Abstract
This study reports the improvement and validation of a colorimetric method to quantify polysorbates (20, 60, 65, and 80) in food by measuring absorbance at 620 nm using ultraviolet–visible spectrophotometry. The method was validated for linearity, limit of detection (LOD), limit of quantitation (LOQ), precision, accuracy, and measurement uncertainty. The coefficient of determination was linear (r2 ≥ 0.9991) over the measured concentration range of 50–1000 mg/L. The LOD and LOQ were 2.3–4.9 and 7.0–15.0 mg/kg, respectively. Intra-day and inter-day accuracy and precision were 91.9–104.1% and 0.1–1.1% RSD, and 91.6–103.8% and 0.4–5.0% RSD, respectively. The result of inter-laboratory recovery was 90.9–99.8% and the measurement uncertainty was < 16% with the compliance of the CODEX recommendation. Sauce, bread, whipped cream, rice cake, ice cream, and various other polysorbate-labeled food products (n = 229, detection range; N.D.–16,442.3 mg/kg) distributed in Korea were analyzed to confirm the applicability of the analytical method.
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs10068-024-01544-w/MediaObjects/10068_2024_1544_Fig1_HTML.png)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs10068-024-01544-w/MediaObjects/10068_2024_1544_Fig2_HTML.png)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs10068-024-01544-w/MediaObjects/10068_2024_1544_Fig3_HTML.png)
Similar content being viewed by others
References
Adamo M, Dick Jr LW, Qiu D, Lee AH, Devincentis J, Cheng KC. A simple reversed phase high-performance liquid chromatography method for polysorbate 80 quantitation in monoclonal antibody drug products. Journal of Chromatography B. 878(21): 1865-1870 (2010)
Association of Official Analytical Chemists. Official methods of analysis of AOAC International. AOAC International. Appendix F: Guidelines for Standard Method Performance Requirements. AOAC International, Gaithersburg (2016)
Cani PD, Everard A. Kee** gut lining at bay: impact of emulsifiers. Trends in Endocrinology and Metabolism. 26: 273-274 (2015)
Chassaing B, Koren O, Goodrich JK, Poole AC, Srinivasan S, Ley RE, Gewirtz. Dietary emulsifiers impact the mouse gut microbiota promoting colitis and metabolic syndrome. Nature. 519: 92-96 (2015)
CODEX Alimentarius Commission. Guidance on Measurement Uncertainty and Uncertainty of Sampling Guidance on Measurement Uncertainty, CX/MAS 08/29/9. Joint FAO/WHO Food Standards Programme CODEX Committee on Methods of Analysis and Sampling. Budapest, Hungary (2008)
CODEX Alimentarius Commission. Codex general standard for food additives (Codex stan 192-1995) (2019)
EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS). Scientific opinion on the re‐evaluation of polyoxyethylene sorbitan monolaurate (E 432), polyoxyethylene sorbitan monooleate (E 433), polyoxyethylene sorbitan monopalmitate (E 434), polyoxyethylene sorbitan monostearate (E 435) and polyoxyethylene sorbitan tristearate (E 436) as food additives. EFSA Journal. 13(7): 4152 (2015)
Ellison SL, Williams A. Quantifying Uncertainty in Analytical Measurements. EURACHEM/CITAC, London, UK, pp. 33-89 (2012)
Engelmaier A, Anderle H, Weber A. Alkaline hydrolysis to increase the selectivity of colorimetric determination of polysorbate. Biologicals. 49: 6-14 (2017)
Evers DH, Schultz-Fademrecht T, Garidel P, Buske J. Development and validation of a selective marker-based quantification of polysorbate 20 in biopharmaceutical formulations using UPLC Qda detection. Journal of Chromatography B. 1157: 122287 (2020)
Feinberg M, Laurentie M. A global approach to method validation and measurement uncertainty. Accreditation and Quality Assurance. 11(1): 3-9 (2006)
Food and Drug Administration (FDA). Food additive status list. Available from: https://www.fda.gov/food/food-additives-petitions/food-additive-status-list. Accessed Feb. 5, 2020 (2019)
Food and Drug Administration. Methods, Method Verification and Validation. Available from: https://www.fda.gov/media/73920/download. Accessed Dec. 28 2021 (2014)
Halmos EP, Mack A, Gibson PR. Review article: emulsifiers in the food supply and implications for gastrointestinal disease. Alimentary Pharmacology and Therapeutics. 49: 41–50 (2019)
Hewitt D, Alvarez M, Robinson K, Ji J, Wang YJ, Kao YH, Zhang T. Mixed-mode and reversed-phase liquid chromatography–tandem mass spectrometry methodologies to study composition and base hydrolysis of polysorbate 20 and 80. Journal of Chromatography A. 1218(15): 2138-2145 (2011)
ICH. Validac ction of analytical procedures: text and methodology. Q2 (R1) 1: 05 (2005)
Ilko D, Braun A, Germershaus O, Meinel L, Holzgrabe U. Fatty acid composition analysis in polysorbate 80 with high performance liquid chromatography coupled to charged aerosol detection. European Journal of Pharmaceutics and Biopharmaceutics. 94: 569-574 (2015)
