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1.
Article in Chinese | WPRIM | ID: wpr-976529

ABSTRACT

Objective To compare the contents variation of six flavonoids includingdaidzin,glycitin, genistin, daidzein, glycitein and genisteinin black beans, semifinished and finished Sojae Semen Praeparatum.Methods The contents of flavonoids were determined by HPLC,the condition were Diamonsil C18 column (4.6×250 mm, 5 μm) , column temperature 30 ℃, detection wavelength 260 nm, mobile phase 0.2% acetic acid water (A) - methanol (B), gradient elution, flow rate 1.0 ml/min.Results The linearity of this method to determine 6 isoflavones was good (r≥0.9993) within the determination range, and the recovery rate met the requirements. The RSD of precision, repeatability and stability experiment was less than 4%, 3%and 3%. The results of HPLC showed that the contents of six flavonoidsin Sojae Semen Praeparatum increased significantly compared with black beans. And, the contents of six flavonoids in finished Sojae Semen Praeparatum were slightly more than those in semifinished Sojae Semen Praeparatum. Conclusion The HPLC method established in this study could accurately determine the content of 6 isoflavones in Sojae Semen Praeparatum. The content of six isoflavones in black beans could be increased by the fermentation, and the combined isoflavones were transformed into free isoflavones during the fermentation process.

2.
Basic & Clinical Medicine ; (12): 1801-1807, 2023.
Article in Chinese | WPRIM | ID: wpr-1018544

ABSTRACT

Objective To investigate the impact of daidzein(DAI)on the pyroptosis of renal tubular epithelial cells induced by high glucose by regulating NOD-like receptor protein 3(NLRP3)/caspase-1 signal pathway.Methods HK-2 cells were divided into control group(NC group)(5.5 mmol/L D-glucose),HG group(30 mmol/L D-glucose),DAI-L,DAI-M,DAI-H groups(HK-2 cells were incubated with 30 mmol/L D-glucose and 25,50,100 μmol/L DAI,respectively),and DAI-H+LPS group(HK-2 cells were incubated with 30 mmol/L D-glucose,100 μmol/L DAI and 1 μg/mL LPS).MTT assay was applied to detect the cytotoxicity and proliferation of HK-2 cells;the apoptosis of HK-2 cells was detected by flow cytometry.The level of interleukin-1β(IL-1β)and inter-leukin-18(IL-18)in HK-2 cells was detected by ELISA.The morphology of pyroptosis cells was ob-served by scanning electron microscope.Immunofluorescence staining was applied to detect pyroptosis related pro-teins.The expression of NLRP3,cleaved casase-1 and GSDMD-N was detected by Western blot.Results In NC group,the cells were spherical with regular boundaries,while in HG group,the cells swelled and became larger with irregular boundaries;the OD value(490 nm)of HK-2 cells in HG group was obviously lower than that in NC group(P<0.05),the apoptosis rate,IL-1β,IL-18 contents,NLRP3,cleaved casase-1,GSDMD-N protein level of HK-2 cells were obviously higher(P<0.05);After DAI treatment,the swelling of cells was alleviated,the A value(490 nm)of HK-2 cells increased significantly(P<0.05),the apoptosis rate of HK-2 cells,IL-1 β,IL-18 content,NLRP3,cleaved-caspase-1 and GSDMD-N protein levels were significantly decreased(P<0.05)and the therapeutic effect of DAI was dose-dependent;LPS eliminated the beneficial effect of DAI-H on high glucose in-duced apoptosis of renal tubular epithelial cells.Conclusions DAI may alleviate pyroptosis of renal tubular epithe-lial cells induced by high glucose through inhibition of NLRP3/caspase-1 signaling pathway.

