Miraghaei Shahram, Mohammadi Bahareh, Babaei Atefeh, Keshavarz Samira, Bahrami Gholamreza
Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran; School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran.
J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Sep 15;1063:118-122. doi: 10.1016/j.jchromb.2017.06.047. Epub 2017 Aug 31.
Although for many analyses tandem mass spectrometry (LC-MS/MS) systems have significant advantage over the high-performance liquid chromatography with diode array detection (HPLC-DAD) however, the HPLC methods are easier, cheaper and more available to perform. As no published method is available for quantitative HPLC analysis of sofosbuvir (SOF), an orally administered anti-hepatitis drug in human serum, this study was aimed to evaluate applicability of the HPLC technique to quantify sofosbuvir and comparison of the two methods for analytical performance. Following extraction of the drug and an internal standard (Hexobarbital), same chromatographic conditions were used for both the systems. After the chromatographic separation on a reverse phase C18 column using a mobile phase consisting of water (containing formic acid 0.5mL/L) and acetonitrile (57:43; v/v) at a flow rate of 0.8mL/min, the eluate was introduced into a DAD detector set at 261nm, then passed through the mass spectrometry system in single ion monitoring mode (SIM). For UV and mass spectrometry detections the calibration curves were linear over a concentration range of 25-3200 and 10-3200ng/mL, respectively and the linearity was over 0.998 for both the systems. Lower limit of quantification (LLOQ) for mass spectrometry and DAD detections were 10 and 25ng/mL, respectively. In conclusion sensitivity of DAD detection is sufficient enough to determine concentrations down to 0.5% of C which achieved in bioequivalence study of sofosbuvir and meet FDA requirements for these types of studies.
尽管对于许多分析而言，串联质谱（LC-MS/MS）系统相对于配有二极管阵列检测的高效液相色谱（HPLC-DAD）具有显著优势，然而，HPLC方法操作起来更简便、成本更低且更易于实现。由于尚无已发表的方法可用于对人血清中口服抗肝炎药物索磷布韦（SOF）进行定量HPLC分析，本研究旨在评估HPLC技术定量索磷布韦的适用性以及比较这两种方法的分析性能。在提取药物和内标（己巴比妥）后，两个系统使用相同的色谱条件。在反相C18柱上进行色谱分离，流动相由水（含0.5mL/L甲酸）和乙腈（57:43；v/v）组成，流速为0.8mL/min，洗脱液引入设置在261nm的DAD检测器，然后以单离子监测模式（SIM）通过质谱系统。对于紫外和质谱检测，校准曲线在浓度范围分别为25 - 3200和10 - 3200ng/mL时呈线性，两个系统的线性度均超过0.998。质谱和DAD检测的定量下限（LLOQ）分别为10和25ng/mL。总之，DAD检测的灵敏度足以测定低至索磷布韦生物等效性研究中达到的C的0.5%的浓度，并符合FDA对这类研究的要求。
