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@#The UPLC fingerprint of colistimethate sodium was established for the study of quality consistency.The chromatographic column was Acquity UPLC? Peptide CSH C18 (2.1 mm × 150 mm, 1.7 μm).The mobile phase A was phosphate buffer-acetonitrile (19∶1), and the mobile phase B was phosphate buffer-acetonitrile (1∶1).The mobile phase was in gradient elution at a flow rate of 0.3 mL/min.The column temperature was set at 30 °C and the detection wavelength was 210 nm.The similarity of the fingerprints was analyzed with the Similarity Evaluation System for Chromatographic Fingerprint of Tradition Chinese Medicine (Version 2012) in combination with content determination of multiple index components to evaluate the quality consistency of imported and domestic bulk drugs.The result showed that both the original and generic bulk drugs met the specified limit requirements in the European Pharmacopoeia standards, and that their UPLC fingerprints were highly similar, indicating that the quality of the two substances was consistent.Establishing a fingerprint for similarity evaluation and combining it with the results of indicator component content determination as a comprehensive evaluation method for the study of drug quality consistency of complex components has the characteristics of fast, accurate, and comprehensive, which is helpful for drug quality evaluation and provides ideas for the evaluation of antibiotic quality consistency of complex components.
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@#A novel method was developed for the content assay and related substances determination of neomycin sulfate by high performance liquid chromatography combined with pulsed amperometric detection(HPLC-PAD). The HPLC was performed on Thermo AcclaimTMAmG C18(4. 6 mm×150 mm, 3 μm). The mobile phase consisted of aqueous solution with 2% trifluoroacetic acid containing 0. 01% pentafluoropropionic acid and 0. 6%NaOH. The pulsed amperometric detector was operated with aquadruple-potential waveform for the detection. Neomycin B, Neomycin C and thirteen related substances were adequately separated by the established HPLC conditions. The limits of detection(LOD)and quantification(LOQ)of neomycin B and neomycin C were both 1. 75 ng and 3. 5 ng, respectively. Good linearities of neomycin B and neomycin C were found in their respective ranges which their correlation coefficients were greater than 0. 998 5. The established method is characterized by high specificity, sensitivity and wide range of linearity which has a good application prospect and provides the basis for improving the standard and quality control of neomycin sulfate.
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Objectives:To investigate in vitro tumor killing effect of recombinant human perforin C terminal truncated 125 amino acid polypeptide (rhPFP C) and N terminal truncated 118 amino acid polypeptide (rhPFP N, 22 139aa) using liver cancer cell line SMMC 7721 as target cells. Methods:Recombinant human PFP C and PFP N peptide were expressed by E.coli in fusion form with glutathione S transferase(GST), and were purified by affinity chromatography with glutathione agarose. Liver cancer cell line SMMC 7721 was treated with fusion proteins in different concentrations for 24 h before surviving cells were measured with microscopy and colorimetric MTT assay. Results:After treatment with rhPFP C or rhPFP N, SMMC 7721 cell membrane damaged, appeared lysis. In MTT assay, optical density values in test group were significantly lower than those in control group. At concentration of 2.5 ?g/ml, the killing activity of rhPFP C and rhPFP N were 33.38% and 5.90% respectively. Conclusions:Both rhPFP C and rhPFP N showed obvious killing effect on liver cancer cell line SMMC 7721.The activity of rhPFP C was great higher than that of rhPFP N.