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Objective: To establish a determination method for kinds of chemical components of iridoids in the roots of Gentiana crassicaulis in transdermal absorption liquid, and research its transdermal permeability, so as to provide scientific basis for transdermal delivery system, clinical medication and reform of the traditional forms of G. crassicaulis. Methods: Based on the results of the previous investigation, in this paper, using the roots of G. crassicaulis as the research subject, a certain concentration solution of G. crassicaulis extract was prepared by the alcohol extraction method. Three kinds of common penetration enhancers, azone, borneol and propylene glycol were used. The effects of single penetration enhancer and dual compound penetration enhancers on the transdermal penetration of five kinds of chemical components of loganic acid, shanzhiside methyl ester, swertimarin, gentiopicroside and sweroside in vitro and the three kinds of chemical components of gentiopicroside, loganic acid and swertimarin in vivo were quantitatively studied by the method of HPLC to investigate the transdermal permeability of G. crassicaulis extract in mice skin model. Results: According to the experimental results, compared to the control group, penetration enhancers significantly increased the absorption of five chemical components of G. crassicaulis in vitro. Transdermal absorption rates (J) of loganic acid, shanzhiside methyl ester, swertimarin, gentiopicroside and sweroside were 12.306 0, 1.248 8, 4.187 5, 153.030 0 and 5.012 6 μg/(cm2∙h), respectively. The transdermal enhancer effects of A (5% azone), B (5% borneol), C (5% propylene glycol), A + B (2.5% azone and 2.5% borneol), A + C (2.5% azone and 2.5% propylene glycol), A + C (2.5% borneol and 2.5% propylene glycol) were 9.73, 2.57, 13.94, 15.92 and 8.08 times faster than the control group, respectively. Among these, the group of A + C had a marked osmotic enhancer effect in vitro. In comparison with the control group, the in vivo percutaneous penetration test indicated that the dual compound penetration enhancers of 2.5% azone and 2.5% propylene glycol had a marked effect for the permeability enhancement. Conclusion: This study showed azone and propylene glycol significantly promoted the percutaneous penetration effect of loganic acid, shanzhiside methyl ester, swertimarin, gentiopicroside and sweroside of G. crassicaulis, and this study laid a foundation for the quality control of percutaneous drug delivery preparation of G. crassicaulis.
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OBJECTIVE： To establish HPLC fingerprints of Nauclea officinalis extract syrup， and to determine the contents of 9 components. METHODS： HPLC method was adopted. The determination was performed on Diamonsil C18（2）column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm， and column temperature was 30 ℃. The sample size was 10 μL. Using strictosamide as reference， HPLC chromatograms of 20 batches of N. officinalis extract syrup were drawn. The similarity of HPLC chromatograms were evaluated by using TCM Fingerprint Similarity Evaluation System （2004A edition） to confirm common peaks. The contents of 9 components were determined by standard curves. RESULTS： There were 26 common peaks in 20 batches of HPLC chromatograms， and the similarity was higher than 0.98. Compared with mixed control， 9 chemical components were identified， such as 3，4-dihydroxybenzoic acid， neochlorogenic acid， loganic acid， chlorogenic acid， cryptochlorogenic acid， swertioside， pumiloside， strictosamide and vincosamide. The linear range of 9 components were 17.24-275.84， 7.56-120.96， 15.40-246.40， 7.84-125.44， 8.64-138.24， 7.96-127.36， 8.40-134.40， 48.56-776.96， 4.16-66.56 μg/mL（all r≥0. 999）， respectively. The limits of detection were 0.043 1， 0.126 0， 0.038 5， 0.130 7， 0.144 0， 0.066 3， 0.070 0， 0.012 1， 0.052 0 μg/mL， respectively. The limits of quantitation were 0.215 5， 0.189 0， 0.077 0， 0.196 0， 0.288 0， 0.132 7， 0.105 0， 0.097 6， 0.138 7 μg/mL， respectively. RSDs of precision， stability and reproducibility tests were all lower than 2.0％ （n＝6）. Average recoveries were 99.6％、106.3％、100.1％、102.0％、98.4％、100.0％、99.3％、100.6％ and 101.2％， and RSDs were 1.20％、0.24％、0.59％、1.00％、0.73％、1.30％、1.10％、1.80％、1.90％（n＝6）. CONCLUSIONS： Established HPLC fingerprints and quantitative determination method of N. officinalis extract syrup are accurate， specific and sensitive. It can provides reference for quality control of N. officinalis extract syrup.
