Abstract
This work was carried out to develop a high-performance liquid chromatography tandem mass spectrometry method for simultaneous determination of 11 marker compounds in phytoestrogen-rich food and dietary supplements (raw materials and formulated products) prepared from various sources: soy, red clover, chastetree, hop, and flax. The method was fully validated and showed detection limits varying from 0.2 to 1.5 μg/mL, average recoveries ranging from 95 to 105%, and relative standard deviation values lower than 4.98%. Evidently lower amounts of phytoestrogens were found in food samples than in the dietary supplements. Calculations for the actual daily dose of total phytoestrogens ranged from 0.01 to 6.61 mg among the different products. All derived total phytoestrogen amounts turned out to be below the total content stated by manufacturers (less than 63% of the claimed specifications). Moreover, two samples did not contained phytoestrogens at all. The most eye-catching batch-to-batch deviations were represented by chasteberry-based products (relative standard deviation higher than 9.8%).
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Almeida IM, Rodrigues F, Sarmento B, Alves RC, Oliveira MB (2015) Isoflavones in food supplements: chemical profile, label accordance and permeability study in Caco-2 cells. Food Funct 6:938–946. https://doi.org/10.1039/c4fo01144a
Andres S, Hansen U, Niemann B, Palavinskas R, Lampen A (2015) Determination of the isoflavone composition and estrogenic activity of commercial dietary supplements based on soy or red clover. Food Funct 6:2017–2025. https://doi.org/10.1039/c5fo00308c
Boniglia C, Carratù B, Gargiulo R, Giammarioli S, Mosca M, Sanzini E (2009) Content of phytoestrogens in soy-based dietary supplements. Food Chem 115:1389–1392. https://doi.org/10.1016/j.foodchem.2009.01.062
Bustamante-Rangel M, Delgado-Zamarreño MM, Pérez-Martín L, Rodríguez-Gonzalo E, Domínguez-Álvarez J (2018) Analysis of isoflavones in foods. Compr Rev Food Sci Food Saf 17:391–411. https://doi.org/10.1111/1541-4337.12325
Chen LJ, Zhao X, Plummer S, Tang J, Games DE (2005) Quantitative determination and structural characterization of isoflavones in nutrition supplements by liquid chromatography-mass spectrometry. J Chromatogr A 1082:60–70. https://doi.org/10.1016/j.chroma.2005.03.066
Clarke DB, Bailey V, Lloyd AS (2008) Determination of phytoestrogens in dietary supplements by LC-MS/MS. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 25:534–547. https://doi.org/10.1080/02652030701658340
Coldham NG, Sauer MJ (2001) Identification, quantitation and biological activity of phytoestrogens in a dietary supplement for breast enhancement. Food and Chemical Toxicology 39:1211–1224. https://doi.org/10.1016/S0278-6915(01)00081-3
Delmonte P, Perry J, Rader JI (2006) Determination of isoflavones in dietary supplements containing soy, Red Clover and kudzu: extraction followed by basic or acid hydrolysis. J Chromatogr A 1107:59–69. https://doi.org/10.1016/j.chroma.2005.11.060
Demarque DP, Crotti AEM, Vessecchi R, Lopes JLC, Lopes NP (2016) Fragmentation reactions using electrospray ionization mass spectrometry: an important tool for the structural elucidation and characterization of synthetic and natural products. Nat Prod Rep 33:432–455. https://doi.org/10.1039/c5np00073d
Fogliatto DK, Barbosa AM, Ferreira VS (2010) Voltammetric determination of the phytoestrogen genistein in soy flours and soy based supplements using cationic surfactant cetyltrimetylammonium bromide. Colloids Surf B Biointerfaces 78:243–249. https://doi.org/10.1016/j.colsurfb.2010.03.007
Fukahori M, Kobayashi S, Naraki Y, Sasaki T, Oka H, Seki M, Masada-Atsumi S, Hakamatsuka T, Goda Y (2014) Quality evaluation of medicinal products and health foods containing chaste berry (Vitex agnus-castus) in Japanese. European and American markets. Chem Pharm Bull. 62:379–385. https://doi.org/10.1248/cpb.c13-00588
Glisic M, Kastrati N, Musa J, Milic J, Asllanaj E, Portilla Fernandez E, Nano J, Ochoa Rosales C, Amiri M, Kraja B, Bano A, Bramer WM, Roks AJM, Danser AHJ, Franco OH, Muka T (2018) Phytoestrogen supplementation and body composition in postmenopausal women: a systematic review and meta-analysis of randomized controlled trials. Maturitas 115:74–83. https://doi.org/10.1016/j.maturitas.2018.06.012
He FJ, Chen JQ (2013) Consumption of soybean, soy foods, soy isoflavones and breast cancer incidence: differences between Chinese women and women in Western countries and possible mechanisms. FSHW 2:146–161. https://doi.org/10.1016/j.fshw.2013.08.002
Krenn L, Pötsch V (2006) An efficient HPLC method for the quantification of isoflavones in soy extracts and soy dietary supplements in routine quality control. Pharmazie 61:582–585
Madeira PJ, Borges CM, Florêncio MH (2010) Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometric and semi-empirical calculations study of five isoflavone aglycones. Rapid Commun Mass Sp 24:3432–3440. https://doi.org/10.1002/rcm.4791
