Highly selective and sensitive determination of several antioxidants in human breast milk using high-performance liquid chromatography based on Ag(III) complex chemiluminescence detection

• We present a sensitive and liable HPLC-CL method to determine AA, UA, GSH and GSSG.
• Luminol-Ag(III) system was first used in the HPLC-CL method.
• A rough correlation between the GSH/GSSG and UA concentration in human colostrums.
• Antioxidant capacity of Chinese women’s breast milk in different times was developed.

Ascorbic acid (AA), uric acid (UA) and glutathione (GSH) are the most important water-soluble antioxidants. The concentrations of GSH and glutathione disulfide (GSSG) and their molar ratio are the indicators of oxidative stress. Little is known about the contents of UA, GSH and GSSG in human milk; a reliable and sensitive method to monitor the concentrations of the four compounds simultaneously in human milk is of critical importance. A new method for separation and quantification of these water-soluble antioxidants by HPLC coupled with Ag(III) chemiluminescence detector has been developed in this work with better recoveries. The antioxidants contents were determined in different times of lactation utilizing this method. The results show that the levels of AA, UA, GSH and GSH/GSSG of human colostrum are significantly higher than those of mature milk (P < 0.05). It is concluded that colostrum contains more water-soluble antioxidants than mature milk.