High-speed counter-current chromatography (HSCCC) was successfully applied to the isolation and purification of four xanthone glycosides from Halenia elliptica, a plant widely used in traditional Tibetan medicine. The introduction of HSCCC greatly improved the efficiency of compounds preparation from Halenia elliptica. The following were obtained from 100 mg of crude sample in one-step separation: 2.5 mg of 1-O-primeverosyl-2,3,4,5,7-pentamethoxyxanthone, 7.0 mg of 1-O-primeverosyl-2,3,4,7- tetramethoxyxanthone, 10.0 mg of 1-O-primeverosyl-2,3,5-trimethoxyxanthone (demethyoxyhaleniaside), and 8.5 mg of 1-O-primeverosyl-2,3,4,5-tetramethoxyxanthone. HPLC analysis showed that each target compound had a purity of over 98%, and UV, 1H NMR, and 13C NMR data confirmed the component chemical structures.
Oxyfadichalcones A and B, two unprecedented chalcone dimers fused through a cyclobutane ring by head-to-tail [2+2] cycloaddition of two chalcones that had never been found previously in nature, along with oxyfadichalcone C, a new head-to-head [2+2] cyclized chalcone dimer, were simultaneously obtained from Oxytropis falcata. Structural elucidation was succeeded by spectroscopic and single-crystal synchrotron radiation analysis. Additionally, the photosynthesis of the chalcone dimers was performed and the plausible biosynthesis was discussed. (C) 2013 Elsevier Ltd. All rights reserved.
The current study focused on the pharmacodynamic activity components of Gentianopsis paludosa against ulcerative colitis (UC) fibrosis including symptoms of intestinal diarrhea and inflammatory. Trinitro-benzene-sulfonic acid induced UC model rats were gavaged with gradient polarity extracts respectively from ethanol-extract of Gentianopsis paludosa. Masson staining and qRT-PCR methods were respectively used to assess the degree of UC fibrosis and detect the mRNA expressions of collagen I, collagen III, a-smooth muscle actin (α-SMA) and E-cadherin in colon tissue. Separated by silica gel column chromatography, further screening was conducted until active components appeared. Infrared, nuclear magnetic resonance, mass spectroscopy and ultraviolet methods were applied to confirm active components' structures. The results indicated that the expression of collagen I, collagen III and α-SMA mRNA in the colon tissues of acetidin group rats was obviously depressed compared with control groups while E-cadherin displayed just opposite. Dyed in blue indicating UC fibrosis degree, the area of acetidin group was less than that other experimental groups. Four components: (1,8-Dihydroxy-3,7-Dimethoxyxanthones, 1-hydroxy-3,7,8-Trimethoxyxanthones, 1,7-Dihydroxy-3,8-Dimethoxyxanthones and 1-hydroxy-3,7-Dimethoxyxanthones), were obtained from acetidin group and all of which have a significant equivalence to Gentianopsis paludosa on the therapeutic effect of UC fibrosis. Our findings revealed the activity components for clinical application history of Gentianopsis paludosa and provided a preliminary foundation for further new drug research and exploitation.
The current study focused on the pharmacodynamic activity components of Gentianopsis paludosa against ulcerative colitis (UC) fibrosis including symptoms of intestinal diarrhea and inflammatory. Trinitro-benzene-sulfonic acid induced UC model rats were gavaged with gradient polarity extracts respectively from ethanol-extract of Gentianopsis paludosa. Masson staining and qRT-PCR methods were respectively used to assess the degree of UC fibrosis and detect the mRNA expressions of collagen I, collagen III, a-smooth muscle actin (α-SMA) and E-cadherin in colon tissue. Separated by silica gel column chromatography, further screening was conducted until active components appeared. Infrared, nuclear magnetic resonance, mass spectroscopy and ultraviolet methods were applied to confirm active components' structures. The results indicated that the expression of collagen I, collagen III and α-SMA mRNA in the colon tissues of acetidin group rats was obviously depressed compared with control groups while E-cadherin displayed just opposite. Dyed in blue indicating UC fibrosis degree, the area of acetidin group was less than that other experimental groups. Four components: (1,8-Dihydroxy-3,7-Dimethoxyxanthones, 1-hydroxy-3,7,8-Trimethoxyxanthones, 1,7-Dihydroxy-3,8-Dimethoxyxanthones and 1-hydroxy-3,7-Dimethoxyxanthones), were obtained from acetidin group and all of which have a significant equivalence to Gentianopsis paludosa on the therapeutic effect of UC fibrosis. Our findings revealed the activity components for clinical application history of Gentianopsis paludosa and provided a preliminary foundation for further new drug research and exploitation.
