Panchal Jignesh, Dhalani Jayesh
Department of Chemistry, School of Science, RK University, Rajkot 360020, Gujarat, India.
J Chromatogr Sci. 2025 Jul 24;63(7). doi: 10.1093/chromsci/bmaf045.
The combination of metformin (MET), teneligliptin (TEN), and pioglitazone (PIO) is newest fixed-dose combination for the treatment type 2 diabetes launched in India. An accurate, rapid, and cost-effective reversed phase ultra-fast liquid chromatography method was developed, validated, and applied for the quantification of MET, TEN, and PIO in bulk and tablet with a very short runtime. Box-Behnken design was implemented to optimize the ultra-fast liquid chromatography conditions. A C18 column (150 × 4.6 mm, 5μ) was utilized with a mobile phase buffer (0.01 M phosphate buffer, pH = 6.2) and acetonitrile in a 51:49 ratio with flow rate of 1.5mL/min at column oven temperature 40°C. Detection was carried out at 255nm by 20 μL injection volume. Retention times were found to be 0.94, 1.36, and 2.07 min, whereas the limit of quantification were 0.209, 0.712, and 57.030 μg/mL for MET, TEN, and PIO, respectively. The linearity of anticipated development was studied in the range of 250-1250 μg/mL (MET, r2 = 0.99974), 10-50 μg/mL (TEN, r2 = 0.99997), and 7.5-37.5μg/mL (PIO, r2 = 0.99999). The relative standard deviation values for method and intermediate precision were <2%. The method was validated as per International Conference on Harmonization guideline for accuracy, precision, linearity, limit of detection, limit of quantification, specificity, robustness, and forced degradation. This is the only study that presents reversed phase ultra-fast liquid chromatography method for the new fixed-dose combination of MET, TEN, and PIO, with the shortest run time of 3.0 min. The proposed method is innovative, efficient, precise, quickest, and economical with high accuracy which makes the study novel.
二甲双胍（MET）、替格列汀（TEN）和吡格列酮（PIO）的组合是印度推出的用于治疗2型糖尿病的最新固定剂量组合。开发、验证并应用了一种准确、快速且经济高效的反相超快速液相色谱法，用于在极短的运行时间内对原料药和片剂中的MET、TEN和PIO进行定量。采用Box-Behnken设计优化超快速液相色谱条件。使用C18柱（150×4.6 mm，5μm），流动相缓冲液（0.01 M磷酸盐缓冲液，pH = 6.2）和乙腈以51:49的比例，在柱温箱温度40°C下流速为1.5 mL/min。通过进样量20μL在255nm处进行检测。保留时间分别为0.94、1.36和2.07分钟，而MET、TEN和PIO的定量限分别为0.209、0.712和57.030μg/mL。预期开发的线性范围为250 - 1250μg/mL（MET，r2 = 0.99974）、10 - 50μg/mL（TEN，r2 = 0.99997）和7.5 - 37.5μg/mL（PIO，r2 = 0.99999）。方法和中间精密度的相对标准偏差值<2%。该方法按照国际协调会议指南进行了准确性、精密度、线性、检测限、定量限、特异性、稳健性和强制降解的验证。这是唯一一项针对MET、TEN和PIO新固定剂量组合提出反相超快速液相色谱法的研究，最短运行时间为3.0分钟。所提出的方法具有创新性、高效性、精确性、快速性和经济性，具有高精度，使该研究具有新颖性。
