HPLC determination of the content for six constituents before and after salt processing of Plantaginis Semen from different habitats / 国际中医中药杂志

Objective:To compare the content change of 6 constituents in Plantaginis Semen from different habitats before and after salt processing. Methods:HPLC method was used to quantitatively analyze 6 ingredients in Plantaginis Semen and processed with salt including geniposidic acid, plantagoguanidinic acid, quercetin, kaempferol, verbascoside and isoverbascoside. Results:The geniposidic acid, plantagoguanidinic acid, quercetin, kaempferol, verbascoside and isoverbascoside were well separated. The linear ranges of which were 0.259 2-3.628 8 μg ( r=0.999 8), 0.054 3-0.760 5 μg ( r=0.999 6), 0.030 0-0.420 6 μg ( r=0.999 4), 0.055 6- 0.777 8 μg ( r=0.999 5), 0.287 0-4.018 0 μg ( r=0.999 8), 0.033 1-0.463 1 μg ( r=0.999 7), respectively. Average recovery rates were 98.68%, 98.46%, 98.87%, 98.99%, 98.34%, 98.75% ( n=6), respectively. There were mild differences in the contents of 6 ingredients of 8 batches of Plantaginis Semen from 5 different habitats. There were no obvious differences between the raw products and the products after salt process in Plantaginis Semen. The content of flavonoids, geniposidic acid and isoverbascoside in Plantaginis Semen were significantly increased after salt process, while the content of verbascoside was reduced. Conclusion:HPLC method to quantitatively analyze the 6 constituents in Plantaginis Semen before and after salt process could provided a reference for the quality change and the material basis for the efficacy of Plantaginis Semen before and after salt process.

Effect of geniposidic acid on hepato-enteric circulation in cholestasis rats through Sirt1-FXR signaling pathway / 中国中药杂志

To investigate the effects of geniposidic acid( GPA) on hepato-enteric circulation in cholestasis rats,and to explore the mechanism based on the sirtuin 1( Sirt1)-farnesol X receptor( FXR) pathway,sixty SD rats were randomly divided into 6 groups:blank control group,ANIT model group,ursodeoxycholic acid group( 100 mg·kg~(-1)·d-1 UDCA),and GPA high,medium and low( 100,50 and 25 mg·kg~(-1)·d-1) dosage groups,10 rats in each group. Corresponding drugs were intragastrically( ig) administered for10 days. After administration on day 8,all rats except blank rats were administered with 65 mg·kg~(-1)α-naphthalene isothiocyanate( ANIT) once. After the last administration,the serum levels of alanine aminotransferase( ALT),glutamine oxalacetate aminotransferase( AST),gamma-glutamyltransferase( γ-GGT),alkaline phosphatase( ALP),total bilirubin( TB) and total bile acid( TBA)were measured,and the mRNA transcription levels of Sirt1,FXR,multidrug resistant associated protein 2( MRP2),bile salt export pump( BSEP),sodium taurocholate contractible polypeptide( NTCP) in liver and apical sodium bile acid transporter( ASBT),ileum bile acid binding protein( IBABP) in ileum were detected by reverse transcription-polymerase chain reaction( RT-PCR). The protein expression levels of Sirt1,FXR and NTCP were detected by Western blot; the expression of MRP2,BSEP in liver and ASBT,IBABP in ileum were determined by immunofluorescence three staining. Primary rat hepatocytes were cultured in vitro to investigate the inhibitory effect of GPA on a potent and selective Sirt1 inhibitor( EX 527),and the mRNA and protein expression levels of Sirt1 and FXR were detected by RT-PCR and Western blot. GPA significantly decreased the levels of ALT,AST,γ-GGT,ALP,TB,TBA in serum( P<0.01) and improved the pathological damage of liver tissues in ANIT-induced cholestasis rats; significantly increased the mRNA and protein expression levels of Sirt1,FXR,MRP2,BSEP,NTCP in liver and ASBT,IBABP in ileum( P< 0.01). In vitro primary hepatocytes experiment indicated that the gene and protein expression levels of FXR and Sirt1 were noticeably improved by GPA in primary hepatocytes inhibited by EX-527( P<0.01). It was found that the improvement of GPA was in a dose-dependent manner. GPA could improve bile acid hepatointestinal circulation and play a liver protection and cholagogu role in cholestasis rats induced by ANIT.The mechanism may be that GPA activated FXR by regulating Sirt1,a key regulator of oxidative stress injury,and then the activated FXR could regulate protein of bile acid hepato-enteric circulation.