3.
China Pharmacy ; (12): 2556-2560, 2023.
Article in Chinese | WPRIM | ID: wpr-997019

ABSTRACT

Osteoporosis, as a systemic bone disease with high incidence rate and high disability rate, has become a research hotspot in recent years. The daidzein in soybean isoflavones can bind with estrogen receptors, simulating the prevention and treatment of osteoporosis with estrogen-like effect. Its mechanism of action includes promoting osteoblast formation and differentiation by activating the Wnt signaling pathway, increasing bone density, and improving bone tissue health; inhibiting osteoclast differentiation and slowing down bone resorption by reducing receptor activator of nuclear factors κB ligand/ osteoprotegerin ratio, downregulating the expression of macrophage colony-stimulating factor (M-CSF); collaborating antioxidant and immune regulation to achieve the goal of preventing and treating osteoporosis. In addition, different doses of daidzein have different effects on bone density and osteoporosis, which may be related to factors such as study design, sample selection, and individual differences.

4.
Chinese Pharmacological Bulletin ; (12): 431-438, 2023.
Article in Chinese | WPRIM | ID: wpr-1013830

ABSTRACT

Aim To investigate the effects of daidzeinDD on the proliferation and apoptosis of non-small cell lung cancer cells,with a focus on the possible role of the p53 signaling pathway in this regard. Methods CCK-8 method and flow cytometry were used to detect the effects of soy isoflavone crude extract and DD on the viability and apoptosis of HELF and H1299 cells. Gene microarray was used to detect the changes in gene expression after treatment of H1299 cells with DD. GSEA and differential analysis were used to screen the major pathways and key genes. RT-qPCR and Western blot were performed to verify the differences in mRNA and protein expression of key genesp53 and CASP9 in the major pathways. After p53 inhibitor Pifithrin-α inhibited the expression of p53,the effect of DD on p53 mRNA and protein expression levels was examined,and the proliferative effect on H1299 cells was observed. Results Soy isoflavone crude extract and DD promoted proliferation and inhibited apoptosis of normal lung cells and inhibited proliferation and promoted apoptosis of lung cancer cells. p53 signaling pathway was significantly enriched in the DD-treated groupNES=1.78,P=0.000,and the expressions of p53 and CASP9 genes were found to be significantly up-regulated in the treated group. Compared with the control group,mRNA expression of CASP9 and p53 significantly increased in both HELF and H1299 cells treated with DDP<0.05,and p53 protein expression also increased in HELF cellsP<0.05. After inhibition of p53 expression,DD significantly increased the mRNA expression of p53 in H1299 and HELF cellsP<0.05 and also markedly increased the expression of p53 protein in H1299 cellsP<0.05,and it was observed that DD inhibited the proliferation of lung cancer cells. Conclusions DD inhibits the proliferation and promotes the apoptosis of lung cancer H1299 cells,and the mechanism mainly involves the p53 signaling pathway.

5.
Zhongguo Zhong Yao Za Zhi ; (24): 5068-5077, 2023.
Article in Chinese | WPRIM | ID: wpr-1008677

ABSTRACT

This study investigated the drug delivery performance of oral co-loaded puerarin(PUE) and daidzein(DAZ) mixed micelles(PUE/DAZ-FS/PMMs) from the perspectives of pharmacokinetics, pharmacodynamics, and tissue distribution. The changes in PUE plasma concentration in rats were evaluated based on PUE suspension, single drug-loaded micelles(PUE-FS/PMMs), and co-loaded micelles(PUE/DAZ-FS/PMMs). Spontaneously hypertensive rats(SHR) were used to monitor systolic blood pressure, diastolic blood pressure, and mean arterial pressure for 10 weeks after administration by tail volume manometry. The content of PUE in the heart, liver, spleen, lung, kidney, brain, and testes was determined using LC-MS/MS. The results showed that compared with PUE suspension and PUE-FS/PMMs, PUE/DAZ-FS/PMMs significantly increased C_(max) in rats(P<0.01) and had a relative bioavailability of 122%. The C_(max), AUC_(0-t), AUC_(0-∞), t_(1/2), and MRT of PUE/DAZ-FS/PMMs were 1.77, 1.22, 1.22, 1.17, and 1.13 times higher than those of PUE suspension, and 1.76, 1.16, 1.08, 0.84, and 0.78 times higher than those of PUE-FS/PMMs, respectively. Compared with the model control group, PUE/DAZ-FS/PMMs significantly reduced systolic blood pressure, diastolic blood pressure, and mean arterial pressure in SHR rats(P<0.05). The antihypertensive effect of PUE/DAZ-FS/PMMs was greater than that of PUE suspension, and even greater than that of PUE-FS/PMMs at high doses. Additionally, the distribution of PMMs in various tissues showed dose dependency. The distribution of PMMs in the kidney and liver, which are metabolically related tissues, was lower than that in the suspension group, while the distribution in the brain was higher than that in the conventional dose group. In conclusion, PUE/DAZ-FS/PMMs not only improved the bioavailability of PUE and synergistically enhanced its therapeutic effect but also prolonged the elimination of the drug to some extent. Furthermore, the micelles facilitated drug penetration through the blood-brain barrier. This study provides a foundation for the development of co-loaded mixed micelles containing homologous components.


