Parabens are esters of p-hydroxybenzoic acid and are widely used as preservatives in cosmetics, pharmaceuticals and foodstuffs. The presences of parabens in infant formulas raise concerns due to their potential to disrupt endocrine function in infants and cause reproductive toxicities.
In this study a new method was developed for extraction and determination of methylparaben in infant formulas using HPLC method and UV detector. Methanol and trichloroacetic acid were used for extraction and isocratic mobile phase comprising equal proportions of glacial acetic acid in water (50:850 v/v) and methanol was used for separation of methylparaben.
Recovery of the extraction procedure was good and interferences between methylparaben and other ingredients peaks in HPLC chromatograms decreased. The average recoveries for methylparaben were about 88–108 %. The limit of detection and limit of quantitation for methylparaben were 0.2 and 0.5 µg/mL, respectively. Results of the method showed good reproducibility (relative standard deviation (RSD) 0.29–1.94 % for within day analysis and 0.84–2.18 % for between day analysis). Results were linear in range of 0.5–20 µg/mL methylparaben. The results of twenty real infant formula samples showed methylparaben was found only in one sample in concentration 0.3 µg/mL.
The new extraction and measurement method was a short-time method and could be applicable for large numbers of samples. This method was fast, sensitive and accurate and was capable of being used in legal laboratory references for determination of methylparaben content.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
The data will not be shared with a reason.
Rastogi SC, Schouten A, De Kruijf N, Weijland JW. Contents of methyl-, ethyl-, propyl-, butyl- and benzylparaben in cosmetic products. Contact Dermatitis. 1995;32(1):28–30.
Soni MG, Taylor SL, Greenberg NA, Burdock GA. Evaluation of the health aspects of methyl paraben: a review of the published literature. Food Chem Toxicol. 2002;40(10):1335–73.
Soni MG, Carabin IG, Burdock GA. Safety assessment of esters of p-hydroxybenzoic acid (parabens). Food Chem Toxicol. 2005;43(7):985–1015.
Zhang Q, Lian M, Liu L, Cui H. High-performance liquid chromatographic assay of parabens in wash-off cosmetic products and foods using chemiluminescence detection. Anal Chim Acta. 2005;537(1):31–9.
Andersen FA. Final amended report on the safety assessment of methylparaben, ethylparaben, propylparaben, isopropylparaben, butylparaben, isobutylparaben, and benzylparaben as used in cosmetic products. Int J Toxicol. 2008;27(Suppl 4):1–82.
Eriksson E, Andersen HR, Ledin A. Substance flow analysis of parabens in Denmark complemented with a survey of presence and frequency in various commodities. J Hazard Mater. 2008;156(1–3):240–59.
El Hussein S, Muret P, Berard M, Makki S, Humbert P. Assessment of principal parabens used in cosmetics after their passage through human epidermis-dermis layers (ex-vivo study). Exp Dermatol. 2007;16(10):830–6.
Ye X, Kuklenyik Z, Bishop AM, Needham LL, Calafat AM. Quantification of the urinary concentrations of parabens in humans by on-line solid phase extraction-high performance liquid chromatography-isotope dilution tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2006;844(1):53–9.
Ye X, Bishop AM, Reidy JA, Needham LL, Calafat AM. Parabens as urinary biomarkers of exposure in humans. Environ Health Perspect. 2006;114(12):1843–6.
Ye X, Tao LJ, Needham LL, Calafat AM. Automated on-line column-switching HPLC-MS/MS method for measuring environmental phenols and parabens in serum. Talanta. 2008;76(4):865–71.
Calafat AM, Ye X, Wong LY, Bishop AM, Needham LL. Urinary concentrations of four parabens in the U.S. population: NHANES 2005–2006. Environ Health Perspect. 2010;118(5):679–85.
Schlumpf M, Kypke K, Wittassek M, Angerer J, Mascher H, Mascher D, et al. Exposure patterns of UV filters, fragrances, parabens, phthalates, organochlor pesticides, PBDEs, and PCBs in human milk: correlation of UV filters with use of cosmetics. Chemosphere. 2010;81(10):1171–83.
Casas L, Fernandez MF, Llop S, Guxens M, Ballester F, Olea N, et al. Urinary concentrations of phthalates and phenols in a population of Spanish pregnant women and children. Environ Int. 2011;37(5):858–66.
Frederiksen H, Jorgensen N, Andersson AM. Parabens in urine, serum and seminal plasma from healthy Danish men determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS). J Expo Sci Environ Epidemiol. 2011;21(3):262–71.
Smith KW, Braun JM, Williams PL, Ehrlich S, Correia KF, Calafat AM, et al. Predictors and variability of urinary paraben concentrations in men and women, including before and during pregnancy. Environ Health Perspect. 2012;120(11):1538–43.
