Bhadoriya Abhaysingh, Shah Priyanka A, Shrivastav Pranav S, Bharwad Kirtikumar D, Singhal Puran
Kadi Sarva Viswavidyalaya, Sector-15, Ghandhinagar, 382715, Gujarat, India.
Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad, 380009, India.
Biomed Chromatogr. 2019 Aug;33(8):e4543. doi: 10.1002/bmc.4543. Epub 2019 Apr 29.
A high-throughput and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed for the determination of terbinafine in human plasma. The method employed liquid-liquid extraction of terbinafine and terbinafine-d7 (used as internal standard) from 100 μL human plasma with ethyl acetate-n-hexane (80:20, v/v) solvent mixture. Chromatography was performed on a BEH C (50 × 2.1 mm, 1.7 μm) column using acetonitrile-8.0 mm ammonium formate, pH 3.5 (85:15, v/v) under isocratic elution. For quantitative analysis, MS/MS ion transitions were monitored at m/z 292.2/141.1 and m/z 299.1/148.2 for terbinafine and terbinafine-d7, respectively, using electrospray ionization in the positive mode. The method was validated according to regulatory guidance for selectivity, sensitivity, linearity, recovery, matrix effect, stability, dilution reliability and ruggedness with acceptable accuracy and precision. The method shows good linearity over the tested concentration range from 1.00 to 2000 ng/mL (r ≥ 0.9984). The intra-batch and inter-batch precision (CV) was 1.8-3.2 and 2.1-4.5%, respectively. The method was successfully applied to a bioequivalence study with 250 mg terbinafine in 32 healthy subjects. The major advantage of this method includes higher sensitivity, small plasma volume for processing and a short analysis time.
已开发出一种高通量、灵敏的超高效液相色谱-串联质谱法（UPLC-MS/MS）用于测定人血浆中的特比萘芬。该方法采用液-液萃取法，从100μL人血浆中用乙酸乙酯-正己烷（80:20，v/v）混合溶剂萃取特比萘芬和特比萘芬-d7（用作内标）。色谱分析在BEH C18柱（50×2.1mm，1.7μm）上进行，使用乙腈-8.0mM甲酸铵，pH 3.5（85:15，v/v）等度洗脱。进行定量分析时，采用正模式电喷雾电离，分别监测特比萘芬和特比萘芬-d7的MS/MS离子跃迁，m/z分别为292.2/141.1和m/z 299.1/148.2。该方法根据法规指南进行了选择性、灵敏度、线性、回收率、基质效应、稳定性、稀释可靠性和耐用性验证，具有可接受的准确度和精密度。该方法在1.00至2000ng/mL的测试浓度范围内显示出良好的线性（r≥0.9984）。批内和批间精密度（CV）分别为1.8 - 3.2%和2.1 - 4.5%。该方法成功应用于32名健康受试者服用250mg特比萘芬的生物等效性研究。该方法的主要优点包括更高的灵敏度、处理所需的血浆量少以及分析时间短。
