Study on UPLC fingerprint and multi-component content determination of Microctis Folium from different origins / 国际中医中药杂志

Objective:To establish the fingerprints of Microctis Folium by ultra high performance liquid chromatography (UPLC); To determine the contents of three flavonoids in the Microctis Folium; To evaluate the quality difference of Microctis Folium from different producing areas. Methods:The fingerprints were performed on Agilent ZORBAX SB C18 column (2.1 mm×150 mm,1.8 μm). The mobile phase was acetonitrile - 0.1 % acetic acid solution with gradient elution at a flow rate of 0.30 ml/min. The column temperature was 30 ℃ and the detection wavelength was 315 nm. The common fingerprint peaks were identified by UPLC-mass spectrometry, and the identification results were confirmed by comparison of reference materials. Waters Cortecs T3 C18 chromatographic column (2.1 mm × 100 mm,1.6 μm) was used for content determination. The mobile phase was methanol-0.1 % formic acid solution with gradient elution at a flow rate of 0.35 ml/min. The column temperature was 30 ℃ and the detection wavelength was 339 nm. The contents of vitexin, isovitexin and narcissoside in 15 batches of Microctis Folium from different habitats were determine. Results:There were 11 common peaks in the fingerprint of Microctis Folium. Identified by mass spectrometry and confirmed by reference substance,10 chemical components were identified, including caffeic acid, p-hydroxycinnamic acid, ferulic acid, vitexin, isovitexin, kaempferol-3-O-rutoside, astragaloside, narcissoside, isorhamnetin-3-O-glucoside and linden glycoside. The similarity between the fingerprints of 15 batches of Microctis Folium and the control fingerprint was greater than 0.95, indicating that the overall similarity of the fingerprints of Microctis Folium from different producing areas was high. The total contents of three active components were 3.23-5.64 mg/g in Yangjiang City, Guangdong, 3.60-5.78 mg/g in Zhanjiang City, Guangdong, 4.68-5.73 mg/g in Yulin City, Guangxi and 3.87-5.21 mg/g in Wuzhishan City, Hainan . There was no significant difference in the content of three active components in different producing areas. Conclusion:The fingerprints and the determination method established in the study are stable and feasible, which can be used for the quality evaluation of Microctis Folium.

Network pharmacology study of Yinqiao Jiedu Soft Capsules in treatment of COVID-19 / 中草药

Objective: To explore the mechanism of Yinqiao Jiedu Soft Capsules in the treatment of coronavirus disease 2019 (COVID-19). Methods: The interactions between 1 418 compounds of Yinqiao Jiedu Soft Capsules and 48 inflammatory target proteins related to COVID-19 were analyzed by molecule docking. The drug-target network was established to clarify the active compounds and potential targets. Results: The network analysis suggested 50 active compounds of Yinqiao Jiedu Soft Capsules, which were mainly flavonoids and triterpenoids, and 37 potential targets, mainly including MTOR, JAK3, ACE, ACE2, PIK3CA, TNF, AKT2, and MAP2K1. The results of molecular docking exhibited that forsythiaside and vitexin 2″-O-rhamnoside had good affinity with SARS-CoV-2 3CL hydrolase, and glycyrrhizic acid had good affinity with ACE2. Conclusion: The molecular mechanism of Yinqiao Jiedu Soft Capsules for COVID-19 may be involved in interfering SARS-CoV-2 replication and regulating the expression of inflammatory signaling pathway and the secretion of inflammatory cytokines.

Preliminary study on Q-markers of anti-tumor activity extracts of Polygonum orientale / 中草药

Objective: Based on the concept of quality marker (Q-marker), the components and the quality of the ethyl acetate extract of Polygonum orientale (POEa) was analyzed and studied. Methods: Firstly, the components of POEa were identified using the UPLC-ESI-HRMS method and standard compounds. Secondly, the main active compounds were determined by HPLC. Antitumor activities of these compounds were reviewed and its Q-marker was predicted. Finally, we evaluated the effects of POEa and the compound of gallic acid, isoquercetin, valerin, vitexin, luteolin, and quercetin on proliferation, apoptosis and migration of A549 cells. Results: A new quality method for simultaneous determining these six compounds of POEa was established. The six chemical ingredients were detected in each sample and the total content was more than 10%. The number of apoptotic cells in A549 cells treated with POEa and six chemical mixtures were all substantial increased, and the migration amount were significantly decreased. Tow groups showed no significantly differeances. Conclusion: The six components are scientific and reasonable to be considered as potential Q-marker represented the anti-tumor activity of POEa. The HPLC method can be used as accurate and stable quality control strategy of POEa.