Subject(s)
Rats , Animals , Micelles , Tissue Distribution , Chromatography, Liquid , Tandem Mass Spectrometry , Rats, Inbred SHR , Isoflavones/pharmacology
6.
Zhongguo Zhong Yao Za Zhi ; (24): 2949-2957, 2023.
Article in Chinese | WPRIM | ID: wpr-981427

ABSTRACT

This study aims to improve the solubility and bioavailability of daidzein by preparing the β-cyclodextrin-daidzein/PEG_(20000)/Carbomer_(940) nanocrystals. Specifically, the nanocrystals were prepared with daidzein as a model drug, PEG_(20000), Carbomer_(940), and NaOH as a plasticizer, a gelling agent, and a crosslinking agent, respectively. A two-step method was employed to prepare the β-cyclodextrin-daidzein/PEG_(20000)/Carbomer_(940) nanocystals. First, the insoluble drug daidzein was embedded in β-cyclodextrin to form inclusion complexes, which were then encapsulated in the PEG_(20000)/Carbomer_(940) nanocrystals. The optimal mass fraction of NaOH was determined as 0.8% by the drug release rate, redispersability, SEM morphology, encapsulation rate, and drug loading. The inclusion status of daidzein nanocrystals was determined by Fourier transform infrared spectroscopy(FTIR), thermogravimetric analysis(TGA), and X-ray diffraction(XRD) analysis to verify the feasibility of the preparation. The prepared nanocrystals showed the average Zeta potential of(-30.77±0.15)mV and(-37.47±0.64)mV and the particle sizes of(333.60±3.81)nm and(544.60±7.66)nm before and after daidzein loading, respectively. The irregular distribution of nanocrystals before and after daidzein loading was observed under SEM. The redispersability experiment showed high dispersion efficiency of the nanocrystals. The in vitro dissolution rate of nanocrystals in intestinal fluid was significantly faster than that of daidzein, and followed the first-order drug release kinetic model. XRD, FTIR, and TGA were employed to determine the polycrystalline properties, drug loading, and thermal stability of the nanocrystals before and after drug loading. The nanocrystals loaded with daidzein demonstrated obvious antibacterial effect. The nanocrystals had more significant inhibitory effects on Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa than daidzein because of the improved solubility of daidzein. The prepared nanocrystals can significantly increase the dissolution rate and oral bioavailability of the insoluble drug daidzein.


Subject(s)
Sodium Hydroxide , Acrylic Resins , Escherichia coli , Nanoparticles
7.
Chinese Pharmacological Bulletin ; (12): 803-809, 2021.
Article in Chinese | WPRIM | ID: wpr-1014439

ABSTRACT

Aim To study the regulatory effect of daidzein on osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) expression in MG-63 cells and its mechanism. Methods RT-PCR, Western blot and siRNA were used to study the regulatory effect of daidzein on OPG and RANKL expression in human osteoblast-like MG-63 cells. Results Daidzein could promote the expression of OPG mRNA and protein in MG-63 cells and inhibit the expression of RANKL mRNA and protein, which could be blocked by ICI 182780. It was confirmed that ERa and ER0 mediated not only the promoting effect of daidzein on OPG expression of MG-63 cells but also the inhibition of RANKL. Conclusions Daidzein promotes OPG gene expression in MG-63 cells and inhibits the expression of RANK gene expression through ERa and ERβ pathways.

