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ObjectiveTo explore the quality differences between steamed products and raw products of Citri Reticulatae Pericarpium（CRP）. MethodThe color of steamed products and raw products of CRP was determined from the perspective of appearance by electronic eye technique， and the quality differences between them was objectively characterized by the luminous value（L*）， yellow-blue value（b*）， red-green value（a*） and total chromatic value（E*ab）. Based on this， ultra-high performance liquid chromatography（UPLC） was used to establish a fingerprint evaluation method with the mobile phase of acetonitrile（A）-0.1% formic acid aqueous solution（B） for gradient elution（0-5 min， 5%A； 5-30 min， 5%-20%A； 30-60 min， 20%-52%A）， detection wavelength at 270 nm， flow rate of 0.3 mL·min-1 and column temperature of 30 ℃. The quality differences between steamed products and raw products of CRP were compared from the perspective of chemical composition， and correlation analysis was used to reveal the correlation between the difference in appearance color and the difference in internal chemical composition. ResultAfter being steamed， L*， b* and E*ab of CRP showed an overall decreasing trend， indicating that the color of the steamed products darkened and deepened from yellow to blue but still tended to be yellow， while a* showed an overall increasing trend， indicating that the color of the steamed products tended to red. A total of 24 peaks were identified in the fingerprint profiles of raw products and steamed products of CRP， and 13 of the main peaks were identified. The precision， stability and repeatability studies showed that compared with the reference peak （peak 14， hesperidin）， the relative standard deviations（RSDs） of the relative peak area and relative retention time of the remaining peaks were<3.0%.The results of chemometric statistical analysis showed that there were some differences between raw products and steamed products of CRP， and 7 main differential components were identified， among which 5-hydroxymaltol（peak 1） and 5-hydroxymethylfurfural（peak 2） were the characteristic components of steamed products. The correlation analysis results showed that， in addition to the above two characteristic components， four components of peak 4， peak 10 （vicenin-2）， peak 23 （tangeretin） and peak 24 （5-demethylnobiletin） also correlated significantly with the color change （E*ab） of the samples （P<0.05， P<0.01）. ConclusionBefore and after steaming， not only the chemical composition changes， but also the color. Comparing the characteristic peaks of chemical composition difference and color difference before and after steaming of CRP， it is found that 5-hydroxymaltol， 5-hydroxymethylfurfural and peak 4 are common characteristic difference components， which can provide a reference for establishing the characteristic quality control method of steamed products， and quickly evaluating the quality difference between raw products and steamed products of CRP.
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Twenty batches of Aurantii Fructus Immaturus(AFI) were collected, with their peel and pulp taken as research objects. Ultra-high performance liquid chromatography(UPLC) fingerprints of peel and pulp of AFI were established with 17 common peaks in peel and 10 in pulp. Six kinds of flavonoids were identified, i.e., narirutin, naringin, rhoifolin, hesperidin, neohesperidin and nobiletin. The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine was employed for similarity analysis, which showed that the chromatographic peaks of peel and pulp were basically similar to their respective reference fingerprints, with all similarities greater than 0.90. The similarity between peel and pulp of the same batch of AFI ranged from 0.850 to 0.983. Cluster analysis(CA), principal component analysis(PCA), and orthogonal partial least squares discriminant analysis(OPLS-DA) were conducted on the common peaks of peel and pulp of AFI with SPSS 17.0 and SIMCA 14.1. Combined with the reference fingerprints, these analyses revealed 12 differential components regarding peel and pulp. Further, the content of the 6 flavonoids and synephrine was determined. The proposed method integrating UPLC fingerprint and multicomponent quantitative analysis is applicable to the quality evaluation of AFI. The results provide a certain basis for the scientific connotation about the appearance characteristic of AFI.