Simultaneous Determination of Contents of 8 Components in Eucommia ulmoides Leaves by HPLC / 中国药房

OBJECTIVE： To establish content determination method for simultaneously determining aucubin，geniposidic acid，catechin，chlorogenic acid，asperuloside，rutin，isoquercitrin and astragalin in Eucommia ulmoides leaves. METHODS：HPLC method was adopted. The determination was performed on Agilent ZORBAX SB-C18 column with the mobile phase consisted of acetonitrile-0.1％ phosphoric acid（gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 203 nm for aucubin and catechin，239 nm for geniposidic acid and asperuloside，220 nm for chlorogenic acid，354 nm for rutin and isoquercitrin，and 266 nm for astragalin. The sample size was 5 μL. RESULTS： The linear range of aucubin， geniposidic acid， catechin， chlorogenic acid， asperuloside， rutin， isoquercitrin and astragalin were 0.812-6.090 μg（r＝0.999 3），0.438-3.285 μg（r＝0.999 2），0.045-0.336 μg（r＝0.999 2），0.882-6.615 μg（r＝0.999 3），0.097-0.726 μg（r＝0.999 1），0.064-0.483 μg（r＝0.999 3），0.048-0.360 μg（r＝0.999 1） and 0.014-0.108 μg（r＝0.999 7），respectively. RSDs of precision， stability and reproducibility tests were all lower than 3.5％（n＝6）. The average recovery rates were 101.60％，103.06％，99.77％，96.93％，98.17％，96.75％，98.97％ and 99.60％，with RSDs of 1.42％，2.65％，2.78％，2.05％，2.26％，0.93％，2.79％ and 3.08％，respectively（n＝6）. The contents of aucubin， geniposide， catechin， chlorogenic acid， plantain， rutin， isoquercetin and astragaloside in 12 batches of E. ulmoides leaves from different collection time and planting varieties were 10.903-17.245， 5.578-7.892， 0.198-0.440， 13.890-19.782， 1.008-1.547， 1.102-2.396， 0.267-0.701， 0.150-0.412 mg/g， respectively. The content fluctuated greatly. The contents of aucubin， geniposide， catechin， chlorogenic acid， rutin and pinoresinol diglucoside in cortex of E. ulmoides were 0.299， 0.123， 0.580， 0.112， 0.026，1.961 mg/g， respectively. CONCLUSIONS： The method is simple， reproducible and accurate. It can be used to evaluate the quality of E. ulmoides leaves. There are obvious differences in composition and content of components in different medicinal parts （cortex， leaves） of E. ulmoides.

Anti-angiogenic activity of iridoids from Galium tunetanum

ABSTRACT The phytochemical study of Galium tunetanum Lam., Rubiaceae, leaves led to the isolation of 13 compounds from the chloroform-methanol and the methanol extracts, including six iridoid glycosides, one non-glycoside iridoid, two p-coumaroyl iridoid glycosides, two phenolic acids, and two flavonoid glycosides. The structural determination of the isolated compounds was performed by mono- and bidimensional NMR spectroscopic data, as well as ESI-MS experiments. All compounds were isolated from this species for the first time. The anti-angiogenic effects of the isolated iridoids were also reported on new blood vessels formation using the chick embryo chorioallantoic membrane as in vivo model. Results showed that among the isolated iridoids tested at the dose of 2 µg/egg, asperuloside (1), geniposidic acid (2), and iridoid V1 (3) reduced microvessel formation of the chorioallantoic membrane on morphological observations using a stereomicroscope. The anti-angiogenic effects of the active compounds, expressed as percentages of inhibition versus control, were 67% (1), 59% (2), and 54% (3), respectively. In addition, the active compounds were able to inhibit angiogenesis in the chorioallantoic membrane assay, in a dose-dependent manner (0.5-2 µg/egg) as compared to the standard retinoic acid.

