人血浆中异丙肌苷与奥普力农两种药物的LC-MSMS测定方法的研究及应用

发布时间：2018-06-16 20:40

本文选题：盐酸奥普力农 + 高效液相串联质谱　；参考：《长春工业大学》2014年硕士论文

【摘要】：盐酸奥普力农属于磷酸二酯酶抑制剂类治疗慢性心衰的正性肌力药物。本文在于建立一种灵敏度高、重现性好、分析时间短、样品处理简便,适用于样品的高通量分析人血浆及尿液中奥普力农的高效液相串联质谱(LC-MS/MS)测定方法,并在该方法的基础上,对健康受试者静脉推注奥普力农盐酸盐后人体药代动力学进行研究。为奥普力农盐酸盐推注液提供临床依据。异丙肌苷是一种免疫增强剂。异丙肌苷有三部分肌苷、对乙酰氨基苯甲酸和N,N一二甲氨基-2-丙醇通过合成方法制备而来。服用异丙肌苷后,其在体内可以迅速水解为肌苷,本文建立了一种快速、灵敏、准确的肌苷LC-MS/MS分析方法,并通过该方法测定健康受试者服用异丙肌苷后,血浆中的肌苷含量。奥普力农采用液液萃取吹干上清复溶后进行LC-MS/MS分析。实验中采用MRM模式,同时检测奥普力农及内标可待因。并对其各种稳定性、提取回收率、基质效应等进行了方法确证,结果表明此方法符合CFDA的相关要求。同时选用12名健康受试者,随机进行三剂量三交叉方式进行静脉推注给药,并于设定时间点进行样品的采集并保存待分析。由于肌苷属于内源性物质,且肌苷在血液中不稳定,本文采用特殊沉淀蛋白的方法对其进行处理：全血取出后置于肝素化试管中,立即低温离心。取出血浆置于另一含有高氯酸的试管中,充分振荡混匀,并低温离心,取上清,于培80℃冷冻待测。本论文建立了人血浆及尿液中盐酸奥普力农的LC-MS/MS分析方法,该方法快速、准确、灵敏,适于奥普力农的生物样品分析。健康人静脉推注低、中、高三剂量后,其Cmax分别为20.86±12.97(mean±SD)、43.92±22.59和67.25±19.37；t1/2分别为0.96±0.26、0.84±0.37和0.99±0.17；AUC0-∞分别为10.93±3.83、22.55±7.53和37.42±7.70。本研究建立LC-MS/MS测定人血浆中肌苷浓度的方法并进行了方法确证。该方法专属性强,样品处理方便,灵敏度高,适用于肌苷临床药动学研究。
[Abstract]:Aupu hydrochloric acid is a positive inotropic drug for the treatment of chronic heart failure with phosphodiesterase inhibitors. This paper is to establish a high throughput analysis method for high throughput analysis of Aupu force farm in human plasma and urine with high sensitivity, good reproducibility, short analysis time, simple sample treatment, and high throughput analysis of LC-MS/MS in human plasma and urine. On the basis of this method, the pharmacokinetics of Aupu in healthy volunteers were studied after intravenous injection of Aupu hydrochloride, which provided a clinical basis for Aupu hydrochloride injection.
Isosinosine is an immunoenhancer. Isosinosine has three parts of inosine, which is prepared by synthesis of acetaminophen and N, N one or two methylamino -2- propanol. After taking isosinosine, it can quickly hydrolyze into inosine in the body. A rapid, sensitive and accurate method for the analysis of inosine LC-MS/MS was established and passed by this method. Methods the inosine content in plasma of healthy subjects after taking isoprenin was determined.
Aupu force farmer uses liquid liquid extraction and blow dry supernatant to carry out LC-MS/MS analysis. In the experiment, MRM model was used, and Aupu force farm and codeine were detected at the same time. All kinds of stability, recovery rate and matrix effect were confirmed by methods. The results showed that this method accords with the requirements of CFDA. At the same time, 12 healthy subjects are selected. Three doses and three crossover methods were randomly administered intravenously, and samples were collected and stored for analysis at set time points.
As inosine is an endogenous substance and inosine is unstable in the blood, this article is treated with a special precipitation protein: after taking out the whole blood and placed in a heparinization test tube, then centrifugation at a low temperature. The plasma is placed in a test tube containing perchloric acid, which is well mixed up and centrifuged at low temperature, Torikami Kiyo, frozen at 80. To be measured.
The method of LC-MS/MS analysis of Aupu hydrochloric acid in human plasma and urine was established in this paper. This method is fast, accurate, sensitive and suitable for the analysis of the biological samples of Aupu. After the low, middle and high dose of healthy human intravenous injection, the Cmax is 20.86 + 12.97 (mean + SD), 43.92 + 22.59 and 67.25 + 19.37 respectively, and t1/2 is 0.96 + 0.26,0.84 + 0.37 respectively. And 0.99 + 0.17; AUC0- infinity were 10.93 + 3.83,22.55 + 7.53 and 37.42 + 7.70. respectively.
In this study, a method for the determination of inosine concentration in human plasma by LC-MS/MS was established and confirmed by method. This method is highly specific, convenient and sensitive. It is suitable for the clinical pharmacokinetic study of inosine.
【学位授予单位】：长春工业大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：O657.63;R927.2

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