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OBJECTIVE： To establish the method for the content determination of gentiopicrin and loganic acid in Gentiana scabra， and to investigate the correlation of their contents with appearance traits and quality gradation criterion. METHODS： HPLC method was adopted. The determination was performed on Ascentis Express C18 column with mobile phase consisted of 0.1％ phosphoric acid-acetonitrile （gradient elution） at the flow rate of 0.4 mL/min. The column temperature was 30 ℃， and detection wavelength was set at 240 nm. The sample size was 1 μL. Taking the length， number and diameter of fibrous roots as indexes， the appearance and morphological characteristics of G. scabra were studied. The relationship of gentiopicrin and loganic acid content with the appearance property of medicinal material was analyzed by SPSS 21.0 software. k-mean clustering analysis was carried out by using SPSS 21.0 software， and gradation standard for G. scabra was established preliminarily. RESULTS： The linear range of gentiopicrin and loganic acid were 0.5-3.0 μg/mL （r＝0.999 9） and 0.05-0.50 μg/mL （r＝0.999 9）. The limit of quantification of gentiopicrin and loganic acid were 0.295， 0.289 μg/mL； the detection limit were 0.082， 0.081 μg/mL； RSDs of precision， stability， repeatability tests were all lower than 2％； the recovery rates were 97.56％- 102.23％ （RSD＝1.56％， n＝6） and 97.58％-102.67％ （RSD＝1.86％， n＝6）. Correlation results showed that there was a significant positive correlation of the length of G. scabra， the number of roots， root diameter， with the contents of gentiopicrin and loganic acid. The order of affecting content was the number of roots ＞length ＞root diameter. k-means clustering analysis showed that 54 batches of G. scabra was divided into two categories； S4-S6，S13，S17-S23，S25，S28，S31-S34 were clustered into a category； S1-S3， S7-S12， S14-S16， S24， S26，S27，S29， S30，S35-S54 were clustered into the other category. The results of gradation showed that 54 batches of G. scabra could be divided into two grades， and the results were consistent with the cluster analysis. CONCLUSIONS： Established method is simple and stable， and can be used for simultaneous determination of gentiopicrin and loganic acid in G. scabra. The more fibrous roots， the longer the length， the thicker the root， the higher the content of gentiopicroside and loganic acid， the better the quality of G. scabra.
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To establish the HPLC fingerprint and determine five index components (loganic acid， chlorogenic acid， loganin， sweroside and asperosaponin Ⅵ) of Zishen Yutai pills by high performance liquid chromatography, and provide a scientific basis for its quality control. The fingerprint chromatogram was analysed by the chromatographic fingerprint similarity evaluation system for tradition Chinese medicine (2012), fifteen common peaks were obtained at the wavelength of 254 nm. Different batches of Zishen Yutai pills showed a similarity of above 0.90 in HPLC fingerprint profiles. For the quantitive analysis method, The separation of five components showed good regression (>0.999 2) with linear ranges, and the mean recoveries were in the range of 97.62%-101.9%, with the RSD (=9) less than 3%. The established fingerprint and quantitative analysis methods are highly specific, simple and accurate, which can reflect the quality of Zishen Yutai pills more comprehensively, and can be used for its quality control.
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OBJECTIVE:To establish a method for the simultaneous determination of loganic acid and isoscoparin in Sanwei longdanhua tablets. METHODS:HPLC method was adopted. The determination was performed on Inertsil ODS-3 column with mobile phase consisted of methanol-0.2% phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm,and column temperature was 30 ℃. The sample size was 10 μL. RESULTS:The linear range were 0.040 08-4.008 0 μg(r＝0.999 9)for loganic acid and 0.021 96-2.196 0 μg(r＝0.999 9)for isoscoparin. The quantitative limits were 0.160 32 and 0.087 8 ng/mL,and detection limits were 0.080 16 and 0.043 92 ng/mL. RSDs of precision,stability and reproducibility tests were all lower than 2%. The recoveries were 103.07%-104.26%(RSD＝0.52%,n＝6) and 95.57%-99.61%(RSD＝1.55%,n＝6). CONCLUSIONS:The method is simple,accurate and suitable for simultaneous determination of loganic acid and isoscoparin in Sanwei longdanhua tablets.
