A "Green" Homogenate Extraction Coupled with UHPLC-MS for the Rapid Determination of Diterpenoids in Croton Crassifolius.

Clerodane diterpenoids are the main bioactive constituents of Croton crassifolius and are proved to have multiple biological activities. However, quality control (QC) research on the constituents are rare. Thus, the major research purpose of the current study was to establish an efficient homogenate extraction (HGE) process combined with a sensitive and specific ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC⁻MS) technique together for the rapid extraction and determination of clerodane diterpenoids in C. crassifolius. All calibration curves showed good linearity (r > 0.9943) within the test ranges and the intra- and inter-day precisions and repeatability were all within required limits. This modified HGE⁻UHPLC⁻MS method only took 5 min to extract nine clerodane diterpenoids in C. crassifolius and another 12 min to quantify these components. The results indicated that the quantitative analysis based on UHPLC⁻MS was a feasible method for QC of clerodane diterpenoids in C. crassifolius, and the findings outlined in the current study also inferred the potential of the method in the QC of clerodane diterpenoids in other complex species of plants.

Synergistic Effects of Combined Anthocyanin and Metformin Treatment for Hyperglycemia In Vitro and In Vivo.

The mechanism underlying the hypoglycemic effect of the simultaneous use of metformin and anthocyanin-rich foods is not yet clear. Hence, the effects and possible mechanisms of action of these substances, alone and in combination, were evaluated in insulin-resistant HepG2 cells and a diabetic mouse model. The results indicated that anthocyanin and metformin had a significant synergistic effect on glucose consumption (CI < 0.9) compared with metformin alone in HepG2 cells. In the mouse model, combined treatment (50 and 100 mg/kg metformin + anthocyanin groups) demonstrated synergistic restorative effects on the blood glucose level, insulin resistance, and organ damage in the liver, pancreas, and ileum. Additionally, combined metformin and anthocyanin treatment suppressed protein tyrosine phosphatase 1B expression and regulated the PI3K/AKT/GSK3ß pathway. Combined treatment also altered the gut microbial composition and structure by increasing the relative abundance of beneficial bacteria and the short-chain fatty acid content. These results suggest that the use of anthocyanins can enhance the efficacy of metformin treatment for hyperglycemia and provide a reference for further clinical research regarding nutrition and supplementary treatment.