新型降糖药米格列醇原料药及其制剂的色谱分离：强制降解及稳定性指示的 HPTLC/密度测定法和 HPLC/UV 法验证。

Chromatographic Separation of the Novel Hypoglycemic Drug Mitiglinide in its Bulk Powder and Pharmaceutical Formulation: Forced Degradation and Validated Stability-Indicating HPTLC/Densitometry and HPLC/UV Methods.

机构信息

Egyptian Russian University, Faculty of Pharmacy, Pharmaceutical Analytical Chemistry Department, Badr City, Cairo, Egypt.

Cairo University, Faculty of Pharmacy, Analytical Chemistry Department, Kasr El-Aini Street, Cairo, Egypt.

出版信息

J AOAC Int. 2020 Jun 1;103(3):736-742. doi: 10.1093/jaocint/qsz004.

DOI:10.1093/jaocint/qsz004

PMID:33241368

Abstract

BACKGROUND

Mitiglinide (MTG) is one of meglitinides group which are used for treatment of type two diabetes mellitus.

OBJECTIVE

Mitiglinide (MTG) is a novel oral hypoglycemic drug. The present work adopts two stability-indicating chromatographic methods for determination of MTG after being exposed to forced degradation using 4 M methanolic HCl for 12 h.

METHODS

The first method is HPTLC/densitometry using methanol:chloroform:acetic acid (5:2.5:0.3 by volume) as the eluting system and silica gel 60 GF254 as the stationary phase; the separated bands were then scanned at 220 nm. The second method is HPLC/UV in which acetonitrile:methanol:0.05 M potassium dihydrogen orthophosphate (pH 3.5) (40:25:35 by volume) was used as the mobile phase and a Zorbax SB-C8 (250 × 4.6 mm, 5 µm) column as a stationary phase, at a flow rate of 1 mL/min and UV detection at 220 nm.

RESULTS

As a result of acid hydrolysis, two degradants were obtained. The first one was benzyl succinic acid to which this study was performed. The second one lacked configuration and was unreadable using UV spectrometry. Linearity was in the range of 8-48 µg/band MTG for HPTLC and 10-80 µg MTG for HPLC. LOD and LOQ values were 1.85 and 5.62 µg/band for the HPTLC method and 2.14 and 6.49 µg/mL for the HPLC method, respectively. The Recovery % was 100.03 ± 1.464 and 99.61 ± 1.44 using the HPTLC and HPLC methods, respectively. The relative standard deviations (RSD, %) for intra- and inter-day assays were 1.111 and 1.430 for the HPTLC method, respectively, and those for the HPLC method were 1.377 and 0.866, respectively. The RSD, %, for robustness testing was 1.162 (saturation time of mobile phase) and 1.592 (change in ratio of methanol content) for the HPTLC method and 1.377 (mobile phase composition), 1.713 (detector wavelength) and 1.770 (mobile phase flow rate) for the HPLC method.

CONCLUSIONS

The adopted methods were successfully applied for the determination of MTG in its pure form, in presence of its acid degradant and in its tablet dosage form.

HIGHLIGHTS

Statistical comparison between the results obtained from the developed methods and those obtained by the reported HPLC method showed no significance difference.

摘要

背景

米格列奈（MTG）是一种用于治疗 2 型糖尿病的新型口服降糖药，属于美格列奈类药物。

目的

米格列奈（MTG）是一种新型的口服降糖药物。本工作采用两种稳定性指示色谱法，在 4M 甲醇盐酸中强制降解 12 小时后测定 MTG。

方法

第一种方法是 HPTLC/密度法，使用甲醇：氯仿：乙酸（5：2.5：0.3 体积比）作为洗脱系统，硅胶 60 GF254 作为固定相；分离的条带在 220nm 处扫描。第二种方法是 HPLC/UV，其中乙腈：甲醇：0.05M 磷酸二氢钾（pH3.5）（40：25：35 体积比）为流动相，Zorbax SB-C8（250×4.6mm，5μm）柱为固定相，流速为 1mL/min，UV 检测波长为 220nm。

结果

酸水解后得到两种降解产物。第一个是苯甲酰琥珀酸，本研究以此为对象。第二个则缺失了构型，用 UV 光谱法无法读取。HPTLC 的线性范围为 8-48μg/条 MTG，HPLC 的线性范围为 10-80μg MTG。HPTLC 法的 LOD 和 LOQ 值分别为 1.85 和 5.62μg/条，HPLC 法的 LOD 和 LOQ 值分别为 2.14 和 6.49μg/mL。HPTLC 和 HPLC 法的回收率分别为 100.03%±1.464%和 99.61%±1.44%。HPTLC 法的日内和日间相对标准偏差（RSD，%）分别为 1.111%和 1.430%，HPLC 法的 RSD，%分别为 1.377%和 0.866%。HPTLC 法的稳健性测试 RSD，%为 1.162%（流动相饱和时间）和 1.592%（甲醇含量比变化），HPLC 法的 RSD，%分别为 1.377%（流动相组成）、1.713%（检测器波长）和 1.770%（流动相流速）。