Food Analytical Methods

, Volume 12, Issue 4, pp 859–868 | Cite as

Simultaneous Determination of Isoflavones and Equol in Egg Yolk Using UPLC-MS/MS

  • Li HuEmail author
  • Keting Jin
  • Baozhan Zheng
  • Xiaofeng Yang
  • Shaorong Lei


A simple and reliable analytical method based on ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was developed and validated for the simultaneous determination of total isoflavone aglycones and its metabolite equol in egg yolk. The seven target analytes were separated on an Acquity ultra performance liquid chromatography (UPLC) system BEH C18 column with gradient elution and detected by electrospray ionization source (ESI) operating in negative subsection acquisition mode under multiple reaction monitoring (MRM) conditions. The results showed that the recoveries of spiked isoflavones and equol from the matrices were satisfactory, ranging between 73.74 and 89.35%, and all with relative standard deviation (RSD) values below 10%. The linearity of their corresponding concentration ranges from 2 to 500 ng/mL with correlation coefficients (r2) all higher than 0.99. The analytes were stable under the experimental conditions and the proposed method exhibited good recovery and reproducibility within only 8 min. The measurement results showed that the tested eggs contained soy isoflavones and equol, which may be beneficial for people who lack estrogens and those with hormone-related disorders. This method can also be useful for the determination of isoflavones and equol in other animal products.


Isoflavones Equol UPLC-MS/MS Enzymolysis Egg yolk 



This work was supported by the Sichuan province financial and genetic engineering special fund project (excellent paper fund) (2013LWJJ-012, 2014LWJJ-012).

Compliance with Ethical Standards

Informed Consent

Not applicable.

Ethical Approval

This article does not contain any studies with animals performed by any of the authors.

Conflict of Interest

Li Hu declare that she has no conflict of interest. Keting Jin declare that she has no conflict of interest. Baozhan Zheng declare that he has no conflict of interest. Xiaofeng Yang declare that she has no conflict of interest. Shaorong Lei declare that he has no conflict of interest.


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Center of Analysis and TestingSichuan Academy of Agricultural SciencesChengduPeople’s Republic of China
  2. 2.Institute of Quality Standards and Testing Technology for Agro-ProductsSichuan Academy of Agricultural SciencesChengduPeople’s Republic of China
  3. 3.College of ChemistrySichuan UniversityChengduChina

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