Formation and inhibition of cholesterol oxidation products in tea-leaf eggs during marinating

The objectives of this study were to develop a GC-MS method for determination of cholesterol oxidation products (COPs) in tea-leaf eggs and study the formation and inhibition of COPs as affected by heating time and various ingredients in marinated juice. The various COPs in egg and juice samples were extracted by a solvent system of chloroform/methanol (2:1, v/v), followed by purification using a silica cartridge and GC-MS for subsequent separation and quantitation, with high recovery ranges from 85.9 to 98.3% and from 83.1-100.1% being obtained for egg and juice, respectively. 5α-Cholestane was shown to be an appropriate internal standard for quantitation. A total of five COPs, including 7-keto, 5,6 β-EP, 7α-OH, 7β-OH, and triol, were formed in tea-leaf eggs during marinating, but not in marinated juice. A peak level of total COPs (2272.2 ng/g) was generated in tea-leaf eggs after 24 h of heating, but reduced to 1068.2 ng/g in 48 h. Both the total phenolic and flavonoid compounds in tea-leaf eggs showed a time-dependent increase during marinating and so did the pH and browning index in tea-leaf eggs and juice. The incorporation of soy sauce or black tea leaf into juice was effective in inhibiting COPs formation in tea-leaf eggs, with the latter being more pronounced than the former. The formation of Maillard reaction products during marinating as well as the presence of total phenolic and total flavonoid in black tea leaf was mainly responsible for COPs reduction in tea-leaf eggs.