Identification of metabolites of Reduning injection in rat plasma, bile, urine and feces after intravenous administration / 药学学报

This study was designed to determine the metabolites of Renduining injection in rats. The ultra-high performance liquid chromatography-LTQ Orbitrap mass spectrometric (UHPLC-LTQ-Orbitrap-MS) and mass defect filter techniques were applied to analyze the metabolites of Reduning injection in rat plasma, bile, urine and feces. As a result, we determined 14 metabolites of geniposide, including oxidation, dehydration, hydroxymethylene loss, hydrolysis, ring-opened, cysteine conjugation and glucuronidation conjugation of aglycone; 9 metabolites of geniposidic acid, consisting of dehydration, ring-opened, double-bond reduction and cysteine conjugation; 6 metabolites of secoxylogain including hydrolysis, hydroxymethylene loss, hydroxylation and ethylation; 12 metabolites of chlorogenic acid, containing decarboxylation, hydrolysis, methylation, acetylation, cysteinylglycine conjugation and glutathione conjugation. It provided information for the therapeutic effect of Reduning in vivo.

Simultaneous Determination of Geniposidic Acid, Caffeic Acid, Acteoside and Isoacteoside in Plantaginis Semen Formula Granules by HPLC / 中国药学杂志

OBJECTIVE: To establish an HPLC method to simultaneously determine the contents of geniposidic acid, caffeic acid, acteoside and isoacteoside in Plantaginis Semen formula granules, in order to provide basis for studying its quality standards. METHODS: An HPLC method was developed. Kinetex C18 column (2.1 mm×100 mm, 2.6 μm)was used and eluted with mobile phase of 0.5% acetic acid-acetonitrile at the flow rate of 0.3 mL·min-1. The wavelength was set at 239 nm for geniposidic acid, 325 nm for caffeic acid, and 330 nm for acteoside and isoacteoside. The column temperature was maintained at 30℃. RESULTS: The good linear relationships between the concentration and peak area were in the range of 0.292 1-2.92 1 μg for geniposidic acid, 0.003 4-0.033 6 μg for caffeic acid, 0.047 6-0.476 μg for acteoside and 0.102 7-1.027 μg for isoacteoside (r≥0.999 8), respectively. The average recoveries were 99.32%, 98.62%, 98.23% and 98.51% with RSDs of 1.47%, 1.36%, 1.62% and 1.53%, respectively. CONCLUSION: The method is simple, feasible and reproducible and can be used for the quality control of Plantaginis Semen formula granules.

Dynamic changes of development and active ingredients content of male flower of Eucommia ulmoides / 中草药

Objective: To establish a method for the determination of six active ingredients in Eucommia ulmoides male flowers, and analyze the dynamic changes of male flower development and active ingredients content of E. ulmoides. Methods: The active ingredients were extracted by the ultrasound-assisted process, the content of total flavonoids was determined by AlCl3 colorimetric method and other six ingredients were determined by HPLC, the samples were separated on a Thermal hypersil gold column (250 mm × 4.6 mm, 5 μm) with gradient elution of methanol-0.5% phosphoric acid at a flow rate of 1.0 mL/min, the UV wavelength was set at 206 nm (0-15 min), 236 nm (15-55 min), and 255 nm (55-100 min). Results: Flower diameter, flower height, fresh weight, dry weight, stamen length and stamen number all reached the maximum at full bloom stage; The contents of total flavonoids, chlorogenic acid and total active ingredients were the highest at the bud stage and the lowest at the initial flowering stage; The content of aucubin showed a tendency of graudal decreasement; The content of geniposidic acid was the lowest at the bud stage and reached the highest at full bloom stage, and decreased at the end of flowering phase; The content of geniposide, isoquercitrin and astragaline were the highest at full bloom stage and the lowest at the initial flowering stage. Conclusion: Flowering stage had a great impact on morphology, yield, active ingredients content and quality of E.ulmoides male flowers, which could provide important information for quality control and product development of E.ulmoides male flowers.