Toxicokinetics of Vitexin Injection in Beagle Dogs / 中国药学杂志

OBJECTIVEP: To investigate the toxicokinetic and tissue distribution of vitexin in Beagle dogs. METHODS: Beagle dogs were randomly divided into three groups, and received vitexin injection at small, medium and big doses of 50, 20 and 8 mg•kg-1. They were given medicine once a day for consecutive 3 months by intravenous drip. The blood samples of Beagle dogs were drawn at different time points on the first and last day of administration, and concentrations in plasma were detected by HPLC method. RESULTS: Intravenous drip the vitexin injection at the doses of 8, 20 and 50 mg•kg-1, the blood concentration of vitexin linearly metabolized in Beagle dogs when given medicine for 1, 22, 44 and 83 d. Vitexin was significantly accumulated in Beagle dogs, and the accumulation was disappeared, and the exposure decreased with the prolonged time at the dose of 50 mg•kg-1; at the dose of 8 and 20 mg•kg-1, vitexin did not accumulate in Beagle dogs, and the exposure decreased with prolonged administration time. CONCLUSION: There is no accumulation of repeated drug delivery in the Beagle dog's body by intravenous drip at the doses of 8 and 20 mg•kg-1.

Studies on chemical constituents from Isodon eriocalyx distributed in Hezhang / 中草药

Objective: To study the chemical constituents of Isodon eriocalyx. Methods The chemical constituents were isolated and purified by chromatography with silica gel, Sephadex LH-20 and semi-preparative HPLC, and their structures were identified by analysis of spectroscopic data and physicochemical properties as well as relevant references. Results :A total of 20 constituents were isolated from I. eriocalyx, plant material extracted with ethyl aceta, and they were elucidated as maoecrystal D (1), isothymusin (2), coetsoidin A (3), maoecrystal A (4), 5,4’-dihydroxy-6,7-dimethoxy flavone (5), vitexin (6), α-amyrin (7), β-daucosterol (8), odinicin (9), cirsimaritin (10), maoecrystal N (11), epi-macoecrystal P (12), pimaric acid (13), laxiflorin R (14), enmelol (15), maoecrystal B (16), pectolarigenin (17), neolaxiflorin V (18), eriocalyxin A (19), and neorabdosin (20). Conclusion: Compounds 2, 5-7, 13-15, 17, 18 are isolated from this plant for the first time, and compounds 5 and 6 are isolated from the plants of Isodon for the first time.

Neuroprotective effect of vitexin on acute cerebral ischemia reperfusion rats and its effect on Th1/Th2 drift / 中草药

Objective: To investigate the neuroprotective effect of vitexin (VT) on acute cerebral ischemia (ACI) reperfusion rats and its effect on helper T cell 1 (Th1)/Th2 drift. Methods: Rat models of ACI reperfusion were established and divided into model group, low, medium and high doses of VT (0.94, 1.88, 3.76 mg/kg) groups and sham operated group. After 1 h of ischemia-reperfusion, rats in different doses of VT were given different concentrations of VT by ip, and rats in sham operated group and model group were given the same amount of saline by ip for three consecutive days. The general state of rats was observed. Longa neurological score before and after administration, and morphological changes of neurons in brain tissue after administration were evaluated and compared. The damage rates of single and double strands breaks of DNA in brain tissue were compared. The levels of Th1 and Th2 markers interferon-γ (INF-γ) and interleukin-4 (IL-4) in brain tissue were measured and the INF-γ/IL-4 was calculated. Results: The success rate of ACI reperfusion model was 88.89%. The mental state of rats in the model group was not good, which was improved in the three doses of VT groups after administration. Compared with model group, the neurological function scores of rats in each dose group of VT were significantly decreased 1 d and 3 d after administration (P < 0.05) in a dose-dependent manner. The results of HE showed that the volume of neurons and nucleus of neurons in model group were reduced, and the injury of neurons in VT groups was alleviated after administration, especially in high dose group. Compared with the sham operated group, the damage rates of single and double strand breaks of DNA, INF-γ level and INF-γ/IL-4 in the model group were significantly higher (P < 0.05), while IL-4 level was significantly lower (P < 0.05). Compared with the model group, the rate of DNA single- and double-strand breakage damage, INF-γ level and INF-γ/IL-4 were significantly decreased (P < 0.05) and the level of IL-4 was significantly increased (P < 0.05) in each dose group of VT in a dose-dependent manner. Conclusion: VT has protective effect on nerve function in ACI reperfusion rats, and with the best protective effect at dose of 3.76 mg/kg, which may be related to regulating the balance of Th1/Th2 cells to shift to Th2 and alleviating DNA damage in brain cells.

