Flavonoids from the leaves and stems of Sedum japonicum var. senanense and their antioxidant activity.

Twenty flavonoids (1-20) were isolated from the leaves and stems of Sedum japonicum var. senanense endemic to Japan. Among them, nine compounds were reported in nature for the first time, and identified as herbacetin 3-O-neohesperidoside-8-O-(2‴-acetylxyloside) (2), gossypetin 8-O-(2″-acetylxyloside) (4), gossypetin 8-O-(3″-acetylxyloside) (5), gossypetin 3-O-glucoside-8-O-(3‴-acetylxyloside) (9), gossypetin 3-O-glucoside-8-O-(2‴,3‴-diacetylxyloside) (10), gossypetin 3-O-neohesperidoside-8-O-xyloside (11), gossypetin 3-O-neohesperidoside-8-O-(2⁗-acetylxyloside) (12), gossypetin 3-O-neohesperidoside-8-O-(3⁗-acetylxyloside) (13) and gossypetin 3-O-glucoside-8-O-xylofuranoside (14) by UV spectral survey, HR-MS, LC-MS, acid hydrolysis, NMR including 1H and 13C NMR, COSY, NOESY, HSQC and HMBC. Moreover, nine major flavonoids were surveyed for antioxidant activity by H-ORAC method. As the results, gossypetin 3-O-glucoside-8-O-(2‴-acetylxyoside) (8) showed the highest antioxidant activity. Conversely, gossypetin 3-O-neohesperidoside-8-O-xyloside (11) and gossypetin 3-O-neohesperidoside-8-O-(2⁗-acetylxyloside) (12) which attach neohesperidose showed the lowest values.

Neuroprotective and cognitive enhancing effects of herbecetin against thioacetamide induced hepatic encephalopathy in rats via upregulation of AMPK and SIRT1 signaling pathways.

Acute liver injury, there is a risky neurological condition known as hepatic encephalopathy (HE). Herbacetin is a glycosylated flavonoid with many pharmacological characteristics. The purpose of this study was to assess the ability of herbacetin to protect against the cognitive deficits associated with thioacetamide (TAA) rat model and delineate the underlying behavioral and pharmacological mechanisms. Rats were pretreated with herbacetin (20 and 40 mg/kg) for 30days. On 30th day, the rats were injected with TAA (i.p. 350 mg/kg) in a single dose. In addition to a histpathological studies, ultra-structural architecture of the brain, liver functions, oxidative stress biomarkers, and behavioral tests were evaluated. Compared to the TAA-intoxicated group, herbacetin improved the locomotor and cognitive deficits, serum hepatotoxicity indices and ammonia levels. Herbacetin reduced brain levels of malodialdeyde, glutamine synthetase (GS), tumor necrosis factor- alpha (TNF-α), interleukin 1 B (IL-1ß), annexin v, and increased brain GSH, Sirtuin 1 (SIRT1), and AMP-activated kinase (AMPK) expression levels. Also, herbacetin improve the histopathological changes and ultra- structure of brain tissue via attenuating the number of inflammatory and apoptotic cells. Herbacetin treatment significantly reduced the toxicity caused by TAA. These findings suggest that herbacetin might be taken into account as a possible neuroprotective and cognitive enhancing agent due to its ability to reduce oxidative stress, inflammation and apoptosis associated with TAA.

Flavonoids from Sedum japonicum subsp. oryzifolium (Crassulaceae).