Although the rhizomes of Rheum nobile Hook. f. et Thoms (Polygonaceae) are widely used in Tibetan medicine, no previous investigations regarding the biological activities and rarely chemical constituents of this plant have been reported. As part of an ongoing search for novel bioactive agents, a phytochemical investigation of R. nobile led to the isolation of two new compounds Rheumone B (1) and piceatannol-4'-O-β-D-(6″-O-acetyl)-glucoside (2), together with 15 known compounds by gel filtration over Sephadex LH-20 and preparative HPLC. Their structures were determined by combined spectroscopic methods. Compounds 1-10 were evaluated for their ability to scavenge 2,2-diphenyl-1-picrylhydzyl (DPPH) radical and compounds 7-10 showed relatively strong scavenging abilities with IC50 values from 2.76 μM to 11.80 μM. In conclusion, naphthalene glycosides, stilbene glycosides, flavanols, especially anthraquinones are main chemical constituents of this plant. The ability to scavenge DPPH radical of compound 8 was the highest among compounds 1-10.
Ethnopharmacological relevanceFrankincense oil and water extracts (FOE, FWE) have long been used for external treatment of inflammation and pain. The present study was conducted to identify the active ingredients responsible for the anti-inflammatory and analgesic effects and to determine the underlying mechanisms.
Materials and methods
The compositions of FOE and FWE were identified and compared by GC–MS. The anti-inflammatory and analgesic activities of the two extracts and their possible active ingredients (α-pinene, linalool, and 1-octanol) were evaluated and compared in a xylene-induced ear edema model and a formalin-inflamed hind paw model. Inflammatory infiltrates and cyclooxygenase-2 (COX-2) expression in hind paw skin were investigated by histological staining.
Results
The contents of α-pinene, linalool, and 1-octanol in FOE were much higher than those in FWE. Mice treated with FOE exhibited greater and faster lessening of swelling and pain than mice treated with FWE. The combination of the three components had more potent pharmacological effects on hind paw inflammation and COX-2 overexpression than the three components used alone.
Conclusions
These findings suggest that topical application of FOE or its active ingredients (including α-pinene, linalool, and 1-octanol) exhibit significantly anti-inflammatory and analgesic effects through inhibiting nociceptive stimulus-induced inflammatory infiltrates and COX-2 overexpression.
OBJECTIVE: In the present study, the antiviral effects of polyphylla saponin I isolated from Parispolyphylla on influenza A virus are investigated both in vitro and in vivo.METHODS: Column chromatography and reversed phase liquid chromatography separation technology were used to extract and purify polyphylla saponin I. The purity of polyphylla saponin I was assayed by high performance liquid chromatography. Methyl thiazolyl tetrazolium assay and analyses of cytopathic effects were performed to examine the antiviral activity of polyphylla saponin I upon MDCK cells infected with influenza A virus. Model mice were made by intranasal inoculation of influenza a virus. Mice infected with influenza A virus were orally administered polyphylla saponin I and oseltamivir twice a day for 5 days to study their antiviral efficacy in vivo.
RESULTS: Polyphylla saponin I had no cytotoxicity on MDCK cells at the concentration of 50 μg/mL. Polyphylla saponin I (6.25, 12.5, 25 and 50 μg/mL) and oseltamivir (40 μg/mL) had remarkable inactivation effects on influenza A virus, prevention effects on influenza A virus adsorption on MDCK cells, and inhibitory effects on the reproduction of influenza A virus in MDCK cells. In addition, polyphylla saponin I (5 and 10 mg/kg), and oseltamivir (3 mg/kg) significantly reduced viral hemagglutination titer, improved the pathologic histology of lung tissues, and decreased the mortality of mice infected with influenza A virus. Polyphylla saponin I (5 and 10 mg/kg) prolonged the survival time of mice from 8.5±0.3 days to 13.2±0.5 days, with the prolonged life rates being 49.4% and 55.3%, respectively.
