This study aims to establish a new method for quality evaluation of Gentianae Macrophyllae Radix by simultaneous determination of five iridoids (loganic acid, 6'-O-beta-D-glucopyranosylgentiopicroside, swertiamarin, gentiopicroside, sweroside), and to detect five iridoids in the root of eight species (Gentiana macrophylla, G. straminea, G. crassicaulis, G. dahurica, G. robusta, G. waltonii, G. lhassica, and G. tibetica). The separation was carried out on a Shiseido SPOLAR C18 (4.6 mm x 250 mm, 5 microm) column eluted with mobile phase of water containing 0.04% formic acid (A) and acetonitrile (B) in a gradient program. The flow rate was 0.8 mL x min(-1). The detect wavelength was set at 240 nm. The column temperature was kept at 30 degrees C. The volume of injection was 5 microL. The five iridoids were well separated with ideal linear correlations. The average recoveries were 97.35% - 106.23%. All the five iridoids were detected in the root of eight species. The contents of same species changed in a somewhat wider range. The contents in root of G. dahurica were lower than that in other species.
China ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Gentianella ; chemistry ; Iridoid Glycosides ; analysis

This study aims to develop an HPLC-DAD method for simultaneous determination of 11 components(6 phenolic acids and 5 iridoids) in Lonicera japonica flowers(LjF) and leaves(LjL), and compare the content differences of LjF at different development stages, LjL at different maturity levels, and between LjF and LjL. One-way ANOVA, principal component analysis(PCA), and orthogonal partial least-squares discriminant analysis(OPLS-DA) were employed to compare the content of the 11 components. The content of total phenolic acids, total iridoid glycosides, and total 11 components in LjF showed an overall downward trend with the development of flowers. The content of total phenolic acids, total iridoid glycosides, and total 11 components in young leaves were higher than those in mature leaves. The results of PCA showed that the samples at different flowering stages had distinguishable differences in component content. The VIP value of OPLS-DA showed that isochlorogenic acid A, chlorogenic acid, and secologanic acid were the main differential components of LjF at different development stages or LjL with different maturity levels. LjF and LjL have certain similarities in chemical composition while significant differences in component content. The content of total phenolic acids in young leaves was significantly higher than that in LjF at various development stages. The content of total iridoid glycosides in young leaves was similar to that in LjF before white flower bud stage. The total content of 11 components in young leaves was significantly higher than that in LjF at green flower bud stage, before and during completely white flower bud stage. LjL have great potential for development. Follow-up research on the pharmacodynamic equivalence of LjF and LjL(especially young leaves) should be carried out to speed up the development and application of LjL.
Chromatography, High Pressure Liquid ; Flowers/chemistry* ; Iridoid Glycosides/analysis* ; Lonicera/chemistry* ; Plant Leaves/chemistry*

OBJECTIVETo develop an HPLC method for the simultaneous quantitation of five constituents in Scrophularia ningpoensis.

METHODSamples were analyzed on an Agilent SB-C18 column(4.6 mm x 250 mm, 5 microm) eluted with acetonitrile and water containing 0.03% phosphate acid as mobile phases in a linear gradient mode. The flow rate was kept at 1.0 mL x min(-1), and the column temperature was set to 30 degrees C. The DAD detector wavelengths were 210, 280, 330 nm.

RESULTThe linear ranges were 50-400 mg x L(-1) for harpagide, 1-40 mg x L(-1) for harpagoside, 1-20 mg x L(-1) for cinnamic acid, 0.5-4.5 mg x L(-1) for acteoside,1-60 mg x L(-1) for angoroside C, respectively. The average recoveries of the five constituents were 100.8% (RSD 0.62%), 101.7% (RSD 0.32%), 98.8% (RSD 0.48%), 99.9% (RSD 1.4%), 99.2% (RSD 1.1%), respectively.

CONCLUSIONThrough the validation, the method was proved to be sensitive, accurate, repeatable, and can be used for quality control of the roots of S. ningpoensis.