Study on HPLC fingerprint of Youguiyin and its multi-component quantitative analysis / 中草药

Objective: To establish the HPLC fingerprint and determine nine components (geniposidic acid, morroniside, chlorogenic acid, geniposide, loganin, pinoresinol diglucoside, liquiritin, rutin, and glycyrrhizic acid) of Youguiyin, so as to provide a scientific basis for the quality control. Methods: HPLC analysis was performed on Pursuit XRs 5 C18 column (250 mm × 4.6 mm, 5 μm). The gradient elution was performed by the mobile phase consisting of acetonitrile and 0.1% formic acid aqueous with the flow rate of 1.0 mL/min, the detection wavelength was set at 230 nm, and the column temperature was 30 ℃. Fingerprints of ten batches of Youguiyin were determined, and the similarities among fingerprints were evaluated. Attributive analysis and identification of common peaks were performed by comparing the retention time and UV spectra among 10 batches of Youguiyin. The formula ingredients and reference substance, and nine components content were determined. Results: The similarities of fingerprints of 10 batches of Youguiyin and reference fingerprints were all greater than 0.904. There were 30 mutual peaks marked in total, which were eight mutual peaks from Eucommia ulmoides, eight mutual peaks from Glycyrrhiza uralensis, six mutual peaks from Cornus officinalis, six mutual peaks from Aconitum carmichaeli, one mutual peaks from Cinnamomum cassia, and none of mutual peaks were originated from Dioscorea opposita and Lycium barbarum, and three of them cannot be originated. Based on the the identification of the common peaks, nine components [geniposidic acid (peaks 7), morroniside (peaks 9), chlorogenic acid (peaks 11), gardenoside (peaks 12), loganin (peaks 13), pinoresinol diglucoside (peaks 14), liquorice glycosides (peaks 18), rutin (peaks 21), and glycyrrhizic acid (peaks 30)] were identified and quantified. The quality faction of nine components were 87.6—119.1 μg/g, 323.6—365.6 μg/g, 108.3—124.1 μg/g, 79.5—85.0 μg/g, 171.7—188.0 μg/g, 163.0—238.3 μg/g, 64.5—53.3 μg/g, 159.8—168.5 μg/g, and 72.8—83.6 μg/g in raw material. Conclusion: The method established in this study is simple, accurate and highly reproducible, and can provide basis for quality control of Youguiyin.

Effects of Different Drying Methods on Contents of 4 Active Ingredients in Leaves of Eucommia ulmoides / 中国药房

OBJECTIVE:To investigate the effects of different drying methods on contents of active ingredients in leaves of Eu-commia ulmoides,and provide reference for establishing its drying methods after habitat harvesting. METHODS:Different drying methods [natural drying in the shade for 72 h,natural drying in the sunlight for 36 h,drying beside or over a fire(60℃6 h,80℃2 h,100℃1 h,120 ℃ 0.5 h),microwave vacuum freeze drying for 12 h,vacuum freeze drying for 12 h] were used for process-ing. HPLC was conducted to determine the contents of aucubin,geniposidic acid,chlorogenic acid and geniposide in samples and compare with the untreated fresh products. RESULTS:Contents of 4 ingredients in samples after the 2 freeze drying were close to these in fresh samples,which were higher than samples after other drying. CONCLUSIONS:Drying methods show significant ef-fects on the effective ingredients in leaves of E. ulmoides. Compared with natural drying in the shade and natural drying sunlight and drying beside or over a fire,microwave vacuum freeze drying and vacuum freeze drying can make better retention of the active ingredients in fresh leaves of E. ulmoides.

Establishment of HPLC fingerprints of Zhihuang Tongfeng Decoction and determination of four constituents / 中成药

AIM To establish HPLC fingerprints of Zhihuang Tongfeng Decoction (Phellodendri chinensis Cortex,Anemarrhenae Rhizoma,Plantaginis Semen,etc.) and to determine the contents of four constituents.METHODS The analysis of aqueous extract of this drug was performed on a 30 ℃ thermostatic Wondasil C18 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.5％ acetic acid in a gradient elution manner,and the detection wavelength was set at 245 nm.RESULTS There were sixteen common peaks in the HPLC fingerprints of ten batches of samples with the similarities of more than 0.9.Berberine hydrochloride,mangiferin,geniposidic acid and salvianolic acid B showed good linear relationships within the ranges of 0.636 2-3.575 μg (R2 =0.9999),0.338-1.425 μg (R2 =0.9990),0.6452-2.7188 μg (R2 =1) and0.938 8-5.275 μg (R2 =0.999 3),whose average recoveries were 100.56％,98.35％,100.25％ and 102.11％ with the RSDs of 2.41％,1.17％,1.24％ and 2.02％,respectively.CONCLUSION This accurate,reliable and specific method can be used for the quality control of Zhihuang Tongfeng Decoction.