Twenty-two flavonoids were isolated from the leaves and stems of Sedum japonicum subsp. oryzifolium (Crassulaceae). Of these compounds, five flavonoids were reported in nature for the first time, and identified as herbacetin 3-O-xyloside-8-O-glucoside, herbacetin 3-O-glucoside-8-O-(2'''-acetylxyloside), gossypetin 3-O-glucoside-8-O-arabinoside, gossypetin 3-O-glucoside-8-O-(2'''-acetylxyloside) and hibiscetin 3-O-glucoside-8-O-arabinoside via UV, HR-MS, LC-MS, acid hydrolysis and NMR. Other seventeen known flavonoids were identified as herbacetin 3-O-glucoside-8-O-arabinoside, herbacetin 3-O-glucoside-8-O-xyloside, gossypetin 3-O-glucoside-8-O-xyloside, quercetin, quercetin 3-O-glucoside, quercetin 3-O-xylosyl-(1→2)-rhamnoside-7-O-rhamnoside, quercetin 3-O-rhamnoside-7-O-glucoside, kaempferol, kaempferol 3-O-glucoside, kaempferol 7-O-rhamnoside, kaempferol 3,7-di-O-rhamnoside, kaempferol 3-O-glucoside-7-O-rhamnoside, kaempferol 3-O-glucosyl-(1→2)-rhamnoside-7-O-rhamnoside, kaempferol 3-O-xylosyl-(1→2)-rhamnoside, kaempferol 3-O-xylosyl-(1→2)-rhamnoside-7-O-rhamnoside, myricetin 3-O-glucoside and cyanidin 3-O-glucoside. Some flavonol 3,8-di-O-glycosides were found in Sedum japonicum subsp. oryzifolium as major flavonoids in this survey. They were presumed to be the diagnostic flavonoids in the species. Flavonoids were reported from S. japonicum for the first time.

Ameliorative effect of herbacetin against cyclophosphamide-induced nephrotoxicity in rats via attenuation of oxidative stress, inflammation, apoptosis and mitochondrial dysfunction.

Herbacetin (HBN) is a glycosylated flavonoid, which possesses numerous pharmacological properties. Cyclophosphamide (CYC) is a chemotherapeutic drug that adversely affects the kidneys. The present investigation aimed to evaluate the curative potential of HBN against CYC-induced nephrotoxicity. Sprague Dawley rats (n = 48) were randomly divided into four groups: control (0.1% DMSO + food), CYC (150 mg/kg b.wt.), CYC+HBN (150 + 40 mg/kg b.wt.), and HBN (40mg/kg b.wt.). CYC treatment significantly decreased the activities of antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GSR) while elevating the concentration of reactive oxygen species (ROS) and malondialdehyde (MDA). Treatment with HBN significantly recovered the activity of CAT, SOD, GPx, and GSR while reducing the concentrations of ROS and MDA. Moreover, an increase in the level of renal functional markers, including Urea, creatinine, kidney injury molecule-1 (KIM-1), and neutrophil gelatinase-associated lipocalin (NGAL), and a decrease in creatinine clearance after CYC administration was recovered to control values by HBN treatment. Furthermore, HBN treatment normalized the increased levels of inflammatory markers such as nuclear factor kappa-B (NF-κB), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) after CYC administration. Besides, HBN administration increased the expression of anti-apoptotic markers (Bcl-2) while decreasing the apoptotic markers (Bax and Caspase-3). Furthermore, HBN decreased the activities of tricarboxylic acid (TCA) cycle enzymes (ICDH, αKGDH, SDH, and MDH) as well as renal mitochondrial respiratory-chain complexes (I-IV) and repolarized mitochondrial membrane potential (ΔΨm). Additionally, HBN administration significantly protected against renal histological damage induced by CYC. In conclusion, CYC-induced toxicity was effectively ameliorated by the HBN administration. These results indicate that HBN might be considered as a potential protective agent against nephrotoxicity. The observed protection may be due to its antioxidant, anti-inflammatory, and anti-apoptotic potential.

Discovery of Flavonoids as Novel Inhibitors of ATP Citrate Lyase: Structure-Activity Relationship and Inhibition Profiles.