Subject(s)
Chromatography, High Pressure Liquid , Citrus , Drugs, Chinese Herbal , SynephrineABSTRACT
To establish ultra performance liquid chromatography( UPLC) fingerprint of Puerariae Lobatae Radix from different habitats and simultaneously determine the contents of six isoflavonoids. The UPLC fingerprint analysis and content determination were performed on a Waters ACQUITY UPLC BEH C_(18)( 2. 1 mm×50 mm,1. 7 μm) chromatographic column,with acetonitrile-0. 05% formic acid as mobile phase for gradient elution. The detection wavelength was set at 250 nm; the flow rate was 0. 2 mL·min~(-1); the column temperature was 30 ℃ and the injection volume was 2 μL. Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine( TCM) was adopted; principal component analysis( PCA) and discriminant analysis by partial least square method( PLS-DA) in Simca-P software were used to identify the differential components in samples from three habitats. The similarity was over 0. 90 in 29 batches of samples,indicating good consistency of the samples. The samples were clustered into 3 categories by PCA and PLS-DA,and six differential components such as puerarin apioside,daidzin,and isoflavoues aglycone were found. The determination results of 6 isoflavones,including 3'-hydroxy puerarin,puerarin,3'-methoxy puerarin,puerarin apioside,daidzin,and isoflavoues aglycone,showed that the content of the same component and the fluctuation range between different components were all different among different habitats. The total content of 6 isoflavones from different regions was Anhui 11. 21% >Henan 10. 97% >Shannxi 9. 38%. The establishment of UPLC fingerprint combined with simultaneous determination of 6 active components provides a more comprehensive reference for quality control and quality evaluation of Puerariae Lobatae Radix.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Chemistry , Ecosystem , Flavonoids , Phytochemicals , Plant Roots , Chemistry , Pueraria , ChemistryABSTRACT
OBJECTIVE: To establish the UPLC fingerprints of G.straminea, and analyze its differences of indications ingredient and genetic diversity. METHODS: The main chemical composition of dry root were measured by UPLC. Similarity evaluation system for chromatographic fingerprint of TCM(Version 2004 A) was used to analyze the relative peak area of common peaks from differently populations,and cluster analysis of common peaks was carried out by SPSS19.0. Genomic DNA of dry leaves collected were extracted and amplified by ISSR markers. The genetic diversity and genetic distance were calculated by Popgen 32. RESULTS: Twelve Common peaks were selected as the fingerprint peaks from 18 populations of G.straminea,and three groups were divided according to the clustering results of chemical composition. It had the highest levels that was gentiopicroside content and total content of five kinds of chemical components in Shandan County, Zhangye City.Six primers produced 73 bands, among which 72 were polymorphic bands, the average percentage of polymorphie bands(PPB) was 98.63%; Shannon's information index() was 0.366 3;Nei's genetic diversity index(H) was 0.231 6. The genetic differentiation coefficient(Gst) and gene flow(Nm) of wild populations were 0.344 1 and 0.953 0. CONCLUSION: UPLC Fingerprint of G.straminea can be used as the quality evaluation system.The genetic diversity among species of G.straminea is at higher level, but the level of genetic diversity within populations is higher than that between populations,and genetic variance mainly existed among populations.
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Objective To establish the analysis method of UPLC fingerprint and simultaneous determination of five active components (gastrodin, 4-hydroxybenzyl alcohol, parishin B, parishin C, and parishin A). Methods The analysis was performed on a Waters Acquity with a Waters ACQUITY UPLC BEH C18 (50 mm × 2.1 mm, 1.7 μm) column. The gradient mobile phase was acetonitrile and 0.1% formic acid-water solution, flow rate was set at 0.3 mL/min, the injection volume was 0.2 μL and the temperature of column was 35 ℃. The detection wavelengths were set at 270 nm (fingerprint), 220 nm (content determination). The 28 batches of samples fingerprint were analyzed by similarity evaluation, principal component analysis (PCA), and partial least squares discriminant analysis (PLS-DA). The identification and content determination of the main common peaks were carried out by comparison with reference. Results We set up the common mode of the fingerprint with 20 common peaks, and six of them were identified, which were gastrodin (1), 4-hydroxybenzyl alcohol (3), p-hydroxybenzaldehyde (7), parishin B (14), parishin C (15), and parishin A (18). The similarities among 28 samples were all above 0.90. PCA was used to divide the samples into five groups, and six different substances were found. The content of the same component and the fluctuation range among different components were all different. The content of the five components of each origin was Guizhou 2.52% > Yunnan 2.49% > Shaanxi 2.33% > Hubei 2.10% > Zhejiang 1.90% > Anhui 1.65%.Conclusion The establishment of UPLC fingerprint and simultaneous determination of five active components provide a reference for quality control of Gastrodia Rhizoma.
