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Lee Fung Ang,Mun Fei Yam,Yvonne Tan Tze Fung,Peh Kok Kiang,Yusrida Darwin 대한약침학회 2014 Journal of pharmacopuncture Vol.17 No.4
Objectives: Quercetin and curcuminoids are importantbioactive compounds found in many herbs. Previouslyreported high performance liquid chromatographyultraviolet (HPLC-UV) methods for the detection ofquercetin and curcuminoids have several disadvantages,including unsatisfactory separation times and lack ofvalidation according the standard guidelines of the InternationalConference on Harmonisation of TechnicalRequirements for Registration of Pharmaceuticals forHuman Use. Methods: A rapid, specific, reversed phase, HPLC-UVmethod with an isocratic elution of acetonitrile and 2%v/v acetic acid (40% : 60% v/v) (pH 2.6) at a flow rate of1.3 mL/minutes, a column temperature of 35°C, and ultraviolet(UV) detection at 370 nm was developed. Themethod was validated and applied to the quantificationof different types of market available Chinese medicineextracts, pills and tablets. Results: The method allowed simultaneous determinationof quercetin, bisdemethoxycurcumin, demethoxycurcuminand curcumin in the concentration ranges of 0.00488 ─ 200 μg/mL, 0.625 ─ 320 μg/mL, 0.07813─ 320 μg/mL and 0.03906 ─ 320 μg/mL, respectively. The limits of detection and quantification, respectively,were 0.00488 and 0.03906 μg/mL for quercetin,0.62500 and 2.50000 μg/mL for bisdemethoxycurcumin,0.07813 and 0.31250 μg/mL for demethoxycurcumin,and 0.03906 and 0.07813 μg/mL for curcumin. Thepercent relative intra day standard deviation (% RSD)values were 0.432 ─ 0.806 μg/mL, 0.576 ─ 0.723 μg/mL, 0.635 ─ 0.752 μg/mL and 0.655 ─ 0.732 μg/mLfor quercetin, bisdemethoxycurcumin, demethoxycurcuminand curcumin, respectively, and those forintra day precision were 0.323 ─ 0.968 μg/mL, 0.805 ─0.854 μg/mL, 0.078 ─ 0.844 μg/mL and 0.275 ─ 0.829μg/mL, respectively. The intra day accuracies were99.589% ─ 100.821%, 98.588% ─ 101.084%, 9.289% ─100.88%, and 98.292% ─ 101.022% for quercetin, bisdemethoxycurcumin,demethoxycurcumin and curcumin,respectively, and the inter day accuracy were99.665% ─ 103.06%, 97.669% ─ 103.513%, 99.569% ─103.617%, and 97.929% ─ 103.606%, respectively. Conclusion: The method was found to be simple, accurateand precise and is recommended for routinequality control analysis of commercial Chinese medicineproducts containing the flour flavonoids as theirprinciple components in the extracts.
Ang, Lee Fung,Yam, Mun Fei,Fung, Yvonne Tan Tze,Kiang, Peh Kok,Darwin, Yusrida KOREAN PHARMACOPUNCTURE INSTITUTE 2014 Journal of pharmacopuncture Vol.17 No.4
Objectives: Quercetin and curcuminoids are important bioactive compounds found in many herbs. Previously reported high performance liquid chromatography ultraviolet (HPLC-UV) methods for the detection of quercetin and curcuminoids have several disadvantages, including unsatisfactory separation times and lack of validation according the standard guidelines of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. Methods: A rapid, specific, reversed phase, HPLC-UV method with an isocratic elution of acetonitrile and 2% v/v acetic acid (40% : 60% v/v) (pH 2.6) at a flow rate of 1.3 mL/minutes, a column temperature of $35^{\circ}C$, and ultraviolet (UV) detection at 370 nm was developed. The method was validated and applied to the quantification of different types of market available Chinese medicine extracts, pills and tablets. Results: The method allowed simultaneous determination of quercetin, bisdemethoxycurcumin, demethoxycurcumin and curcumin in the concentration ranges of $0.00488-200{\mu}g/mL$, $0.625-320{\mu}g/mL$, $0.07813-320{\mu}g/mL$ and $0.03906-320{\mu}g/mL$, respectively. The limits of detection and quantification, respectively, were 0.00488 and $0.03906{\mu}g/mL$ for quercetin, 0.62500 and $2.50000{\mu}g/mL$ for bisdemethoxycurcumin, 0.07813 and $0.31250{\mu}g/mL$ for demethoxycurcumin, and 0.03906 and $0.07813{\mu}g/mL$ for curcumin. The percent relative intra day standard deviation (% RSD) values were $0.432-0.806{\mu}g/mL$, $0.576-0.723{\mu}g/mL$, $0.635-0.752{\mu}g/mL$ and $0.655-0.732{\mu}g/mL$ for quercetin, bisdemethoxycurcumin, demethoxycurcumin and curcumin, respectively, and those for intra day precision were $0.323-0.968{\mu}g/mL$, $0.805-0.854{\mu}g/mL$, $0.078-0.844{\mu}g/mL$ and $0.275-0.829{\mu}g/mL$, respectively. The intra day accuracies were 99.589%-100.821%, 98.588%-101.084%, 9.289%-100.88%, and 98.292%-101.022% for quercetin, bisdemethoxycurcumin, demethoxycurcumin and curcumin, respectively, and the inter day accuracy were 99.665%-103.06%, 97.669%-103.513%, 99.569%-103.617%, and 97.929%-103.606%, respectively. Conclusion: The method was found to be simple, accurate and precise and is recommended for routine quality control analysis of commercial Chinese medicine products containing the flour flavonoids as their principle components in the extracts.