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HPLC Method for Simultaneous Quantitative Detection of Quercetin and Curcuminoids in Traditional Chinese Medicines

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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.

Abstract

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.

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