ATP citrate lyase (ACLY) is a key enzyme in glucolipid metabolism and its aberrantly high expression is closely associated with various cancers, hyperlipemia and atherosclerotic cardiovascular diseases. Prospects of ACLY inhibitors as treatments of these diseases are excellent. To date, flavonoids have not been extensively reported as ACLY inhibitors. In our study, 138 flavonoids were screened and 21 of them were subjected to concentration-response curves. A remarkable structure-activity relationship (SAR) trend was found: ortho-dihydroxyphenyl and a conjugated system maintained by a pyrone ring were critical for inhibitory activity. Among these flavonoids, herbacetin had a typical structure and showed a non-aggregated state in solution and a high inhibition potency (IC50 = 0.50 ± 0.08 µM), and therefore was selected as a representative for the ligand-protein interaction study. In thermal shift assays, herbacetin improved the thermal stability of ACLY, suggesting a direct interaction with ACLY. Kinetic studies determined that herbacetin was a noncompetitive inhibitor of ACLY, as illustrated by molecular docking and dynamics simulation. Together, this work demonstrated flavonoids as novel and potent ACLY inhibitors with a remarkable SAR trend, which may help design high-potency ACLY inhibitors. In-depth studies of herbacetin deepened our understanding of the interactions between flavonoids and ACLY.

Discovery of Herbacetin as a Novel SGK1 Inhibitor to Alleviate Myocardial Hypertrophy.

Cardiac hypertrophy is a pivotal pathophysiological step of various cardiovascular diseases, which eventually leads to heart failure and death. Extracts of Rhodiola species (Ext.R), a class of commonly used medicinal herbs in Europe and East Asia, can attenuate cardiac hypertrophy both in vitro and in vivo. Serum/glucocorticoid regulated kinase 1 (SGK1) is identified as a potential target of Ext. R. By mass spectrometry-based kinase inhibitory assay, herbacetin (HBT) from Ext.R is identified as a novel SGK1 inhibitor with IC50 of 752 nmol. Thermal shift assay, KINOMEscan in vitro assay combined with molecular docking proves a direct binding between HBT and SGK1. Site-specific mutation of Asp177 in SGK1 completely ablates the inhibitory activity of HBT. The presence of OH groups at the C-3, C-8, C-4' positions of flavonoids is suggested to be favorable for the inhibition of SGK1 activity. Finally, HBT significantly suppresses cardiomyocyte hypertrophy in vitro and in vivo, reduces reactive oxygen species (ROS) synthesis and calcium accumulation. HBT decreases phosphorylation of SGK1 and regulates its downstream forkhead box protein O1 (FoxO1) signaling pathway. Taken together, the findings suggest that a panel of flavonoids structurally related to HBT may be novel leads for developing new therapeutics against cardiac hypertrophy.

Metabolism of flavonoids and lignans by lactobacilli and bifidobacteria strains improves the nutritional properties of flaxseed-enriched beverages.

Flaxseed (Linum usitatissimum L.) is of interest as functional food because of the presence of compounds in its composition with potential health benefits, such as fatty acid omega-3, fiber, lignans and flavonoids. The bioactivity of lignans and flavonoids depends greatly on bacterial metabolism. Previously, lactobacilli and bifidobacteria strains were described to produce enterolignans and bioactive flavonoids (herbacetin, quercetin, quercetagetin, kaempferol, naringenin and eriodictyol) from flaxseed extracts and/or from secoisolariciresinol (SECO) in culture medium. In this work, cow's milk and soy beverage were supplemented with flaxseed extracts and fermented with selected lactobacilli and bifidobacteria strains. Lacticaseibacillus rhamnosus INIA P224, Limosilactobacillus mucosae INIA P508 and Lactiplantibacillus plantarum ESI 144 were capable of producing enterolactone (ENL) in both beverages supplemented with flaxseed, in addition to matairesinol and the flavonoids daidzein, genistein, glycitein, quercetin, naringenin, kaempferol and eriodictyol. On the other hand, Bifidobacterium breve INIA P367, Bifidobacterium pseudocatenulatum INIA P815 and Bifidobacterium pseudocatenulatum INIA P946 were able to produce quercetin, quercetagetin and high concentrations of herbacetin and SECO, in addition to pinoresinol, matairesinol, daidzein, genistein, naringenin, kaempferol and eriodictyol. The co-incubation of Lacticaseibacillus paracasei INIA P74 and Ligilactobacillus salivarius INIA P183 with Lactococcus lactis MG1363 harboring the food grade vector pLEB590.gly913, facilitated the production of ENL in soy beverage enriched with flaxseed. In this work, it is demonstrated how lactobacilli and bifidobacteria strains can improve the nutritional properties of flaxseed-enriched beverages, providing metabolites of great interest for human health.