Effects of Different Drying Methods on Contents of 4 Active Ingredients in Leaves of Eucommia ulmoides / 中国药房

OBJECTIVE:To investigate the effects of different drying methods on contents of active ingredients in leaves of Eu-commia ulmoides,and provide reference for establishing its drying methods after habitat harvesting. METHODS:Different drying methods [natural drying in the shade for 72 h,natural drying in the sunlight for 36 h,drying beside or over a fire(60℃6 h,80℃2 h,100℃1 h,120 ℃ 0.5 h),microwave vacuum freeze drying for 12 h,vacuum freeze drying for 12 h] were used for process-ing. HPLC was conducted to determine the contents of aucubin,geniposidic acid,chlorogenic acid and geniposide in samples and compare with the untreated fresh products. RESULTS:Contents of 4 ingredients in samples after the 2 freeze drying were close to these in fresh samples,which were higher than samples after other drying. CONCLUSIONS:Drying methods show significant ef-fects on the effective ingredients in leaves of E. ulmoides. Compared with natural drying in the shade and natural drying sunlight and drying beside or over a fire,microwave vacuum freeze drying and vacuum freeze drying can make better retention of the active ingredients in fresh leaves of E. ulmoides.

Establishment of HPLC fingerprints of Zhihuang Tongfeng Decoction and determination of four constituents / 中成药

AIM To establish HPLC fingerprints of Zhihuang Tongfeng Decoction (Phellodendri chinensis Cortex,Anemarrhenae Rhizoma,Plantaginis Semen,etc.) and to determine the contents of four constituents.METHODS The analysis of aqueous extract of this drug was performed on a 30 ℃ thermostatic Wondasil C18 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.5％ acetic acid in a gradient elution manner,and the detection wavelength was set at 245 nm.RESULTS There were sixteen common peaks in the HPLC fingerprints of ten batches of samples with the similarities of more than 0.9.Berberine hydrochloride,mangiferin,geniposidic acid and salvianolic acid B showed good linear relationships within the ranges of 0.636 2-3.575 μg (R2 =0.9999),0.338-1.425 μg (R2 =0.9990),0.6452-2.7188 μg (R2 =1) and0.938 8-5.275 μg (R2 =0.999 3),whose average recoveries were 100.56％,98.35％,100.25％ and 102.11％ with the RSDs of 2.41％,1.17％,1.24％ and 2.02％,respectively.CONCLUSION This accurate,reliable and specific method can be used for the quality control of Zhihuang Tongfeng Decoction.

Serum pharmacochemistry of stem bark of Eucommia ulmoides / 中草药

Objective: To investigate the bioactive constituents in the stem bark of Eucommia ulmoides and study on their serum pharmacchemistry. Methods: The UHPLC-Q-TOF-MS was used to analyze the ingredients in the serum samples in rats, and the chromatogram was compared among the peaks of extracts in the stem bark of E. ulmoides, rat serum with drug and blank serum sample was used to determine them after the administration of extracts by comparing the finger-print. Results: Seven compounds absorbed into blood were be detected, and had the same retention time with those in the finger-print of extracts, after repeated experiments, which were original constituents of the extracts. The extracts were identified by comparing the retention times with standard substance, five compounds were obtained and identified as geniposidic acid, protocatechuic acid, geniposide, pinoresinol diglucoside, and pinoresinol monoglucoside. Through review of the literature, two compounds may be 1-hydroxypinoresinol glucoside, and eucommiol. Conclusion: The compounds are absorbed into the blood are the effective constituents, and the research provides a scientific fundament for material basis of effectiveness of E. ulmoides.