Vitexin induces apoptosis through mitochondrial pathway and PI3K/Akt/mTOR signaling in human non-small cell lung cancer A549 cells

BACKGROUND: Currently, the prognosis of patients with non-small cell lung cancer (NSCLC) remains dismal; hence, it is critical to identify effective anti-NSCLC agents with limited side effects. This study aimed to evaluate the therapeutic potential of flavonoid compound vitexin in human NSCLC cells and the underlying mechanisms. RESULTS: The experimental results indicated that vitexin reduced the viability of A549 cells in a dose-dependent manner with nearly no toxicity against normal human bronchial epithelial 16HBE cells. Vitexin also dose-dependently increased A549 cell apoptosis, accompanied by the decreased Bcl-2/Bax ratio and the increased expression of cleaved caspase-3. Moreover, the in vivo anticancer activity of vitexin was further determined in nude mice bearing A549 cells. In addition, vitexin induced the release of cytochrome c from the mitochondria to the cytosol and the loss of mitochondrial membrane potential. Vitexin also significantly reduced the levels of p-PI3K, p-Akt and p-mTOR, and the pro-apoptotic effect of vitexin on A549 cells was partly blocked by SC79, an Akt activator. CONCLUSIONS: Accordingly, we believed that vitexin could be used as a potential therapeutic agent for the treatment of NSCLC in the future.

Quantitative research on multiple components in Qingreling Granules based on UHPLC-ESI-HRMSn / 中草药

Objective To establish a quantitative analysis method of multiple active components liquiritin, vitexin, baicalin, rutin, cryptochlorogenic acid, chlorogenic acid, quercetin, and kaempferol in Qingreling Granules (QG) based on UHPLC-ESI-HRMSn, in order to provide a comprehensive evaluation for the quality control of QG. Methods The chromatographic separation was carried on BEH C18 (100 mm × 2.1 mm, 1.7 μm) column with methanol-0.1% formic acid water as mobile phase at the flow rate of 0.3 mL/min. Full scan mode with an electrospray ionization (ESI) source was used for the detection. The quantitative determination results were calculated by the pattern recognition function of the software SIMCA 14.1 to evaluate the quality of QG. Results Liquiritin, baicalin, rutin, vitexin, quercetin, chlorogenic acid, cryptochlorogenic acid, and kaempferol all showed good liners relationship (r ≥ 0.999 0) in the ranges of 570-9 127, 10 032-160 500, 293-4 690, 1 625-26 000, 40.5-645, 41-1 325, 44-1 413, and 13-209 ng/mL, respectively. The precision, repeatability, and stability were all up to the standards. The recoveries of standard addition was 98.83% to 100.65% with precision of below 3% RSD (n = 5). The average mass fractions of liquiritin, baicalin, rutin, vitexin, quercetin, chlorogenic acid, cryptochlorogenic acid, and kaempferol in five batches of QG were 202.07-438.15, 10 258.03-11 046.56, 56.09-87.7, 689.19-818.56, 4.95-6.0, 8.87-18.37, 22.49-42.12, 3.21-4.11 μg/g, respectively. The data analyzed by SIMCA 14.1 showed that the quality deviation of five batches of QG were within ± 2. Conclusion The method established in this study is simple, rapid, sensitive and accurate. The results of methodology conform to the relevant requirements and the method can be used as a quantitative method for the active ingredients in QG. The research also provides a new basis for the quality control at the same time.