A comprehensive review of herbacetin: From chemistry to pharmacological activities.

ETHNOPHARMACOLOGICAL RELEVANCE: Herbacetin is an active constituent of traditional Chinese medicines such as Ephedra sinica Stapf (MaHuang) and Sedum roseum (L.). Scop. (Hong JingTian). MaHuang was used to treat cough, asthma, fever, and edema for more than 5000 years, while Hong JingTian was used to treat depression, fatigue, cancers, and cardiovascular disease. Recent studies indicate that herbacetin and its glycosides play a critical role in the pharmacological activities of these herbs. However, currently, no comprehensive review on herbacetin has been published yet. AIM OF THE STUDY: This review aimed to summarize information on the chemistry, natural sources, and pharmacokinetic features of herbacetin, with an emphasis on its pharmacological activities and possible mechanisms of action. MATERIALS AND METHODS: A literature search was performed on the Web of Science, PubMed, and China Knowledge Resource Integrated databases (CNKI) using the search term "herbacetin" ("all fields") from 1935 to 2020. Information was also obtained from classic books of Chinese herbal medicine, Chinese pharmacopeia, and the database "The Plant List" (www.theplantlist.org). Studies have been analyzed and summarized in this review if they dealt with chemistry, taxonomy, pharmacokinetic, and pharmacological activity. RESULTS: Herbacetin is distributed in various plants and can be extracted or synthesized. It showed diverse pharmacological activities including antioxidant, antiviral, anti-inflammatory, anticancer, antidiabetic, and anticholinesterase. It is thought to have great potential in cancer treatment, especially colon and skin cancers. However, the bioavailability of herbacetin is low and the toxicity of herbacetin has not been studied. Thus, more studies are required to solve these problems. CONCLUSIONS: Herbacetin shows promising pharmacological activities against multiple diseases. Future research should focus on improving bioavailability, further studying its pharmacological mechanism, evaluating its toxicity and optimal dose, and performing the clinical assessment. We hope that the present review will serve as a guideline for future research on herbacetin.

Isolation, X-ray crystal structure of the new diterpene and identification of others lignans and flavonoids from the fresh needles of Pinus massoniana.

Three new compounds, namely massonside C (1), massonianoside F (2), and 3, 8-dimethyl- herbacetin-7-O-ß-D-glucopyranoside (3), together with five known compounds (4-8), were isolated from the fresh needles of Pinus massoniana. Their structures were established by 1D, 2D NMR, HRMS and comparison with the literature data. The absolute configuration of 1 was confirmed by a combination of X-ray single crystal analysis. All isolated compounds were evaluated for the protective effect of human umbilical vein endothelial cells against oxidative damage.

Herbacetin suppressed MMP9 mediated angiogenesis of malignant melanoma through blocking EGFR-ERK/AKT signaling pathway.

Malignant melanoma remains a challenge for clinical practice and novel therapeutic strategies are urgently needed. Herbacetin, a natural flavonoid compound that has multiple pharmacological activities, exerts anticancer effects on several human tumors. In this study, the anti-angiogenesis effect of Herbacetin in human malignant melanoma was investigated. The results indicated that Herbacetin treatment significantly suppressed tumor growth and angiogenesis of malignant melanoma both in vitro and in vivo. In melanoma A375 and Hs294T cells, Herbacetin treatment suppressed both EGF-induced and constitutive phosphorylation of EGFR, accelerated the internalization and degradation of EGFR, and subsequently suppressed the activation of the downstream kinases (AKT and ERK). Moreover, MMP9 was determined as a key angiogenic factor in Herbacetin treated melanoma cells. Knockdown of MMP9 suppressed the in vitro angiogenesis while overexpression of MMP9 in Herbacetin treated melanoma cells restored the angiogenesis ability. We concluded that Herbacetin suppressed melanoma angiogenesis through blocking EGFR-ERK/AKT-MMP9 signaling pathway and Herbacetin may be developed as a potential drug for melanoma treatment.

