗪經(jīng)AscetnisC18柱（4.6×150mm I.D.，5μm粒徑），在流動(dòng)相為甲醇-0.1%甲酸水（80:20，v/v）的色譜系統(tǒng)中予以分離之后，再通過(guò)高效液相質(zhì)譜進(jìn)行分析檢測(cè)。選用MRM模式，即多重反應(yīng)監(jiān)測(cè)技術(shù)，選擇ESI (電噴霧離子化源)作為離子源，正離子模式掃描阿那格雷和內(nèi)標(biāo)格列吡嗪，分別以m/z256.3m/z199.0和m/z446.1m/z321.0為定量分析的離子反應(yīng)。 為了確保建立的阿那格雷定量分析方法有效可靠，試驗(yàn)對(duì)如下指標(biāo)進(jìn)行了全面的方法學(xué)確證：包括其特異性、線性范圍、最低定量下限、準(zhǔn)確度、精密度、基質(zhì)效應(yīng)、提取回收率和穩(wěn)定性等。 對(duì)招募的十二名健康志愿者，口服給予鹽酸阿那格雷膠囊各1mg以及多次口服1mg劑量后，為測(cè)出不同時(shí)刻采集的血漿中阿那格雷的濃度，使用已經(jīng)建立好的HPLC-MS/MS定量方法來(lái)進(jìn)行分析。為了了解阿那格雷在人體內(nèi)的藥代動(dòng)力學(xué)所具備的特點(diǎn)，我們使用DAS3.0和SPSS17.0軟件算出相應(yīng)的藥代動(dòng)力學(xué)參變量并用統(tǒng)計(jì)學(xué)方法闡明其藥代動(dòng)力學(xué)參變量的特征。 研究結(jié)果： 為了測(cè)出人體血漿中阿那格雷的濃度，此次實(shí)驗(yàn)創(chuàng)建了快速靈敏、準(zhǔn)確可靠、重現(xiàn)性極好且穩(wěn)定的HPLC-MS/MS測(cè)定方法。結(jié)果表明：在待測(cè)物和內(nèi)標(biāo)格列吡嗪出峰時(shí)間處均沒(méi)有外源性和內(nèi)源性物質(zhì)影響的情況下，阿那格雷血漿樣品的定量線性范圍是0.1-100ng/mL，LLOQ是0.1ng/mL，本方法線性良好(r=0.9971)，且定量范圍較寬。日內(nèi)精密度(日內(nèi)RSD)5.92，日間精密度(日間RSD)10.4，準(zhǔn)確度為3.11%-4.04%，以上值均±15%。阿那格雷低、中、高三個(gè)濃度回收率分別是96.1±3.6%、99.5±1.2%、104±0.91%。內(nèi)標(biāo)格列吡嗪回收率也穩(wěn)定，結(jié)果精密可重現(xiàn)。阿那格雷低、中、高三濃度基質(zhì)效應(yīng)分別是95.4±3.9%、97.1±5.0%、91.1±5.6%以及格列吡嗪基質(zhì)效應(yīng)是95.6±4.0%，即基質(zhì)效應(yīng)干擾不了阿那格雷和格列吡嗪的測(cè)定，能夠滿(mǎn)足測(cè)定的基本要求。經(jīng)過(guò)反復(fù)3次-20C凍融，血漿樣品置于室溫4h后，提取處理后室溫放置4h后，-20C冰凍放置80d后，儲(chǔ)備液室溫放置6h和4C放置7d后都很穩(wěn)定。測(cè)定阿那格雷血漿樣本的分析方法可用于阿那格雷人體藥代動(dòng)力學(xué)試驗(yàn)。 單次服藥和多次服藥后，得到的藥代動(dòng)力學(xué)參變量分別是：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。 經(jīng)過(guò)統(tǒng)計(jì)分析，，在給藥劑量為1mg時(shí)，阿那格雷單次口服與連續(xù)給藥的t1/2、Cmax、CLz/F、Vz/F（P0.05）等主要藥代動(dòng)力學(xué)參變量沒(méi)有明顯性差別。即藥物的連續(xù)口服不會(huì)改變藥物在體內(nèi)的藥代動(dòng)力學(xué)特征。阿那格雷在多次給藥后，AUC下降，MRT變小，而阿那格雷的t1/2小，CLz/F較大，表明多次給藥后阿那格雷快速地被代謝并排出到體外，它與組織親和力小，不會(huì)導(dǎo)致藥物濃度偏高而在體內(nèi)產(chǎn)生蓄積。全程試驗(yàn)周期內(nèi)，自愿者均沒(méi)有不適反應(yīng)現(xiàn)象出現(xiàn)，說(shuō)明阿那格雷具備非常好的安全性能。
[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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R969.1

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相關(guān)期刊論文 前3條

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本文編號(hào)：2511994