Analysis on pharmacokinetics-pharmacodynamics correlation of effective ingredients of Qing'e wan / 中国中药杂志

To study the pharmacokinetics of three active ingredients in Qing'e wan, namely geniposidic acid, psoralen and isopsoralen, in rats, in order to investigate their correlation in the anti-osteoporotic effect. The rats were taken blood from their eye sockets at different time points after being orally administered with raw and salt-processed Qing'e wan. Geniposidic acid, psoralen and isopsoralen in rats plasma were determined by means of UHPLC-MS/MS to draw the concentration-time curve. The proliferation rate of osteoblasts was taken as the pharmacodynamic index, and determined by MTT method to draw effect-time curve. In comparison between the effect-time curve and the concentration-time curve, the blood concentrations of geniposidic acid and psoralen were close to the peak when the cell proliferation rate reached its peak, indicating a good correlation between them. The peak blood concentration of isopsoralen was slightly lagging behind the peak of efficacy. According to the correlation analysis after fitting the effect-time curve and the concentration-time curve, salt-processed Qing'e wan had a better correlation than the raw one. The above experimental results showed that the effect-time curve and the concentration-time curve of geniposidic acid and psoralen had a good correlation, and the correlation of salt-processed Qing'e wan was better than the raw one.

Chemical constituents of Aganosma marginata / 中草药

Objective: To investigate the chemical constituents from Aganosma marginata and to provide the material basis for the quality control. Methods: The chemical constituents were separated and purified by silica gel Sephadex LH-20, MCI. Their structures were determined by physicochemical properties and spectral data analyses. Results: Twenty three compounds were isolated from A. marginata and identified as: periseoside C (1), 3-O-β-D-glucopyranosyl-3β,15α-dihydroxypregn-5-en-20-one (2), 3β,20α-dihydroxy- 5-en-pregane (3), 28-methylstigmast-5-en-3-ol (4), 29-norcycloart-23-ene-3,25-diol (5), 29-norcycloartan-3-ol (6), geniposidic acid (7), syringaresinol (8), syringaresinol-4,4'-O-bis-β-D-glucopyranoside (9), syringic acid 4-O-β-D-glucopyranoside (10), salicylic acid (11), scopoletin (12), azelaic acid (13), 3-O-[β-D-xylopyranosyl]-(1→4)-β-D-allopyranoside-14-hydroxycard-20 (22)-enolide (14), bis (2-ethylhexyl) phthalate (15), kaempferol-3-O-α-L-rhamnopyranosyl-(l→4)-α-L-glucopyranoside (16), kaempferol-3-O-α-L- glucopyranoside-(1→2)-β-D-glucopyranoside (17), kaempferol-3-O-α-L-rhamnopyranosyl-(1→4)-[α-L-rhamnopyranosyl (1→4)]-β- D-glucopyranoside (18), kaempferol-3-O-α-L-rhamnopyranosyl-(1→4)-[β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside (19), hexacosanoicacid 1-carbonate (20), 5,8,12-trihydroxy-9-octadecenoic acid (21), (2S,3S,4R)-phytosphingosine (22), and nebularine (23). Conclusion: All the compounds are obtained from plants of Aganosma G. Don for the first time.

Separation and enrichment of geniposidic acid from leaves of Eucommia ulmoides by aqueous two-phase flotation / 中草药

Objective: Ethanol/NaH2PO4 aqueous two-phase gas solvent sublation coupled with response surface methodology was developed for the separation/enrichment and analysis of geniposidic acid from the leaves of Eucommia ulmoides. Methods: For the aqueous two-phase flotation project, the flotation effects of different solvent, salts, flotation rate, flotation time, amount of crude extract and so on were deeply investigated; Furthermore, a Box-Behnken central composite test design was studied to determine the best flotation process conditions of GPA, and three factors including the mass fraction of NaH2PO4, the mass fraction of ethanol, and the flotation rate were selected based on the single factor test in the design. A 100 times scale-up experiment also was tested. Results: The optimal conditions of the flotation were as follows: the mass fraction of NaH2PO4 was 25%, the mass fraction of ethanol was 20%, the amount of crude extract was 5 g, the flotation rate was 30 mL/min, and the flotation time was 20 min. Under the optimal condition, the flotation efficiency was 97.88%, and the enrichment factor was 27.34; The 100 times scale-up results showed that the flotation efficiency of GPA was 95.60%, and the RSD value was 0.77%. Conclusion: The method with ethanol/NaH2PO4 aqueous two-phase gas solvent sublation is suitable for the separation and enrichment of the active ingredients from E. ulmoides because of its high distribution coefficient and large enrichment factor.