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OBJECTIVE:To optimize the extraction technology of Yigu granule. METHODS:L9(34)orthogonal test was used, using comprehensive scores of transfer rate of icariin,gentiopicroside,loganic acid and yield rate as evaluation indexes,alcohol volume fraction,the amount of solvent,extraction times and extraction time as investigation factors,extraction technology of Yigu granule was optimized,and the verification test was conducted. RESULTS:The optimized extraction technology was 10-fold 60%ethanol reflux extraction for twice,1 h once. Results of verification test showed,average transfer rates of icariin,gentiopicroside and loganic acid were 81.28%(RSD=1.54%,n=3),48.71%(RSD=2.37%,n=3)and 59.82%(RSD=2.52%,n=3);aver-age yield rate was 31.48%(RSD=1.97%,n=3). CONCLUSIONS:The optimized extraction technology for Yigu granule is sta-ble and feasible with good reproducibility,which can provide basis for the follow-up study of production process.
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Objective: To identify and determine the main chemical constituents of Lonicera maackii, and to provide data support for the development and utilization of L. maackii. Methods: The chemical constituents of L. maackii were identified by HPLC-DAD-ESI-Q-TOF/MS and the main chemical constituents were determined by HPLC-DAD. Results: A total of 31 chromatographic peaks were detected and 19 chemical constituents were identified from L. maackii, and the contents of the 10 compounds (logaric acid, chlorogenic acid, loganin, morroniside, Secoxyloganin, rutin, hyperoside, luteoloside, chlorogenic acid A, and chlorogenic acid C) were determined. Conclusion: The contents of the main chemical constituents in the flower buds of L. maackii were higher than those in the blooming stage and yellow flowering stage. The contents of main components in L. maackii were similar to those of Lonicerae Japonicae Flos. The results of this study provide data support for the development and utilization of L. maackii.
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Objective: An ultra performance liquid chromatographic (UPLC) method was developed for simultaneously determining seven constituents, such as loganic acid, swertiamarin, 6'-O-β-D-glucosyl gentiopicroside, gentiopicroside, sweroside, isoorientin, and isovitexin, from the medicinal plants of Gentiana macrophylla. Methods: The separation was performed on an Acquity UPLC® BEH C18 column (50 mm ×2.1 mm, 1.7 μm) through a gradient elution of methanol-0.04% aqueous phosphorite at a flow rate of 0.3 mL/min. The detection wavelength was 242 nm, and the column temperature was set at 30℃. Results: For the seven analytes, loganic acid, swertiamarin, 6'-O-β-D-glucosyl gentiopicroside, gentiopicroside, sweroside, isoorientin, and sovitexin, a good linearity (r≥0.9995) was obtained in the range of 2.100-537.100, 1.050-270.000, 0.920-236.000, 11.100-2830.000, 0.750-192.000, 0.167-102.000, and 0.216-52.800 μg, respectively. Their average recoveries (n=6) were 97.83%-100.08%, respevtively, with RSD values less than or equal to 3.76%. Conclusion: The UPLC method is simpler and more effective than HPLC, and can be used for the simultaneous determination of seven indicative constituents in medicinal plants of G. macrophylla.
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Objective: To study the chemical constituents of Yao medicine Zhongliuteng, the canes of Pileostegia tomentella. Methods: The chemical constituents were isolated and purified by silica gel chromatography repeatedly from the canes of P. tomentella and their structures were identified by spectral analysis and chemical methods. Results: Sixteen compounds were isolated from the canes of P. tomentella and their structures were identified as loganic acid (1), 4-O-α-L-arabinofuranosyl-(1→6)-β-D-glucopyranoside (4S)-4-hydroxyl-β-ionone (2), resorcinol (3), daphnin (4), skimmin (5), umbelliferone (6), vogeloside (7), diethyl phthalate (8), secologanin (9), secologanin dimethyl acetal (10), sweroside (11), foliasalacioside B (12), benzyl-O-β-D-apiofuranosyl-(1″→6')-β-D-glucopyranoside (13), maltose (14), glucose (15), and daucosterol (16). Conclusion: All compounds are obtained from the plant for the first time, and all compounds are also obtained from the plants of Pileostegia Hook. f. et Thoms. for the first time except compounds 5, 6 and 16.
