Validation of analytical procedures using HPLC‐ELSD to determine six sesquiterpene lactones in Eremanthus species
BIOMEDICAL CHROMATOGRAPHY(2016)
摘要
Eremanthus species display sesquiterpene lactones with therapeutic potential. We are proposing the development of a new analytical method that has been completely validated to qualify Eremanthus species and its main lactones in raw material using HPLC-ELSD. For the sample preparation, 10.0mg of powdered Eremanthus leaves was extracted with a 5mL MeOH/H2O (9:1v/v) solution containing scopoletin at 140 mu g/mL as the internal standard. For the separation of eight compounds, six of which were lactones, one internal (IS) and one secondary standard were performed utilizing monolithic columns with a nonlinear gradient. The selectivity, stability, precision and matrix effects parameters showed values of RSD of <10%. The six lactones and scopoletin (IS) were recovered with a proportion between 74 and 90% with accuracy represented by error at -25.41%. The linear dynamic range was obtained between 10.0 and 310.0 mu g/mL for all compounds with r(2)>0.9987. The limits of detection and quantitation ranged from 2.00 to 6.79 mu g/mL and from 6.00 to 20.40 mu g/mL, respectively. Assessing the robustness study, this method can be used in inter-laboratory studies. Using the HPLC-ELSD method, six sesquiterpene lactones including 4,5-dihydro-2,3-dihydroxy-15-deoxy-goyazensolide, goyazensolide, 4,5-dihydro-2,3-epoxy-15-deoxy-goyazensolide, centratherin, 4,5-dihydro-15-deoxy-goyazensolide and lychnofolide, were detected and quantified from distinct Eremanthus species, which were collected in different regions. Copyright (c) 2015 John Wiley & Sons, Ltd.
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关键词
Eremanthus,sesquiterpene lactones,HPLC-ELSD,standardization,validation
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