Chromatography, High Pressure Liquid ; methods ; Cinnamates ; analysis ; Coumaric Acids ; analysis ; Glucosides ; analysis ; Glycosides ; analysis ; Iridoid Glycosides ; Phenols ; analysis ; Pyrans ; analysis ; Scrophularia ; chemistry ; Trisaccharides ; analysis

OBJECTIVETo establish an HPLC method for simultaneous determination of eight iridiods in Gardeniae Fructus.

METHODKromasil C18 column (4. 6 mm x 250 mm, 5 microm) was adopted, with acetonitrile-water-trifluoroacetic acid (6:94: 0.05) as the mobile phase at the flow rate of 1.0 mL x min(-1). The detection wavelength was set at 238 nm, and the column temperature was 40 degrees C.

RESULTThe linear ranges of geniposide, gardoside, shanzhiside, geniposidic acid, deacetyl asperulosidic acid methyl ester, gardenoside, scandoside methyl ester, and genipin gentiobioside were 1.5036 - 15.036, 0.04256 - 0.4256, 0.1038 - 1.038, 0.00992 - 0.0992, 0.02332 - 0.2332, 0.4128 - 4.128, 0.02040 - 0.2040 and 0.4656 - 4.656 microg, respectively. Their average recoveries were 99.6% , 100.6% , 101.2%, 99.5%, 100.3% , 98.7%, 99.8% and 100.1%, respectively.

CONCLUSIONThe method shows good separation and it is so simple, accurate and highly repeatable that it can be used for providing basis for quality control of Gardeniae Fructus.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Gardenia ; chemistry ; Iridoid Glycosides ; analysis ; isolation & purification

OBJECTIVETo develop an RP-HPLC method for the determination of five iridoid glycosides in Phlomis younghusbandii.

METHODHPLC analysis was performed on a Symmetry C18 (4.6 mm x 150 mm, 5 microm, Waters) column eluted with acetonitrile (A) and water (B) in gradient elution. The gradient program was as follows: 0-5 min kept 7% A; 5-10 min changed to 12% A; 10-40 min kept 12% A. The flow rate was 1.0 mL x min(-1). The column temperature was 20 degrees and the detection wavelength was 235 nm.

RESULTThe linear ranges of sesamoside, shanzhiside methyl ester, 7, 8-dehydropenstemoside, penstemoside and 8-O-acetylshanzhiside methyl ester were 0.050-0.650 (r = 0.999 3), 0.050-0.350 (r = 0.999 5), 0.040-0.280 (r = 0.999 4), 0.010-0.070 (r = 0.999 6), 0.040-0. 280 (r = 0.999 7) g x L(-1), respectively. The average recoveries (n = 6) of them were all between 96% and 104%, RSD < 5.0%.

CONCLUSIONThe method is simple, accurate, repeatable and stable, which can be used for quality control of P. younghusbandii.

Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Iridoid Glycosides ; analysis ; Phlomis ; chemistry ; Quality Control ; Reproducibility of Results

This present study aims to establish a UPLC method for simultaneously determining eleven components such as new chlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,artichoke,isochlorogenic acid A,isochlorogenic acid B,isochlorogenic acid C,rutin,hibisin and loganin in Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica and comparing the differences in the contents of phenolic acids,flavonoids and iridoid glycosides of Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.The method was carried out on an ACQUITY UPLC BEH C18column(2.1 mm×100 mm,1.7 μm) by a gradient elution using acetonitrile and 0.1% phosphoric acid.The flow rate was 0.3 mL·min-1.The column temperature was maintained at 30 ℃.The sample room temperature was 8 ℃.The wavelength was set at 326 nm for new chlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,artichoke,isochlorogenic acid A,isochlorogenic acid B and isochlorogenic acid C,352 nm for rutin and lignin,and 238 nm for loganin.The injection volume was 1 μL.The eleven components has good resolution and was separated to baseline.Each component had a wide linear range and a good linear relationship(r≥0.999 6),the average recovery rate(n=9) was 98.96%,100.7%,97.24%,97.06%,99.53%,96.78%,98.12%,95.20%,95.12%,100.2%,98.61%and with RSD was 2.5%,1.4%,1.9%,2.1%,1.7%,1.9%,1.6%,2.0%,1.4%,2.2%,2.0%,respectively.Based on the results of the content determination,the chemometric methods such as cluster analysis and principal component analysis were used to compare the Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.The results showed that Lonicerae Japonicae Flos and leaves of Lonicera japonica were similar in the chemical constituents,but both showed chemical constituents difference compored to Lonicerae Japonicae Caulis.The established multi-component quantitative analysis method can provide a reference for the quality control of Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Flavonoids ; analysis ; Flowers ; chemistry ; Hydroxybenzoates ; analysis ; Iridoid Glycosides ; analysis ; Lonicera ; chemistry ; Phytochemicals ; analysis ; Plant Leaves ; chemistry ; Quality Control

