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A novel method has been established for the rapid separation and determination of free fatty acids from 37 different varieties of raspberry. In this study, a new fluorescent labeling reagent for fatty acids, 2-(4-amino)-phenyl-1-hydrogen-phenanthrene [9, 10-d] imidazole (PIA), has been synthesized and successfully applied to the high-performance liquid chromatography (HPLC) determination of fatty acids in raspberry. The novel method has been optimized by HPLC with fluorescence detection and online mass spectrometry identification (HPLC-FLD-MS/MS). The 22 main fatty acids (FAs) present in raspberry were derivatized by PIA and separated on a reversed-phase Hypersil GOLD column with gradient elution. The main experimental parameters affecting extraction efficiency and derivatization yield were investigated and optimized by response surface methodology (RSM) combined with Box-Behnken design (BBD). Under the optimum conditions, the method was successfully applied for the analysis of 22 fatty acids in 37 different varieties of raspberry. Good linear correlations were observed for all fatty acids with correlation coefficients of > 0.9978. Limits of detection and quantification (LOD and LOQ) were in the range of 0.12 to 0.49 ng/mL and 1.07 to 2.81 ng/mL, respectively. Furthermore, the results indicated that the raspberries were rich in fatty acids, but the contents of the fatty acids varied among the different varieties.

A method that involved the combination of pH-zone-refining counter-current chromatography and semipreparative reversed-phase liquid chromatography has been established for the preparative separation of alkaloids from Hypecoum leptocarpum. From 1.2 g of crude sample, 31 mg N-feruloyltyramine, 27 mg oxohydrastinine, 47 mg hydroprotopine, 25 mg leptopidine, and 18 mg hypecocarpine have been obtained. The structure of the new compound, hypecocarpine, is confirmed based on the analysis of spectroscopic data, including NMR, UV, and IR spectroscopy and positive electrospray ionization mass spectrometry. The known chemical structures were characterized on the basis of (1) H and (13) C NMR spectroscopy. The purities of the five alkaloids are all over 92.7% as determined by high-performance liquid chromatography. The alkaloids' cytotoxicity in breast cancer cells is assessed by using a Cell Counting Kit assay and their inhibitory effect on fatty acid synthase expression is assessed by a Western blot assay. These results suggest that leptopidine could suppress growth and induce cytotoxicity in breast cancer cells and that the cytotoxicity of leptopidine may be related to its inhibitory effect on fatty acid synthase expression.

Fenugreek is a well known annual herb widely used in both medicine and food. Four flavonoid glycosides have been separated from fenugreek seeds in our previous study. In this study, the effects of the four flavonoid glycosides on regulating glycolipid metabolism and improving mitochondrial function were investigated. Isoorientin showed a very significant activity among these flavonoid glycosides. First, isoorientin decreased the accumulation of lipid droplets in 3T3-L1 preadipocytes by reducing the expression of adipokines including PPARγ, C/EBPα, and FAS. Second, isoorientin restored insulin-stimulated glucose uptake in dexamethasone-induced insulin-resistant 3T3-L1 adipocytes by reactivating Akt and AMPK. Finally, isoorientin improved mitochondrial dysfunction induced by dexamethasone in 3T3-L1 adipocytes. Isoorientin also reversed dexamethasone-induced decrease in mitochondrial membrane potential (MMP) and intracellular ATP production, reduced accumulation of intracellular reactive oxygen species (ROS), and protected mitochondrial DNA (mtDNA) from oxidative damage. At the same time, mitochondrial biogenesis is promoted. Therefore, isoorientin may be an attractive candidate as a glucose-lowering and insulin-resistance-improving agent for the treatment of diabetes.

<i>Potentilla parvifolia </i>Fisch. (Rosaceae) is a traditional medicinal plant in P. R. China. In this study, seven flavonoids, ayanin (<b>1</b>), tricin (<b>2</b>), quercetin (<b>3</b>), tiliroside (<b>4</b>), miquelianin (<b>5</b>), isoquercitrin (<b>6</b>), and astragalin (<b>7</b>), were separated and purified from ethyl acetate extractive fractions from ethanol extracts of <i>P. parvifolia</i> using a combination of sevaral chromatographic methods. The human neuroblastoma SH-SY5Y cells were differentiated with all trans-retinoic acid and treated with okadaic acid to induce tau protein phosphorylation and synaptic atrophy, which could establish an Alzheimer's disease cell model. The neuroprotective effects of these flavonoids in cellular were evaluated <i>in vitro</i> by this cell model. Results from the Western blot and morphology analysis suggested that compounds <b>3</b> and <b>4</b> had the better neuroprotective effects.