8.
Chinese Pharmacological Bulletin ; (12): 245-249, 2020.
Article in Chinese | WPRIM | ID: wpr-857024

ABSTRACT

Aim To investigate the effect of Daidzein ( Daid ) on proliferation and migration of non-small cell lung cancer cell lines A549 and HI299 and its possible mechanism. Methods CCK-8 was adopted to detect cell proliferation of A549 and HI299 cells inhibited by Daid (0, 5, 10, 25, 50, 100, 200

9.
Braz. J. Pharm. Sci. (Online) ; 56: e18388, 2020. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1089192

ABSTRACT

Parkinson disease is a neurodegenerative disorder characterised by the cardinal symptoms of stiffness, resting tremor, slowness (bradykinesia) and reduction of movement (hypokinesia). Involvement of oxidative damage has been reported in the pathophysiology of Parkinson disease and its related complications. The purpose of this study was to examine the effect of daidzein to quench the free radicals produced as a result of the increased oxidative stress in Parkinson disease.Parkinson disease is induced by administration of reserpine (5 mg/kg/day, i.p) for 5 consecutive days. The symptoms of PD such as tremors, akinesia and rigidity were evaluated. The effect was evaluated by assessing various behavioral parameters (grip strength and locomotor activity), biochemical parameters (lipid peroxidation, and reduced glutathione), as well as histopathological parameters in brain tissue. Daidzein (an antioxidant) was administered at the dose of 50 and 100 mg/kg, p.o. once daily for 5 days. Reserpine significantly causes tremor, rigidity, akinesia and oxidative damage which were reversed by daily administration of daidzein when compared toreserpine group. There was a significant histological improvement in the neuronal degeneration in brain tissue with daidzein. So, the results indicated the protective effect of daidzein against PD.

10.
Article in Chinese | WPRIM | ID: wpr-798492

ABSTRACT

Objective:To prepare daidzein nanosuspension capsules,and to investigate intestinal absorption and oral bioavailability by comparing with commercial daidzein capsules. Method:Daidzein nanosuspensions were prepared by precipitation method combined with high pressure homogenization,orthogonal design method was utilized to optimize its formulation.Daidzein nanosuspensions was characterized by X-ray powder diffraction(XRPD),Fourier transform infrared spectroscopy(FT-IR),transmission electron microscope(TEM),and indexes including mean particle size,polydispersity index(PDI),and Zeta potential.Intestinal absorption study was carried out to compare the accumulative permeated amount of daidzein from daidzein nanosuspensions and commercial daidzein capsules.Biodistribution of daidzein in gastrointestinal tract was investigated,and oral bioavailability was examined through pharmacokinetic study by HPLC. Result:The in vitro small intestinal absorption enhancement ratio of daidzein nanosuspension capsules was approximately 2.49-fold higher than that of commercial capsules(PConclusion:Daidzein nanosuspensions prepared by combined method can be applied to the production of capsules,which is beneficial to increase the absorption of drug in small intestine and improve its bioavailability after oral administration.