OBJECTIVETo develop a method for the determination of harpagide and harpagoside in Scrophulariae Radix (Xuanshen) by HPLC-UV under double wavelength, and to study the changes of these two constituents during processing, and to set the limitation of harpagide and harpagoside contents in crude drug and sliced pieces of Xuanshen.

METHODThe analyses were performed on an Agilent Technologies ZORBAX SB-C18 (4.6 mm x 250 mm, 5 microm) eluted with acetonitrile-water (containing 0.03% phosphoric acid) in gradient model. The flow rate was 1.0 mL x min(-1) . The column temperature was 25 degrees C. The UV detector wavelength was set at 210 nm before 13 min and then changed to 280 nm.

RESULTHarpagide and harpagoside were separated well. The linear calibration curves were obtained over of 0.0549 - 1.46 microg for harpagide (r = 0.9999, n =7) ,0.0225 - 0.900 microg for harpagoside (r = 0.9998, n = 9). The recoveries ( +/- RSD)% were 98.1 (+/- 2.4)% for harpagide and 98.8 (+/- 4.3)% for harpagoside. The contents of harpagide were 0. 277% - 0.620%, harpagoside were 0.078% - 0.362% in Xuanshen, and harpagide were 0.276% - 1.059%, harpagoside were 0. 059% - 0.183% in sliced Xuanshen, respectively. After the processing of Scrophulariae Radix, the content of harpagide increases 13.7% - 96.0%, while harpagoside decreases 11.0%-73.9%.

CONCLUSIONThis method is simple, accurate, and can be used for the quality control of Scrophulariae Radix. We propose that the total content of harpagide and harpagoside in either crude drug or sliced pieces of Scrophulariae Radix should not be less than 0.45%.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Glycosides ; analysis ; isolation & purification ; Iridoid Glycosides ; Magnoliopsida ; chemistry ; Plant Roots ; chemistry ; Pyrans ; analysis ; isolation & purification ; Spectrophotometry, Ultraviolet ; methods

In order to improve the quality control level of Ligustri Lucidi Fructus(LLF) and to explore the changes of chemical components after processing,the HPLC method for fingerprint and simultaneous determination of the major polar components in LLF were established. The octadecylsilane bonded silica gel was used as the stationary phase,with acetonitrile as the mobile phase A and0. 2% formic acid as the mobile phase B in a gradient elution procedure at a flow rate of 1. 0 m L·min-1. The detection wavelength was set at 280 nm and the column temperature was 25 ℃. There were 22 common peaks,20 of which were selected from the fingerprint of LLF and its wine-steamed product,respectively,and 14 chromatographic peaks were identified with reference substances. With the same chromatographic conditions,seven components were quantitatively analyzed and the results of system adaptability and methodology investigation all met the requirements of content determination. Compared with the crude LLF,the content of 5-hydroxymethyl furfural and salidroside significantly increased in wine-steamed LLF,while the contents of iridoid glycosides generally decreased. The method provided a basis for quality control of LLF and its processed products as well as the related preparations.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Furaldehyde ; analogs & derivatives ; Glucosides ; Iridoid Glycosides ; Ligustrum ; chemistry ; Phenols ; Phytochemicals ; analysis

AIMTo establish a sensitive and specific HPLC method for controlling the quality of total glycosides from Swertia franchetiana H. Smith.

