A Validated Method for Quantification of Dolutegravir Using Ultra Performance Liquid Chromatography Coupled With UV Detection
A Validated Method for Quantification of Dolutegravir Using Ultra Performance Liquid Chromatography Coupled With UV Detection
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DOI：
10.1097/ftd.0000000000000286
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发表时间：
2016-06-01
影响因子：
2.5
通讯作者：
Boffito, Marta
中科院分区：
文献类型：
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作者：
Wang, Xinzhu;Penchala, Sujan Dilly;Boffito, Marta
背景：度鲁特韦（Dolutegravir，DTG）是一种整合酶链转移抑制剂，是一种新批准的抗逆转录病毒药物，用于治疗HIV感染初治和经验者。DTG药理学的许多方面仍有待研究。我们的目的是开发和充分验证一个强大的分析方法，用于定量DTG在血浆中使用液相色谱法结合UV detection.Methods：一个简单而快速的蛋白质沉淀法被用于从100 μ L血浆中提取分析物。在C8反相分析柱上，采用50 mmol/L甲酸和50 mmol/L乙酸铵水溶液（移动的相A）和100%乙腈（移动的相B）梯度洗脱，流速为0.3 mL/min，总运行时间为10 min，实现分离。结果：在0.25 ~ 10 mcg/mL浓度范围内线性关系良好（r > 0.9999，n = 6）。试验内变异性范围为3.3%-6.1%，试验间变异性范围为4.5%-5.7%。3种不同浓度质控样品的总体准确度范围为90.7%-97.7%。萃取回收率为94.3%~ 100%。该方法是高度选择性的，没有干扰，从共同伴随的抗逆转录病毒药物或内源性metabolites.Conclusions：所描述的方法是简单的，强大的，选择性，准确，精密度，和成本效益。因此，该测定法可以容易地转移并在临床环境中实施，并用于药代动力学研究和治疗药物监测程序。
Background:Dolutegravir (DTG) is an integrase strand transfer inhibitor, which is a newly approved antiretroviral drug used for the treatment of HIV-infected naive and experienced individuals. Many aspects of DTG pharmacology remain to be studied. Our aim was to develop and fully validate a robust analytical method for the quantification of DTG in plasma using liquid chromatography coupled with UV detection.Methods:A simple and rapid protein precipitation method was used for analyte extraction from 100 mu L plasma. The separation was achieved on a C8 reverse-phase analytical column using a gradient elution with 50 mmol/L formic acid and 50 mmol/L ammonium acetate in water (mobile phase A), and 100% acetonitrile (mobile phase B) and at a flow rate of 0.3 mL/min and a total run time of 10 minutes. The detector wavelength was set at 258 nm.Results:The linearity of the calibration curve (r > 0.9999, n = 6) was validated over a concentration range of 0.25-10 mcg/mL. Intra-assay variability ranged from 3.3% to 6.1% and inter-assay variability ranged from 4.5% to 5.7%. The overall accuracy ranged from 90.7% to 97.7% for the 3 different concentrations of quality control samples. Recovery efficiency of extraction ranged from 94.3%-100%. This method is highly selective with no interferences from commonly concomitant antiretroviral drugs or endogenous metabolites.Conclusions:The described method is simple, robust, selective, accurate, precise, and cost-effective. Thus, this assay can be readily transferred and implemented in clinical settings and used for pharmacokinetic studies and therapeutic drug monitoring programs.