Research and determination of related substances in flumazenil
10.16438/j.0513-4870.2023-1265
- VernacularTitle:氟马西尼有关物质的研究与测定
- Author:
Xue-yan MIAO
1
,
2
;
Yuan YANG
3
;
Si-si LU
1
;
Jin-mei MO
1
;
Lin-kai HUANG
1
;
Jia-jun WEI
1
;
Yi-ping GU
1
,
2
Author Information
1. School of Public Health, Guilin Medical University, Guilin 541199, China
2. The Guangxi Key Laboratory of Environmental Exposomics and Entire Lifecycle Heath, Guilin 541199, China
3. NMPA Key Laboratory for Quality Research and Control of Drug Products, Wuhan Institute for Drug and Medical Device Control, Wuhan 430075, China
- Publication Type:Research Article
- Keywords:
flumazenil;
correction factor;
related substance;
egradation product;
HPLC
- From:
Acta Pharmaceutica Sinica
2024;59(6):1765-1772
- CountryChina
- Language:Chinese
-
Abstract:
A high performance liquid chromatography (HPLC) method utilizing correction factors was established for the quantitative detection of related substances in flumazenil. Separation was achieved using an Agilent Pursuit XRs C18 column (250 mm × 4.6 mm, 5 μm) with an isocratic elution of dilute phosphoric acid, methanol, and tetrahydrofuran as the mobile phases. Correction factors calculated from a standard curve method were applied to determine the impurity content. The quantification of impurities in flumazenil was conducted using both external standard and correction factor methods, followed by validation and comparison of the two. For the identification of degradation products, a forced degradation approach was employed to prepare a flumazenil degradation solution, and the resulting impurities were confirmed by LC-MS analysis. The separation of flumazenil and its impurities was found to be efficient. The limits of quantification for impurities A, B, D, and E were established at 0.169 9, 0.314 7, 0.143 9, and 0.270 8 ng, respectively, with the limits of detection at 0.055 8, 0.096 9, 0.048 8, and 0.089 0 ng. These impurities demonstrated a strong linear relationship across the concentration ranges of 0.034 9-7.847 0, 0.038 7-8.710 7, 0.034 6-7.794 1, and 0.032 4-7.292 8 µg·mL-1, respectively (n = 7). The method achieved average recoveries between 98.25% and 99.42%, with an RSD of less than 2.0% (n = 9), indicating high accuracy. The external standard and correction factor methods were used to determine the related substances in flumazenil, and the results of the two methods were consistent. The established HPLC method is characterized by its high accuracy, sensitivity, and repeatability, and is suitable for determining related substances in flumazenil.