Chemical pattern recognition on contents of five flavonoids and anti-oxidant activity of Spirodelae Herba from different growing areas / 中草药

Objective: To determine the contents of five flavonoids in Spirodelae Herba from different growing areas, and evaluate its anti-oxidant activity. Methods: Contents of orientin, vitexin, cynaroside, apigenin-7-glucoside, and luteolin in 17 batches of Spirodelae Herba from eight origins were determined by HPLC with acetonitrile and 0.1% formic acid as mobile phase, and DPPH method was used to compare their anti-oxidant activities, together with cluster analysis and principal component analysis, the quality of Spirodelae Herba was evaluated. Results: The contents of five flavonoids in Spirodelae Herba from different growing areas exist certain differences, which can be divided into three different categories by cluster analysis and principal component analysis. The contents of orientin, vitexin, and cynaroside were significantly higher in samples from Shaanxi and Shanxi provinces than those from other origins, as well as their oxidation clearance rates were relatively high. The correlation analysis showed there was a significant positive correlation between the rate of oxidative elimination and flavonoid content. Conclusion: Spirodelae Herba from Shaanxi and Shanxi provinces has higher contents of flavonoids with strong antioxidant capacity. The analytical method is stable and reliable, which can provide the reference for enhancing the quality control of Spirodelae Herba.

Analysis on components exposing in rat plasma after oral administration of Baoerkang Powder / 中草药

Objective: To analyze the components exposing in rat plasma after oral administration of Baoerkang Powder, and to study the mechanism of Baoerkang Powder and pharmacokinetic behavior. Methods: Components absorbed in rat plasma after oral administration of Baoerkang Powder to rats were analyzed by UPLC tandem High-resolution mass spectrometer. The structures of Baoerkang Powder in rat plasma identified by comparing the retention time, molecular weight, and CID fragmentation patterns with their corresponding compounds reported in the literatures. Results: Twenty-three components in rat plasma were identified after oral administration of Baoerkang Powder to rats for 1 h, including five components confirmed by comparing retention time and information of mass with their reference substances, components confirmed were vitexin, liquiritigenin, hesperetin, glycyrrhetic acid, and ursolic acid. Conclusion: It is suggested that the method could be applied to quick analysis of the components exposing in rat plasma after oral administration of Baoerkang Powder, which is beneficial to studying its mechanism and pharmacokinetic behavior.

Chemical Constituents from Stem Barks of Pandanus tectorius / 中国药学杂志

OBJECTIVE: To study the chemical constituents in the stem barks of Pandanus tectorius Soland. METHODS: The compounds were isolated by repeated chromatography. Their structures were elucidated by spectroscopic methods. RESULTS: Ten compounds were isolated and identified as 1'-O-benzyl-α-L-rhamnopyranosyl-(1″→6')-β-D-glucopyranoside(1), dihydrodehydrodiconiferyl alcohol(2), isovitexin(3), vitexin(4), 3,5-di-O-caffeoylquinic acid methyl ester(5), 3,5-di-O-caffeoylquinic acid ethyl ester(6), 3,4-di-O-caffeoylquinic acid methyl ester(7),(+)-lyoniresinol 3a-O-β-glucopyranoside(8),(-)-lyoniresinol 3a-O-β-glucopyranoside(9), and benzyl-β-D-glucopyranoside(10). CONCLUSION: All compounds are isolated from this plant for the first time.

Contents Determination of Chlorogenic Acid and Vitexin in Lophatherum gracile by HPLC / 中国药房

OBJECTIVE:To establish a method for the simultaneous determination of chlorogenic acid and vitexin in Lophather-um gracile. METHODS:HPLC was performed on the column was Waters Atlantis C18 with mobile phases of acetonitrile- water (gradient elution)at a flow rate of 1.0 ml/min,detection wavelength was 280 nm,column temperature was 35 ℃,and the injec-tion volume was 10 μl. RESULTS:The linear range was 0.041 0-1.228 8 μg for chlorogenic acid(r=0.999 8)and 0.264 0-7.920 0μg for vitexin(r=0.999 9);RSDs of precision, stability and reproducibility tests were lower than 2%;recoveries were 97.6%-102.3%(RSD=1.85%,n=9) and 97.1%-101.3%(RSD=1.19%,n=9),respectively. CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous determination of chlorogenic acid and vitexin in L. gracile.

