BackgroundCommon farming environmental elements, such as longitude, latitude, and altitude, and physiological conditions, such as age and body weight, are thought to influence medicinal animal homeostasis and material quality by altering endocrine functions for primary and secondary metabolite formation. However, the currently available methods for evaluating complex components of traditional Chinese animal medicines have insufficient sensitivity and specificity. PurposeCharacterizing the primary/secondary metabolomes of medicinal animals is essential for understanding their material basis, controlling product quality, and reflecting on distribution interactions. Therefore, this study aimed to screen ecological- and physiological-related metabolites in captive Moschus berezovskii throughout the collection period based on the quality marker (Q-marker) concept. Study design and methodsFifty-one musk deer samples from 12 different distribution farms ranging in age from 2 to 11 years were enrolled. Differentially expressed musk metabolites were assessed via chromatography-tandem mass spectrometry technologies. A metabolome that mapped connections among these factors was established using chemometric and topological calculations. ResultsStatistical analysis revealed that muscone, cis-9-hexadecenal, antioxidant 2264, prasterone-3-sulfate, androstan-17-one, and 1,2-benzenedicarboxylic acid showed significantly altered expression. Partial least squares (PLS) regression analysis of qualified data for these 6 secondary metabolites (active components) demonstrated that age is the most important factor underlying the varying levels of muscone, androstan-17-one and 1,2-benzenedicarboxylic acid. Furthermore, weight was the most important factor for cis-9-hexadecenal, longitude was important for antioxidant 2264, latitude was important for prasterone-3-sulfate, and altitude was important for antioxidant 2264, androstan-17-one and 1,2-benzenedicarboxylic. Metabolite analysis within the MetaboAnalyst (MetPA) suite showed that 18 candidate biomarker metabolites were screened, including allantoin, glycine, serine, creatine, alanine, taurine, lactate, 2-oxoglutarate (2-OG), fumarate, proline, xanthine, cytosine, carnitine, arginine, threonine, aspartate, and urea. Metabolic network analysis showed 4 important pathways that were involved: arginine and proline metabolism, the urea cycle, aspartate metabolism, and glycine, serine and threonine metabolism. ConclusionUsing this combined metabolomic and chemometric approach, this study was successful in screening Q-markers for musk quality control and provided new insights into correlations among “ecological & physiological factors→Q-markers→metabolites”, which potentially provides crucial information for musk breeding and material quality control.
Read full abstract