Determination of Camel Hide Gelatin in Donkey Hide Gelatin Based on Enzymatic Digestion by Ultra-performance Liquid Chromatography-Tandem Mass Spectrometry

The commercial value and diversified applications of donkey hide gelatin (DHG) have precipitated sophisticated adulteration practices, particularly with camel-hide gelatin substitutes. We present a mass spectrometry-based strategy to authenticate species-specific collagen peptides in commercial DHG....

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Main Authors: Qiangsheng Xie, Weijian Wang, Tianyang Gao, Feiyan Diao, Wenyue Hu, Xiaoyun Wu, Silong Li, Feng Shi, Liping Gong, Qiyan Li, Guangxi Zhai
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Journal of Food Protection
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Online Access:http://www.sciencedirect.com/science/article/pii/S0362028X2500105X
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author Qiangsheng Xie
Weijian Wang
Tianyang Gao
Feiyan Diao
Wenyue Hu
Xiaoyun Wu
Silong Li
Feng Shi
Liping Gong
Qiyan Li
Guangxi Zhai
author_facet Qiangsheng Xie
Weijian Wang
Tianyang Gao
Feiyan Diao
Wenyue Hu
Xiaoyun Wu
Silong Li
Feng Shi
Liping Gong
Qiyan Li
Guangxi Zhai
author_sort Qiangsheng Xie
collection DOAJ
description The commercial value and diversified applications of donkey hide gelatin (DHG) have precipitated sophisticated adulteration practices, particularly with camel-hide gelatin substitutes. We present a mass spectrometry-based strategy to authenticate species-specific collagen peptides in commercial DHG. A new qualitative and quantitative methodology is constructed for camel hide gelatin(CHG) material analysis in DHG. It consists of enzyme-mediated digestion, ultra-high-performance liquid chromatography (UHPLC), and triple quadrupole mass spectrometry (MS/MS) with multireaction monitoring (MRM) mode. In brief, the samples were digested with trypsin and subjected to efficient and quick separation by UHPLC, and MS/MS performed CHG identification and determination analysis. Two peptides (Camel peptide A, CPA, and Camel peptide B, CPB) were identified as CHG-specific peptides. A specificity test was then used to verify these two peptides. The method performance showed that these two peptides could distinguish CHG from other animal hide gelatins. The CPA and CPB detection limits were 5 μg/kg and 3 μg/kg, respectively. LC-MS/MS analysis detected CPA and CPB in 4.5% of market-collected DHG samples (n = 157), confirming the prevalence of this emerging adulteration practice. Thus, the present protocol is a sensitive, accurate, quick, and suitable application of species-specific peptide biomarkers, ensuring the quality of DHG products and making them authentic and traceable to protect consumers from potential health risks and food frauds.
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publisher Elsevier
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spelling doaj-art-d1bc443b9d9346be9d5695217019d2e82025-08-20T03:22:42ZengElsevierJournal of Food Protection0362-028X2025-06-0188710055310.1016/j.jfp.2025.100553Determination of Camel Hide Gelatin in Donkey Hide Gelatin Based on Enzymatic Digestion by Ultra-performance Liquid Chromatography-Tandem Mass SpectrometryQiangsheng Xie0Weijian Wang1Tianyang Gao2Feiyan Diao3Wenyue Hu4Xiaoyun Wu5Silong Li6Feng Shi7Liping Gong8Qiyan Li9Guangxi Zhai10Health Food and Cosmetic Department, Shandong Institute for Food and Drug Control, Jinan, Shandong 250101, People's Republic of China; Department of Pharmaceutics School of Pharmaceutical Sciences, Shandong University Jinan 250101, People's Republic of China; Industrial Technology Foundation Public Service Platform, Jinan, Shandong, 250101, People's Republic of ChinaHealth Food and Cosmetic Department, Shandong Institute for Food and Drug Control, Jinan, Shandong 250101, People's Republic of China; Industrial Technology Foundation Public Service Platform, Jinan, Shandong, 250101, People's Republic of ChinaHealth Food and Cosmetic Department, Shandong Institute for Food and Drug Control, Jinan, Shandong 250101, People's Republic of ChinaHealth Food and Cosmetic Department, Shandong Institute for Food and Drug Control, Jinan, Shandong 250101, People's Republic of ChinaHealth Food and Cosmetic Department, Shandong Institute for Food and Drug Control, Jinan, Shandong 