CONCLUSION: Polyphylla saponin I has antiviral activity on influenza A virus both in vitro and in vivo.
Introduction: Xanthones, the primary constituents of <i>Swertia mussotii</i>, are known to possess a variety of biological activities, including anti-depressant, anti-leukaemic, anti-tumour, anti-tubercular, choleretic, diuretic, anti-microbial, anti-fungal, anti-inflammatory, anti-viral, cardiotonic and hypoglycemic properties. However, high performance, environmentally friendly methods for isolating and purifying xanthones from <i>S. mussotii</i> are not currently available.<br>Objective: To develop a high performance and environmentally friendly method for the preparative separation of xanthones methylswertianin, swerchirin and decussatin from <i>S. mussotii</i> using high-speed counter-current chromatography (HSCCC).<br>Methodology: A solvent system composed of <i>n</i>-hexane:ethyl acatate:methanol:water (5:5:10:4, v/v/v/v) was developed for the separation method. The upper phase was used as the stationary phase, and the lower phase was used as the mobile phase at a flow rate of 1.5 mL/min, a rotation speed of 800 rpm and a temperature of 25 °C.<br>Results: Using the described method, 8 mg of methylswertianin, 21 mg of swerchirin and 11 mg of decussatin with purities of over 98% could be isolated from a 150 mg crude sample. They were identified by ¹H-NMR and <sup>13</sup> C-NMR analysis.<br>Conclusion: Three xanthones in <i>Swertia mussotii</i> could be systematically isolated and purified using HSCCC. Copyright © 2011 John Wiley & Sons, Ltd.
Abstract Ethnopharmacological relevance Qiwei Tiexie capsule (QWTX) is a representative prescription of Tibetan medicine, which is widely used for long-term treatment of chronic liver disease and nonalcoholic fatty liver disease (NAFLD). Aim of the study This study explored the effects and mechanism of QWTX on 3T3-L1 adipocytes and NAFLD. Materials and methods The 3T3-L1 preadipocytes and NAFLD rat model were used in the study. In 3T3-L1 cells, the cytotoxicity of QWTX was tested by CKK-8, and glucose uptake and fat acid oxidation were assessed by 2-deoxy-D-[3H] glucose and [1–14C] palmitic acid, respectively. The expression levels of carnitine palmitoyltransferase-1 (CPT-1), liver X receptor α (LXRα), peroxisome proliferator-activated receptor (PPAR) γ, inducible nitric oxide synthase (iNOS), ikappa B α (IκBα), and AKT were determined by PCR and western blot. NAFLD was established by the administration of fat emulsion and sucrose for 9 weeks. The effects of QWTX on lipid metabolism, liver function, and hepatic morphology were observed in NAFLD rats by HE and transmission electron microscope. Serum level of nitric oxide (NO) and fee fatty acid (FFA), superoxide dismutase (SOD) and malondialdehyde (MDA) contents in the liver, as well as the expression levels of Cytochrome P450 2E1 (CYP2E1), NF-κB, monocyte chemoattractant protein 1 (MCP-1), CPT-1, LXRα, PPARα, PPARβ/δ, PPARγ, and iNOS were all detected. Results QWTX showed no cell cytotoxicity in 3T3-L1 preadipocyte cells, and increased the 14CO 2 production rate to 4.15, which indicated the reducing the fatty accumulation. In NAFLD, QWTX attenuated liver steatosis, fat vacuoles and inflammation from the HE staining and electron micrograph tests. For the oxidative stress biomarkers, serum FFA level was reduced and serum NO level was enhanced after QWTX treatment. In liver tissue, SOD was decreased and MDA was significantly increased in NAFLD, and both of them were restored by QWTX. NF-κB and CYP2E1 were also upregulated in NAFLD, while downregulated by QWTX. Downregulation of LXRα, PPARγ and iNOS by QWTX were both observed in the 3T3-L1 adipocytes and NAFLD model. Conclusions QWTX protected the liver injury in differentiated 3T3-L1 adipocytes and NAFLD by regulating the LXRα, PPARγ, and NF-κB-iNOS-NO signal pathways. Graphical abstract Image 1 [ABSTRACT FROM AUTHOR]
Chebulae Fructus immaturus, a traditional Tibetan medicine, originated from the immature fruit of Terminalia chebula Retz., has been proven to have antioxidat function. However, its protection to injury liver cell caused by carbon tetrachloride (CCl4) has not been clarified. This study evaluated the effect of phenolic acid from Chebulae Fructus immaturus (PATC) on CCl4-induced acute liver injury in mice and related molecular mechanisms. Our data showed that PATC had convincing protective effects on the CCl4-induced acute liver injury by enhancing the anti-oxidative defense system, ameliorating inflammation and inhibiting the hepatocyte apoptosis.
Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease with a broad spectrum of liver injury. Oxidant stress is believed to be the pathogenesis of NAFLD as the "second hit". Hydrogen peroxide is widely used as an oxidant reagent to induce the oxidant injury of cells and larval zebrafish. Recently, cichoric acid is being studied extensively for its obesity attenuating, hepatic steatosis reduction and anti-oxidant effects. In this study, to identify whether CRA could protect the H2O2 induced oxidant injury via anti-oxidant impact by using L02 and HepG2 hepatocytes as in vitro and larval zebrafish as in vivo injury models, and evaluated the protective and anti-oxidant effects of CRA by pretreated it on both in vitro and in vivo models. CRA was found to reduce the production of ROS and MDA, activate the anti-oxidant enzymes SOD and GSH-px, and pathways Keap1-Nrf2 and HO-1. These results demonstrated that CRA might protect the liver injury by its anti-oxidant effect, which could be a potential therapeutic agent of NAFLD.
In this work, a hyphenated technique of dual ultrasound-assisted dispersive liquid-liquid microextraction combined with microwave-assisted derivatization followed by ultra high performance liquid chromatography tandem mass spectrometry has been developed for the determination of phytosterols in functional foods and medicinal herbs. Multiple reaction monitoring mode was used for the tandem mass spectrometry detection. A mass spectrometry sensitive reagent, 4'-carboxy-substituted rosamine, has been used as the derivatization reagent for five phytosterols, and internal standard diosgenin was used for the first time. Parameters for the dual microextraction, microwave-assisted derivatization, and ultra high performance liquid chromatography tandem mass spectrometry were all optimized in detail. Satisfactory linearity, recovery, repeatability, accuracy and precision, absence of matrix effect, extremely low limits of detection (0.005-0.015 ng/mL) and limits of quantification (0.030-0.10 ng/mL) were achieved. The proposed method was compared with previously reported methods. It showed better sensitivity, selectivity, and accuracy. The matrix effect was also significantly reduced. The proposed method was successfully applied to the determination of five phytosterols in vegetable oil (sunflower oil, olive oil, corn oil, peanut oil), milk and orange juice (soymilk, peanut milk, orange juice), and medicinal herbs (Ginseng, Ganoderma lucidum, Cordyceps, Polygonum multiflorum) for the quality control of functional foods and medicinal herbs.
