Quantitative Analysis of Two Isoflavones in Pueraria Lobata Flowers from Eleven Chinese Provinces Using High Performance Liquid Chromatography

Background: Pueraria lobata flower (Gehua) is a medicinal herb to treat intoxication, hepatic and gastrointestinal tractlesion induced by alcohol. This study aims to develop a new HPLC method for the determination of two majorisoflavones in P. lobata flowers, namely tectoridin and 6"-O-xylosyl-tectoridin.Methods: A high performance liquid chromatography (HPLC) method with a C18 column (250 mm × 4.6 mm, 5 μm)was developed for the quantitative analysis of tectoridin and 6"-O-xylosyl-tectoridin, the main isoflavone componentsin P. lobata flower. A simple gradient of acetonitrile/water (0 min 15:85; 35 min 50:50; 36 min 15:85; 40 min 15:85; v/v)was used, and 265 nm was selected as detection wavelength. Tectoridin and 6"-O-xylosyl-tectoridin were used as theexternal standards in quality control of P. lobata flower for the first time. The method was applied to practical use inquality assessment of eleven batches of P. lobata flower samples in Chinese herbal medicine market.Results: The peak area response was linear for tectoridin in the 11.8-236.4 μg/mL range with a correlation coefficient of0.9996 (P < 0.001), and for 6"-O-xylosyl-tectoridin in the 10.33-185.99 μg/mL range with a correlation coefficient of0.9984 (P < 0.001) respectively. The average recoveries were 102.7-103.7% for tectoridin and 95.7-103.2% for 6"-Oxylosyl-tectoridin (RSDs < 3%), and the intra-day and inter-day RSDs of the two components were less than 2%. ThisHPLC method was applied to assess the quality of P. lobata flower from eleven provinces in China. P. lobata flowers fromnorthern China contained 26.46-43.28 mg/g of tectoridin and 30.90-48.23 mg/g of 6"-O-xylosyl-tectoridin comparingto 10.00-19.81 mg/g of tectoridin and 11.08-37.03 mg/g of 6"-O-xylosyl-tectoridin in those from southern China.Conclusion: The results showed that P. lobata flowers from northern China contained more tectoridin and 6"-Oxylosyl-tectoridin than those from southern China

Quantitative analysis of two isoflavones in Pueraria lobata flowers from eleven Chinese provinces using high performance liquid chromatography

<p>Abstract</p> <p>Background</p> <p><it>Pueraria lobata </it>flower (<it>Gehua</it>) is a medicinal herb to treat intoxication, hepatic and gastrointestinal tract lesion induced by alcohol. This study aims to develop a new HPLC method for the determination of two major isoflavones in <it>P. lobata </it>flowers, namely tectoridin and 6"-O-xylosyl-tectoridin.</p> <p>Methods</p> <p>A high performance liquid chromatography (HPLC) method with a C₁₈column (250 mm × 4.6 mm, 5 μm) was developed for the quantitative analysis of tectoridin and 6"-O-xylosyl-tectoridin, the main isoflavone components in <it>P. lobata </it>flower. A simple gradient of acetonitrile/water (0 min 15:85; 35 min 50:50; 36 min 15:85; 40 min 15:85; v/v) was used, and 265 nm was selected as detection wavelength. Tectoridin and 6"-O-xylosyl-tectoridin were used as the external standards in quality control of <it>P. lobata </it>flower for the first time. The method was applied to practical use in quality assessment of eleven batches of <it>P. lobata </it>flower samples in Chinese herbal medicine market.</p> <p>Results</p> <p>The peak area response was linear for tectoridin in the 11.8-236.4 μg/mL range with a correlation coefficient of 0.9996 (P < 0.001), and for 6"-O-xylosyl-tectoridin in the 10.33-185.99 μg/mL range with a correlation coefficient of 0.9984 (P < 0.001) respectively. The average recoveries were 102.7-103.7% for tectoridin and 95.7-103.2% for 6"-O-xylosyl-tectoridin (RSDs < 3%), and the intra-day and inter-day RSDs of the two components were less than 2%. This HPLC method was applied to assess the quality of <it>P. lobata </it>flower from eleven provinces in China. <it>P. lobata </it>flowers from northern China contained 26.46-43.28 mg/g of tectoridin and 30.90-48.23 mg/g of 6"-O-xylosyl-tectoridin comparing to 10.00-19.81 mg/g of tectoridin and 11.08-37.03 mg/g of 6"-O-xylosyl-tectoridin in those from southern China.</p> <p>Conclusion</p> <p>The results showed that <it>P. lobata </it>flowers from northern China contained more tectoridin and 6"-O-xylosyl-tectoridin than those from southern China.</p

Human Ecology, Process Philosophy and the Global Ecological Crisis

This paper argues that human ecology, based on process philosophy and challenging scientific materialism, is required to effectively confront the global ecological crisis now facing us