Magiera S, Baranowska I, Lautenszleger A (2015) UHPLC-UV method for the determination of flavonoids in dietary supplements and for evaluation of their antioxidant activities. J Pharm Biomed Anal 102:468–475. https://doi.org/10.1016/j.jpba.2014.10.004
Mari A, Montoro P, Pizza C, Piacente S (2012) Liquid chromatography tandem mass spectrometry determination of chemical markers and principal component analysis of Vitex agnus-castus L. fruits (Verbenaceae) and derived food supplements. J Pharm Biomed Anal 70:224–230. https://doi.org/10.1016/j.jpba.2012.07.001
Messina M (2014) Soy foods, isoflavones, and the health of postmenopausal women. Am J Clin Nutr 100:423S–430S. https://doi.org/10.3945/ajcn.113.071464
Mornar A, Sertić M, Nigović B (2013) Development of a rapid LC/DAD/FLD/MSnmethod for the simultaneous determination of monacolins and citrinin in red fermented rice products. J Agric Food Chem 61:1072–1080. https://doi.org/10.1021/jf304881g
Mornar A, Sertić M, Amidžić Klarić D, Klarić I, Stipanović K, Nigović B (2016) Evaluation of alcohol content and metal impurities in liquid dietary supplements by sHSS-GC-FID and GFAAS techniques. Food Chem 211:285–293. https://doi.org/10.1016/j.foodchem.2016.05.068
Nakamura Y, Tsuji S, Tonogai Y (2000) Determination of the levels of isoflavonoids in soybeans and soy-derived foods and estimation of isoflavonoids in the Japanese daily intake. J AOAC Int 83:635–650
Neacsu M, Vicas S, Socaciu C (2008) HPLC analysis coupled with diode array detector used in determination of isoflavone aglycones from different Romanian varieties of soybeans. Bulletin UASVM Agriculture 65:310–315. https://doi.org/10.15835/buasvmcn-agr:915
Popova IE, Hall C, Kubátová A (2009) Determination of lignans in flaxseed using liquid chromatography with time-of-flight mass spectrometry. J Chromatogr A 1216:217–229. https://doi.org/10.1016/j.chroma.2008.11.063
Prokudina EA, Havlíček L, Al-Maharik N, Lapčík O, Strnad M, Gruzb J (2012) Rapid UPLC–ESI–MS/MS method for the analysis of isoflavonoids and other phenylpropanoids. J Food Compost Anal 26:36–42. https://doi.org/10.1016/j.jfca.2011.12.001
Rizzo G, Baroni L (2018) Soy, soy foods and their role in vegetarian diets. Nutrients 20:1–51. https://doi.org/10.3390/nu10010043
Romani A, Vignolin P, Tanini A, Pampaloni B, Heimler D (2010) HPLC/DAD/MS and antioxidant activity of isoflavone-based food supplements. Nat Prod Commun. 5:1775–1780
Rybak ME, Parker DL, Pfeiffer CM (2008) Determination of urinary phytoestrogens by HPLC-MS/MS: a comparison of atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI). J Chromatogr B 861:145–150. https://doi.org/10.1016/j.jchromb.2007.11.013
Sertić M, Mornar A, Nigović B (2014) A rapid profiling of hypolipidemic agents in dietary supplements by direct injection tandem mass spectrometry. J Food Compost Anal 34:68–74. https://doi.org/10.1016/j.jfca.2014.03.001
Sowa I, Wójciak-Kosior M, Strzemski M, Rokicka K, Blicharski T, Kocjan R (2014) Analysis of compounds with phytoestrogenic activity in dietary supplements with use of HPTLC-densitometry method. Acta Pol Pharm 71:265–269
Thompson LU, Boucher BA, Cotterchio M, Kreiger N, Liu Z (2007) Dietary phytoestrogens, including isoflavones, lignans, and coumestrol, in nonvitamin, nonmineral supplements commonly consumed by women in Canada. Nutr Cancer 59:176–184. https://doi.org/10.1080/01635580701420616
Uifălean A, Farcaş A, Ilieş M, Hegheş SC, Ionescu C, Iuga CA (2015) Assessment of isoflavone aglycones variability in soy food supplements using a validated HPLC-UV method. Clujul Med 88:373–380. https://doi.org/10.15386/cjmed-468
Yanaka K, Takebayashi J, Matsumoto T, Ishimi Y (2012) Determination of 15 isoflavone isomers in soy foods and supplements by high-performance liquid chromatography. J Agric Food Chem 60:4012–4016. https://doi.org/10.1021/jf205154x
Yilmazer M, Stevens JF, Deinzer ML, Buhler DR (2001) In vitro biotransformation of xanthohumol, a flavonoid from hops (Humulus lupulus), by rat liver microsomes. Drug Metab Dispos 29:223–231
Funding
This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of Interest
Ana Mornar declares that she has no conflict of interest. Tajana Buhač declares that she has no conflict of interest. Daniela Amidžić Klarić declares that she has no conflict of interest. Ilija Klarić declares that he has no conflict of interest. Miranda Sertić declares that she has no conflict of interest. Biljana Nigović declares that she has no conflict of interest.
Ethical Approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Informed Consent
Not applicable.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Mornar, A., Buhač, T., Klarić, D.A. et al. Multi-targeted Screening of Phytoestrogens in Food, Raw Material, and Dietary Supplements by Liquid Chromatography with Tandem Mass Spectrometry. Food Anal. Methods 13, 482–495 (2020). https://doi.org/10.1007/s12161-019-01653-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12161-019-01653-x