250101, People's Republic of ChinaHealth Food and Cosmetic Department, Shandong Institute for Food and Drug Control, Jinan, Shandong 250101, People's Republic of ChinaHealth Food and Cosmetic Department, Shandong Institute for Food and Drug Control, Jinan, Shandong 250101, People's Republic of ChinaHealth Food and Cosmetic Department, Shandong Institute for Food and Drug Control, Jinan, Shandong 250101, People's Republic of ChinaHealth Food and Cosmetic Department, Shandong Institute for Food and Drug Control, Jinan, Shandong 250101, People's Republic of ChinaHealth Food and Cosmetic Department, Shandong Institute for Food and Drug Control, Jinan, Shandong 250101, People's Republic of China; Industrial Technology Foundation Public Service Platform, Jinan, Shandong, 250101, People's Republic of China; Corresponding authors.Department of Pharmaceutics School of Pharmaceutical Sciences, Shandong University Jinan 250101, People's Republic of China; Corresponding authors.The commercial value and diversified applications of donkey hide gelatin (DHG) have precipitated sophisticated adulteration practices, particularly with camel-hide gelatin substitutes. We present a mass spectrometry-based strategy to authenticate species-specific collagen peptides in commercial DHG. A new qualitative and quantitative methodology is constructed for camel hide gelatin(CHG) material analysis in DHG. It consists of enzyme-mediated digestion, ultra-high-performance liquid chromatography (UHPLC), and triple quadrupole mass spectrometry (MS/MS) with multireaction monitoring (MRM) mode. In brief, the samples were digested with trypsin and subjected to efficient and quick separation by UHPLC, and MS/MS performed CHG identification and determination analysis. Two peptides (Camel peptide A, CPA, and Camel peptide B, CPB) were identified as CHG-specific peptides. A specificity test was then used to verify these two peptides. The method performance showed that these two peptides could distinguish CHG from other animal hide gelatins. The CPA and CPB detection limits were 5 μg/kg and 3 μg/kg, respectively. LC-MS/MS analysis detected CPA and CPB in 4.5% of market-collected DHG samples (n = 157), confirming the prevalence of this emerging adulteration practice. Thus, the present protocol is a sensitive, accurate, quick, and suitable application of species-specific peptide biomarkers, ensuring the quality of DHG products and making them authentic and traceable to protect consumers from potential health risks and food frauds.http://www.sciencedirect.com/science/article/pii/S0362028X2500105XCamel hide gelatinEnzymatic hydrolysisLiquid chromatographyMass spectrometryMethod validation
spellingShingle Qiangsheng Xie
Weijian Wang
Tianyang Gao
Feiyan Diao
Wenyue Hu
Xiaoyun Wu
Silong Li
Feng Shi
Liping Gong
Qiyan Li
Guangxi Zhai
Determination of Camel Hide Gelatin in Donkey Hide Gelatin Based on Enzymatic Digestion by Ultra-performance Liquid Chromatography-Tandem Mass Spectrometry
Journal of Food Protection
Camel hide gelatin
Enzymatic hydrolysis
Liquid chromatography
Mass spectrometry
Method validation
title Determination of Camel Hide Gelatin in Donkey Hide Gelatin Based on Enzymatic Digestion by Ultra-performance Liquid Chromatography-Tandem Mass Spectrometry
title_full Determination of Camel Hide Gelatin in Donkey Hide Gelatin Based on Enzymatic Digestion by Ultra-performance Liquid Chromatography-Tandem Mass Spectrometry
title_fullStr Determination of Camel Hide Gelatin in Donkey Hide Gelatin Based on Enzymatic Digestion by Ultra-performance Liquid Chromatography-Tandem Mass Spectrometry
title_full_unstemmed Determination of Camel Hide Gelatin in Donkey Hide Gelatin Based on Enzymatic Digestion by Ultra-performance Liquid Chromatography-Tandem Mass Spectrometry
title_short Determination of Camel Hide Gelatin in Donkey Hide Gelatin Based on Enzymatic Digestion by Ultra-performance Liquid Chromatography-Tandem Mass Spectrometry
title_sort determination of camel hide gelatin in donkey hide gelatin based on enzymatic digestion by ultra performance liquid chromatography tandem mass spectrometry
topic Camel hide gelatin
Enzymatic hydrolysis
Liquid chromatography
Mass spectrometry
Method validation
url http://www.sciencedirect.com/science/article/pii/S0362028X2500105X
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