Japan External Trade Organization (JETRO). Specifications and standards for foods, food additives, etc. Under the food sanitation act (Abstract). Available from: https://www.jetro.go.jp/ext_images/en/reports/regulations/pdf/foodext2010e.pdf Accessed Dec. 23, 2021 (2010)
Jie M. Chinese standards for food additives GB2760-2015. Global agricultural information network. Giant report number (FAIRS) 15013. Bei**g, China. pp. 10–34 (2015)
Kato H, Nagai Y, Yamamoto K, Sakabe Y. Determination of polysorbates in foods by colorimetry with confirmation by infrared spectrophotometry, thin-layer chromatography, and gas chromatography. Journal of the Association of Official Analytical Chemists. 72(1): 27-29 (1989)
Lind TK, Nilsson EJ, Wyler B, Scherer D, Skansberger T, Morin M, Kocherbitov V, Engblom J. Effects of ethylene oxide chain length on crystallization of polysorbate 80 and its related compounds. Journal of Colloid and Interface Science. 592: 468-484 (2021)
Ministry of Health, Labour, Welfare (MHLW). Food additives analysis methods. Available from:https://www.ffcr.or.jp/tsuuchi/upload/sikenhoH200430.pdf. Accessed Dec. 23, 2021 (2008)
Mondal B, Kote M, Lunagariya C, Patel M. Development of a simple high performance liquid chromatography (HPLC)/evaporative light scattering detector (ELSD) method to determine Polysorbate 80 in a pharmaceutical formulation. Saudi Pharmaceutical Journal. 28(3): 325-328 (2020)
Moore J. Final report on the safety assessment of polysorbates 20, 21, 40, 60, 61, 65, 80, 81, and 85. Journal of the American College of Toxicology. 3: 1-82 (1984)
Nair LM, Stephens NV, Vincent S, Raghavan N, Sand PJ. Determination of polysorbate 80 in parenteral formulations by high-performance liquid chromatography and evaporative light scattering detection. Journal of Chromatography A. 1012(1): 81-86 (2003)
Nayak VS, Tan Z, Ihnat PM, Russell RJ, Grace MJ. Evaporative light scattering detection based HPLC method for the determination of polysorbate 80 in therapeutic protein formulations. Journal of Chromatographic Science. 50(1): 21-25 (2012)
Nguyen NT. Acinetobacter soli SP2 capable of high-efficiency degradation of food emulsifier polysorbate 80. Current Microbiology. 75(7): 896-900 (2018)
Sannaningannavar FM, Patil SN, Melavanki RM, Navati BS, Ayachit NH. Ultrasonic study of thermo-acoustic parameters of the polysorbate 20, 40, 60 and 80 liquid surfactants at different temperatures. Journal of Molecular Liquids. 196: 244-248 (2014)
Savjani N, Babcock E, Khor HK, Raghani A. Use of ferric thiocyanate derivatization for quantification of polysorbate 80 in high concentration protein formulations. Talanta. 130: 542-546 (2014)
Shende N, Karale A, Bhagade S, Gulhane A, Bore P, Marathe P, Chakraborty S, Mallya AD, Dhere RM. Evaluation of a sensitive GC–MS method to detect polysorbate 80 in vaccine preparation. Journal of Pharmaceutical and Biomedical Analysis. 183: 113126 (2020)
Shi S, Chen Z, Rizzo JM, Semple A, Mittal S, Cheung JK, Richardson D, Antochshuk V, Shameem M. A highly sensitive method for the quantitation of polysorbate 20 and 80 to study the compatibility between polysorbates and m-cresol in the peptide formulation. Journal of Analytical and Bioanalytical Techniques. 6(3): 1 (2015)
Titapiccolo GI, Corredig M, Alexander M. Modification to the renneting functionality of casein micelles caused by nonionic surfactants. Journal of Dairy Science. 93(2): 506-514 (2010)
Udomrati S, Cheetangdee N, Gohtani S, Surojanametakul V, Klongdee S. Emulsion stabilization mechanism of combination of esterified maltodextrin and Tween 80 in oil-in-water emulsions. Food Science and Biotechnology. 29(3): 387-392 (2020)
Wei Z, Bilbulian S, Li J, Pandey R, O'Connor E, Casas‐Finet J, Cash PW. Universal method for the determination of nonionic surfactant content in the presence of protein. Journal of Separation Science. 38(8): 1318-1325 (2015)
Zhang Q, Wang A, Meng Y, Ning T, Yang H, Ding L, **ao X, Li X. NMR method for accurate quantification of polysorbate 80 copolymer composition. Analytical Chemistry. 87(19): 9810-9816 (2015)
Acknowledgements
This study was supported by a grant from the Ministry of Food and Drug Safety, Republic of Korea in 2021.
Funding
Funding was provided by Ministry of Food and Drug Safety (Korea).
Author information
Authors and Affiliations
Contributions
JWP: Formal analysis, Writing—original draft. GYL: Formal analysis, Writing—original draft. CIY: Conceptualization, Methodology, Investigation, Writing—review and editing. JWS: Formal analysis, Writing—original draft. YJK: Conceptualization, Validation, Writing—review and editing, Funding acquisition.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Park, JW., Lee, G., Shin, JW. et al. Validation, measurement uncertainty, and determination of polysorbate-labeled foods distributed in Korea. Food Sci Biotechnol (2024). https://doi.org/10.1007/s10068-024-01544-w
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s10068-024-01544-w