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A UHPLC-MS/MS method for profiling multifunctional steroids in human hair

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Abstract

It is important to profile steroids in many physiological and pathological processes. Recently, hair has been used for the long-term measurement of endogenous steroid hormones. Analyzing hair has advantages of being noninvasive and time sequential compared with other bio-specimens. Liquid chromatography-mass spectrometry (LC-MS) techniques have been widely used over the past decades; however, it is challenging to profile estrogens in hair by LC-MS, and more comprehensive steroid profiling is required. In this paper, an ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed to simultaneously profile 28 multifunctional steroids, including corticosteroids (n = 6), estrogens (n = 13), androgens (n = 5) and progestogens (n = 4), in human scalp hair in a single run. To optimize the sample preparation procedure, we evaluated extraction time, post-incubation purification and hair fragment length; 30 mg hair samples were washed with hexane, cut into 5 mm pieces and incubated in methanol for 18 h at 25 °C. Methanol extraction derivatized using Girard P and dansyl chloride reagent was analyzed within 25 min using an automated injection program combined with a diverter valve switch and step analysis (AIDSA). The method was well validated in terms of linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, matrix effect and recovery, and was successfully applied to a steroid profile from male and female hairs. Significant differences were observed between genders. In addition, steroids showed a declining trend from the proximal to more distal hair segments; thus, care should be taken when obtaining hair samples for analysis to account for this difference in steroid levels along the length of hair.

The workflow of the estabished UHPLC-MS/MS method

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Acknowledgments

This work was supported by the Ministry of Science and Technology of the People’s Republic of China (2016ZX09101017).

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Authors and Affiliations

  1. State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100050, China

    Zhen Dong, Caihong Wang, Jinlan Zhang & Zhe Wang

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  1. Zhen Dong
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  2. Caihong Wang
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  3. Jinlan Zhang
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  4. Zhe Wang
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Correspondence to Jinlan Zhang.

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The authors declare that there are no conflicts of interest.

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Informed consent was obtained from all individual participants who provided hair samples. Ethics approval for this study has been obtained from the Ethics Committee, Peking Union Medical College Hospital, China.

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Dong, Z., Wang, C., Zhang, J. et al. A UHPLC-MS/MS method for profiling multifunctional steroids in human hair. Anal Bioanal Chem 409, 4751–4769 (2017). https://doi.org/10.1007/s00216-017-0419-2

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  • DOI: https://doi.org/10.1007/s00216-017-0419-2

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