In the treatment of hepatitis C, direct-acting antivirals (DAA) are highly efficient and well tolerated with a series of DAA combinations available for treatment. A sensitive high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed and validated for the simultaneous quantification of Sofosbuvir (SOF) and Daclatasvir (DAC) in human plasma. Sofosbuvir D6 (SOF D6) and Daclatasvir ¹³C₂²H₆ (DAC ¹³C₂²H₆₎ are used as internal standard (IS). Quantification for both the analytes has been attained with MS-MS detection in positive ion mode using an Acquity UPLC system (Waters) equipped with Waters Xevo TQ MS system with a Gemini NX 5µ C18 (50 × 2.0mm) (Phenomenex) column, and a gradient mobile phase consisting 5 mM Ammonium Formate buffer: Acetonitrile at a flow rate of 0.300 mL/min is used as mobile phase to separate the analytes and detection is performed by electrospray ionization technique using the mass spectrometer. Full validation is performed for bio-analytical methods with respect to linearity, precision, accuracy, selectivity, carry-over, stability and dilution integrity. Linearity is obtained over a concentration range of 10.002 -3000.488 and 10.004 -3001.218 ng mL^-1 for SOF and DAC respectively by applying weighted least-squares linear regression method (1/x2). The developed method was applied successfully in bioequivalence and/or clinical studies in 48 male subjects for the simultaneous quantification of SOF and DAC.

中文翻译：

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通过LCMS / MS同时定量达拉他韦和索非布韦在人血浆中的含量和药代动力学研究

在丙型肝炎的治疗中，直接作用的抗病毒药物（DAA）高效且对一系列可用的DAA组合具有很好的耐受性。已开发出灵敏的高效液相色谱-串联质谱（UPLC-MS / MS）方法，并已验证可同时定量人血浆中的Sofosbuvir（SOF）和Daclatasvir（DAC）。Sofosbuvir D6（SOF D6）和Daclatasvir ¹³ C ₂ ² H ₆（DAC ¹³ C ₂ ² H _6）用作内部标准（IS）。使用配备了带有Gemini NX 5µ C18（50×2.0mm）（Phenomenex）色谱柱的Waters Xevo TQ MS系统的Acquity UPLC系统（Waters），通过阳离子模式的MS-MS检测，可以对两种分析物进行定量。使用由5 mM甲酸铵缓冲液组成的梯度流动相：以0.300 mL / min的流速的乙腈作为流动相分离分析物，并使用质谱仪通过电喷雾电离技术进行检测。对生物分析方法的线性，精密度，准确性，选择性，残留，稳定性和稀释完整性进行了全面验证。在10.002 -3000.488和10.004 -3001.218 ng mL ^-1的浓度范围内获得线性SOF和DAC分别采用加权最小二乘线性回归方法（1 / x 2）。所开发的方法已成功应用于48位男性受试者的生物等效性和/或临床研究中，用于同时定量SOF和DAC。