Qualitative and Quantitative Analysis of Ejiao-Related Animal Gelatins through Peptide Markers Using LC-QTOF-MS/MS and Scheduled Multiple Reaction Monitoring (MRM) by LC-QQQ-MS/MS

Abstract

Donkey-hide gelatin, also called Ejiao (colla corii asini), is commonly used as a food health supplement and valuable Chinese medicine. Its growing popular demand and short supply make it a target for fraud, and many other animal gelatins can be found as adulterants. Authentication remains a quality concern. Peptide markers were developed by searching the protein database. However, donkeys and horses share the same database, and there is no specific marker for donkeys. Here, solutions are sought following a database-independent strategy. The peptide profiles of authentic samples of different animal gelatins were compared using LC-QTOF-MS/MS. Fourteen specific markers, including four donkey-specific, one horse-specific, three cattle-specific, and six pig-specific peptides, were successfully found. As these donkey-specific peptides are not included in the current proteomics database, their sequences were determined by de novo sequencing. A quantitative LC-QQQ multiple reaction monitoring (MRM) method was further developed to achieve highly sensitive and selective analysis. The specificity and applicability of these markers were confirmed by testing multiple authentic samples and 110 batches of commercial Ejiao products, 57 of which were found to be unqualified. These results suggest that these markers are specific and accurate for authentication purposes.

Effects of E’Jiao on Skeletal Mineralisation, Osteocyte and WNT Signalling Inhibitors in Ovariectomised Rats

“…E’Jiao, also known as Colla corii asini, is a traditional Chinese medicine derived from donkey-hide gelatine [ 20 , 21 , 22 ]. Modern pharmacology studies demonstrated that E’Jiao could significantly increase haematopoietic stem cells, burst-forming unit-erythroid and colony-forming unit granulocyte-macrophages in the bone marrow.…”

A review on proteomic and genomic biomarkers for gelatin source authentication: Challenges and future outlook

“…The dietary laws and requirements followed by Muslims (Halal food) and Jews (Kosher food) need to ensure that food products are free from porcine products and by-products, while Hindu customers demand food products of no bovine origin [ 3 , 11 , 12 ]. On the other hand, gelatin is also utilized as a supplement or traditional medicine, such as donkey-hide gelatin, a traditional Chinese medicine manufactured from donkey skin [ 13 , 14 ].…”