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To establish a content determination method for 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside (TSG) of the crude/processed root of Polygonum multiflorum from different habitats in China and set up the fingerprint by using UPLC. Various samples were pretreated by macro-porous resin. Then UPLC analysis was performed on Waters ACQUITY UPLC@BEH C18 chromatographic column (2.1 mm×50 mm, 1.7 μm) at (25±5) ℃. A binary gradient elution system was composed of acetonitrile (phase A) and 0.5% acetic acid solution (phase B). Detection was performed at the wavelength of 254 nm, and the mobile flow rate was set at 0.3 mL•min⁻¹. Results showed that the yield of extraction of the 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside from root of P. multiflorum was all over 25.0% after macro-porous resin separation; an exclusive UPLC fingerprint method of the crude/processed root of P. multiflorum from different habitats was successfully set up and 17 chromatographic peaks were calibrated. Cluster analysis can not entirely distinguish the crude one from the processed one, while principal component analysis absolutely can. 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside is the composition that has largest differences in variable importance in projection (VIP) between crude and processed root of P. multiflorum. The separating method can gain high-purity 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside, and the determination method is simple, sensitive, reliable and can be used in fast identifying the crude/processed root of P. multiflorum or as a method for overall quality control of root of P. multiflorum.
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OBJECTIVE: To predict the potential geographical distribution of Gentiana rhodantha and study the relationship between species distribution, chemical compounds content and ecological factors in Yunnan-Guizhou Plateau. METHODS: MaxEnt modeling combined with geographic information system (GIS) was used to predict the potential geographical distribution of G. rhodantha. Ultra performance liquid chromatography (UPLC)was used to establish the UPLC fingerprints of different parts of medicinal materials. Pearson correlation analysis and stepwise regression analysis were used to analyze the relationship between various chemical compounds and ecological factors. RESULTS: The AUC of the ROC curves of the training data and test data were 0.919 and 0.915, respectively, which indicated that the predictive results with the maximum model were highly precise and exact. The results of species distribution modeling showed that the main environmental factors determining the potential distribution were annual temperature range (the most suitable range: 16.0-27.0℃), mean diurnal range (the most suitable range: 8.5-11.6℃), average monthly precipitation of June (the most suitable range: 200-400 mm), average monthly precipitation of September (the most suitable range: 90-125 mm), average monthly maximum temperature of June (the most suitable range: 21.0-27.0℃), temperature seasonality (the most suitable range: 4 000-5 200), average monthly precipitation of October (the most suitable range: 65-110℃), average monthly minimum temperature of July (the most suitable range: 14.5-20.5℃), average monthly maximum temperature of January (the most suitable range: 14.5-25.5℃), subsoil pH (the most suitable range: pH<5), and average monthly precipitation of April (the most suitable range: 14.5-25.5 mm). Correlation analysis showed that the contents of mangiferin, total compounds and their ratios of aerial part and underground part had significant relationship with temperature, precipitation and hysico-chemical properties of top soil (P<0.05 or P<0.01) and had significant negative (P<0.05) or most significant negative relationship with habitat suitability (P<0.01). The variation of temperature and precipitation of June to August and subsoil pH subsoil CEC were the key ecological factors of accumulation of the chemical compounds in G.rhodantha. CONCLUSION: The best growing areas for G. rhodantha are mainly located in Middle and West Yunnan, Southeast and South Guizhou.