11.
Zhongcaoyao ; Zhongcaoyao;(24): 5753-5759, 2019.
Article in Chinese | WPRIM | ID: wpr-850668

ABSTRACT

Objective: In order to provide a scientific basis for the quality control of Yuquan Pills (YP), HPLC fingerprint strategy was established and six components were determined. Methods: The HPLC analysis was performed on Symmetery C18 column (250 mm × 4.6 mm, 5 μm), using acetonitrile and 0.1% phosphoric acid solution as the mobile phase at a flow rate of 1.0 mL/min, and the column temperature was 30 ℃. The fingerprints of 11 batches of YP were established and evaluated by the similarity evaluation system of TCM (version 2012A), clustering analysis and principal component analysis. Furthermore, the content of puerarin, daidzein, verbascoside, schisandrin, glycyrrhizic acid, and glycyrrhizin was determined. Results: The HPLC fingerprint with 12 common peaks of YP was established, and the similarities of samples were over 0.9. After validating the multiple component quantitative analysis condition through methodology, the average recoveries (n = 9) were between 92.06% and 109.34%, and the RSD were in the range of 0.22%-2.76%. The content of puerarin, daidzein, verbascoside, schisandrin, glycyrrhizic acid, and glycyrrhizin in 11 batches of YP were in the range of 0.838-2.777, 0.550-1.014, 0.312-0.618, 0.023-0.092, 1.154-1.674, 0.035-0.052 mg/g, respectively. Conclusion: The combination methods of HPLC fingerprint and simultaneous determinations of multiple components are rapid, simple and reproducible, which can provide methodological reference for the quality control of YP.

12.
Article in Chinese | WPRIM | ID: wpr-857314

ABSTRACT

Aim To study the effects of daidzein on sodium channel current ( /Na) in ventricular myocytes of rats and the mechanism of its antiarrhythmia. Methods The effect of daidzein on the viability of ventricular myocytes was delected by MTT assay; single ventricular myocytes from rats were isolated by single enzymatic hydrolysis; the changes of /N, and its dynamic characteristics in rat ventricular myocytes before and after administration of daidzein were observed, recorded and analyzed by cell patch clamp technique. Results MTT experiments showed that the ICjo of daidzein was 30 to 100 jjimol • L"1 ,so the concentration of 0. 3 - 10 jimol • L"1 was chosen for the subsequent experiments. When daidzein was given 0. 3,1,3,10 pjnol • L"',the /Nb amplitude of ventricular myocytes in rats showed a concentration-dependent inhibition. The concentration of daidzein 0. 3 imol • L"1 also had certain effect on the time course of /Nt. The /Nl, peak decreased gradually over time. The 1,3,10 jimol • L"1 daidzein raised the I-U curve obviously. Under the same condition, the activation curve moved to the direction of depolarization. The steady-state inactivation curve shifted toward hyperpolarization, and the t value of the recovery curve was prolonged after inactivated state. Conclusions Daidzein significantly inhibited the Na∗ channel of ventricular myocardium in rats, which may l)e one of its mechanisms of anti-arrhythmia.

13.
Zhongcaoyao ; Zhongcaoyao;(24): 2993-3000, 2018.
Article in Chinese | WPRIM | ID: wpr-851860

ABSTRACT

Objective: To establish simultaneous determination method of eight main flavonoids from Puerariae Lobatae Radix (PLR) by using UPLC, and to study the effect of ultrafine grinding technology on the content of flavonoids in PLR for optimizing its production process. Methods: Response surface method was used to investigate the crushing time, sampling capacity, and initial particle size in the superfine pulverizing technology of PLR. Using the particle size distribution (D50, D90), and the content of eight flavonoids (3′-hydroxy puerarin, puerarin, 3′-methoxy puerarin, daidzin, genistin, daidzein, genistein, and formononetin) from PLR powders with various particle sizes (10—40, 40—65, 300 mesh) determined by UPLC, we selected these 10 elements as the response factors to evaluate the ultrafine powders technology for PLR. Results: Eight flavonoids from PLR had a good linear relationship with the linear range of 75.8—242.7, 205.6—658.0, 147.3—417.3, 10.2—163.3, 11.3—182.0, 1.2—18.8, 0.25—4.00, 0.35—5.37 μg/mL, and the average recovery ranged from 98.86%, 99.25%, 99.90%, 100.17%, 100.21%, 101.40%, 100.73%, and 101.42%. In addition, the repeatability, stability, and precision RSD were all less than 3%. UPLC result showed that, compared with the PLR powders with other particle sizes, the ultrafine powder of PLR (300 mesh) had a higher flavonoids contents. Results: of response surface method showed the optimized preparing parameters of PLR ultrafine powder technology as follows: 80 mesh [(180 ± 7.6) μm] of particle size, 247 g of sampling amount, and 26 min of crushing time. Conclusion: Optimization of ultrafine powders process from PLR by response surface method is simple and accurate, which can obtain PLR ultrafine powder with higher content of flavonoids. This process can provide reference for the ultrafine grinding technology of PLR.

