New Method for Analyzing Bioactive Compounds in Goat Serum Using UHPLC–MS/MS

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A newly developed UHPLC–MS/MS method allows for the accurate analysis of bioactive compounds, including apigenin, apigenin 7-glucoside, and chlorogenic acid, in goat serum.

A novel method utilizing ultrahigh-pressure liquid chromatography (UHPLC)–tandem mass spectrometry (MS/MS) has been developed for the accurate determination of bioactive compounds, namely chlorogenic acid (CA), apigenin (AP), and apigenin 7-glucoside (AP7G), in goat serum. The study, published in the journal Chromatographia, was conducted by researchers from Poznan University of Medical Sciences in Poland, employed a Kinetex C18 analytical column and a mobile phase with gradient elution consisting of 0.1% formic acid in water and methanol (1). Detection of the analytes was performed using a triple quadrupole mass spectrometer with electrospray ionization in positive mode.

Chlorogenic acid (CA), apigenin (AP), and apigenin 7-glucoside (AP7G) are important compounds in goat serum due to their potential health benefits and bioactivity. Chlorogenic acid is a polyphenolic compound known for its antioxidant and anti-inflammatory properties, which contribute to its potential protective effects against oxidative stress-related diseases. Apigenin, a flavonoid, has been associated with various health benefits such as anti-cancer, anti-inflammatory, and antioxidant activities. Its presence in goat serum suggests its absorption from dietary sources or endogenous synthesis. Apigenin 7-glucoside, a glycosylated form of apigenin, is a bioactive metabolite that can exert similar biological effects and has shown potential in various physiological processes. The presence of these compounds in goat serum highlights their bioavailability and suggests their potential contribution to the health-promoting properties of goat-derived products.

The validated method demonstrated excellent sensitivity, with a lower limit of quantitation of 0.5 ng mL-1 for AP and AP7G, and 2.5 ng mL-1 for CA. The precision of the method, expressed as relative standard deviation, ranged from 1.22% to 14.5%, while the accuracy, expressed as relative error, ranged from 0.147% to 14.6%. The extraction recoveries of the compounds ranged from 76.2% to 89.5%.


To showcase the applicability of the method, the researchers analyzed the serum of sixty goats, half of which were fed with fodder supplemented with an herbal mixture. The results revealed higher concentrations of all analyzed compounds in the serum of the herb-fed animals compared to the control group that received standard fodder.

This innovative UHPLC–MS/MS method offers a reliable and sensitive means of quantifying bioactive compounds in goat serum. By accurately measuring the levels of AP, AP7G, and CA, researchers can gain insights into the presence and concentrations of these important compounds in goat blood. This analytical approach opens up new possibilities for studying the bioavailability, pharmacokinetics, and potential health benefits associated with the consumption of bioactive compounds in goat-derived products. The application of this method has the potential to advance our understanding of the bioactivity and therapeutic potential of these compounds, paving the way for the development of functional foods and nutraceuticals with improved health benefits derived from goat sources.


(1) Karaźniewicz-Łada, M.; Wójtowski, J. A.; Główka, F.; Danków, R.; Pikul, J.; Gryszczyńska, A.; Foksowicz-Flaczyk, J.; Mikołajczak, P. L. Application of UPLC-MS/MS Method for Analysis of Apigenin, Apigenin 7-Glucoside and Chlorogenic Acid in Goat Serum. Chromatographia 2023, 86, 401–411. DOI: