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Journal of Integrative Agriculture  2016, Vol. 15 Issue (9): 2163-2174    DOI: 10.1016/S2095-3119(16)61386-2
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Analysis of 13 kinds of steroid hormones in raw milk using modified QuEChERS method combined with UPLC-QTOF-MS
AN Xin-tong1, LI Zeng-mei2, 3, DENG Li-gang2, 3, ZHAO Shan-cang2, 3, WANG Ming-lin1
1 Academy of Food Science and Engineering, Shandong Agricultural University, Tai’an 271018, P.R.China
2 Institute of Agricultural Quality Standards and Testing Technology Research, Shandong Academy of Agricultural Sciences, Jinan 250100, P.R.China
3 Key Laboratory of Test Technology on Food Quality and Safety of Shandong Province, Jinan 250100, P.R.China
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Abstract      Thirteen kinds of steroid hormones in raw milk (cow, goat and buffalo milk) were analyzed with ultra performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-QTOF-MS) after extraction and cleanup with the modified QuEChERS method. These steroid hormones included 17β-estradiol, estriol, estrone, diethylstilbestrol, progesterone, melengestrol acetate, megestrol acetate, chlormadinone acetate, 19-nortestosterone, metandienone, boldenone, epitestosterone, and testosterone. The limits of detection for the raw milk basing on 3 times the signal to noise ratios (S/N=3) was in range of 0.07-0.51 µg kg–1, and the limits of quantification (basing on S/N=10 method) covered the ranges from 0.23 to 1.7 µg kg–1. With matrix external standard method, the substances presented recoveries over the range 74.2–99.7%. Qualitative analysis was also done in the mass/mass spectrum (MS/MS) mode and each debris structure of 13 kinds of steroid hormones was achieved. The methodology was then applied in real raw milk samples which were collected in several areas of China and the progesterone was detected with high level.
Keywords:  UPLC-QTOF-MS        raw milk        QuEChERS        steroid hormones        acidic alumina  
Received: 04 November 2015   Accepted:

This research was supported by the Natural Science Foundation of Shandong Province, China (ZR2015CM015) and the Special Fund for Agro-Scientific Research in the Public Interest, China (201403071). The authors show great appreciation to Shandong Academy of Agricultural Sciences, China, for supporting the laboratory equipments.

Corresponding Authors:  WANG Ming-lin, Tel: +86-538-8246029, Fax: +86-538-8249157, E-mail:   
About author:  TAN Xin-tong, E-mail:;

Cite this article: 

TAN Xin-tong, LI Zeng-mei, DENG Li-gang, ZHAO Shan-cang, WANG Ming-lin. 2016. Analysis of 13 kinds of steroid hormones in raw milk using modified QuEChERS method combined with UPLC-QTOF-MS. Journal of Integrative Agriculture, 15(9): 2163-2174.

Anastassiades M, Lehotay S J, Stajnbaher D, Schenck F J. 2003. Fast easy multiresidue method employing acetonitrile extraction/partitioning, “dispersive solid-phase extraction” for the determination of pesticide residues in produce. Journal of Aoac International, 86, 412-431.

Avila-Poveda O H, Montes-Pérez R C, Benitez-Villalobos F, Rosas C. 2013. Development and validation of a solid-phase radioimmunoassay for measuring progesterone and testosterone in octopus gonad extracts. Malacologia, 56, 121–134.

Berry D A. 2008. The science of doping. Nature, 454, 692–693.

Cie?lik E, Sadowska-Rociek A, Ruiz J M M, Surma-Zadora M. 2011. Evaluation of QuEChERS method for the determination of organochlorine pesticide residues in selected groups of fruits. Food Chemistry, 125, 773–778.

Chen X H, Sheng X F, Jin M C. 2010. Simultaneous determination of eight progesterone derivatives residues in milk by ultra-fast liquid chromatography coupled with tandem mass spectrometry. Chinese Journal of Health Laboratory Technology, 11, 2671–2675.  (in Chinese)

Danaceau J P, Scott Morrison M, Slawson M H. 2008. Quantitative confirmation of testosterone and epitestosterone in human urine by LC/Q-ToF mass spectrometry for doping control. Journal of Mass Spectrometry, 43, 993–1000.

Farke C, Rattenberger E, Roiger S U, Meyer H H. 2011. Bovine colostrum: Determination of naturally occurring steroid hormones by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Journal of Agricultural and Food Chemistry, 59, 1423–1427.

Furlani R P Z, Marcilio K M, Leme F M, Tfouni S A V. 2011. Analysis of pesticide residues in sugarcane juice using QuEChERS sample preparation and gas chromatography with electron capture detection. Food Chemistry, 126, 1283–1287.

Gao W, Stalder T, Foley P, Rauh M, Deng H, Kirschbaum C. 2013. Quantitative analysis of steroid hormones in human hair using a column-switching LC-APCI-MS/MS assay. Journal of Chromatography (B), 928, 1–8.

GB/T 22973-2008. 2009. Determination of melengestrol acetate,chlormadinone acetate and megestrol acetate residues in milk and milk powder—HPLC-MS-MS method. China Standards Press. pp. 20-21

Hansen M, Jacobsen N W, Nielsen F K, Björklund E, Styrishave B, Halling-Sørensen B. 2011. Determination of steroid hormones in blood by GC-MS/MS. Analytical and Bioanalytical Chemistry, 400, 3409–3417.

Ingrand V, Herry G, Beausse J, de Roubin M R. 2003. Analysis of steroid hormones in effluents of wastewater treatment plants by liquid chromatoraphy-tandem mass spectrometry. Journal of Chromatography (A), 1020, 99–104.

James R E, Lukanova A, Dossus L, Becker S, Rinaldi S, Tjønneland A, Olsen A, Overvad K, Mesrine S, Engel P. 2011. Postmenopausal serum sex steroids and risk of hormone receptor-positive and -negative breast cancer: a nested case-control study. Cancer Prevention Research, 4, 1626–1635.

Key T, Appleby P, Reeves G, Roddam A, Helzlsouer K, Alberg A, Rollison D, Dorgan J, Brinton L, Overvad K. 2011. Circulating sex hormones and breast cancer risk factors in postmenopausal women: Reanalysis of 13 studies. British Journal of Cancer, 105, 709–722.

Koesukwiwat U, Sanguankaew K, Leepipatpiboon N. 2008. Rapid determination of phenoxy acid residues in rice by modified QuEChERS extraction and liquid chromatography-tandem mass spectrometry. Analytica Chimica Acta, 626, 10–20.

Li Y F, Qiao L Q, Li F W, Ding Y, Yang Z J, Wang M L. 2014. Determination of multiple pesticides in fruits and vegetables using a modified quick, easy, cheap, effective, rugged and safe method with magnetic nanoparticles and gas chromatography tandem mass spectrometry. Journal of Chromatography (A), 1361, 77–87.

Pareja L, Cesio V, Heinzen H, Fernández-Alba A R. 2011. Evaluation of various QuEChERS based methods for the analysis of herbicides and other commonly used pesticides in polished rice by LC-MS/MS. Talanta, 83, 1613–1622.

Pérez R A, Albero B, Tadeo J L, Molero E, Sánchez-Brunete C. 2014. Analysis of steroid hormones in water using palmitate-coated magnetite nanoparticles solid-phase extraction and gas chromatography-tandem mass spectrometry. Chromatographia, 77, 837–843.

Reckelhoff J F, Zhang H, Granger J P. 1998. Testosterone exacerbates hypertension and reduces pressure-natriuresis in male spontaneously hypertensive rats. Hypertension, 31, 435–439.

Stanczyk F Z, Clarke N J. 2010. Advantages and challenges of mass spectrometry assays for steroid hormones. The Journal of Steroid Biochemistry and Molecular Biology, 121, 491–495.

Tso J, Aga D S. 2010. A systematic investigation to optimize simultaneous extraction and liquid chromatography tandem mass spectrometry analysis of estrogens and their conjugated metabolites in milk. Journal of Chromatography (A), 1217, 4784–4795.

Wu C C, Schwartzman M L. 2011. The role of 20-HETE in androgen-mediated hypertension. Prostaglandins & Other Lipid Mediators, 96, 45–53.
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