14.
Zhongcaoyao ; Zhongcaoyao;(24): 2013-2018, 2018.
Article in Chinese | WPRIM | ID: wpr-851993

ABSTRACT

Objective To study the ethyl acetate-soluble chemical constituents of Callicarpa kwangtungensis. Methods The chemical constituents were isolated by column chromatography on silica gel, ODS, Sephadex LH-20, and MPLC. Their chemical structures were elucidated on the basis of special analysis. Results Sixteen compounds were isolated and identified as carpeside B (1), loliolide (2), (3S,6E,10R)-10-β-D-glucopyranosyloxy-3,11-dihydroxy-3,7,11-trimethyldodeca-1,6-dine (3), lanceolatin A (4), daidzein (5), secroisolariciresinol (6), (7S,8R)-4,9,9'-trihydroxy-3,3'-dimethoxy-7,8-dihydrobenzofuran-1'-propylneolignan (7), silybin A (8), isosilybin A (9), isosilybin B (10), 2,3-dehydrosilyhin (11), silychristin A (12), silychristin B (13), isosilychristin (14), silydianin (15), and nigaichigoside F2 (16). Conclusion Compounds 4-15 are isolated from the genus Callicarpa for the first time.

15.
Zhongcaoyao ; Zhongcaoyao;(24): 1667-1676, 2018.
Article in Chinese | WPRIM | ID: wpr-852084

ABSTRACT

Objective: To determine the components of 3'-hydroxy puerarin, puerarin, daidzin, daidzein, genistein glycosides, and genistein from Pueraria Radix and to study the quality of medicinal materials combining with anti-oxidant activity. Methods: The contents of six main components from Pueraria Radix samples were determined by using UPLC-DAD method, which performed on AgiLent Zorbax SB-C18 (5 mm×2.1 mm, 1.8 μm), acetonitrile and 0.1% acetic acid aqueous solution as mobile phase, column temperature 30 ℃, and the flow rate of 0.4 mL/min. The anti-oxidant activity evaluation of DPPH was accomplished by using UV spectrophotometry. Comprehensive weighted evaluation was based on seven indicators, combined six kinds of ingredients with the IC50 of medicine, and made cluster analysis. Results: Studies had shown that the quality of No. 8, 9, 50, and 51 samples of Pueraria thomsonii was better than others, which collected from Jianyang and Jiangyou. And the quality of No. 19, 20, 29, 30, and 31 samples of cultivated-planting Pueraria omeiensis was poor. The quality of No. 33 ~ 37, 65 ~ 69 samples of Pueraria Radix was better than other places in Sichuan Province, which collected from Pingwu, Beichuan, and Qingchuan. Conclusion: The UPLC wavelength switching technology multicomponent simultaneous determination method is simple and convenient, which has good reproducibility, strong specificity. Comprehensive weighted evaluation method can analyze the quality of medicinal materials better, which provides references for comprehensive evaluation on the main varieties of Pueraria.

