Gohar Muhammad Saqib, Rahman Taj Ur, Bahadur Ali, Ali Ashraf, Alharthi Sarah, Al-Shaalan Nora Hamad
Department of Chemistry, Mohi-Ud-Din Islamic University, Nerian Sharif, Azad Jammu & Kashmir 12080, Pakistan.
Department of Chemistry, School of Natural Sciences (SNS), National University of Science and Technology (NUST), H-12, Islamabad 46000, Pakistan.
Pharmaceuticals (Basel). 2024 Apr 15;17(4):505. doi: 10.3390/ph17040505.
In the present work, an efficient isocratic HPLC method was developed for the precise and accurate estimation of vitamin D in tablet form. The chromatographic conditions comprised an L3 silica column (5 µm in particle size, 4.6 mm × 250 mm) with a mobile phase n-hexane/ethyl acetate (85:15 /) with a flow rate of 2.0 mL/min and a detection wavelength of 292 nm. The new methodology was validated for accuracy, precision, specificity, robustness, and quantification limits according to an official monograph of USP/BP and ICH guidelines. The peak areas of the six replicates of the homogeneous sample were recorded. The mean value obtained was 67,301, and the relative standard deviation (RSD) was 0.1741. The linearity and range were in the acceptable bounds, i.e., 0.999, which was calculated using regression line analysis. The results show that the method is truly acceptable as the RSD, as the flow rate was 0.81%, while for the mobile phase composition, it was 0.72%, which lies in the acceptable range. The limit of detection (LOD) and the limit of quantification (LOQ) values were 0.0539 µg/mL and 0.1633 µg/mL, respectively. The % RSD of the intra and inter-day precision of the method was deemed acceptable according to the international commission for harmonization guidelines. The developed method has potential to be used for the detection and quantification of vitamin D during routine analysis for tablets in dosage form.
在本研究中，开发了一种高效等度高效液相色谱法，用于精确准确地测定片剂形式的维生素D。色谱条件包括L3硅胶柱（粒径5 µm，4.6 mm×250 mm），流动相为正己烷/乙酸乙酯（85:15 /），流速为2.0 mL/min，检测波长为292 nm。根据美国药典/英国药典官方专论和国际协调会议指南，对新方法的准确性、精密度、特异性、稳健性和定量限进行了验证。记录了均匀样品六次重复进样的峰面积。得到的平均值为67301，相对标准偏差（RSD）为0.1741。线性和范围在可接受范围内，即0.999，这是使用回归线分析计算得出的。结果表明，该方法确实可接受，因为RSD，流速为0.81%，而流动相组成的RSD为0.72%，均在可接受范围内。检测限（LOD）和定量限（LOQ）值分别为0.0539 µg/mL和0.1633 µg/mL。根据国际协调委员会指南，该方法日内和日间精密度的%RSD被认为是可接受的。所开发的方法有潜力用于剂型片剂常规分析中维生素D的检测和定量。
