Determination of flavonol aglycones in Ginkgo biloba dietary supplement crude materials and finished products by high-performance liquid chromatography: single laboratory validation.
A single laboratory validation (SLV) was completed for a method to determine the flavonol aglycones quercetin, kaempferol, and isorhamnetin in Ginkgo biloba products. The method calculates total glycosides based on these aglycones formed following acid hydrolysis. Nine matrixes were chosen for the study, including crude leaf material, standardized dry powder extract, single and multiple entity finished products, and ethanol and glycerol tinctures. For the 9 matrixes evaluated as part of this SLV, the method appeared to be selective and specific, with no observed interferences. The simplified 60 min oven heating hydrolysis procedure was effective for each of the matrixes studied, with no apparent or consistent differences between 60, 75, and 90 min at 90 degrees C. A Youden ruggedness trial testing 7 factors with the potential to affect quantitative results showed that 2 factors (volume hydrolyzed and test sample extraction/hydrolysis weight) were the most important parameters for control during sample preparation. The method performed well in terms of precision, with 4 matrixes tested in triplicate over a 3-day period showing an overall repeatability (relative standard deviation, RSD) of 2.3%. Analysis of variance testing at alpha = 0.05 showed no significant differences among the within- or between-group sources of variation, although comparisons of within-day (Sw), between-day (Sb), and total (St) precision showed that a majority of the standard deviation came from within-day determinations for all matrixes. Accuracy testing at 2 levels (approximately 30 and 90% of the determined concentrations in standardized dry powder extract) from 2 complex negative control matrixes showed an overall 96% recovery and RSD of 1.0% for the high spike, and 94% recovery and RSD of 2.5% for the low spike. HorRat scores were within the limits for performance acceptability, ranging from 0.4 to 1.3. Based on the performance results presented herein, it is recommended that this method progress to the collaborative laboratory trial.
Midwest Research Institute, 425 Volker Blvd, Kansas City, MO 64110-2299, USA. dgray@mriresearch.org
Dean
D
Gray
Kerri
K
LeVanseler
Meide
M
Pan
D002138:Calibration; D002623:Chemistry Techniques, Analytical / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D000431:Ethanol / Q000737:chemistry; D044948:Flavonols / Q000032:analysis / Q000737:chemistry*; D020441:Ginkgo biloba / Q000737:chemistry* / Q000378:metabolism*; D005990:Glycerol / Q000737:chemistry; D006868:Hydrolysis; D044949:Kaempferols / Q000032:analysis*; D008956:Models, Chemical; D010936:Plant Extracts / Q000032:analysis; D018515:Plant Leaves / Q000378:metabolism; D011208:Powders; D011794:Quercetin / Q000032:analysis* / Q000031:analogs & derivatives; D015203:Reproducibility of Results; D013696:Temperature; D013997:Time Factors
D044948:Flavonols; D044949:Kaempferols; D010936:Plant Extracts; D011208:Powders; C047368:3-methylquercetin; D000431:Ethanol; C006552:kaempferol; D011794:Quercetin; D005990:Glycerol