Determination of plasma protein binding rate of five components in Eucommia ulmoides extract / 中国药理学通报

Aim To determine the plasma protein binding rate of five components of Eucommia ulmoides extract. Methods The equilibrium dialysis method was used to study the plasma protein binding rate. The plasma samples were extracted by protein precipitation with methanol. With the use of puerarin as the internal standard, UPLC-MS/MS was carried out to determine the concentration of the five compounds in and out of the dialysis membrane. Results The average plasma protein binding rates of five compounds on the area of the concentration which was determinate were as fol-lows, respectively: geniposidic acid was ( 25. 77 ± 2. 68 )%, protocatechuic acid was ( 57. 54 ± 3. 79)%, chlorogenic acid was (53. 91 ± 3. 00)%, pinoresinol diglucoside was (24. 15 ± 4. 92)%, and pinoresinol singleglucoside was (49. 78 ± 3. 61)%. Conclusions The results show that the binding percentage of geniposidic acid and pinoresinol diglucoside is relatively low, but the binding rate of the others with rat plasma protein is moderate.

Chemical constituents from the fruits of Akebiae quinata / 中国药学杂志

OBJECTIVE: To study the chemical constituents of the fruits of Akebiae quinata. METHODS: Various column chromatography techniques including silica gel, Sephadex LH-20, and macroporous adsorption resin column chromatography were used for fractionization and purification.The structures were identified on the basis of their physicochemical and spectroscopic evidence. RESULTS: Fifteen compounds were obtained, and their structures were identified as geniposidic acid(1), 10-O-acetylgeniposidic acid (2), vomifoliol(3), p-hydroxybenzoic acid(4), protocatechuic acid(5), caffeic acid(6), tyrosol(7), palmitic acid(8), 15-nona-cosanol(9), stigmasterol(10), stigmasterol-3-O-β-D-glycopyranoside(11), β-sitosterol(12), β-daucosterol(13), ursolic acid(14), and oleanolic acid(15). CONCLUSION: Compounds 1-7, 9 and 14 were isolated from the fruits oi Akebiae quinata for the first time.

Effect of different drying and storing methods on quality control for leaves of Eucommia ulmoides / 中草药

Objective: To study the effect of different drying and storing methods on the content of main components in the leaves of Eucommia ulmoides. Methods: The contents of chlorogenic acid, geniposide acid, and geniposide in the leaves of E. ulmoides after being treated with different drying and storing methods were determined by HPLC. Results: The contents of geniposide acid and geniposide were higher in the fresh leaves of E. ulmoides and the content of chlorogenic acid was higher in the shade dried leaves of E. ulmoides after dried, microwave heated, and stored under low temperature. The content of main components in the leaves of E. ulmoides stored in darkness at low temperature decreased slowly. Conclusion: The contents of main components in the leaves of E. ulmoides collected in the best harvest time remain almost the same after fixed, dried, microwave heated, sealed, stored at low temperature in darkness for one year.

Comparison on HPLC fingerprint of Eucommiae Folium obtained from different habitats and by various original processing techniques / 中草药

Objective: To illustrate the influences of various original processing techniques on chemical composition of Eucommiae Folium from different habitats by comparing HPLC fingerprint profiles, so as to select the suitable techniques for processing Eucommiae Folium. Methods: To evaluate the quality of Eucommiae Folium with the various processing techniques by HPLC fingerprint. The analysis was performed on the Promosil C18(250 mm × 4.6 mm, 5 μm) column. Acetonitrile-0.1% H3PO4 was used as the mobile phase with gradient elution. The scanned wavelength was from 200 to 400 nm, and the detection wavelength was at 208 nm. The flow rate was set at 0.8 mL/min. The column temperature was set at 25°C. Results: The HPLC fingerprint profiles of Eucommiae Folium by the various processing techniques which were systematically researched were investigated. A total of main 22 common peaks were found, but the areas of the peaks were significantly different. The contents of chlorogenic acid and geniposidic acid obtained by the various processing techniques had the evident differences. Conclusion: HPLC fingerprint technique with the application to the content determination of the active ingredients has the good reproducibility and maneuverability. It is one of the potential process analytical techniques to explore the ingredient differences in the processed Eucommiae Folium from different habitats.