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PloS one2022; 17(8); e0273021; doi: 10.1371/journal.pone.0273021

Species-specific identification of donkey-hide gelatin and its adulterants using marker peptides.

Abstract: Donkey-hide gelatin is an important traditional Chinese medicine made from donkey skin. Despite decades of effort, identifying the animal materials (donkeys, horses, cattle and pigs) in donkey-hide gelatin remains challenging. In our study, we aimed to identify marker peptides of donkey-hide gelatin and its adulterants and develop a liquid chromatography-tandem mass spectrometry method to identify them. Theoretical marker peptides of four animals (donkeys, horses, cattle and pigs) were predicted and verified by proteomic experiments, and 12 species-specific marker peptides from donkey-hide gelatin and its adulterants were identified. One marker peptide for each gelatin was selected to develop the liquid chromatography-tandem mass spectrometry method. The applicability of the method was evaluated by investigating homemade mixed gelatin samples and commercial donkey-hide gelatin products. Using the liquid chromatography-tandem mass spectrometry method, the addition of cattle-hide gelatin and pig-hide gelatin to donkey-hide gelatin could be detected at a level of 0.1%. Horse-hide gelatin was detected when added at a level of 0.5%. Among 18 batches of donkey-hide gelatin products, nine were identified as authentic, and eight of the remaining samples were suspected to be adulterated with horse materials. These results provide both a practical method to control the quality of donkey-hide gelatin and a good reference for quality evaluations of other medicinal materials and foods containing protein components.
Publication Date: 2022-08-12 PubMed ID: 35960756PubMed Central: PMC9374224DOI: 10.1371/journal.pone.0273021Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

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The research aims to develop a method to accurately identify the animal sources used in the manufacture of donkey-hide gelatin, a popular traditional Chinese medicine. It does this by using marker peptides and a refined liquid chromatography-tandem mass spectrometry method to detect the presence of donkey, horse, cattle, and pig materials.

Use of Marker Peptides

  • The researchers first identified marker peptides pertinent to four different animals – donkeys, horses, cattle, and pigs. These peptides were then used to distinguish between the different types of gelatin.
  • Marker peptides are unique molecular signatures found within proteins that can be used to differentiate between different species.
  • The researchers verified the identified marker peptides through proteomic experiments, which involve analyzing proteins to understand their structures and functions.

Liquid Chromatography-Tandem Mass Spectrometry Method

  • After the successful identification of marker peptides, the researchers chose one marker peptide for each type of gelatin to develop the liquid chromatography-tandem mass spectrometry method.
  • This method allows the separation and identification of these marker peptides in a sample, effectively determining the source of the gelatin.
  • Liquid Chromatography-Tandem Mass Spectrometry (LC-MSMS) is a technique often used in biochemistry to analyze the mass and composition of individual molecules in a mixture. It is a highly precise tool and can detect even trace levels of certain compounds.

Method Validation

  • The researchers tested the applicability of their method by examining homemade and commercial donkey-hide gelatin samples.
  • This LC-MSMS method could detect even small percentages (0.1%) of cattle-hide and pig-hide gelatins when added to donkey-hide gelatin. Horse-hide gelatin was detectable when added at a slightly higher level of 0.5%.
  • Among the 18 batches of commercial donkey-hide gelatin tested, the researchers found that half were authentic, while the others were suspected to be adulterated with horse material.

Implications of the Study

  • This research provides a reliable method to control the quality of donkey-hide gelatin and ensures that products on the market are authentic.
  • Further, this methodology could also serve as a model for evaluating the quality of other medicinal materials and food items that contain protein components.

Cite This Article

APA
Zhang J, Wu M, Ma Z, Zhang Y, Cao H. (2022). Species-specific identification of donkey-hide gelatin and its adulterants using marker peptides. PLoS One, 17(8), e0273021. https://doi.org/10.1371/journal.pone.0273021

Publication

ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 17
Issue: 8
Pages: e0273021
PII: e0273021

Researcher Affiliations

Zhang, Jinju
  • College of Pharmacy, Research Center for Traditional Chinese Medicine of Lingnan (Southern China), National Engineering Research Center for Modernization of Traditional Chinese Medicine Lingnan Resources Branch, Jinan University, Guangzhou, Guangdong, China.
  • Guangdong Provincial Key Laboratory of Traditional Chinese Medicine Informatization (2021B1212040007), Guangzhou, Guangdong, China.
Wu, Menghua
  • College of Pharmacy, Research Center for Traditional Chinese Medicine of Lingnan (Southern China), National Engineering Research Center for Modernization of Traditional Chinese Medicine Lingnan Resources Branch, Jinan University, Guangzhou, Guangdong, China.
  • Guangdong Provincial Key Laboratory of Traditional Chinese Medicine Informatization (2021B1212040007), Guangzhou, Guangdong, China.
Ma, Zhiguo
  • College of Pharmacy, Research Center for Traditional Chinese Medicine of Lingnan (Southern China), National Engineering Research Center for Modernization of Traditional Chinese Medicine Lingnan Resources Branch, Jinan University, Guangzhou, Guangdong, China.
  • Guangdong Provincial Key Laboratory of Traditional Chinese Medicine Informatization (2021B1212040007), Guangzhou, Guangdong, China.
Zhang, Ying
  • College of Pharmacy, Research Center for Traditional Chinese Medicine of Lingnan (Southern China), National Engineering Research Center for Modernization of Traditional Chinese Medicine Lingnan Resources Branch, Jinan University, Guangzhou, Guangdong, China.
  • Guangdong Provincial Key Laboratory of Traditional Chinese Medicine Informatization (2021B1212040007), Guangzhou, Guangdong, China.
Cao, Hui
  • College of Pharmacy, Research Center for Traditional Chinese Medicine of Lingnan (Southern China), National Engineering Research Center for Modernization of Traditional Chinese Medicine Lingnan Resources Branch, Jinan University, Guangzhou, Guangdong, China.
  • Guangdong Provincial Key Laboratory of Traditional Chinese Medicine Informatization (2021B1212040007), Guangzhou, Guangdong, China.

MeSH Terms

  • Animals
  • Biomarkers / analysis
  • Cattle
  • Chromatography, Liquid / methods
  • Equidae
  • Gelatin / chemistry
  • Horses
  • Peptides / chemistry
  • Proteomics
  • Swine

Conflict of Interest Statement

The authors have declared that no competing interests exist.

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