Quantitative determination of vitexin in Passiflora foetida Linn. leaves using HPTLC

Objective: To establish a simple, rapid, precise and accurate high performance thin layer chromatography (HPTLC) method with densitometric detection for the determination of vitexin in Passiflora foetida Linn. (P. foetida). Methods: Ethanolic extract of the plant leaf powder was used for the experimental work. Separation was performed on silica gel 60 F

Study on the First-Pass Effects of Vitexin-4′-O-Glucoside in Rats in vivo / 中国药房

OBJECTIVE:To investigate the first-pass effect and mechanism of vitexin-4′-O-glucoside(VG)in rats so as to pro-vide a basis for new drug development. METHODS:10 SD rats were divided into a group of hepatic portal venous administration and a group of femoral venous administration,which respectively received VG iv at superior mesenteric vein and femoral vein,and then metabolic rate was calculated by finding out the AUC of VG in the rats’livers. 15 SD rats were divided into a group of gastric infusion,a group of intestinal infusion and a group of hepatic portal venous infusion,which respectively received VG by infusion at gastric fundus and duodenum and iv at superior mesenteric vein,and then metabolic rate was calculated by finding out the AUC of VG in the rats’stomachs and intestines. 15 SD rats were divided into a group of intestinal infusion,a group of femoral venous administration and a group of normal saline. At 10 min before administration,the former two groups were given by infusion vera-pamil injection(60 ml/kg),the substrate of CYP3A and P-glycoprotein(P-gp);and the group of normal saline were given by infu-sion of isometric normal saline,and then the rats were given VG as above to observe the effect of verapamil on intestinal absorp-tion of VG. RESULTS:The metabolic rates of VG in the liver,stomach and intestine were 54.9%,1.7% and 91.9% respectively. After infusion of verapamil,slight increase in AUC of VG was found in the rats in the group of intestinal infusion. CONCLU-SIONS:The first-pass effects in the liver and intestine are the main factors related to the low bioavailability of VG. Based on pre-liminary judgment,VG is the substrate of intestinal CYP3A and/or P-gp.

Research progress on neuroprotective effects of vitexin and its mechanisms / 中国药理学通报

Vitexin, a natural flavonoid compound, has extensive pharmacological activity. In recent years, many studies have re-vealed that vitexin has significant protective effects on central nervous system and peripheral nervous system, including anti-memory impairment, anti-epilepsy, anti-ischemic hypoxic brain damage, anti-depression, analgesia, etc. Vitexin exerts neuro-protective effects through the mechanisms of multiple pathways and multi-targets, such as reducing free radical level, inhibiting neuronal apoptosis, modulating inflammatory factors and related pathways, and regulating neurotransmitters and related recep-tors. This review mainly discusses the neuroprotective effects of vitexin and its underlying mechanisms.

Virtual screening for potential COX-inhibiting constituents from Mimosa pudica.

Developing a new agent in the anti-inflammatory and analgesic field, plants secondary metabolites can be a good source for the Non-Steroidal Anti-inflammatory Drugs (NSAID) drug development. For this purpose we subjected the active compounds of Mimosa pudica Linn. to reveal its potentiality by molecular docking analysis to find out its potent compound against COX which was done by GOLD docking analysis. Docking studies by GOLD showed that vitexin of Mimosa pudica had the highest fitness score against the COX-1 which is 60.43 and 63.49 for COX-2 enzyme. Vitexin of Mimosa pudica detected with significant fitness score and hydrogen bonding against COX-1 and COX-2 which may be a potent analgesic compound.

Lifespan Extending and Stress Resistant Properties of Vitexin from Vigna angularis in Caenorhabditis elegans

Several theories emphasize that aging is closely related to oxidative stress and disease. The formation of excess ROS can lead to DNA damage and the acceleration of aging. Vigna angularis is one of the important medicinal plants in Korea. We isolated vitexin from V. angularis and elucidated the lifespan-extending effect of vitexin using the Caenorhabditis elegans model system. Vitexin showed potent lifespan extensive activity and it elevated the survival rates of nematodes against the stressful environments including heat and oxidative conditions. In addition, our results showed that vitexin was able to elevate antioxidant enzyme activities of worms and reduce intracellular ROS accumulation in a dose-dependent manner. These studies demonstrated that the increased stress tolerance of vitexin-mediated nematode could be attributed to increased expressions of stress resistance proteins such as superoxide dismutase (SOD-3) and heat shock protein (HSP-16.2). In this work, we also studied whether vitexin-mediated longevity activity was associated with aging-related factors such as progeny, food intake, growth and movement. The data revealed that these factors were not affected by vitexin treatment except movement. Vitexin treatment improved the body movement of aged nematode, suggesting vitexin affects healthspan as well as lifespan of nematode. These results suggest that vitexin might be a probable candidate which could extend the human lifespan.

