Department of Chemistry (UG & PG), Dr. A P J Abdul Kalam Central Research Laboratory (CRL), Sir C R Reddy Autonomous College, Eluru, India.
D-216, Discovery Lab, Analytical Chemistry Division, Indian Institute of Chemical Technology, Tarnaka, India.
Chirality. 2022 Jul;34(7):989-998. doi: 10.1002/chir.23448. Epub 2022 Apr 7.
The direct separation of dipeptidyl peptidase IV (DPP-4) inhibitors such as Sitagliptin (STG), Linagliptin (LIG), and Saxagliptin (SAG) enantiomers in normal phase conditions have been achieved on immobilized polysaccharide-based chiral stationary phases (CSPs), as well as on the macrocyclic glycopeptide vancomycin chiral stationary phase (Chirobiotic V2) under polar ionic mode. The enantiomers of these targets could be separated completely (resolution factor Rs > 2) using the Chirobiotic V2 column in polar ionic mode with the mobile phase (MeOH/AcOH/TEA 100/0.3/0.1 v/v/v) in an isocratic elution at 1.0 ml min . The effect of the mobile phase composition on separation, including buffer salts, acid-base modifiers, and analyte structures, was evaluated. The developed technique was validated in the polar ionic mode according to the International Conference on Harmonization (ICH) Q2R1 guidelines in terms of accuracy, precision, selectivity, linearity, limit of detection (LOD), and limit of quantification (LOQ). The calibration curve was linear in a concentration range from LOQ to 3.75 μg/ml. The LOD and LOQ of STG, LIG, and SAG were 0.15 and 0.45, 0.15 and 0.50, 0.16 and 0.50, respectively. The proposed method is said to be selective, accurate, and precise. Finally, the validated method was used successfully for the quantitative determination of DPP-4 enantiomers in pharmaceutical analytes.
在正相条件下，已成功实现了二肽基肽酶 IV（DPP-4）抑制剂（如西他列汀（STG）、利格列汀（LIG）和沙格列汀（SAG））对映体的直接分离，该分离是在固定化多糖手性固定相（CSP）上，以及在大环糖肽万古霉素手性固定相（Chirobiotic V2）上，在极性离子模式下进行的。在极性离子模式下，使用 Chirobiotic V2 柱，以等度洗脱方式，流动相（MeOH/AcOH/TEA 100/0.3/0.1 v/v/v），流速为 1.0 ml/min，可以完全分离这些目标物的对映体（分离度 Rs>2）。评估了流动相组成（包括缓冲盐、酸碱调节剂和分析物结构）对分离的影响。根据国际协调会议（ICH）Q2R1 指南，该技术在极性离子模式下进行了验证，在准确度、精密度、选择性、线性、检测限（LOD）和定量限（LOQ）方面均符合要求。在 LOQ 至 3.75 μg/ml 的浓度范围内，校准曲线呈线性。STG、LIG 和 SAG 的 LOD 和 LOQ 分别为 0.15 和 0.45、0.15 和 0.50、0.16 和 0.50。该方法被认为具有选择性、准确性和精密度。最后，该验证方法成功用于药物分析物中 DPP-4 对映体的定量测定。