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Objective: To develope a method for UV-Vis and UPLC fingerprint on various medicinal parts of Gentiana rhodantha. The chemical compounds variation in roots, steams, leaves, and flowers were studied by using fingerprint data combined with multivariate analysis. Methods: Using Shim-pack XR-ODS III liquid chromatographic column (150 mm × 2.0 mm, 2.2 μm) for gradient elution, mobile phase was water with 0.1% formic acid and acetonitrile, temperature was 40℃, detection wavelength was 242 nm, injection value was 0.3 μL, and flow rate was 1.00 mL/min. Electrospray ionization (ESI) source and MRM model, the interface voltage was set to 3.5 kV. Desolation line (DL) temperature was 250℃. Nebulizing gas and drying gas were nitrogen at a flow rate of 3.0 and 15.0 L/min, respectively. Collision gas was 0.15 mL/min. UV-Vis detection wavelength range at 200 - 500 nm, slit was 1.0 nm, step was 0.5 nm. Multivariate analysis methods including partial least squares discriminant analysis (PLS-DA), variable importance (VIP), and hierarchical cluster were used for this projection. Results: The investigation of UPLC and UV-Vis showed RSD was lower than 2.00% for precision, repeatability, and stability. The recoveries of loganin acid, mangiferin, and sweroside were 97.89% - 102.71% and RSD was 1.09% - 2.88%. Pretreatment by 11 smooth + first order was the best data processing method for PLS-DA model (R2cal = 0.9315, RMSEE = 0.302, R2val = 0.901, and RMSEP = 0.341). UV-Vis spectra of roots, steams, leaves, and flowers had the significant fingerprint characteristic. Similarity analysis showed the chemical compounds in the leaves and flowers were similar and those in the stems and roots were similar too. Similarity index of root medicinal material has a widely range. The quality of the roots was not stable. The contents of loganic acid and mangiferin were the highest in the leaves and flowers [(1.46 ± 0.42) and (51.59 ± 15.45) mg/g]. The content of sweroside was the hightest in the roots [(4.41 ± 3.24) mg/g]. The total content of compounds of the leaves was higher than other medicinal part, the three compounds, loganic acid, mangiferin, and sweroside were very used for the discrimination of different parts of G. rhodantha. Cluster anslysis showed there were geography characteristics of the constituents. Sample collected from high altitude was higher than samples collected from lower altitude. And they were classified in different groups. Conclusion: UPLC and UV-Vis fingerprint could describe the variation of chemical compounds in the roots, steams, leaves, and flowers. All the results could provide the evaluation method and basic theory of G. rhodantha resource.
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Objective: In order to identify the authenticity and control the quality of Xinjiang chrysanthemum, the fingerprinting of it was established using UPLC and discussed in this paper. Methods: Thirteen batches of Xinjiang chrysanthemum from different origins were analyzed by UPLC with catechin, luteoloside, quercetin, and 3, 5-di-O-caffeoyl quinic acid as reference substances. The chromatographic condition was performed on Waters ACQUITY UPLC HSS T3 C18 (150 mm × 2.1 mm, 1.8 μm) and eluted with mobile phase containing acetonitrile-pH 3.0 phosphoric acid solution. The flow rate was 0.2 mL/min with detection wavelength 280 nm in the temperature of 30℃. Results: There were 17 peaks in the UPLC fingerprint of Xinjiang chrysanthemum and three components were identified by three peaks determined using reference substance. Conclusion: The similarity of 13 batches is little difference, but is more than 0.90. The results showed that the UPLC fingerprint chromatogram could be used as a valuable method for the quality evaluation of Xinjiang chrysanthemum.
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OBJECTIVE: To establish a UPLC fingerprint of Ginkgo biloba leaves and determine the contents of eleven flavonoid glycosides. METHODS: A UPLC method was established for the first time to simultaneously quantify the eleven major flavonoid glycosides in Ginkgo biloba leaves. The analysis was performed on an Agilent ZORBAX Eclipse Plus C18 column (4.6 mm × 50 mm, 1.8 μm), the mobile phase A was acetonitrile, the mobile phase B was 0.4% phosphoric acid, gradient elution was performed at the flow rate of 0.6 mL · min-1, and the detection was carried out at 360 nm. Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine(2004 A) was used in data analysis. RESULTS: The UPLC fingerprint of Ginkgo biloba leaves was established. Total 21 peaks were selected as the characteristic common peaks and 11 of them were identified. There were some differences in the fingerprint chromatograms between 32 batches of Ginkgo biloba leaf samples. The validation result showed that the 11 flavonoid glycosides had good linearity and recoveries. CONCLUSION: The established UPLC method is simple, rapid, accurate, reliable, and sensitive, and can be applied to the identification and quality control of Ginkgo biloba leaves.