Herbacetin treatment remitted LPS induced inhibition of osteoblast differentiation through blocking AKT/NF-κB signaling pathway.

Inflammation, a common situation during the process of bone healing, is reported to play a negative role in bone regeneration. Up to date, therapeutic strategies for inflammation triggered inhibition of osteoblast differentiation are still limited. The aim of this study was to explore the potential roles and molecular mechanisms of Herbacetin in the process of osteoblast differentiation under LPS-mediated inflammatory environment. By using MC3T3-E1, C2C12 and primary mouse calvarial osteoblast (PMCO) cells as experimental models, we observed that LPS stimulation suppressed osteoblast differentiation via inhibiting alkaline phosphatase (ALP) activity and the expression of several osteoblastic genes (osterix, runx2 and osteocalcin). However, the negative role of LPS during osteoblast differentiation could be restored by Herbacetin treatment. Mechanistical studies revealed that Herbacetin treatment suppressed AKT activation and in turn blocked NF-κB signaling pathway. Furthermore, reactivating AKT by a selective PTEN inhibitor SF1670 suppressed the effect of Herbacetin. These data suggested that Herbacetin might play a protective role in osteoblast differentiation in MC3T3-E1/C2C12/PMCO cells under LPS stimulation.

Herbacetin, a flaxseed flavonoid, ameliorates high percent dietary fat induced insulin resistance and lipid accumulation through the regulation of hepatic lipid metabolizing and lipid-regulating enzymes.

Healthy plants and their constituents have been considered as a safe remedy for the treatment of obesity and obesity associated diseases. Herbacetin is a dietary flavonoid that has been explored for many pharmacological activities; but, the anti-hyperglycaemic and anti-hyperlipidemic properties of herbacetin have not yet been explored. The present study was performed to evaluate the ameliorative effect of herbacetin on high-fat diet-induced hyperglycaemia and hyperlipidemia in 57BL/6 J mice. Obesity associated insulin resistance was induced by continuously feeding the mice with high-fat diet for 10 weeks. Afterwards, mice were subjected to intragastric administration of herbacetin (different doses) daily along with high-fat diet for the next 5 weeks. At the end of 106th day, changes in body weight, blood glucose, insulin, HOMA-IR, and lipids profiles and lipid-regulating enzymes were evaluated. Herbacetin significantly reduced the body weight, plasma glucose, plasma insulin, and HOMA-IR activity in obesity associated insulin resistant mice (OIR). In addition, herbacetin administration significantly reduced the plasma and hepatic total cholesterol, triglycerides, and free fatty acids in OIR mice. Moreover, herbacetin significantly improved the altered hepatic lipid metabolizing and lipid-regulating enzymes such as SREBP-1c, and 2, fatty acid synthase (FAS), fatty acid ß-oxidation (ß-oxidation), malic enzyme, glucose 6-phosphate dehydrogenase (G6PD), and carnitine palmitoyltransferase (CPT) when compared to OIR control mice. Histopathological examination clearly showed that herbacetin decreases lipid droplets in the liver tissue. Thus, observed results strongly indicate that herbacetin provides remarkable protection against the harmful effects of chronic high-fat diet consumption because of its anti-hyperglycaemic and anti-hyperlipidemic properties through the regulation of hepatic lipid metabolizing and lipid-regulating enzymes.