• Simultaneously identified and quantified 18 phenolic compounds from LR fruit by UPLC-Q-Orbitrap MS. • Catechin, naringenin and 9 phenolic acids are the first time to conduct qualitative and quantitative analysis in LR. • Total phenolics content and total anthocyanin content were determined. • The antioxidant activities in vitro of the LR were also evaluated.<br><b>Lycium ruthenicum</b> Murray (LR) is a functional food, and it has long been used in traditional folk medicine. However, detailed qualitative and quantitative analyses related to its phenolic compounds remains scarce. This work reports, for the first time, the establishment of a rapid method for simultaneous identification and quantification of 25 phenolic compounds by UPLC-quadrupole-Orbitrap mass spectrometry (UPLC-Q-Orbitrap MS). This method was validated by LODs, LOQs, precision, repeatability, stability, mean recovery, recovery range and RSD. The confirmed method was applied to the analysis of phenolic compounds in LR. Finally, 18 phenolic compounds in LR were qualitatively and quantitatively analyzed. Among them, 11 constituents were detected for the first time, which included two flavonoids (catechin and naringenin) and seven phenolic acids (gallic acid, vanillic acid, 2,4-dihydroxybenzoic acid, veratronic acid, benzoic acid, ellagic acid and salicylic acid). Moreover, Phloretin and protocatechuate, belonging to the dihydrochalcone flavonoid and protocatechuic acid respectively, were also identified and quantified. The total phenolics content (20.17 ± 2.82 mg/g) and the total anthocyanin content (147.43 ± 1.81 mg/g) were determined. In addition, the antioxidant activities of the LR extract were evaluated through 2,2-azinobis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging activity, ferric reducing antioxidant power (FRAP) and total antioxidant activity (T-AOC) assays.
This book will expand the horizon of higher education, helping students, faculty and administrators to return to their roots and be in touch with their whole being. This book stresses that learning is much more than just accumulating knowledge and skills. Learning includes knowing ourselves—mind, body, and spirit. The learning of compassion, care, and service are as crucial or even more important in higher education in order for universities to address students’ individual needs and the society’s needs. Higher education must contribute to a better world. The book acknowledges that knowing not only comes from outside, but also comes from within. Wisdom is what guides students to be whole, true to themselves while learning. There are many ancient and modern approaches to gaining wisdom and wellness. This book talks about contemplative methods, such as meditation, qigong, yoga, arts, and dance, that help people gain wisdom and balance in their lives and enhance their ability to be reflective and transformative educators and learners.
This book will expand the horizon of higher education, helping students, faculty and administrators to return to their roots and be in touch with their whole being. This book stresses that learning is much more than just accumulating knowledge and skills. Learning includes knowing ourselves—mind, body, and spirit. The learning of compassion, care, and service are as crucial or even more important in higher education in order for universities to address students’ individual needs and the society’s needs. Higher education must contribute to a better world. The book acknowledges that knowing not only comes from outside, but also comes from within. Wisdom is what guides students to be whole, true to themselves while learning. There are many ancient and modern approaches to gaining wisdom and wellness. This book talks about contemplative methods, such as meditation, qigong, yoga, arts, and dance, that help people gain wisdom and balance in their lives and enhance their ability to be reflective and transformative educators and learners.
This book will expand the horizon of higher education, helping students, faculty and administrators to return to their roots and be in touch with their whole being. This book stresses that learning is much more than just accumulating knowledge and skills. Learning includes knowing ourselves—mind, body, and spirit. The learning of compassion, care, and service are as crucial or even more important in higher education in order for universities to address students’ individual needs and the society’s needs. Higher education must contribute to a better world. The book acknowledges that knowing not only comes from outside, but also comes from within. Wisdom is what guides students to be whole, true to themselves while learning. There are many ancient and modern approaches to gaining wisdom and wellness. This book talks about contemplative methods, such as meditation, qigong, yoga, arts, and dance, that help people gain wisdom and balance in their lives and enhance their ability to be reflective and transformative educators and learners.
The investigation aims to better understand the resource status of Rhodiola kirilowii, analysis the suitable habitat of wild Rh. kirilowii and protect the wild resources of Rh. Kirilowii, promoting the sustainable utilization of Rh. kirilowii resources. In this paper, we investigated the wild resources of Rh. kirilowii in 16 counties of Sichuan, Qinghai, Gansu and Yunnan by means of investigation and sampling investigation combined with interview. The results showed that the population densities of wild Rh. kirilowii in 4 provinces were very different and the reserve of wild resources decreased gradually in many areas. According to the survey results, the current total reserve of Rh. kirilowii in four provinces was about 1 100 t. The reserve of wild Rh. kirilowii in Sichuan province was the largest. Simultaneously, the Rh. kirilowii had a certain ecological value. We found that a sand control base with planting Rh. kirilowii was set up in Hongyuan County of Sichuan Aba Tibetan and Qiang Autonomous Prefecture. The investigation provides a scientific basis for the development and sustainable utilization of Rh. kirilowii resources.
