Dataset Information

Quantitative 1H Nuclear Magnetic Resonance Method for Assessing the Purity of Dipotassium Glycyrrhizinate.

ABSTRACT: A simple, rapid, accurate, and selective quantitative method based on 1H nuclear magnetic resonance (qNMR) was successfully established and developed for assessing the purity of dipotassium glycyrrhizinate (KG). In this study, using potassium hydrogen phthalate and fumaric acid as internal standard (IS), several important experimental parameters, such as relaxation delay and pulse angle, were explored. Reliability, specificity, linearity, limit of quantification, precision, stability, and accuracy were also validated. Calibration results obtained from qNMR were consistent with those obtained from HPLC coupled with ultraviolet detection. The proposed method, independent of the reference standard substance, is a useful, reliable, and practical protocol for the determination of KG and glycyrrhizin analogs.

SUBMITTER: Zhang YY

PROVIDER: S-EPMC8230393 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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Quantitative <sup>1</sup>H Nuclear Magnetic Resonance Method for Assessing the Purity of Dipotassium Glycyrrhizinate.

Zhang Yuan-Yuan YY Zhang Jie J Zhang Wen-Xuan WX Wang Yue Y Wang Ying-Hong YH Yang Qing-Yun QY Wu Song S

Molecules (Basel, Switzerland) 20210610 12

A simple, rapid, accurate, and selective quantitative method based on <sup>1</sup>H nuclear magnetic resonance (qNMR) was successfully established and developed for assessing the purity of dipotassium glycyrrhizinate (KG). In this study, using potassium hydrogen phthalate and fumaric acid as internal standard (IS), several important experimental parameters, such as relaxation delay and pulse angle, were explored. Reliability, specificity, linearity, limit of quantification, precision, stability, ...[more]

PMID: 34200734

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Project description:Glycyrrhizic acid (GA), a natural compound isolated from licorice (Glycyrrhiza glabra), has exhibited anti-inflammatory and anti-tumor effects in vitro. Dipotassium glycyrrhizinate (DPG), a dipotassium salt of GA, also has shown an anti-tumor effect on glioblastoma cell lines, U87MG and T98G. The study investigated the DPG effects in the melanoma cell line (SK-MEL-28). MTT assay demonstrated that the viability of the cells was significantly decreased in a time- and dose-dependent manner after DPG (IC50 = 36 mM; 24 h). DNA fragmentation suggested that DPG (IC50) induced cellular apoptosis, which was confirmed by a significant number of TUNEL-positive cells (p-value = 0.048) and by PARP-1 [0.55 vs. 1.02 arbitrary units (AUs), p-value = 0.001], BAX (1.91 vs. 1.05 AUs, p-value = 0.09), and BCL-2 (0.51 vs. 1.07 AUs, p-value = 0.0018) mRNA compared to control cells. The proliferation and wound-healing assays showed an anti-proliferative effect on DPG-IC50-treated cells, also indicating an inhibitory effect on cell migration (p-values < 0.001). Moreover, it was observed that DPG promoted a 100% reduction in melanospheres formation (p-value = 0.008). Our previous microRNAs (miRs) global analysis has revealed that DPG might increase miR-4443 and miR-3620 expression levels. Thus, qPCR showed that after DPG treatment, SK-MEL-28 cells presented significantly high miR-4443 (1.77 vs. 1.04 AUs, p-value = 0.02) and miR-3620 (2.30 vs. 1.00 AUs, p-value = 0.01) expression compared to control cells, which are predicted to target the NF-kB, CD209 and TNC genes, respectively. Both genes are responsible for cell attachment and migration, and qPCR revealed significantly decreased CD209 (1.01 vs. 0.54 AUs, p-value = 0.018) and TNC (1.00 vs. 0.31 AUs, p-value = 2.38 × 10-6) mRNA expression levels after DPG compared to untreated cells. Furthermore, the migration of SK-MEL-28 cells stimulated by 12-O-tetradecanoylphorbol-13-acetate (TPA) was attenuated by adding DPG by wound-healing assay (48 h: p-value = 0.004; 72 h: p-value = 7.0 × 10-4). In addition, the MMP-9 expression level was inhibited by DPG in melanoma cells stimulated by TPA and compared to TPA-treated cells (3.56 vs. 0.99 AUs, p-value = 0.0016) after 24 h of treatment. Our results suggested that DPG has an apoptotic, anti-proliferative, and anti-migratory effect on SK-MEL-28 cells. DPG was also able to inhibit cancer stem-like cells that may cause cerebral tumor formation.