In vitro anti-Helicobacter pylori activity and the underlining mechanism of an empirical herbal formula – Hezi Qingyou

BackgroundHelicobacter pylori (H. pylori) is thought to primarily colonize the human stomach and lead to various gastrointestinal disorders, such as gastritis and gastric cancer. Currently, main eradication treatment is triple or quadruple therapy centered on antibiotics. Due to antibiotic resistance, the eradication rate of H. pylori is decreasing gradually. Therefore, searching for anti-H. pylori drugs from herbal sources has become a strategy for the treatment. Our team proposed a Hezi Qingyou Formula (HZQYF), composed of Chebulae Fructus, Ficus hirta Vahl and Cloves, and studied its anti-H. pylori activity and mechanism.MethodsChemical components of HZQYF were studied using UHPLC–MS/MS and HPLC. Broth microdilution method and agar dilution method were used to evaluate HZQYF’s antibacterial activity. The effects of HZQYF on expression of adhesion genes (alpA, alpB, babA), urease genes (ureE, ureF), and flagellar genes (flaA, flaB) were explored using Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) technology. Effects on morphology and permeability of the extracellular membrane were studied using scanning electron microscopy (SEM) and N-phenylnaphthalen-1-amine (NPN) uptake. Effect on urease activity was studied using a urease kinetics analysis in vitro. Immunofluorescence staining method was used to examine the effect on adhesion. Western blot was used to examine the effect on cagA protein.ResultsMinimum inhibitory concentration (MIC) values of the formula against H. pylori clinical strains and standard strains were 80–160 μg/mL, and minimum bactericidal concentration (MBC) values were 160–320 μg/mL. The formula could down-regulate the expression of adhesion genes (alpA, alpB, babA), urease genes (ureE, ureF) and flagellar genes (flaA, flaB), change the morphology of H. pylori, increase its extracellular membrane permeability, and decrease its urease activity.ConclusionPresent studies confirmed that HZQYF had promising in vitro anti-H. pylori activities and demonstrated its possible mechanism of action by down-regulating the bacterial adhesion, urease, and flagellar gene expression, which provided scientific bases for further clinical investigations

Simultaneous Determination and Pharmacokinetics Study of Six Triterpenes in Rat Plasma by UHPLC-MS/MS after Oral Administration of <i>Sanguisorba officinalis</i> L. Extract

A selective and sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for the determination of ziyuglycoside I (I), 3&#946;,19&#945;-dihydroxyurs-12-en-28-oic-acid 28-&#946;-d-glucopyranosyl ester (II), 3&#946;-[(&#945;-l-arabinopyranosyl) oxy]-urs-12,18(19)-dien-28-oic acid &#946;-d-glucopyranosyl ester (III), rosamultin (IV), 1&#946;-hydroxyeuscaphic acid (V) and alpinoside (VI) in rats after oral administration of Sanguisorba officinalis L. (S. officinalis) extract. The 3&#946;,19&#945;-dihydroxyurs-12-en-28-oic-acid 28-&#946;-d-glucopyranosyl ester, 3&#946;-[(&#945;-l-arabinopyranosyl) oxy]-urs-12,18(19)-dien-28-oic acid &#946;-d-glucopyranosyl ester, rosamultin, 1&#946;-hydroxyeuscaphic acid and alpinoside in rat plasma were the first report in the pharmacokinetics study in the present study. The analytes were quantified using the multiple reaction monitoring (MRM) mode with the electrospray ion source in positive electrospray ionization. Plasma was extracted with ethyl acetate via liquid&#8315;liquid extraction. Bifendate was used as internal standard (IS). The current method was validated for linearity, intra-day and inter-day precisions, accuracy, extraction recovery, matrix effect and stability. The lower limits of quantification of ziyuglycoside I, 3&#946;,19&#945;-dihydroxyurs-12-en-28-oic-acid 28-&#946;-d-glucopyranosyl ester, 3&#946;-[(&#945;-l-arabinopyranosyl) oxy]-urs-12,18(19)-dien-28-oic acid &#946;-d-glucopyranosyl ester, rosamultin, 1&#946;-hydroxyeuscaphic acid and alpinoside were 6.1, 4.9, 1.3, 3.8, 1.5 and 5.7 ng/mL, respectively. Intra-day and inter-day precision and the accuracy of the assay were in range from &#8722;9.48 to 12.74%. The extraction recoveries of analytes and bifendate (IS) from rat plasma ranged from 77.17% to 92.48%. Six compounds could be rapidly absorbed into blood (Tmax, 0.58&#8315;1.58 h), and then eliminated relatively slowly (t1/2, 6.86&#8315;11.63 h). The pharmacokinetic results might contribute to further guide the clinical application of S. officinalis

