Comprehensive metabolomic variations of hawthorn before and after insect infestation based on the combination analysis of (1)H NMR and UPLC-MS

Hawthorn, the sliced and dried ripe fruits of Crataegus pinnatifida Bge. Var. Major N. E. Br. (Rosaceae), is an edible and medicinal substance with a variety of health-promoting benefits. Hawthorn needs to be stored in warehouses after harvesting to meet people's perennial demand. However, it is easily infested by insects of Plodia interpunctella and Tribolium castaneum during storage, which inevitably leads to poor quality and causes adverse effects on people's health. So far, there has been no report on insect-infested hawthorn. In this study, we analyzed the changes of metabolites in hawthorn before and after insect infestation and screened out potential biomarkers to effectively and quickly detect the occurrence of insect infestation. A combination analysis of (1)H nuclear magnetic resonance (NMR) and ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) was used to identify the primary and secondary metabolites. By the comparison of hawthorn and insect-infested hawthorn samples, it was found that the differences were mainly manifested in the content of metabolites. The metabolites of 32 and 1463 were identified by (1)H NMR and UPLC-MS analysis, respectively. According to the parameters of VIP >1 and P < 0.05, 10 differential metabolites were screened from (1)H NMR analysis. Based on the parameters of VIP >1.0, P < 0.05, and (FC) > 1 or < 1, 47 differential metabolites were screened from UPLC-MS analysis. Therefore, a total of 57 differential metabolites were considered as differential biomarkers. The heat map analysis showed that the content of some differential biomarkers with significant pharmacological activities decreased after insect infestation. Through receiver operating characteristic (ROC) curve assessment, 52 differential biomarkers (6 of (1)H NMR analysis and 46 of UPLC-MS analysis) were screened to distinguish whether insect infestation occurred in hawthorn. This is the first report on the changes of metabolites between hawthorn and insect-infested hawthorn and on the screening of differential biomarkers for monitoring insects. These results contributed to evaluate quality of hawthorn and ensure food safety for consumers. It also laid a foundation for further research on the infestation mechanism and safe storage monitoring in hawthorn.