16.
Article in Chinese | WPRIM | ID: wpr-700388

ABSTRACT

Monocarboxylate transporter-8 (MCT8) is a specific thyroid hormone transporter, essential for the uptake of thyroid hormone into target tissues. Mutations in the MCT8 gene have been identified as the cause of Allan-Herndon-Dudley syndrome (AHDS). It has been reported that soy isoflavones influence thyroid hormone system and can interact with thyroid hormone transporter proteins. Therefore, the present study aimed to find out whether soy isoflavones (genistein, daidzein and glycitein) can be used as a natural inhibitor to target MCT8 in AHDS. Docking studies were performed for soy isoflavones in order to evaluate their binding affinity to MCT8 protein using AutoDock4 (version 4.2.6) and AutoDock Vina. After docking, the ligands were ranked according to their binding energy and the best lead compound was selected based on the least binding energy. The docking results indicated that daidzein possesses the lowest binding energy against MCT8. Moreover, it was found that the residues PRO-338, HIS-341, and GLU-348 were involved in hydrogen bond interactions with genistein and daidzein. This study suggests that daidzein is a promising natural inhibitor to target MCT8 in AHDS.

17.
Zhongcaoyao ; Zhongcaoyao;(24): 4167-4173, 2017.
Article in Chinese | WPRIM | ID: wpr-852447

ABSTRACT

Objective To establish the HPLC fingerprint of Liujing Toutong Tablets (LTT) and multi-wavelength method for determination of the contents of 3’-hydroxy puerarin, puerarin, daidzin, nuezhenide, and daidzein, so as to provide a reference for the quality control. Methods Using the method of HPLC, the fingerprint chromatography of high polarity and low polarity were established, with a mobile phase of acetonitrile-0.1% phosphoric acid and Diamonsil C18 column (250 mm × 4.6 mm, 5 μm). Furtherly, the Diamonsil C18 column was used with a mobile phase of acetonitrile-0.1% acetic acid gradient elution to determine the five contents. Results The fingerprint chromatography of high and low polarities with 11 batches of LTT were established, and the similarities of 11 batches were all over 0.90. The low polarity fingerprint chromatography included 16 mutual peaks, 12 of which belonged to herbs: No. 11, 12, 13 belonged to Angelicae Dahuricae Radix, No. 1, 2, 3, 7 belonged to Puerariae Lobatae Radix, No. 8 belonged to Ligustri Lucidi Fructus, No. 5, 6 belonged to Chuanxiong Rhizoma, No. 15 belonged to Ligustici Rhizoma et Radix and No. 4 belonged to Chuanxiong Rhizoma and Ligustici Rhizoma et Radix. Seven peaks including puerarin (No. 1), 3’-methoxy puerarin (No. 2), daidzin (No. 3), ferulic acid (No. 4), daidzein (No. 7), imperatorin (No. 11), and isoimperatorin (No. 13) were verified by standard compounds. The high polarity fingerprint chromatography included 10 mutual peaks, among them No. 2-9 belonged to Puerariae Lobatae Radix, No. 1, 10 belonged to Ligustri Lucidi Fructus. Five peaks including 3’-hydoxypuerarin (No. 2), puerarin (No. 3), 3’-methoxypuerarin (No. 4), daidzin (No. 7) and specnuezhenide (No. 10) were verified by standard compounds. The five active components were well separated and showed good lineaeity, such as 3’-hydroxy puerarin 71.60-716.00 ng (r = 0.999 1), puerarin 407.78-4 077.80 ng (r = 0.999 4), daidzin 90.72-907.20 ng (r = 0.999 9), nuezhenide 95.80-958.00 ng (r = 0.999 1), daidzein 21.98-219.80 ng (r = 0.999 9). The average recoveries and corresponding RSD values were 101.1% (1.38%), 97.8% (0.72%), 99.1% (0.75%), 97.8% (2.75%), and 98.7% (0.70%). The contents of 3’-hydoxypuerarin, puerarin, daidzin, specnuezhenide and daidzein in the 10 batches of sample were 3.08-3.84 mg/mL, 17.71-21.24 mg/mL, 3.51-4.71 mg/mL, 1.40-5.69 mg/mL, and 0.66-0.86 mg/mL, respectively. Conclusion The HPLC fingerprint combined with multi index determination method can reflect the intrinsic quality of LTT, which is stability, accurate, and reproducible, providing a scientific reference for quality control.