Bioconcentration and Metabolism of Emodin in Zebrafish Eleutheroembryos

Emodin is a major active anthraquinone of various herbal laxatives, which can exert many pharmacological effects. However, chronic use of anthranoid laxatives, even at low dosages, may cause melanosis coli (MC). It has been suggested that the accumulation of anthraquinones is a risk factor in the MC process. To investigate the accumulation of emodin, we conducted a bioconcentration study of emodin in zebrafish eleutheroembryos. Based on the economic cooperation and development (OECD) 305 test, zebrafish eleutheroembryos were exposed to emodin at a constant concentration for 48 h, before the test media were replaced by the blank medium for 24 h of depuration. To eliminate the effect of metabolism of emodin for assessment of the bioconcentration factor (BCF), we also conducted a modified test for which zebrafish eleutheroembryos were exposed to the non-renewed test media, whose emodin concentration decreased with time. At different exposure time points, zebrafish eleutheroembryos and exposure media were sampled for analysis of emodin concentration using HPLC-MS/MS. The results showed rapid accumulation of emodin in zebrafish eleutheroembryos to reach a steady-state concentration within 24 h. Meanwhile, emodin was actively metabolized by zebrafish eleutheroembryos to result in 29.5–40.7% of its elimination. In the groups with high or low concentrations of emodin, the standardized BCF (sBCF) values in the standard test were 24.0 and 20.0, while those in the modified test were 50.4 and 52.0. These results showed that emodin could accumulate in zebrafish eleutheroembryos when used for 48 h and beyond, suggesting that the accumulation of anthraquinones may be a risk factor in the MC process. Accordingly, emodin should be unsuitable for long-term use due to its accumulation

Syzygium aromaticum enhances innate immunity by triggering macrophage M1 polarization and alleviates Helicobacter pylori-induced inflammation

Previous studies suggested that Syzygium aromaticum dried buds (S. aromaticum), which is an edible and medicinal herb, possess potential effect on innate immunity and Helicobacter pylori (H. pylori) infection-related inflammation, but the specific action and especially the molecular mechanism have not been well elucidated. The objective of this work was to further explore the effects of Syzygium aromaticum aqueous extract (AE) on innate immunity and H. pylori related-inflammation, especially the potential molecular mechanisms involved. In order to accomplish this, analyses of the phagocytic rate, NO content, ROS level, as well as ELISA, RT-qPCR, and Western blotting et al. assays were carried out. Results indicated that AE increased phagocytic rate, ROS, and cytokine expression levels, meanwhile activated TLR4/MyD88 mediated NF-κB and MAPK pathways and decreased Nrf2/HO-1 signaling. AE also prevented the release of pro-inflammatory substances made by H. pylori-infected RAW 264.7 and GES-1 cells. All things considered, it was shown that AE's ability to boost innate immunity involves inducing macrophage M1 polarization by increasing TLR4/MyD88-mediated NF-κB and MAPK pathways and blocking Nrf2/HO-1 signaling. Additionally, by lowering the production of pro-inflammatory molecules, AE can lessen the inflammation brought on by H. pylori. Considering that S. aromaticum is an edible herbal medicine with immune-enhancing activity and good safety, it deserves further study and utilization for treating diseases including H. pylori infection-related disorders

Generation of Chicken IgY against SARS-COV-2 Spike Protein and Epitope Mapping

This new decade has started with a global pandemic of COVID-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), precipitating a worldwide health crisis and economic downturn. Scientists and clinicians have been racing against time to find therapies for COVID-19. Repurposing approved drugs, developing vaccines and employing passive immunization are three major therapeutic approaches to fighting COVID-19. Chicken immunoglobulin Y (IgY) has the potential to be used as neutralizing antibody against respiratory infections, and its advantages include high avidity, low risk of adverse immune responses, and easy local delivery by intranasal administration. In this study, we raised antibody against the spike (S) protein of SARS-CoV-2 in chickens and extracted IgY (called IgY-S) from egg yolk. IgY-S exhibited high immunoreactivity against SARS-CoV-2 S, and by epitope mapping, we found five linear epitopes of IgY-S in SARS-CoV-2 S, two of which are cross-reactive with SARS-CoV S. Notably, epitope SIIAYTMSL, one of the identified epitopes, partially overlaps the S1/S2 cleavage region in SARS-CoV-2 S and is located on the surface of S trimer in 3D structure, close to the S1/S2 cleavage site. Thus, antibody binding at this location could physically block the access of proteolytic enzymes to S1/S2 cleavage site and thereby impede S1/S2 proteolytic cleavage, which is crucial to subsequent virus-cell membrane fusion and viral cell entry. Therefore, the feasibility of using IgY-S or epitope SIIAYTMS-specific IgY as neutralizing antibody for preventing or treating SARS-CoV-2 infection is worth exploring