Analysis of streptokinase by validated liquid chromatography methods and correlation with an in vitro bioassay
| Autor: | Bruna Xavier, Rafaela Ferreira Perobelli, Sérgio Luiz Dalmora, Clovis Dervil Appratto Cardoso, Francielle Santos da Silva, Fernanda Pavani Stamm Maldaner |
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| Rok vydání: | 2016 |
| Předmět: |
Quality Control
Calibration curve Streptokinase Size-exclusion chromatography Filtration and Separation 030204 cardiovascular system & hematology 01 natural sciences Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Sodium sulfate medicine Bioassay Potency Enzyme Assays Chromatography 010401 analytical chemistry Reproducibility of Results Reversed-phase chromatography 0104 chemical sciences chemistry Chromatography Gel Biological Assay Sodium acetate medicine.drug Chromatography Liquid |
| Zdroj: | Journal of separation science. 40(2) |
| ISSN: | 1615-9314 |
| Popis: | Reversed-phase and size-exclusion liquid chromatography methods were validated for the assessment of Streptokinase. The reversed-phase method was carried out on a Jupiter C4 column (250 mm × 4.6 mm i.d.) maintained at 25°C. The mobile phase consisted of 50 mM sodium sulfate solution pH 7.0 and methanol (90:10, v/v), run isocratically at a flow rate of 0.8 mL/min. The size-exclusion method was carried out on a Protein KW 802.5 column (300 mm × 8.0 mm i.d.), at 25°C. The mobile phase consisted of 40 mM sodium acetate solution pH 7.0, run isocratically at a flow rate of 1.0 mL/min. Retention times were 19.3 min, and 14.1 min, and calibration curves were linear over the concentration range of 0.25–250 μg/mL (25.75–25 750 IU/mL) (r2 = 0.9997) and 5–80 μg/mL(515–8240 IU/mL) (r2 = 0.9996), respectively, for reversed-phase and size exclusion, with detection at 220 and 204 nm. Chromatographic methods were employed in conjunction with the in vitro bioassay for the content/potency assessment of Streptokinase, contributing to improve the quality control and ensure the efficacy of the biotherapy. This article is protected by copyright. All rights reserved |
| Databáze: | OpenAIRE |
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