ETHNOPHARMOCOLOGICAL RELEVANCE: Herbo-metallic preparations have a long history in the treatment of diseases, and are still used today for refractory diseases, as adjuncts to standard therapy, or for economic reasons in developing countries.AIM OF THE REVIEW: This review uses cinnabar (HgS) and realgar (As4S4) as mineral examples to discuss their occurrence, therapeutic use, pharmacology, toxicity in traditional medicine mixtures, and research perspectives.
MATERIALS AND METHODS: A literature search on cinnabar and realgar from PubMed, Chinese pharmacopeia, Google and other sources was carried out. Traditional medicines containing both cinnabar and realgar (An-Gong-Niu-Huang Wan, Hua-Feng-Dan); mainly cinnabar (Zhu-Sha-An-Shen Wan; Zuotai and Dangzuo), and mainly realgar (Huang-Dai Pian; Liu-Shen Wan; Niu-Huang-Jie-Du) are discussed.
RESULTS: Both cinnabar and realgar used in traditional medicines are subjected to special preparation procedures to remove impurities. Metals in these traditional medicines are in the sulfide forms which are different from environmental mercurials (HgCl2, MeHg) or arsenicals (NaAsO2, NaH2AsO4). Cinnabar and/or realgar are seldom used alone, but rather as mixtures with herbs and/or animal products in traditional medicines. Advanced technologies are now used to characterize these preparations. The bioaccessibility, absorption, distribution, metabolism and elimination of these herbo-metallic preparations are different from environmental metals. The rationale of including metals in traditional remedies and their interactions with drugs need to be justified. At higher therapeutic doses, balance of the benefits and risks is critical. Surveillance of patients using these herbo-metallic preparations is desired.
CONCLUSION: Chemical forms of mercury and arsenic are a major determinant of their disposition, efficacy and toxicity, and the use of total Hg and As alone for risk assessment of metals in traditional medicines is insufficient.
ETHNOPHARMACOLOGICAL RELEVANCE: Rhodiola crenulata, a traditional Tibetan medicine, has shown promise in the treatment of hypobaric hypoxia (HH)-induced brain injury. However, the underlying mechanisms remain unclear. This study investigated the protective effects of R. crenulata aqueous extract (RCAE) on HH-induced brain injury in rats.MATERIALS AND METHODS: An animal model of high-altitude hypoxic brain injury was established in SD rats using an animal decompression chamber for 24 h. Serum and hippocampus levels of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), oxidized glutathione (GSSG), and lactate dehydrogenase (LDH) were then determined using commercial biochemical kits. Neuron morphology and vitality were also evaluated using H&E and Nissl staining, and TUNEL staining was used to examine apoptosis. Gene and protein expression of HIF-1α, microRNA 210, ISCU1/2, COX10, Apaf-1, cleaved Caspase-3, Caspase-3, Bax, Bcl-2, and Cyto-c were determined by western blot, immunohistochemical and qRT-PCR analysis.
RESULTS: RCAE administration attenuated HH-induced brain injury as evidenced by decreased levels of MDA, LDH, and GSSG, increased GSH and SOD, improvements in hippocampus histopathological changes, increased cell vitality and ATP level, and reduced apoptotic cell numbers. RCAE treatment also enhanced HIF-1α, ISCU1/2, COX10, and Bcl-2 protein expression, while dramatically inhibiting expression of Apaf-1, Bax, Cyto-c, and cleaved Caspase-3. Treatment also increased gene levels of HIF-1α, microRNA 210, ISCU1/2, and COX10, and decreased Caspase-3 gene production.
CONCLUSIONS: RCAE attenuated HH-induced brain injury by regulating apoptosis and mitochondrial energy metabolism via the HIF-1α/microRNA 210/ISCU1/2 (COX10) signaling pathway.
