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Establishment Of Indirect Detection Method For Captopril And Determination Of GSH As Well As Total Thiols (-SH) In Okra

Posted on:2017-03-13Degree:MasterType:Thesis
Country:ChinaCandidate:Y J LiuFull Text:PDF
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Thiol compounds in organism play an important physiological role, while many thiol-containing compounds as a drug widely used in clinical treatment, so accurately detect thiol compounds of different samples in the life sciences, pharmacology, clinical medicine and other fields, which has practical significance. In this study, use the active free thiol groups in low molecular weight thiol compounds interact with 5, 5’-dithiobis-nitrobenzoic acid (DTNB) through disulfide interaction. Measured the derivation product 2-nitro-4-thiol-benzoic acid (NTB) of the derivative reagents and thiols by HPLC. Established indirect measurement method for the active ingredient content of captopril in thiol-containing drugs captopril (CTP) tablets, while detected the glutathione (GSH) and total sulfhydryl contents of the okra seeds by the same principle determinate.More content detection methods of sulfhydryl containing drug were reported in the literature, but no matter what kind of detection methods are required standard, and it is not easy to get. In this study, starting from the operation and economic point of view to simplify the experiment, through the CTP and alternative standards GSH react with DTNB, and quantitatively released derivative product NTB that can be detected by HPLC. Meanwhile GSH as standard determined the captopril in CTP tablets with thiol derivative reagents DTNB, thereby avoiding the thoil-containing drug determination on the issue of standard. Recovery of the assay was 100.4%, the detection limit was 6.18 μmol/L. While use UV spectroscopy to contrast, both measurement results are consistent, preliminary determine the feasibility of detection method.The biggest detecting problem intracellular thoil compounds in biological samples are that thoils are easily oxidized which can lead to reduce sensitivity in sample pre-treatment process even missed detection. So many plant samples are lack of important information about biological activity on the content of thiol compounds. In this study, using extraction with cell disruption and derivatization synchronization methods, which avoided thoils were oxidized. The contents of GSH and total thoils were determined by HPLC. The results showed that glutathione content in okra seeds was 1.47 μmol/g, RSD 1.18%; total thoils (-SH) containing an amount of 4.85 umol/g, RSD 0.74%. Detection limits were 0.1584 μmol/g and 0.1837 μmol/g, the recoveries were 100.38% and 100.7%.This experiment not only provides a simple and accurate analytical method that is economic and efficient for the analysis and quality testing drugs sulfhydryl, thoil compounds for screening to detect sulfhydryl rich biological samples and biological samples to solve difficult problems. Provide basis of methodology for qualitative and quantitative analysis about thoil group-containing compounds.
Keywords/Search Tags:Captopril, Glutathione, sulfhydryl drugs, 5,5’-dibromo-nitrobenzoic acid, High performance liquid chromatography, Spectrometry, Derivatized
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