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Isolation and simultaneous determination of bioactive components from cuscuta chinensis

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Dept. of Pharmacy/석사
Increased in life span of populations in the world has led to an elevated frequency of neurodegenerative diseases. A gradual and progressive neuronal cell death occurs as a consequence of increased nitrosative stress and an uncontrolled neuroinflammatory response. This process involves the activation of microglial cells and leads to the initiation and progression of various neurodegenerative diseases. Activation of microglia and consequent release of different proinflammatory factors such as interleukin-1 beta, IL-6, tumor necrosis factor-alpha, nitric oxide, reactive oxygen species , inducible nitric oxide synthase and cyclooxygenase-2 contribute to neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease by increasing inflammations in the brain.The seeds of Cuscuta chinensis (Conbolvulaceas) are commonly used as a medicinal plant for the treatment of the aching and weakness of the loins and knees as well as tonifying the defects of liver and kidney. It was also used for the treatment of diarrhea caused to hypofunction of kidney and spleen. To the best of the knowledge, lignans, flavonoids and other phenolic components are known to be its chemical constituents.In this study, one new α-pyrone, 5-hydroxy-6-methyl-2H-phran-2-one methyl-2H-phran-2-one (1), along with seventeen known compounds were isolated from the CHCl3, EtOAc and n-BuOH fractions of C. chinensis by various chromatographic techniques. The known compounds were identified as methyl paraben (2), p-hydroxycinnamate (3), p-coumaric acid (4), ferulic acid (5), chlorogenic acid (6), β-sitosterol (7), daucosterol (8), (-)-asarinin (9), 4-hydroxysesamin (10), (+)-pinoresinol (11), (+)-epipinoresinol (12), (+)-medioresinol (13), kaempferol (14), quercetin (15), isorhamnetin (16), hyperoside (17), and rutin (18) by using various spectroscopic data and comparing them with literatures. Based on the previous study of nitric oxide inhibitory activity of Cuscuta chinensis extract in BV-2 microglia, these isolated compounds were evaluated for anti-neuroinflammatory activities against the lipopolysaccharide-stimulated BV-2 microglial cells. The EtOAc fraction of C.chinensis inhibited lipopolysaccharide-induced nitric oxide production in BV-2 microglial cells. Among the isolated compounds from EtOAc fraction, compound 14, 15 and 16 showed significant anti-inflammatory activity.In the present study, a new HPLC-DAD method for simultaneous determination of isolated compounds that has been majorly detected in EtOAc fraction, namely chlorogenic acid (6), (+)-pinoresinol (11), kaempferol (14), quercetin (15), and hyperoside (17) was developed and validated. The method was validated in terms of specificity, linearity (R2 > 0.9999), precision [relative standard deviation < 0.97%] and accuracy (98.5–107.3%). The limit of detection and limit of quantification were < 8.96 and 122.05 ng for all analytes, respectively. The validated method was successfully applied to quantify five compounds in C.chinensis.
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2. Thesis / Dissertation (학위논문) > 5. Others (기타) > Others (기타)
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