Glucuronidation is the dominating in-vivo metabolism pathway of herbacetin:elucidation of herbacetin pharmacokinetics after intravenous and oral administration in rats / 中国药理学与毒理学杂志

OBJECTIVE To map a comprehensive metabolic pathway of herbacetin in rats, specifically, to elucidate the biotransformation of herbacetin in vivo and to simultaneously monitor the pharmacokinetic process of both parent drug and its major metabolites. METHODS liquid chromatography/ion trap mass spectrometry (LC/MSn) and ultra-liquid chromatography coupled with mass spectrometry (UPLC/MS) were combined in the current study for qualitative and quantitative determinations of herbacetin and its metabolites in bile, urine and feces after both oral and intravenous administration of herbacetin to rats. Enzyme kinetic studies on the intestinal and hepatic metabolism of herbacetin were further conducted to elucidate metabolic profiles of herbacetin in rat tissues and organs. Additionally, plasma concentration profiles of herbacetin and its metabolites in rats were obtained to characterize the overall pharmacokinetic behavior of herbacetin. RESULTS It was found that herbacetin was excreted primarily from rat urine in the form of glucuronide-conjugations. Subsequent in vitro enzyme kinetic studies and in vivo pharmacokinetic investigations suggested an extensive hepatic metabolism of herbacetin and the high exposure of herbacetin- glucuronides in systemic circulation. The clearance, half- life and bioavailability of herbacetin in rats were determined as (16.4±1.92)mL·kg-1·min-1, (11.9±2.7)min, and 1.32%, respectively. On basis of these findings, a comprehensive metabolic pathway of herbacetin in rats was composed. In addition, a physiology based pharmacokinetic (PBPK) model was successfully developed with the aid of the GastroPlus to simulate the pharmacokinetic process of herbacetin in rats. Application of the PBPK modeling can provide a useful starting point to understand and extrapolate pharmacokinetic parameters among different species, populations, and disease states. CONCLUSION After oral administration, herbacetin was subjected to colonic degradation and extensive first pass metabolism, with glucuronidation as its dominating in vivo metabolic pathway.

Efficiently prepared ephedrine alkaloids-free Ephedra Herb extract: a putative marker and antiproliferative effects.

Ephedrine alkaloids (EAs) have been considered the main pharmacologically active substances in Ephedra Herb (, Mao; EH) since they were first identified by Prof. N. Nagai, and are known to induce palpitation, hypertension, insomnia, and dysuria as side effects. Therefore, the administration of drugs containing EH to patients with cardiovascular-related diseases is severely contraindicated. While our previous studies suggest that some of the effects of EH may not be due to EAs, considering their side effects would be expedient to develop a new EAs-free EH extract (EFE). Here, we established a preparation method for EFE and revealed its chemical composition, including the content of herbacetin, a flavonoid aglycon present in EH and a potential putative marker for EFE quality control. In addition, we showed the antiproliferative effects of EFE against the H1975 non-small cell lung cancer (NSCLC) cell line. EFE was prepared from EH extract using the ion exchange resin SK-1B. LC/Orbitrap MS analysis revealed the removal of EAs, 6-methoxykynurenic acid, and 6-hydroxykynurenic acid from the original extract. Quantitative analysis of herbacetin using LC/MS in acid-hydrolyzed EFE showed that its content was 0.104 %. Although several alkaloidal constituents were removed from EH extract, the antiproliferative effect of EFE against H1975 cells was comparable to that of EH extract. These results indicate that EFE retained the anticancer effect of EH and demonstrated its potential for future development as a new herbal medicine with reduced side effects.

Herbacetin inhibits RANKL-mediated osteoclastogenesis in vitro and prevents inflammatory bone loss in vivo.