Project description:UnlabelledDengue, due to its global burden, is the most important arthropod-borne flavivirus disease, and early detection lowers fatality rates to below 1%. Since the metabolic resources crucial for viral replication are provided by host cells, detection of changes in the metabolic profile associated with disease pathogenesis could help with the identification of markers of prognostic and diagnostic importance. We applied (1)H nuclear magnetic resonance exploratory metabolomics to study longitudinal changes in plasma metabolites in a cohort in Recife, Brazil. To gain statistical power, we used innovative paired multivariate analyses to discriminate individuals with primary and secondary infection presenting as dengue fever (DF; mild) and dengue hemorrhagic fever (DHF; severe) and subjects with a nonspecific nondengue (ND) illness (ND subjects). Our results showed that a decrease in plasma low-density lipoprotein (LDL) and very-low-density lipoprotein (VLDL) discriminated dengue virus (DENV)-infected subjects from ND subjects, and also, subjects with severe infection even presented a decrease in lipoprotein concentrations compared to the concentrations in subjects with mild infection. These results add to the ongoing discussion that the manipulation of lipid metabolism is crucial for DENV replication and infection. In addition, a decrease in plasma glutamine content was characteristic of DENV infection and disease severity, and an increase in plasma acetate levels discriminated subjects with DF and DHF from ND subjects. Several other metabolites shown to be altered in DENV infection and the implications of these alterations are discussed. We hypothesize that these changes in the plasma metabolome are suggestive of liver dysfunction, could provide insights into the underlying molecular mechanisms of dengue virus pathogenesis, and could help to discriminate individuals at risk of the development of severe infection and predict disease outcome.ImportanceDengue, due to its global burden, is the most important mosquito-borne viral disease. There is no specific treatment for dengue disease, and early detection lowers fatality rates to below 1%. In this study, we observed the effects of dengue virus infection on the profile of small molecules in the blood of patients with mild and severe infection. Variations in the profiles of these small molecules reflected the replication of dengue virus in different tissues and the extent of tissue damage during infection. The results of this study showed that the molecules that changed the most were VLDL, LDL, and amino acids. We propose that these changes reflect liver dysfunction and also that they can be used to discriminate subjects with mild dengue from those with severe dengue.

Project description:BACKGROUND:Objective and reliable methods to measure dietary exposure and prove associations and causation between diet and health are desirable. OBJECTIVE:The aim of this study was to investigate if 1H-nuclear magnetic resonance (1H-NMR) analysis of serum samples may be used as an objective method to discriminate vegan, vegetarian, and omnivore diets. Specifically, the aim was to identify a metabolite pattern that separated meat-eaters from non-meat-eaters and vegans from nonvegans. METHODS:Healthy volunteers (45 men and 75 women) complying with habitual vegan (n = 43), vegetarian (n = 24 + vegetarians adding fish n = 13), or omnivore (n = 40) diets were enrolled in the study. Data were collected on clinical phenotype, body composition, lifestyle including a food-frequency questionnaire (FFQ), and a 4-d weighed food diary. Serum samples were analyzed by routine clinical test and for metabolites by 1H-NMR spectroscopy. NMR data were nonnormalized, UV-scaled, and analyzed with multivariate data analysis [principal component analysis, orthogonal projections to latent structures (OPLS) and OPLS with discriminant analysis]. In the multivariate analysis volunteers were assigned as meat-eaters (omnivores), non-meat-eaters (vegans and vegetarians), vegans, or nonvegans (lacto-ovo-vegetarians, vegetarians adding fish, and omnivores). Metabolites were identified by line-fitting of 1D 1H-NMR spectra and the use of statistical total correlation spectroscopy. RESULTS:Although many metabolites differ in concentration between men and women as well as by age, body mass index, and body composition, it was possible to correctly classify 97.5% of the meat-eaters compared with non-meat-eaters and 92.5% of the vegans compared with nonvegans. The branched-chain amino acids, creatine, lysine, 2-aminobutyrate, glutamine, glycine, trimethylamine, and 1 unidentified metabolite were among the most important metabolites in the discriminating patterns in relation to intake of both meat and other animal products. CONCLUSIONS:1H-NMR serum metabolomics appears to be a possible objective tool to identify and predict habitual intake of meat and other animal products in healthy subjects. These results should be confirmed in larger cohort studies or intervention trials. This trial was registered at clinicaltrials.gov as NCT02039609.

Dataset Information

Quantitative 1H Nuclear Magnetic Resonance Method for Assessing the Purity of Dipotassium Glycyrrhizinate.

Publications

Quantitative <sup>1</sup>H Nuclear Magnetic Resonance Method for Assessing the Purity of Dipotassium Glycyrrhizinate.

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