HPLC Simultaneous Determination of Vitexin and Isovitexin inMelastoma Dodecandrum / 世界科学技术-中医药现代化

This study was aimed to develop HPLC for determination of vitexin and isovitexin inM. dodecandrum. The HPLC column was SunFireTM C18 (4.6 mm× 150 mm, 5μm). The detection wavelength was 365 nm. The mobile phase was methanol-0.2% formic acid aqueous solution. The column temperature was 40℃. The flow rate was 1.0 mL·min-1. The results showed that the regression equations of vitexin and isovitexin wereY = 1× 106X– 14 396, Y = 1× 106X– 13 900, respectively. The linear ranges were 0.210μg - 1.050μg (r = 0.999) and 0.186μg - 0.930μg (r = 1.000), respectively. The recovery rates were 97.48% and 104.64%, respectively. The RSD were 2.32%and 1.51%, respectively. The sample contents of vitexin and isovitexin were 1.25 and 1.86 mg·g1, respectively. It was concluded that the method was simple, feasible and reproducible for the content determination of vitexin and isovitexin inM. dodecandrum.

Simultaneous determination of six constituents in rukuaixiao granules by UHPLC-MS/MS / 中国药学杂志

OBJECTIVE: To establish a UHPLC-MS/MS method for simultaneous determination of tangertin, hesperetin, 5-demethylnobiletin, nobiletin, vitexin, and narirutin in Rukuaixiao granules. METHODS: The analysis method was performed on a Waters ACQUITY UPLC® BEH C18 eolumn (2.1 mm×50 mm, 1.7 μm) with gradient elution of acetonitrile (containing 0.1% formic acid)-0.1% formic acid at a flow rate of 0.6 mL·min-1, and the column temperature was set at 50℃. Electrospray ionization (ESI) source with positive mode and the detector with multiple reaction monitoring (MRM) mode were adopted to determine the analytes by mass spectormeter. RESULTS: Satisfactory linearities were obtained for the six constituents in the investigated ranges (r>0.9958): The lower limit of quantitation (LOQ) was 0.80, 1.10, 1.30, 0.66, 2.00, 7.10 and 0.70 ng·mL-1, respectively. All the recoveries of samples were between 95.78%-101.35% with RSDs less than 2.1%. CONCLUSION: The method is simple, rapid, sensitive, and highly reproducible. It can be applied to the quality control of Rukuaixiao granules.

Separation and Content Comparison of Vitexin from Anemarrhenae Rhizoma Before and After Salt Processing / 中国中医药信息杂志

Objective To study the chemical components of Anemarrhenae Rhizoma;To establish a method to determine the content of vitexin;To compare the content difference of vitexin from Anemarrhenae Rhizoma before and after salt processing. Methods 70% ethanol extract of Anemarrhenae Rhizoma was separated and purified by silica gel column chromatography technique, and compound was identified by physicochemical properties and spectral data. HPLC was used with Ecosil column (4.6 mm×150 mm, 5μm), mobile phase of acetonitrile-0.2%acetic acid solution (14.5∶85.5), velocity of 1.0 mL/min, and determine wavelength of 340 nm. Results The compound was vitexin, a good linearity (r=0.999 8) in the range of 0.039 8-1.99μg. The average recovery rate was 98.02%, RSD=0.21%. Vitexin content was 0.008 5% in crude Anemarrhenae Rhizoma, 0.008 1% in processed products. Conclusion This is the first time separation of vitexin from Anemarrhenae Rhizoma. The method for content determination of vitexin is accurate, specific, and highly sensitive in the present experiment. Content of vitexin decreases slightly in processed products of Anemarrhenae Rhizoma compared with crude Anemarrhenae Rhizoma.