Background: We hypothesized that specific amino acids or acylcarnitines would have benefits for the differential diagnosis of diabetes. Thus, a targeted metabolomics for amino acids and acylcarnitines in patients with diabetes and its complications was carried out. Methods: A cohort of 54 normal individuals and 156 patients with type 2 diabetes mellitus and/or diabetic complications enrolled from the First Affiliated Hospital of Jinzhou Medical University was studied. The subjects were divided into five main groups: normal individuals, impaired fasting glucose, overt diabetes, diabetic microvascular complications, and diabetic peripheral vascular disease. The technique of tandem mass spectrometry was applied to obtain the plasma metabolite profiles. Metabolomics multivariate statistics were applied for the metabolic data analysis and the differential metabolites determination. Results: A total of 10 cross-comparisons within diabetes and its complications were designed to explore the differential metabolites. The results demonstrated that eight comparisons existed and yielded significant metabolic differences. A total number of 24 differential metabolites were determined from six selected comparisons, including up-regulated amino acids, down-regulated medium-chain and long-chain acylcarnitines. Altered differential metabolites provided six panels of biomarkers, which were helpful in distinguishing diabetic patients. Conclusion: Our results demonstrated that the biomarker panels consisted of specific amino acids and acylcarnitines which could reflect the metabolic variations among the different stages of diabetes and might be useful for the differential diagnosis of prediabetes, overt diabetes and diabetic complications.
To explore metabolism mechanism of paeoniflorin in the liver and further understand intact metabolism process of paeoniflorin, a rapid, convenient and effective assay is described using ultra-performance liquid chromatography coupled with hybrid quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS). The strategy was confirmed in the following primary processes: firstly, different concentration of paeoniflorin, rat liver microsomes, coenzymes and different incubated conditions were optimized to build a biotransformation model of rat liver microsomes in vitro by high performance liquid chromatography with diode array detection (HPLC-DAD); secondly, the metabolites of paeoniflorin in rat liver microsomes were detected and screened using UPLC-Q-TOF-MS/MS by comparing the total ion chromatogram (TIC) of the experimental group with those of control groups; finally, the molecular formulae and corresponding chemical structures of paeoniflorin metabolites were identified by comparing the MS and MS/MS spectra with the self-constructed database and simulation software. Based on this analytical strategy, 20 metabolites of paeoniflorin were found and 6 metabolites (including four new compounds) were tentatively identified. It was shown that hydrolysis and oxidation were the major metabolic pathways of paeoniflorin in rat liver microsomes, and the main metabolic sites were the structures of pinane and the ester bond. These findings were significant for a better understanding of the metabolism of paeoniflorin in rat liver microsomes and the proposed metabolic pathways of paeoniflorin might provide fundamental support for the further research in the pharmacological mechanism of Paeoniae Radix Rubra (PRR).
Fluorescent carbon dots are a promising drug carrier and bio‐sensor material. The study between human serum albumin and carbon dots could explore the possible interaction mechanism. In this paper, melamine was adopted as passivating agent and citric acid as carbon source, high fluorescence nitrogen doped carbon dots (N‐CDs) were synthetized by one‐pot solid phase pyrolysis successfully. The obtained blue emissive N‐CDs with an average particle size of 2.8 ± 0.7 nm displayed an excitation‐dependent photoluminescence behavior, and the fluorescence quantum yield was estimated to be 12.8%. Moreover, the N‐CDs contained abundant hydroxyl, carboxyl, and amine groups. The molecular interaction between as‐prepared N‐CDs and HSA was systematically investigated in vitro through several spectroscopic methods associated with cyclic voltammetry (CV) and molecular docking. The quenching mechanism of HSA by N‐CDs was static through forming the HSA‐N‐CDs complex, and both hydrogen bonding and van der Waals forces were the fundamental driving forces in the spontaneous binding process. The distance between N‐CDs and HSA was 2.2 nm on the basis of Förster's theory. The competitive binding experiment showed that the binding of N‐CDs was primarily in site I of HSA. Through the analysis of circular dichroism (CD), synchronous fluorescence, and three‐dimensional fluorescence spectra, the secondary structure of HSA was altered in the presence of N‐CDs and the polarity around the amino acid residues of HSA was not changed. Besides, molecular simulation was applied to determine the possible binding situation. Our study explored the interaction mechanism of N‐CDs with HSA, which provided some valuable information for understanding the structure and function of proteins as well as the transport and metabolism of N‐CDs.
Anthocyanins are a sort of important natural pigments in red raspberry which have many pharmacological effects. First, the ultrasound-assisted extraction (UAE) of total anthocyanins was optimized by Box–Behnken design (BBD) in this study. The detailed optimized method which has the advantages of high extraction rate and short time was as follows: the ultrasonic extraction temperature was 20 °C, the solvent to sample ratio was 26 mL/g, and the pH value was 3. The total anthocyanin content extracted from thawed red raspberry was 9.178 mg/100 g. Then, the crude extracts obtained from red raspberry fruits with 80% ethanol were isolated with a macroporous resin column (AB-8) and eluted with ethanol in gradient elution. Third, the system of a two-phase solvent, which consisted of n-butanol, methyl tert-butyl ether (MTBE), acetonitrile, water, and trifluoroacetic acid (5:1:1:4:0.001, v/v), was applied for high-speed counter-current chromatography (HSCCC). Finally, three anthocyanin compounds were identified as cyanidin-3-O-sophoroside, cyanidin-3-O-(2G-O-glucosylrutinoside), and cyanidin-3-O-glucoside by UV, MS, and NMR analyses, and their purities were 95.35%, 91.31%, and 99.12%, respectively. This study demonstrated an effective procedure of extraction and purification for anthocyanins from red raspberry fruits, which could provide interesting information for the in-depth study of bioactive compounds of red raspberry fruits.
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