Development and Validation of Rapid and Sensitive HPLC Method for the Determination of Methotrexate in Human Serum

Stamford Journal of Pharmaceutical Sciences Pub Date : 2010-08-12 DOI:10.3329/SJPS.V2I1.1693

M. Nagulu, V. Kiran, Y. N. Reddy, D. Krishna

{"title":"Development and Validation of Rapid and Sensitive HPLC Method for the Determination of Methotrexate in Human Serum","authors":"M. Nagulu, V. Kiran, Y. N. Reddy, D. Krishna","doi":"10.3329/SJPS.V2I1.1693","DOIUrl":null,"url":null,"abstract":"Methotrexate competitively inhibits dihydrofolic acid reductase and thereby inhibits DNA synthesis and cellular replication.This study describes a simple and fast high-performance liquid chromatography method for the determination of methotrexate [MTX] in serum.samples were collected from adult cancer patients receiving high dose MTX at Mahathma Gandhi Memorial hospital (Warangal,AP.India) at various time intervals after the end of each infusion. Serum was deproteinized with trichloroacetic acid and the supernatant was injected into a 250×4.6 mm octadecylsilane column. Mobile phase was made of TRIS-phosphate buffer (pH 5.7): methanol: acetonitrile (70:20:10) with a flow rate of 1ml/min. Ultraviolet detection was done at 313 nm and at ambient temperature. Para aminoacetophenone was used as internal standard. Methotrexate and internal standard retention times were 4.6 and 9.5 minutes, respectively. Results showed that reproducibility (precision) of method within a day was 2.6 to 6 percent and between days was 5.5 to 9.5 percent. The recovery of the method was between 61.5 and 72.7 percent. The quantitation limit of the method for methotrexate was 0.1μM. This method is suitable for quantitation of methotrexate after infusion of high doses of this drug and has good accuracy, precision and quantitation limit. Key Words: Methotrexate; HPLC; Serum Concentration. DOI: 10.3329/sjps.v2i1.1693 Stamford Journal of Pharmaceutical Sciences Vol.2(1) 2009: 8-13","PeriodicalId":21823,"journal":{"name":"Stamford Journal of Pharmaceutical Sciences","volume":"20 1","pages":"8-13"},"PeriodicalIF":0.0000,"publicationDate":"2010-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"7","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Stamford Journal of Pharmaceutical Sciences","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.3329/SJPS.V2I1.1693","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}

引用次数: 7

Abstract

Methotrexate competitively inhibits dihydrofolic acid reductase and thereby inhibits DNA synthesis and cellular replication.This study describes a simple and fast high-performance liquid chromatography method for the determination of methotrexate [MTX] in serum.samples were collected from adult cancer patients receiving high dose MTX at Mahathma Gandhi Memorial hospital (Warangal,AP.India) at various time intervals after the end of each infusion. Serum was deproteinized with trichloroacetic acid and the supernatant was injected into a 250×4.6 mm octadecylsilane column. Mobile phase was made of TRIS-phosphate buffer (pH 5.7): methanol: acetonitrile (70:20:10) with a flow rate of 1ml/min. Ultraviolet detection was done at 313 nm and at ambient temperature. Para aminoacetophenone was used as internal standard. Methotrexate and internal standard retention times were 4.6 and 9.5 minutes, respectively. Results showed that reproducibility (precision) of method within a day was 2.6 to 6 percent and between days was 5.5 to 9.5 percent. The recovery of the method was between 61.5 and 72.7 percent. The quantitation limit of the method for methotrexate was 0.1μM. This method is suitable for quantitation of methotrexate after infusion of high doses of this drug and has good accuracy, precision and quantitation limit. Key Words: Methotrexate; HPLC; Serum Concentration. DOI: 10.3329/sjps.v2i1.1693 Stamford Journal of Pharmaceutical Sciences Vol.2(1) 2009: 8-13

查看原文本刊更多论文

高效液相色谱法测定人血清中甲氨蝶呤的快速、灵敏方法的建立与验证

甲氨蝶呤竞争性地抑制二氢叶酸还原酶，从而抑制DNA合成和细胞复制。本研究建立了一种简便、快速的高效液相色谱法测定血清中甲氨蝶呤的方法。在每次注射结束后的不同时间间隔，从Mahathma Gandhi Memorial hospital (Warangal,AP.India)接受高剂量MTX的成年癌症患者中收集样本。血清用三氯乙酸脱蛋白，上清液注入250×4.6 mm十八烷基硅烷柱。流动相为tris -磷酸缓冲液(pH 5.7):甲醇:乙腈(70:20:10)，流速为1ml/min。紫外检测在313 nm，室温下进行。以对氨基苯乙酮为内标。甲氨蝶呤保留时间为4.6 min，内标保留时间为9.5 min。结果表明，该方法日内重现性(精密度)为2.6% ~ 6%，日间重现性(精密度)为5.5% ~ 9.5%。该方法的回收率为61.5% ~ 72.7%。方法的定量限为0.1μM。本方法适用于高剂量输注甲氨蝶呤后甲氨蝶呤的定量，具有良好的准确度、精密度和定量限。关键词:甲氨蝶呤;高效液相色谱法;血清浓度。DOI: 10.3329/sjps.v2i1.1693斯坦福德药物科学杂志Vol.2(1) 2009: 8-13

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Stamford Journal of Pharmaceutical Sciences

自引率

0.00%

发文量