Miller D, Wheals BB, Beresford N, Sumpter JP. Estrogenic activity of phenolic additives determined by an in vitro yeast bioassay. Environmental health perspectives. 2001;109(2):133–8.
Okubo T, Yokoyama Y, Kano K, Kano I. ER-dependent estrogenic activity of parabens assessed by proliferation of human breast cancer MCF-7 cells and expression of ERalpha and PR. Food Chem Toxicol. 2001;39(12):1225–32.
Byford JR, Shaw LE, Drew MG, Pope GS, Sauer MJ, Darbre PD. Oestrogenic activity of parabens in MCF7 human breast cancer cells. J Steroid Biochem Mol Biol. 2002;80(1):49–60.
Darbre PD, Harvey PW. Paraben esters: review of recent studies of endocrine toxicity, absorption, esterase and human exposure, and discussion of potential human health risks. J Appl Toxicol. 2008;28(5):561–78.
Boberg J, Taxvig C, Christiansen S, Hass U. Possible endocrine disrupting effects of parabens and their metabolites. Reprod Toxicol. 2010;30(2):301–12.
Witorsch RJ, Thomas JA. Personal care products and endocrine disruption: a critical review of the literature. Crit Rev Toxicol. 2010;40(Suppl 3):1–30.
Routledge EJ, Parker J, Odum J, Ashby J, Sumpter JP. Some alkyl hydroxy benzoate preservatives (parabens) are estrogenic. Toxicol Appl Pharmacol. 1998;153(1):12–9.
Pedersen KL, Pedersen SN, Christiansen LB, Korsgaard B, Bjerregaard P. The preservatives ethyl-, propyl- and butylparaben are oestrogenic in an in vivo fish assay. Pharmacol Toxicol. 2000;86(3):110–3.
Oishi S. Effects of propyl paraben on the male reproductive system. Food Chem Toxicol. 2002;40(12):1807–13.
Oishi S. Effects of butyl paraben on the male reproductive system in mice. Arch Toxicol. 2002;76(7):423–9.
Toxicological evaluation of certain food additives with a review of general principles and of specifications. Seventeenth report of the joint FAO-WHO Expert Committee on Food Additives. 1974/01/01 ed1974. 1–40 p.
Scientific committee on consumer safety. SCCS/1446/11 Clarification on Opinion SCCS/1348/10 in the light of the Danish clause of safeguard banning the use of parabens in cosmetic products intended for children under three years of age. 10 October 2011.
Liao C, Liu F, Kannan K. Occurrence of and dietary exposure to parabens in foodstuffs from the United States. Environ Sci Technol. 2013;47(8):3918–25.
Martin CR, Ling P-R, Blackburn GL. Review of infant feeding: key features of breast milk and infant formula. Nutrients. 2016;8(5):279.
Lemaire M, Le Huërou-Luron I, Blat S. Effects of infant formula composition on long-term metabolic health. J Dev Orig Health Dis. 2018;9(6):573–89.
Wu H, Wu L-H, Wang F, Gao C-J, Chen D, Guo Y. Several environmental endocrine disruptors in beverages from South China: occurrence and human exposure. Environ Sci Pollut Res Int. 2019;26(6):5873–84.
Chiesa LM, Pavlovic R, Panseri S, Arioli F. Evaluation of parabens and their metabolites in fish and fish products: a comprehensive analytical approach using LC-HRMS. Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2018;35(12):2400–13.
Del Olmo A, Calzada J, Nuñez M. Benzoic acid and its derivatives as naturally occurring compounds in foods and as additives: Uses, exposure, and controversy. Crit Rev Food Sci Nutr. 2017;57(14):3084–103.
Gao Y, Wang Y, Yan Y, Tang K, Ding CF. Self-assembly of poly(ionic liquid) functionalized mesoporous magnetic microspheres for the solid-phase extraction of preservatives from milk samples. J Sep Sci. 2020;43(4):766–73.
Sivaraman T, Kumar TK, Jayaraman G, Yu C. The mechanism of 2,2,2-trichloroacetic acid-induced protein precipitation. J Protein Chem. 1997;16(4):291–7.
We are grateful Food and Drug Control Reference Laboratory, Iranian Food and Drug Administration for their assistance and providing some reagents.
The research was conducted with the internal funding of the Food and Drug Laboratory Research Center, Food and Drug Administration and no grants were used.
Conflict of interest
All authors declare that they have no conflict of interest.
This article does not contain any studies with human or animals.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Khansari, N., Adib, N., Alikhani, A. et al. Development and validation of a new method for determination of methylparaben in Iran market infant formulae by HPLC. J Environ Health Sci Engineer (2021). https://doi.org/10.1007/s40201-021-00628-7
- Infant formula
- Iran market