Application of UPLC-MS/MS Method for Analysis of Apigenin, Apigenin 7-Glucoside and Chlorogenic Acid in Goat Serum

Abstract

UPLC-MS/MS method was developed for determination of bioactive compounds including chlorogenic acid (CA), apigenin (AP) and apigenin 7-glucoside (AP7G) in goat serum. The analytes were separated on Kinetex C18 analytical column and a mobile phase consisting 0.1% formic acid in water and methanol with gradient elution. Detection of analytes was performed using a triple quadrupole mass spectrometer with electrospray ionization in positive mode. The lower limit of quantitation was 0.5 ng mL–1 for AP and AP7G and 2.5 ng mL–1 for CA. The inter- and intra-day precision of the method expressed as relative standard deviation was in the range of 1.22–14.5%. The accuracy of the method expressed as the relative error was in the range of 0.147–14.6%. The extraction recoveries of the compounds were 76.2–89.5%. The validated method was successfully applied for analysis of the compounds in the serum of sixty goats, half of which were fed with fodder with the addition of an herbal mixture. The higher concentration values of all analyzed compounds were reported in the serum of the animals fed with herbs compared to the control group receiving standard fodder.