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OBJE CTIVE To establish a method for quality evaluation of Xin ’an capsule by combining fingerprint ， multi-component quantitative analysis and chemical pattern recognition analysis. METHODS High performance liquid chromatography（HPLC）combined with Similarity Evaluation System of TCM Chromatogram Fingerprint （2012 edition）were used to establish the fingerprints of 24 batches of Xin ’an capsules and evaluate the similarity. The common peaks were determined. The contents of glucosylvitexin ，rhamnosylvitexin，vitexin，hyperoside and isoquercetin in Xin ’an capsules were determined by the same HPLC method. Taking the common peak area of fingerprint as the variable ，MetaboAnalyst 5.0 tool was used to draw the cluster analysis （CA）heat map. SIMCA 14.1 software was used to perform principle component analysis （PCA）and partial least squares-discriminant analysis （PLS-DA）. RESULTS Twelve common peaks were identified with the similarity greater than 0.97. Six common peaks were identified as chlorogenic acid ，glucosylvitexin，rhamnosylvitexin，vitexin，hyperoside and isoquercetin.The linear range of glucosylvitexin ，rhamnosylvitexin，vitexin， hyperoside and isoquercetin were 2.36-151.35，9.15-585.20， 1.20-76.50， 0.68-43.20， 0.44-27.90 µg/mL（all r＞0.999）.RSDs of precision ，repeatability and stability （24 h）tests were 163.com all less than 2.00％ . The average recoveries were 95.80％（RSD＝0.96％ ，n＝6），102.10％ （RSD＝0.93％ ，n＝6）， 103.26％（RSD＝1.28％，n＝6），103.89％（RSD＝0.73％，n＝6） and 102.09％（RSD＝1.79％，n＝6），respectively. The contents of the five components were 0.988 8-1.559 1，4.336 6-11.220 1， 0.065 1-0.830 5，0.043 8-0.692 5 and 0.023 2-0.427 2 mg/grain，respectively. The results of CA and PCA showed that 24 batches of samples could be divided into three categories ，i.e. S 1-S15，S16-S18 and S 19-S24. PLS-DA showed that variable importance in projection values of the corresponding component of peak 6 and glucosylvitexin （peak 7），rhamnosylvitexin（peak 8），hyperoside （peak 10） and isoquercetin （peak 11） were greater than 1. CONCLUSIONS The established HPLC fingerprint and multi-component quantitative method are simple and feasible. Combined with chemical pattern recognition analysis ，it can be used for the quality control of Xin ’an capsules. Glucosylvitexin ，rhamnosylvitexin and other components may be differentital markers affecting the quality of each batch of samples.
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The medicinal and edible Polygonatum cyrtonema is one of the original species of Polygonati Rhizoma. In this study,HPLC fingerprints for 25 batches of P. cyrtonema from 6 provinces were established. A total of 14 common peaks were identified and the similarities of the fingerprints were in the range of 0. 939-0. 999. In additon, the partial least squares-discriminant analysis(PLSDA) demonstrated that the samples had low discriminability except for JX-1 and most components of them had no significant correlation with environmental factors such as longitude, latitude, and altitude. Thus, chemical composition specificity of P. cyrtonema in natural distribution areas had no obvious regularity and their variation might be induced by the local environment. This conclusion explained the lack of records about Dao-di area of Polygonati Rhizoma. However, JX-1 boasted significantly higher content of 5-hydroxymethylfurfural(HMF) and 4',5,7-trihydroxy-6,8-dimethylhomoisoflavone( HIF), thick and long inflorescence and rhizome, and extremely high yield. Therefore, excellent variety of P. cyrtonema might have great potential to improve the quality and yield of Polygonati Rhizoma. Moreover, three components of HMF, polygonalline A(PA), and HIF were identified in the fingerprint. Among them, HMF has the activities of blood rheology improvement, antioxidation, and anti-myocardial ischemia and PA is an indolizine alkaloid with potential anti-inflammatory activity. HIF, the characteristic homoisoflavone in Polygonatum, has the pharmacological activities of regulating blood glucose and anti-tumor. A quantitative analysis method can provide a theoretical basis for the improvement of the quality evaluation of Polygonati Rhizoma.
Subject(s)
Antioxidants , Chromatography, High Pressure Liquid , Polygonatum , RhizomeABSTRACT
OBJECTIVE: To establish a RP-HPLC method for the simultaneous determination of phenolic glycosides in Curculigins Rhizoma. METHODS: Angilent 1100 HPLC on a ZORBAX SB C18 column(4.6 mm × 250 mm, 5 μm) with Extend C18 guard column(4.6 mm × 12.5 mm, 5 μm) were used. The mobile phase consisted of acetonitrile(A)-0.1% phosphoric acid aqueous(B). The gradient elution program was as follows: 0-25 min, 4%-6% A; 25-58 min, 6% - 17% A; 58-85 min, 17%-22% A The flow rate was kept at 1 mL · min-1, and the column temperature was set at 30°C. The detection wavelength was 230 nm. RESULTS: The concentrations and peak areas of the 8 phenolic glycosides including 5-hydroxymethylfurfural(1), 2-hydroxy-5-(2-hydroxyethyl) phenyl-β-D-glucopyranoside(2), anacardoside (3), orcinol glucoside (4), orcinol-1-O-β-D-apiofuranosyl-(1 → 6) -β-D-glucopyrano-side(5), 2, 6-dimethoxybenzoic acid(6), curculigoside(7) and curculigine A(8) were in good linear relationship. The average recoveries of the 8 constituents met the requirement for determination. CONCLUSION: The method is simple, sensitive, reproductive, and suitable for simultaneous determination of a variety of phenolic glycosides in Curculigins Rhizoma. Copyright 2012 by the Chinese Pharmaceutical Association.
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ObjectiveTo develop a quantitative method for simultaneously determining multi-components in Rhei Radix et Rhizoma using one chemical reference substance.MethodsThe contents of multi-components were calculated by the UV relative correction factors (RCFs) of chrysophanol,physcion,and rhein to emodin.ResultsThe values of RCFs at 274 nm for rhein,chrysophanol,and physcion to emodin were 0.712,0.674,and 1.051.The calibration curves were linear over the ranges of 0.02-4.08,0.02-4.12,0.07- 12.92,and 0.02-3.68 μg/mL for rhein,emodin,chrysophanol,and physcion,respectively.The contents of emodin in 18 samples were determined by the extemal standard method,and the contents of the other three anthraquinone aglycones were calculated according to their RCFs.ConclusionNo significant difference is found in comparison with the classical method,indicating that the RCFs have high reliability within their linear ranges and could be used in quality control of Rhei Radix et Rhizoma.The quantitative analysis of multi-component with a single marker is especially suitable for herbal medicines containing unstable or hard to be purified components as quality control markers.