This study presents an efficient strategy for separation of three phenolic compounds with high molecular weight from the crude extract of Terminalia chebula Retz. by ultrasound-assisted extraction and high-speed counter-current chromatography. The ultrasound-assisted extraction conditions were optimized by response surface methodology and the results showed the target compounds could be well enriched under the optimized extraction conditions. Then the crude extract was directly separated by high-speed counter-current chromatography without any pretreatment using n-hexane/ethyl acetate/methanol/water (1:7:0.5:3, v/v/v/v) as the solvent system. In 180 min, 13 mg of A, 18 mg of B, and 9 mg of C were obtained from 200 mg of crude sample. Their structures were identified as Chebulagic acid (A, 954 Da), Chebulinic acid (B, 956 Da), and Ellagic acid (C) by (1) H NMR spectroscopy.
This study is to develop an UPLC-PDA method for determination of 10 major components in Pterocephalus. The UPLC-PDA assay was performed on a Waters Acquity UPLCR BEH C₁₈（2.1 mm ×100 mm,1.7 μm）, and the column temperature was at 30 ℃. The mobile phase consists of water containing 0.2% phosphoric acid (A) and acetonitrile (B) in gradient elution at a flow rate of 0.4 mL•min⁻¹. The detection wave length was set at 237 and 325 nm, and the injection volume was 1 μL in the UPLC system. The linear range of 10 detected compounds were good (r≥0.999 7), and the overall recoveries ranged from 96.30% to 103.0%, with the RSD ranging from 0.72% to 2.9%. The method was simple, accurate and reproducible, which can be used for the simultaneous determination of the content of ten major components in P. hookeri.
• In situ derivatization-UADLLME was firstly reported for NTs in rat brain microdialysates. • Lissamine rhodamine B sulfonyl chloride was firstly used as derivatization reagent. • The method was simple, rapid, green, efficient, sensitive and low matrix effect. • This method was successfully applied for Parkinson’s rat brain microdialysates.<br>Simultaneous monitoring of several neurotransmitters (NTs) linked to Parkinson’s disease (PD) has important scientific significance for PD related pathology, pharmacology and drug screening. A new simple, fast and sensitive analytical method, based on in situ derivatization-ultrasound-assisted dispersive liquid-liquid microextraction (in situ DUADLLME) in a single step, has been proposed for the quantitative determination of catecholamines and their biosynthesis precursors and metabolites in rat brain microdialysates. The method involved the rapid injection of the mixture of low toxic bromobenzene (extractant) and acetonitrile (dispersant), which containing commercial Lissamine rhodamine B sulfonyl chloride (LRSC) as derivatization reagent, into the aqueous phase of sample and buffer, and the following in situ DUADLLME procedure. After centrifugation, 50 μL of the sedimented phase (bromobenzene) was directly injected for ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) detection in multiple reaction monitoring (MRM) mode. This interesting combination brought the advantages of speediness, simpleness, low matrix effects and high sensitivity in an effective way. Parameters of in situ DUADLLME and UHPLC-MS/MS conditions were all optimized in detail. The optimum conditions of in situ DUADLLME were found to be 30 μL of microdialysates, 150 μL of acetonitrile containing LRSC, 50 μL of bromobenzene and 800 μL of NaHCO3-Na2CO3 buffer (pH 10.5) for 3.0 min at 37 °C. Under the optimized conditions, good linearity was observed with LODs (S/N > 3) and LOQs (S/N > 10) of LRSC derivatized-NTs in the range of 0.002-0.004 and 0.007-0.015 nmol/L, respectively. It also brought good precision (3.2-12.8%, peak area CVs%), accuracy (94.2-108.6%), recovery (94.5-105.5%) and stability (3.8-8.1%, peak area CVs%) results. Moreover, LRSC derivatization significantly improved chromatographic resolution and MS detection sensitivity of NTs when compared with the reported studies through the introduction of a permanent charged moiety from LRSC into NTs. Taken together, this in situ DUADLLME method was successfully applied for the simultaneous determination of six NTs in biological samples.