Obesity, a major health problem worldwide, is a complex multifactorial chronic disease that increases the risk for insulin resistance, type 2 diabetes, coronary heart disease, and hypertension. In this study, we assessed methods to isolate hypaphorine, a potent drug candidate for obesity and insulin resistance. Semi-preparative reversed-phase liquid chromatography (semi-preparative RPLC) was established as a method to separate three compounds, adenosine, l-tryptophan, and hypaphorine, from the crude extracts of <i>Caragana korshinskii </i>Kom. Due to its specific chemical structure, the effect of hypaphorine on differentiation and dexamethasone (DXM) induced insulin resistance of 3T3-L1 cells was investigated. The structures of the three compounds were confirmed by UV, ¹H-NMR, and <sup>13</sup>C-NMR analysis and compared with published data. The activity results indicated that hypaphorine prevented the differentiation of 3T3-L1 preadipocytes into adipocytes by down-regulating hormone-stimulated protein expression of peroxisome proliferator activated receptor <i>γ</i> (PPAR<i>γ</i>) and CCAAT/enhancer binding protein (C/EBP<i>α</i>), and their downstream targets, sterol regulatory element binding protein 1 c (SREBP1c) and fatty acid synthase (FAS). Hypaphorine also alleviated DXM-induced insulin resistance in differentiated 3T3-L1 adipocytes <i>via</i> increasing the phosphorylation level of Akt2, a key protein in the insulin signaling pathway. Taken together, we suggest that the method can be applied to large-scale extraction and large-quantity preparation of hypaphorine for treatment of obesity and insulin resistance.

Fenugreek (Trigonella foenum-graecum L.) is a well-known annual plant that is widely distributed worldwide and has possessed obvious hypoglycemic and hypercholesterolemia characteristics. In our previous study, three polyphenol stilbenes were separated from fenugreek seeds. Here, we investigated the effect of polyphenol stilbenes on adipogenesis and insulin resistance in 3T3-L1 adipocytes. Oil Red O staining and triglyceride assays showed that polyphenol stilbenes differently reduced lipid accumulation by suppressing the expression of adipocyte-specific proteins. In addition, polyphenol stilbenes improved the uptake of 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG) by promoting the phosphorylation of protein kinase B (AKT) and AMP-activated protein kinase (AMPK). In present studies, it was found that polyphenol stilbenes had the ability to scavenge reactive oxygen species (ROS). Results from adenosine triphosphate (ATP) production and mitochondrial membrane potentials suggested that mitochondria play a critical role in insulin resistance and related signaling activation, such as AKT and AMPK. Rhaponticin, one of the stilbenes from fenugreek, had the strongest activity among the three compounds in vitro. Future studies will focus on mitochondrial biogenesis and function.

A method of using high-speed counter-current chromatography (HSCCC) and semi-preparative reversed-phase liquid chromatography (semi-preparative RPLC) to preparatively separate flavone glucosides and lignan from the crude extracts of <i>Caragana korshinskii</i> has been established for the first time in this study. Five flavone glucosides, including 9 mg of kaempferol 3-O-{β-<i>d</i>-glucopyranosyl(1→2)-[α-<i>l</i>-rhamonopyranosyl(1 → 6)]-β-<i>d</i>-galactopyranoside}, 21 mg of kaempferol 3-O-α-<i>l</i>-rhamnopyranosyl(1→6)-β-<i>d</i>-galactopyranoside-7-O-α-<i>l</i>-rhamnopyranoside, 34 mg of kaempferol 3-O-β-<i>d</i>-galactopyranoside-7-O-α-<i>l</i>-rhamnopyranoside, 27 mg of kaempferol 3-O-β-<i>d</i>-glucopyranosyl-7-O-α-<i>l</i>-rhamnopyranoside, and 14 mg of calycosin 7-O-β-<i>d</i>-glucopyranoside, and one lignan, 16 mg of alangilignoside B, were successfully isolated from 1.8 g of the crude sample through the combination of HSCCC with a two-phase solvent system composed of ethyl acetate-<i>n</i>-butanol-0.5 % formic acid (4:1:5, <i>v/v/v</i>) and semi-preparative RPLC with a mobile phase of methanol-water (20:80, <i>v/v</i>). The purities of the six compounds are all over 95 % as determined by HPLC and the structures are confirmed by the analysis of UV, <sup>1</sup>H-NMR, and <sup>13</sup>C-NMR and compared with published data.

AIM: The objective of this study was to evaluate the effects of seed oil of Caragana korshinskii Kom. against Trichophyton mentagrophytes on an in vivo guinea pig model of dermatophytosis.METHODS: The skin of albino guinea pigs was infected with T. mentagrophytes, and the animals were divided into five groups: negative control (NC group), positive control (PC group), vehicle control, CK50% group (received topical 50% seed oil of C.korshinskii), and CK100% group (received topical 100% seed oil of C.korshinskii). Evaluation of clinical efficacy was performed 72 h after the completion of a 10-day treatment regimen. Skin biopsy samples were processed for histopathological examination. RESULTS: The infected untreated control guinea pigs showed patches of hair loss and ulcerated or scaly skin. Lower clinical scores indicate improved efficacy compared with NC. The lesion scores significantly declined in the CK50%, CK100%, and PC groups in comparison with the NC group. The CK50% group (45.31%) and the CK100% group (75%) showed clinical efficacy compared with the PC group (78.13%). In addition, no fungal elements, inflammation, or tissue destruction was observed in any of the PAS-stained sections of the infected skin in the groups treated with CK100% or 1% terbinafine. CONCLUSION: Seed oil of C.korshinskii demonstrated high antifungal efficacy in experimental dermatophytosis.