Herbacetin is an active flavonol (a type of flavonoid) that has various biologic effects such as antioxidant, antitumor, and anti-inflammatory activities. However, one of its novel effects remains to be investigated, that is, the induction of osteoclastogenesis by the receptor activator of nuclear factor-κB ligand (RANKL). In this study, we examined the effects and mechanisms of action of herbacetin on osteoclastogenesis in RANKL-treated bone marrow-derived macrophages (BMMs) and murine macrophage RAW264.7 cells in vitro and on lipopolysaccharide (LPS)-induced bone destruction in vivo. Herbacetin significantly inhibited RANKL-induced osteoclast formation and differentiation in BMMs and RAW264.7 cells in a dose-dependent manner. Moreover, the suppressive effect of herbacetin resulted in a decrease in osteoclast-related genes, including RANK, tartrate-resistant acid phosphatase, cathepsin K, and matrix metalloproteinase-2 and -9 (MMP-9). Consistent with mRNA results, we confirmed that herbacetin treatment downregulated protein expression of MMP-9 and cathepsin K. Herbacetin also decreased induction of the osteoclastogenic transcription factor c-Fos and nuclear factor of activated T cells c1 (NFATc1) and blocked RANKL-mediated activation of Jun N-terminal kinase (JNK) and nuclear factor-κB. Herbacetin clearly inhibited the bone resorption activity of osteoclasts on plates coated with fluorescein-labeled calcium phosphate. More importantly, the application of herbacetin significantly reduced LPS-induced inflammatory bone loss in mice in vivo. Taken together, our results indicate that herbacetin has potential for use as a therapeutic agent in disorders associated with bone loss.

Quantification of six active ingredients in Rhodiola extract by UPLC-MS/MS / 国际药学研究杂志

Objective To develop and fully validate an UPLC-MS/MS method for simultaneous quantification of six active ingredients in Rhodiola extract including salidroside, rhodiosin, rhodionin, herbacetin, kaempferol and quercetin. Methods The chromatographic separation of the analytes was achieved by using Zorbax Eclipse plus C18 column (2.1 mm×100 mm, 3.5 μm). The mobile phase consisted of 0.2% aqueous formic acid and acetonitrile. Analytes were separated using a linear gradient with a flow rate of 0.4 ml/min. The column temperature was set at 25 °C. The mass spectrometric conditions were electrospray negative ionization (ESI) and the scanning mode was multi reaction monitoring (MRM). Results The calibration curves of the six active ingredients were linear over 1020000, 10-4000, 10-4000, 5-2000, 10-2000, and 5-2000 ng/ml concentration ranges, respectively. The RSD of the instrument precision was within 3.31%. The current assay was stable and reproducible in quantification of all analytes with RSD within 5.07% and 4.05%, respectively. The average recovery of all analytes was in a range from 99.27% to 108.91%, with an acceptable RSD of no more than 4.92%. Conclusion The UPLC-MS/MS method could be successfully applied to simultaneously quantifying six major active ingredients in Rhodiola extract with the acceptable specificity, sensitivity and reproducibility.

Chemical constituents from rhizomes of Rhodiola wallichiana var. cholaensis and their protective effects on myocardium / 中草药

Objective: To investigate the chemical constituents from the rhizomes of Rhodiola wallichiana var. cholaensis and their protective effects on myocardium of H9c2 cells. Methods: The compounds were isolated by dynamic axial compression chromatography, silica gel column chromatography, and HPLC. Their structures were identified on the basis of spectral data analysis. The protective effects of all isolated compounds on myocardium were determined. Results: Sixteen compounds were isolated from the rhizomes of R. wallichiana var. cholaensis and identified as salidroside (1), gallic acid (2), methyl gallate (3), quercetin (4), pyrogallol (5), 6″-O-galloylsalidroside (6), ethyl gallate (7), kaemnpferol-7-O-α-L-rhamnopyranoside (8), herbacertin-7-O-β-D-glucopyranoside (9), rhodiosin (10), kaemnpferol-3-O-β-D-glucopyranoside (11), herbacetin (12), herbacertin-7-α-L-rhamnopyranoside (13), tricin (14), rutin (15), and kaemnpferol-3-O-(2″-O-β-D-xylosyl)-β-D-glucopyranoside (16). Compounds 2, 6, 8, 12, and 15 showed the significant protective effects against H9c2 cells at the concentration of 25 μg/mL and the protective ratios were 20.40%, 31.54%, 67.61%, 44.27%, and 47.84%. Conclusion: Compound 5 is isolated from the species of genus Rhodiola L. for the first time, and compounds 7-16 are obtained from the plant for the first time. Compounds 2, 6, 8, 12, and 15 could protect the myocardium of H9c2 cells to some extent.

