{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T15:30:09Z","timestamp":1789140609146,"version":"build-2803163510"},"reference-count":52,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2025,1,23]],"date-time":"2025-01-23T00:00:00Z","timestamp":1737590400000},"content-version":"vor","delay-in-days":22,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J of Separation Science"],"published-print":{"date-parts":[[2025,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>The objective of this study is to develop an HPLC\u2010UV method for the cost\u2010effective and quantitative determination of vitamin D3 in food, even in the presence of vitamin D2, with a specific focus on egg yolk. During method development, the performance of three stationary phases in resolving the peak of vitamin D2 from that of vitamin D3 was investigated. The physicochemical properties of these phases differed particularly in the extent of hydrophobicity and silanophilic activity, including a GraceSmart RP C18 column without silanol endcapping, a Robusta RP C18 column with silanol endcapping, and a Waters Xbridge RP C18 column <jats:italic>with ethylene\u2010bridged hybrid (BEH) particle technology<\/jats:italic>. The Xbridge C18 stationary phase exhibited the most favorable performance, leading to an <jats:italic>R<\/jats:italic><jats:sub>S<\/jats:sub> of 1.6 under the following nonaqueous reversed\u2010phase (NARP) experimental conditions: mobile phase, acetonitrile, methanol, and trifluoroacetic acid in a (99\/1\/0.1, v\/v\/v) ratio; column temperature, 15\u00b0C. The developed chromatographic method does not require preanalytical purification steps and is also compatible with mass spectrometry. <jats:italic>The identity of the vitamin D3 peak observed in the HPLC analysis was verified via GC\u2013MS. The NARP\u2010HPLC\u2010UV method was partially validated<\/jats:italic>, demonstrating satisfactory linearity, precision, accuracy, limit of quantification, and robustness. The HPLC method was then successfully applied to the analysis of real egg yolk samples, revealing average concentrations of vitamin D3 of 4\u20135\u00a0\u00b5g\/g of wet weight sample.<\/jats:p>","DOI":"10.1002\/jssc.70087","type":"journal-article","created":{"date-parts":[[2025,1,23]],"date-time":"2025-01-23T06:34:01Z","timestamp":1737614041000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["A Cost\u2010Effective Nonaqueous Reversed\u2010Phase High\u2010Performance Liquid Chromatography Method to Measure Vitamin D3 in Hen's Egg Yolk"],"prefix":"10.1002","volume":"48","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4905-7115","authenticated-orcid":false,"given":"Ina","family":"Varfaj","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences University of Perugia  Perugia Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alice Cartoni","family":"Mancinelli","sequence":"additional","affiliation":[{"name":"Department of Agricultural Food and Environmental Sciences University of Perugia  Perugia Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Migni","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences University of Perugia  Perugia Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0644-9461","authenticated-orcid":false,"given":"Laura","family":"Mercolini","sequence":"additional","affiliation":[{"name":"Department of Pharmacy and Biotechnology (FaBiT) Alma Mater Studiorum\u2010University of Bologna  Bologna Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cesare","family":"Castellini","sequence":"additional","affiliation":[{"name":"Department of Agricultural Food and Environmental Sciences University of Perugia  Perugia Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"Galli","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences University of Perugia  Perugia Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Desir\u00e9e","family":"Bartolini","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences University of Perugia  Perugia Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9856-0230","authenticated-orcid":false,"given":"Roccaldo","family":"Sardella","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences University of Perugia  Perugia Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,1,23]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.3390\/nu13103328"},{"key":"e_1_2_10_3_1","first-page":"346","article-title":"Vitamin D and Chronic Diseases","volume":"2","author":"Hanmin W.","year":"2017","journal-title":"Aging and Disease"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.31138\/mjr.33.1.7"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s12276-018-0038-9"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1111\/nyas.13219"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1093\/ajcn\/80.6.1678S"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.3390\/nu12061743"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/jbm4.10453"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1017\/jns.2022.11"},{"key":"e_1_2_10_11_1","first-page":"115","article-title":"Dietary Sources of Vitamin D, Vitamin D Supplementation, and Its Bio\u2010Viability","volume":"14","author":"Ku\u010dan R.","year":"2018","journal-title":"Central European Journal of Paediatrics"},{"key":"e_1_2_10_12_1","first-page":"1","article-title":"The Effect of Sunlight Exposure on Vitamin D Status in Countries of Low and High Latitudes: A systematic Literature Review","volume":"12","author":"Lizarralde O. 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