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Objective: To establish an HPLC method for simultaneously determining seven components, such as loganic acid, 6′-O-β-D-glucopyranosyl gentiopicroside, swertiamarine, gentiopicroside, swertiamarin, isoorientin, and isovitexin, from the flowers of Gentiana tibetica and G. dendrology. Methods: Chromatographic analysis was achieved on an Agilent Zorbax ODS column (150 mm × 4.6 mm, 5 μm) by gradient elution of acetonitrile-0.4% acetic acid in water at 30℃. The flow rate was 0.4 mL/min and the detection wavelength was 254 nm. Results: The calibration curves of all the seven constituents showed good linearity in a relatively wide concentration range. The linear ranges of loganic acid, 6′-O-β-D-glucopyranosyl gentiopicroside, swertiamarine, gentiopicroside, gentiopicroside, isoorientin, and isovitexin were 0.228-2.280, 0.680-6.800, 0.220-2.20, 1.476-14.760, 0.200-2.000, 0.436-4.360, and 0.276-2.760 μg (R2 ≥ 0.999 2), respectively. The recoveries (n = 9) of loganic acid, 6′-O-β-D-glucopyranosyl gentiopicroside, swertiamarine, gentiopicroside, gentiopicroside, isoorientin, and isovitexin were 100.9%, 99.4%, 101.0%, 105.7%, 103.1%, 98.4%, 104.2%. Conclusion: This method is simple, accurate, and specific, and can be used for the determination of seven constituents in the flowers of G. tibetica and G. dendrologi.
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Objective: To establish a method for the determination of loganic acid, chiratin, and loganin in Dipsaci Radix and improve the quality evaluation by analyzing Dipsaci Radix from different habitats. Methods: The analysis was performed on Venusil MP C18 column (250 mm×4.6 mm, 5 μm) with mobile phase consisting of 0.1% H3PO4-acetonitrile for gradient elution: 0-10 min, 8%-9% B; 10-30 min, 9%-11% B; 30-35 min, 11%-100% B. The flow rate was 1 mL/min and the column temperature was 25℃. Results: The results showed that loganic acid, chiratin, and loganin were well separated with the good linearity in 18.4-368.2, 2.02-40.4, and 17.5-349.6 μg/mL, respectively. The average recoveries of the three iridoid glycosides were 99.34%, 99.19%, and 101.61%. Conclusion: Loganic acid, chiratin, and loganin are the main iridoid glycosides in Dipsaci Radix. The method can easily be applied to the content determination of loganic acid, chiratin, and loganin in Dipsaci Radix quickly. The content ranges of loganic acid, chiratin, and loganin are 20.4-186.8, 26.4-177.7, and 1.9-13.2 mg/g, respectively, and the sum of the three iridoid glycosides shows no significant differences in various Dipsaci Radix from different habitats, and proposes to evaluate the quality of Dipsaci Radix.
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OBJECTIVE: To establish the HPLC fingerprint of Semen Strychni by chemometrics and the method of linear calibration using two reference substances (LCTRS). METHODS: The fingerprint of Semen Strychni was established by HPLC, using similarity analysis, 2-dimensional clustering analysis and principal component analysis for quality evaluation. LCTRS was used for identification of chromatographic peak. RESULTS: Fourteen chromatographic peaks were selected as common peaks in the fingerprints of 10 batches of Semen Strychni, and the chromatographic peaks of loganic acid, chlorogenic acid, strychnine and brucine were selected as characteristic peaks. By means of chemometrics evaluation, 10 batches of Semen Strychni were classified by their origins. The precision of predicting retention time by LCTRS was superior to the relative retention time method. CONCLUSION: LCTRS is more accurate for predicting retention time and suitable for more kinds of chromatographic columns than relative retention time method. In addition, the established fingerprint is specific, and can be used for the quality evaluation of Semen Strychni.