METHODSHPLC method was applied for quality and quantitative assessment of the pharmaceutical extracts from Swertia franchetiana H. Smith. The preparation of sample, the HPLC column, mobile phase, elution mode (isocratic or gradient) and gradient program were optimized in order to obtain HPLC profile. The HPLC system consisted of a SPD-1OAvp pump, SPD-M1OAVP photodiode-array detector (PAD), SIL-10ADVP auto injector. Data were acquired and processed with the CLASS-VP6.1 workstation. HPLC analysis was performed on a Kromasil C18 column (250 mm x 4. 6 mm ID, 5 microm) with methanol and water as mobile phase. The column temperature was set up at 40 degrees C and the flow-rate was 1 mL x min(-1). The reference solution of chemical standards and sample were injected into HPLC system, separately.

RESULTSThe HPLC chromatographic fingerprinting of the total glycosides, showing 16 characteristic peaks which were partitioned into three parts: one peak in 0-10 min of retention time, nine peaks containing main 1-7 peaks in 10-15 min of retention time, 6 peaks in 15-30 min of retention time, was established from 10 lots of their products. By comparison of the retention time and the on-line UV spectra and their molecule weights of chemical standards, peak 1-7 were identified as swertiamarin (1), gentiopicroside (2), sweroside (3), isoorientin (4), swertisin (5), isoswertisin (6) and swetianolin (7), respectively. The ratios of peak area between 1-16 were in their extent. Moreover, comparison of the HPLC profiles of the total glycosides, the extracts prepared using another process and the plant indicated that they were closely related to each other.

CONCLUSIONThe HPLC profiles and quantitative assessment of the total glycosides from Swertia franchetiana H. Smith with high specificity can be used to control their quality and assure lot to lot consistency.

Chromatography, High Pressure Liquid ; methods ; Glucosides ; analysis ; chemistry ; Glycosides ; analysis ; chemistry ; Iridoid Glucosides ; Iridoids ; analysis ; chemistry ; Luteolin ; analysis ; chemistry ; Molecular Structure ; Plants, Medicinal ; chemistry ; Pyrans ; analysis ; chemistry ; Pyrones ; analysis ; chemistry ; Quality Control ; Swertia ; chemistry

OBJECTIVETo study the chemical constituents of Halenia elliptica.

METHODThe air-dried whole plants of Halenia elliptica were extracted with 90% EtOH. The EtOH extract was condensed to a small amount of volume and extracted with petroleum ether, EtOAc and n-BuOH, successively. The compounds were isolated and purified by column chromatography from the EtOAc fraction, and identified based on spectral analyses (MS, 1H-NMR, 13C-NMR).

RESULT12 compounds were isolated from H. elliptica, and characterized as 8-hydroxy-2-methylchromone (1), 5-methoxy-2-methylchromone (2), 7-epi-vogeloside (3), coniferl aldehyde (4), sinapaldehyde (5), norbellidifolin (6), 1-hydroxyl-2,3,4,6-tetramethoxyxanthone (7), 1-hydroxyl-2,3,4,7-tetramethoxyxanthone (8), 1-hydroxyl-2,3,5-trimethoxyxanthone (9), together with azelaic acid, beta-sitosterol, and oleanolic acid.

CONCLUSIONCompounds 1, 2 were new natural compounds and compounds 3-6, 10 were obtained from H. elliptica for the first time and compound 6 showed inhibitory activities against HBsAg and HBeAg secretion with IC50 value of 0.77 and < 0.62 mmol x L(-1), respectively.

Acrolein ; analogs & derivatives ; analysis ; isolation & purification ; Chromatography ; Dicarboxylic Acids ; analysis ; isolation & purification ; Gentianaceae ; chemistry ; Iridoid Glycosides ; isolation & purification ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Oleanolic Acid ; analysis ; isolation & purification ; Plant Extracts ; isolation & purification ; Sitosterols ; analysis ; isolation & purification ; Xanthones ; analysis ; isolation & purification