Abstract Ethnopharmacological relevance Srolo Bzhtang (SBT), a traditional Tibetan medicine formula, was composed of three herbs, Solms-Laubachia eurycarpa , Bergenia purpurascens , Glycyrrhiza uralensis , and one lac, and was first documented in the ancient Tibetan medical work Four Medical Tantras (rGyud-bzhi) in the eighth century AD. It has been widely used to treat lung "phlegm-heat" syndromes such as chronic bronchitis and chronic obstructive pulmonary disease (COPD). Objective The aim of this study was to evaluate the potential influences of aqueous extract of SBT on airway inflammation and mucus secretion and to reveal the underlying mechanism in a rat model of cigarette smoke (CS)-induced chronic bronchitis (CB). Materials and methods Sixty male Sprague-Dawley rats were randomly divided to six groups: control (room air exposure), model (CS exposure), DEX (CS exposure and 0.2 mg/kg/day dexamethasone), and three SBT (CS exposure and 1.67, 2.50, and 3.34 g/kg/day SBT) groups. DEX and the three doses of SBT were administered by oral gavage every day for eight weeks. Pathological changes and mucus expression in the lung tissue were determined by hematoxylin and eosin (H&E), Alcian blue-periodic acid-Schiff (AB-PAS) and immunohistochemical staining. The levels of cytokines in bronchoalveolar lavage fluid (BALF) were assessed by ELISA. Western blot analysis and qRT-PCR were performed to explore the effects of SBT on the expression of IL-13, STAT6 and MUC5AC. Results Pretreatment with SBT attenuated the TNF-α, IL-8, IL-13 expression levels in BALF and the inflammatory cell infiltration in bronchial walls and peribronchial lung tissue. SBT exhibited a dose-dependent downregulation of MUC5AC expression as assessed by AB-PAS and immunohistochemical staining. The protein and mRNA levels of IL-13, STAT6/p-STAT6 and MUC5AC were also downregulated by SBT preconditioning. Conclusion These results for the first time demonstrated that SBT exhibited protective effects on CS-induced airway inflammation and MUC5AC hypersecretion, which might be related to the downregulation of the IL-13/STAT6 signaling pathway. Graphical abstract fx1 [ABSTRACT FROM AUTHOR]
Four common traditional tibetan medicine prescription preparations "Anzhijinghuasan, Dangzuo, Renqingchangjue and Rannasangpei" in tibetan areas were selected as study objects in the present study. The purpose was to try to establish a kind of wet digestion and flow injection-hydride generation-atomic absorption spectrometry (FI-HAAS) associated analysis method for the content determinations of lead and arsenic in traditional tibetan medicine under optimized digestion and measurement conditions and determine their contents accurately. Under these optimum operating conditions, experimental results were as follows. The detection limits for lead and arsenic were 0.067 and 0.012 µg · mL(-1) respectively. The quantification limits for lead and arsenic were 0.22 and 0.041 µg · mL(-1) respectively. The linear ranges for lead and arsenic were 25-1,600 ng · mL(-1) (r = 0.9995) and 12.5-800 ng · mL(-1) (r = 0.9994) respectively. The degrees of precision(RSD) for lead and arsenic were 2.0% and 3.2% respectively. The recovery rates for lead and arsenic were 98.00%-99.98% and 96.67%-99.87% respectively. The content determination results of lead and arsenic in four traditional tibetan medicine prescription preparations were as fol- lows. The contents of lead and arsenic in Anzhijinghuasan are 0.63-0.67 µg · g(-1) and 0.32-0.33 µg · g(-1) in Anzhijinghua- san, 42.92-43.36 µg · g(-1) and 24.67-25.87 µg · g(-1) in Dangzuo, 1,611. 39-1,631.36 µg · g(-1) and 926.76-956.52 µg- g(-1) in Renqing Changjue, and 1,102.28-1,119.127 µg-g(-1) and 509.96-516.87 µg · g(-1) in Rannasangpei, respectively. This study established a method for content determination of lead and arsenic in traditional tibetan medicine, and determined the content levels of lead and arsenic in four tibetan medicine-prescription preparations accurately. In addition, these results also provide the basis for the safe and effective use of those medicines in clinic.
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