Determination of cholic acid and derivative residues in eggs by UPLC-MS/MS

Abstract

Objective: The pretreatment method of through-type solid-phase extraction was improved to be coupled with ultra performance liquid chromatography tandem mass spectrometry for detection of cholic acid，deoxycholic acid and dehydrochloric in eggs. Methods: The samples were extracted by methanol and purified by PRiME HLB，then separated on C18（100 mm×2.1 mm，3.0 μm） column with methanol-0.1% ammonia as the mobile phase for gradient elution. Meanwhile，the samples were quantified by external standard with negative ion select reaction monitoring （SRM） mode. Results: The results showed that the linear relationships of cholic acid （CA），deoxycholic acid （DCA） and dehydrochloric acid（DHCA） were good in the range from 0.5 ng/mL to 500 ng/mL（R2＞0.999）. The detection limits and quantitative limits of CA，DCA and DHCA were 1.0 μg/kg and 3.3 μg/kg. The average recovery of CA，DCA and DHCA from 1.0~10 μg/kg were 93.7%~102.5%，with the RSD of 1.8%~9.2%. 30 batches of randomly selected eggs were tested, and cholic acid was detected in all 28 batches of samples. Conclusion: This method is rapid with high sensitivity，accuracy，and it can be used as a method for the determination of cholic acid and derivative residues in eggs.

Keywords

• cholic acid
• deoxycholic acid
• dehydrochloric acid
• egg
• ultra performance liquid chromatography-tandem mass spectrometry