Determination of six flavonoids in Rhodiolae Crenulatae Radix et Rhizoma by HPLC-DAD / 中草药

Objective: To establish a determination method for the contents of ternatumoside II, rhodiosin, rhodionin, herbacetin, kaempferol, and rhodiolin in Rhodiolae Crenulatae Radix et Rhizoma and to compare the content differences of the six flavonid compounds in Rhodiolae Crenulatae Radix et Rhizoma from different sources. Methods: Using HPLC-DAD method and Kromasil C18 column (250 mm × 4.6 mm, 5 μm), with acetonitrile and water as mobile phase, gradient elution at flow rate of 1.0 mL/min and column temperature of 35℃. The detection wavelength was 382 nm. Results: Ternatumoside II, rhodiosin, rhodionin, herbacetin, kaempferol, and rhodiolin had good linearity in the ranges of 0.015 08-0.301 6, 0.123 2-2.464, 0.109 5-2.190 8, 0.007 8-0.156, 0.021 46-0.429 2, and 0.006 48-0.129 6 μg, respectively. The average recoveries of the six constituents were 98.17%-100.52% and RSD values were 1.64%-1.98%. Conclusion: The method is simple and accutate. It provides the reference for comprehensive quality control of Rhodiolae Crenulatae Radix et Rhizoma.

Quantification of six active ingredients in Rhodiola extract by UPLC-MS/MS / 国际药学研究杂志

Objective To develop and fully validate an UPLC-MS/MS method for simultaneous quantification of six active in?gredients in Rhodiola extract including salidroside,rhodiosin,rhodionin,herbacetin,kaempferol and quercetin. Methods The chro?matographic separation of the analytes was achieved by using Zorbax Eclipse plus C18 column(2.1 mm×100 mm,3.5μm). The mobile phase consisted of 0.2%aqueous formic acid and acetonitrile. Analytes were separated using a linear gradient with a flow rate of 0.4 ml/min. The column temperature was set at 25℃. The mass spectrometric conditions were electrospray negative ionization(ESI)and the scan?ning mode was multi reaction monitoring(MRM). Results The calibration curves of the six active ingredients were linear over 10-20000,10-4000,10-4000,5-2000,10-2000,and 5-2000 ng/ml concentration ranges,respectively. The RSD of the instrument precision was within 3.31%. The current assay was stable and reproducible in quantification of all analytes with RSD within 5.07%and 4.05%,respectively. The average recovery of all analytes was in a range from 99.27%to 108.91%,with an acceptable RSD of no more than 4.92%. Conclusion The UPLC-MS/MS method could be successfully applied to simultaneously quantifying six major active in?gredients in Rhodiola extract with the acceptable specificity,sensitivity and reproducibility.

Network pharmacology-based prediction of the active ingredients and potential targets of Mahuang Fuzi Xixin decoction for application to allergic rhinitis.

ETHNOPHARMACOLOGICAL RELEVANCE: Certain herbal formulae from Traditional Chinese Medicine (TCM) are effective for treating and preventing diseases in clinical practice. Mahuang fuzi Xixin Decoction (MFXD) is a TCM that is used to treat allergic rhinitis (AR); however, the active ingredients and potential targets of its action against AR remain unclear. Therefore, further investigation is required. METHODS: A network pharmacology approach comprising drug-likeness evaluation, oral bioavailability prediction, multiple drug target prediction, and network analysis has been used in this study. RESULTS: The comprehensive systematic approach was successfully to indentify 41 bioactive ingredients in MFXD, while 37 potential targets hit by these ingredients related to AR. Moreover, wherein four predicted ingredients possess anti-inflammatory effects were found by this technique. CONCLUSIONS: Our works successfully predict the active ingredients and potential targets of MFXD for application to allergic rhinitis and helps to illustrate mechanism of action on a systematic level. This study not only provides new insights into the chemical basis and pharmacology of MFXD but also demonstrates a feasible method for discovering potential drugs from herbal medicine.