18.
Zhongcaoyao ; Zhongcaoyao;(24): 777-781, 2017.
Article in Chinese | WPRIM | ID: wpr-852986

ABSTRACT

Objective: To establish a method for simultaneously determination of five isoflavones (daidzein, daidzin, puerarin, mirificin, and 6″-O-xylosidepuerarin) in Pueraria Radix by single marker (QAMS), which is feasible and accurate. Methods: Puerarin was taken as internal standard substance to establish the relative correction factor (RCF) for quantitative analysis of multi-components with QAMS. Thus, the contents of mirificin, 6″-O-xylosidepuerarin, daidzin, and daidzein were calculated. The contents in 10 batches of samples were determined by external standard method and QAMS. The scientificalness and feasibility of the methods were evaluated by comparison of the quantitative results between external standard method and QAMS. Results: The reproducibility of RCF was perfect. The results calculated with QAMS were consistent with the results by the external standard method. Conclusion: QAMS is accurate and feasible to evaluate the quality of Pueraria Radix.

19.
Article in Chinese | WPRIM | ID: wpr-508732

ABSTRACT

Objective To observe the effect of daidzein on extracellular matrix of uremic rats and to discuss its mechanism.Methods Uremic rat model were established by 5/6 nephrectomized. Model rats were devided into daidzein group, control group. Rats with sham-operation were regarded as the normal control. At time of baseline, 4th and 8th week after operation, urinary protein and biochemical detection were measured. The pathologic changes, fibronectin (FN) and typeⅣcollagen (ColⅣ) were investigated at 8th week. The Western-Blot and RT-PCR were used to measure protein expression and mRNA transcription of TGF-β1. Results At 8th week after operation, the urinary protein (12.35 ± 2.13 mg/24 hvs. 19.93 ± 3.19 mg/24 h), serum urea (10.11 ± 0.65 mmol/Lvs.12.09 ± 0.78 mmol/L) and creatinine (68.10 ± 2.51μmol/Lvs.77.63 ± 3.20μmol/L) in the daidzein group decreased significantly than those in the control group (P<0.01). The deposition of ColⅣ (16.33% ± 2.14%vs. 24.68% ± 3.97%) and FN (19.17 ± 2.68 vs. 29.35 ± 4.15) in the daidzein group decreased significantly than those in the control group (P<0.01). Compared with control group, the pathological lesion in the daidzein group was less serious. The mRNA transcription (0.37 ± 0.06vs. 0.64 ± 0.08) and protein expression of TGF-β1 (0.28 ± 0.09vs. 1.40 ± 0.13) in the daidzein group were attenuated significantly than those in the control group (P<0.05). ConclusionsDaidzein had a beneficial effect on uremic rats. It may be associated with a decrease of extracellular matrix accumulation.

20.
Article in Chinese | WPRIM | ID: wpr-512391

ABSTRACT

To improve the water solubility of daidzein, solid inclusion complexes of daidzein with two amino-modified β-cyclodextrins (ACDs), i.e., mono-6-amino-6-deoxy-β-cyclodextrin (NCD) and mono-6-ethylenediamino-6-deoxy-β-cyclodextrin (ENCD), were prepared by the saturated solution method in water under the preparation conditions as follows: the ratio of daidzein/ACD was 3∶1 and the stirring time was 72 h (83% and 67% yields, respectively).The formation of two inclusion complexes was confirmed by x-ray diffractometry (XRD) and themogravimetric (TG) analysis.The inclusion stoichiometry of the inclusion complexes was 1∶1 from the Job plot and their complexation stability constants (KS) were 899.2 and 203.8 L/mol from fluorescence titration, respectively.After formation of inclusion complexes with NCD and ENCD, the water solubility of daidzein was dramatically raised from 8.31 μg/mL to 15.2 and 13.2 mg/mL at 25℃, increasing by 1800-fold and 1500-fold.

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