Jehangir Muhammad, Ahmed Mahmood, Imtiaz Shafiq Muhammad, Samad Abdul
Department of Chemistry, Forman Christian College (A Chartered University), Lahore, Pakistan.
NovaMed Pharmaceuticals, Lahore, Pakistan.
J Anal Methods Chem. 2017;2017:1208753. doi: 10.1155/2017/1208753. Epub 2017 Aug 21.
A newly developed method based on ultrahigh performance liquid chromatography (UHPLC) was optimized for the simultaneous determination of vitamin D and menaquinone-7 (MK-7) in tablet formulation in the present study. UHPLC separation of vitamin D and MK-7 was performed with ACE Excel 2 C18-PFP column (2 m, 2.1 × 100 mm) at 0.6 mL min flow rate, whereas the mobile phase consisted of methanol/water (19 : 1, v/v, phase A) and isopropyl alcohol (99.9%, phase B) containing 0.5% triethylamine. Isocratic separation of both the analytes was performed at 40°C by pumping the mobile phases A and B in the ratio of 50 : 50 (v/v, pH, 6.0). Both analytes were detected at a wavelength of 265 nm and the injection volume was 1.0 L. The overall runtime per sample was 4.5 min with retention time of 1.26 and 3.64 min for vitamin D and MK-7, respectively. The calibration curve was linear from 5.0 to 100 g mL for vitamin D and MK-7 with a coefficient of determination () ≥ 0.9981, while repeatability and reproducibility (expressed as relative standard deviation) were lower than 1.46 and 2.21%, respectively. The proposed HPLC method was demonstrated to be simple and rapid for the determination of vitamin D and MK-7 in tablets.
在本研究中，对一种新开发的基于超高效液相色谱（UHPLC）的方法进行了优化，用于同时测定片剂制剂中的维生素D和甲基萘醌-7（MK-7）。维生素D和MK-7的UHPLC分离在ACE Excel 2 C18-PFP柱（2μm，2.1×100mm）上进行，流速为0.6mL/min，而流动相由甲醇/水（19:1，v/v，A相）和含0.5%三乙胺的异丙醇（99.9%，B相）组成。两种分析物的等度分离在40℃下进行，通过以50:50（v/v，pH 6.0）的比例泵入流动相A和B。两种分析物均在265nm波长处检测，进样体积为1.0μL。每个样品的总运行时间为4.5分钟，维生素D和MK-7的保留时间分别为1.26分钟和3.64分钟。维生素D和MK-7的校准曲线在5.0至100μg/mL范围内呈线性，决定系数（R²）≥0.9981，而重复性和再现性（以相对标准偏差表示）分别低于1.46%和2.21%。所提出的HPLC方法被证明用于测定片剂中的维生素D和MK-7简单且快速。
