Application Of Metabolomics In Quality Control Of Raw Materials And Products Of Gelatin

RATIONALES:Colla Corii Asini was recorded in "Shen Nong’s Herbal Classic" for the first time. Its blood enriching and blood activating function has long been accepted in the field of medicine. But, the existence of inferior and adulterants products on the Colla Corii Asini market worried customers, so numerous scholars has been committing to finding a simple and exclusive method to achieve the identification of Colla Corii Asini.Our research group studied local quality standards of donkey-hide(the raw material of Colla Corii Asini) and used a biological method to identify donkey-hide and adulterants previously. Based on preliminary results, this study intends to make a research on raw materials, gelatins and compound preparations of Colla Corii Asini.Objective:Raw materials, gelatins and compound preparations of Colla Corii Asini was studied in combination with the newer systems biology approach-’H-NMR metabolomics technology to realize authenticity, quality difference and uniformity study, to find their differential constituents, and to provide a basis for quality control study of raw materials and gelatin preparations of Colla Corii Asini.Methods:Firstly, water-soluble components of raw material and adulterants of Colla Corii Asini, acid-hydrolysis components of Colla Corii Asini of different manufacturers and different batches, and acid-hydrolysis components gelatin compound preparations (granules) of different batches were analyzed by using ’H-NMR techniques. And then multivariate statistical analysis techniques such as PCA, HCA, PLS-DA, OPLS-DA were applied to study the experimental spectra obtained in the first step.Results:(1)’H-NMR metabolomic study results of water-soluble components of raw donkey-hide and adulterants showed that:42 water-soluble components were pointed out in their 1H-NMR spectra. PCA analysis showed that the donkey-hide and horse-hide, mule-hide are quite different. OPLS-DA analysis showed that, compared with donkey-hide, horsehide contains relatively more acetate, leucine, valine, isoleucine and relatively less lactate, serine, pyroglutamate, creatine; while mule-hide contains relatively more lactate, creatine, alanine, choline and less glycerol, acetic acid, and they all have a significant difference (P<0.05).(2)1H-NMR metabolomics study results of acid-hydrolysis components of Colla Corii Asini from different manufacturers showed that:17 components were recognized from their 1H-NMR spectra. Colla Corii Asini of manufacturer A, C, D, E were separately compared to manufacturer B and the corresponding differential components, including isoleucine, hydroxyproline, arginine, leucine, phenylalanine, valine, lysine, levulinate and so on were figured out.(3)1H-NMR metabolomics study results of acid-hydrolysis components of Colla Corii Asini of different batches from one same manufacturer showed that:PCA and PLS-DA analysis results showed no obvious difference between different batches, so they have a good quality uniformity.(4)1H-NMR metabolomics study results of acid-hydrolysis components of compound granular of different batches from one same manufacturer showed that: their PLS-DA analysis showed no obvious difference. Consquently, they also have a good quality uniformity.Conclusions: 1H-NMR metabolomics technology enables fast identification of donkey-hide and counterfeits, and the corresponding differential components can be found at the same time. And 1H-NMR metabolomics technology realizes the quality difference and uniformity study of Colla Corii Asini and its compound granule preparation from different manufacturers and different batches of the same manufacturer.