Validation of the Analytical Method for Chlorogenic Acid in Blueberry Yeast Fermented Powder for Standardization as a Functional Health Ingredient

摘要

This study validated an analytical method for chlorogenic acid in blueberry yeast fermented powder (BYFP) for standardization as a functional health ingredient. Measurements of the yield according to the concentration of the extraction solvent for analyzing the level chlorogenic acid before validation showed that the extraction yield was highest at 2,040.14 ug/mL when extracted with 50% ethanol. The quantitative determination method of chlorogenic acid was optimized by HPLC analysis using a Capceilpak-C18 column with a gradient elution of 1% acetic acid in water and acetonitrile. This method was validated according to the specificity, linearity, accuracy, precision test, and recovery test. The method showed reliable linearity of the calibration curve with a R~2 of 0.9999, and the limit of quantitation and the detection limit were 5.38 ug/mL and 16.32 pg/mL, respectively. The relative standard deviation (RSD) from the intra- and inter-day precision according to the chlorogenic acid concentration ranged from 1.34 to 2.5% and 0.92 to 2.17%, respectively. The recovery test ranged from 103.03% to 103.36% with RSD values from 0.83 to 1.22%. Therefore, chlorogenic acid as a marker compound in BYFP was analyzed. The amount of chlorogenic acid in BYFP wasapproximately 2,032.66 pg/mL after triplicate analysis, and the RSD was 0.58% using the validated method. These results suggest that the method is very useful for evaluating the marker compounds in BYFP to develop a health functional food material.