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Objective:To establish an analytical method for the determination of furazolidone and indometacin in furazolidone, in-dometacin and cuscohygrinolis α-acetylbenzoacetate suppositories by HPLC. Methods: The separation was performed on a ZORBAX Extend-C18 (250 mm × 4. 6 mm,5μm) column. The mobile phase was acetonitrile and 0. 035 mol·L-1 potassium phosphate monobas-ic aqueous solution (adjusting pH to 3. 0 with acetic acid) with gradient elution. The flow rate was 1. 0 ml·min-1, and the detection wavelengths were set at 364 nm and 318 nm. The column temperature was 30℃ and the injection volume was 20 μl. Results:Under the selected chromatographic conditions, the two components showed good linearity within the range of 0.005-0.05 mg·ml-1(r=0. 9999). The limit of detection was 20 ng·ml-1 and 26 ng·ml-1, respectively. The limit of quantitation was 70 ng·ml-1 and 90 ng·ml-1, respectively. The average recovery was 99. 4％(RSD=0. 6％, n=9)and 99. 4％(RSD=0. 3％,n=9),respectively. Conclusion:The method is simple, rapid and specific, and the results are accurate and reliable. The method can be used for the de-termination of the two components in furazolidone, indometacin and cuscohygrinolis α-acetylbenzoacetate suppositories.
RESUMEN
OBJECTIVE: To compare several common staining and detection methods using NFS-60 cells for biological activity test of recombinant human granulocyte colony stimulating factor (rhG-CSF). METHODS: The biological activity of rhG-CSF was detected using some common methods, named NFS-60 cells/MTT staining, NFS-60 cells/MTS staining, NFS-60 cells/CCK-8 staining, and NFS-60 cells/fluorescence staining. The biological activity was detected using the NFS-60 cells/MTT method using dual wavelength (570 nm detection, 630 nm reference) and single wavelength (570nm detection and 630nm detection). The biological activity was detected using NFS-60 cells/MTS dynamic detection method and NFS-60 cell/CCK-8 dynamic method. Then, the results were analyzed and compared. RESULTS: The different methods were not significantly different (P>0.05); the difference between the dual wavelength detection and single wavelength detection of NFS-60 cells/MTT was not significant (P>0.05). The results from NFS-60 cells/MTS dynamic detection method, NFS-60 cells/CCK-8 dynamic method and NFS-60 cells/MTT method had not significant difference (P>0.05). CONCLUSION: The biological activity determination results of the tested methods using NFS-60 cells are consistent. This study provides basis for utilization of results from different laboratories using different methods, support for expansion of the biological activity detection method of rhG-CSF in Chinese Pharmacopoeia, and reference for expansion of the biological activity detection method of other cytokines.