Establishment of an HPLC-based method to identify key proteases of proteins in vitro

Wei, Qingqing; Tian, Hong; Zhang, Fan; Sai, Wenbo; Ge, Yang; Gao, Xiangdong; Yao, Wenbing

doi:10.1016/j.ab.2019.02.030

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2021

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“…1 and 2. The HPLC [3][4] analysis for related substance of Nebivolol bulk drug shown the presence of impurity at 0.88 RRT was up to 0.1% level.…”

Section: Introductionmentioning

confidence: 99%

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2021

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“…1 and 2. The HPLC [3][4] analysis for related substance of Nebivolol bulk drug shown the presence of impurity at 0.88 RRT was up to 0.1% level.…”

Section: Introductionmentioning

confidence: 99%

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2021

IJPSR

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Isolation and Structural Elucidation of Impurity in Sulfonamide Intermediate of Glyburide

Gadani

Tak

Mehta³

et al. 2021

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A reproducible isolation method by Reverse Phase (RP) preparative HPLC technique for the isolation of one crucial impurity at 1.65 RRT (Relative Retention Time) in sulfonamide stage of Glyburide API (Active Pharmaceuticals Ingredient) was developed. Preparative chromatography was done on Luna C8, 10µm (250 mm x 21.2mm) preparative HPLC column with acetonitrile: water in 70:30 % v/v proportion as a mobile phase and 8 ml/min as a flow rate. This impurity was detected at 300 nm UV-wavelength maximum. This impurity was isolated from synthesized crude impurity of sulfonamide stage of Glyburide substance by preparative HPLC by injecting 50 mg/ml concentration over 5 ml fixed loop. Isolated impurity was elucidated as N-methyl impurity of sulfonamide intermediate of Glyburide API by means of chromatographic and spectral data. Structural elucidation carried out by spectral data was reviewed. This impurity was analyzed by reverse phase HPLC for purity analysis. A Inertsil C8 (250 x 4.6) mm, 5µ particle size was employed for separation. The mobile phase consisted of Water: Acetonitrile: Methanol in the ratio of 60:15:25 % v/v. The flow rate was set at 1 ml/min. Detection was carried out at 300 nm. 10µL of 2 mg/ml concentration of sample in methanol was injected. The column oven temperature was at 25°C.

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Establishment of an HPLC-based method to identify key proteases of proteins in vitro

Cited by 2 publications

References 40 publications

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Isolation and Structural Elucidation of Impurity in Sulfonamide Intermediate of Glyburide

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