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OBJECTIVE：To establish the UPLC fingerprint of Polygonum cuspidatum ，and to determine the contents of four active ingredients and to provide reference for the quality evaluation of P. cuspidatum . METHODS ：The determination was performed on Waters BEH C 18 column（100 mm×2.1 mm，1.7 μm）with mobile phase consisted of acetonitrile- 0.2％ formic acid （gradient elution ）at flow rate of 0.4 mL/min. The column temperature was 40 ℃，and detection wavelength was 290 nm. The sample size was 1 μL. The fingerprints were evaluated by similarity calculation，cluster analysis and orthogonal partial least square discriminant analysis （OPLS-DA）. Using polydatin as internal standard ，relative calibration factors of resveratrol ，emodin-8-O- β-D-glucoside and emodin were determined to develop a method of QAMS. The contents of 4 above components in 15 batches of P. cuspidatum were calculated by relative calibration factors. The results of QAMS were compared with those of external standard. RESULTS：UPLC fingerprints of 15 batches of P. cuspidatum were established ，and 12 common peaks were confirmed. Five components were identified ，i.e. polydatin ，resveratrol，emodin-8-O-β-D-glucoside，emodin，emodin methyl ether. The fingerprint similarity of 15 batches of P. cuspidatum was in the range of 0.865-0.976. According to cluster analysis ，15 batches of P. cuspidatum were classified into 4 categories，showing certain regularity of origin. Seven markers were identified by OPLS-DA method. The order of difference significance was peak 7＞emodin-8-O-β-D-glucoside＞resveratrol＞peak 8＞polydatin＞peak 1＞ peak 10. The relative deviation among the contents of resveratrol ，emodin-8-O-β-D-glucoside and emodin in 15 batches of P. cuspidatum determined by QAMS and external standard method was less than 5.0％，indicating that there was no significant difference between the two methods. CONCLUSIONS ：UPLC fingerprint combined with QAMS method is convenient and reliable for the quality evaluation of P. cuspidatum ；the quality of P. cuspidatum produced in Chongqing and Anhui province is better.
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OBJECTIVE：To provide reference for the identification and proces sing end-point determination of raw Morus alba and its processed products （honey-processed M. alba ）. METHODS ：UPLC method was adopted. The determination was performed on Waters BEH Shield RP C 18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid solution （gradient elution ） at the flow rate of 0.30 mL/min. The column temperature was set at 30 ℃. The program wavelengths were set at 280 nm（0-4 min） and 320 nm（4-35 min）. Similarity Evaluation System for Chromatogram Fingerprint of TCM （2012 edition）was used to establish UPLC fingerprint and carry out similarity evaluation of 13 batches of M. alba and honey-processed M. alba . The chromatographic peaks were identified with reference substance fingerprint. The colorimetric value （L，a，b） of 13 batches of M. alba and honey-processed M. alba powder were determined ，and average total colorimetric value （E）was calculated. OPLS-DA and cluster analysis were adopted to analyze the differences in fingerprints and colorimetric values of M. alba before and after processing. At the same time ，the dynamic change rule of fingerprint and colorimetric value of honey-processed M. alba at different processing time points were analyzed to determine the processing end-point. RESULTS ：There were obvious differences in fingerprints before and after processing ，and the similarity of 13 batches of M. alba and honey-processed M. alba were all higher than 0.9. Totally 21 common peaks were calibrated for M. alba ，and 23 common peaks for honey-processed M. alba ；peak 1 and peak 2 were newly produced compounds of honey-processed M. alba . Peak 2，peak 7，peak 14 and peak 19 were identified as 5-hydroxymethylfurfural， mulberry glucoside A ，oxidized resveratrol ，mulberry flavonoids G. Results of OPLS-DA showed that the peak area-sample quantity ratio of peak 1，peak 2，peak 18，peak 20 and the chromaticity values （L，a，b）were the most important factors affecting the difference of raw and processed products of M. alba . When the E ranged 75.84-80.88 as the processing end-point of honey-processed M. alba ，the processing time was determined as 22-34 min. CONCLUSIONS ： The established UPLC fingerprint and colorimetric value determination method can be used to identify the raw and processed products of M. alba as well as determine the processing end-point of honey-processed M. alba .
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OBJECTIVE：To e stablish UPLC characteristic chrom atograms of Euodia rutaecarpa ，processed E. rutaecarpa decoction piece ，water decoction and formula granules ，and to compare its relationship and difference. METHODS ：UPLC method was used. The determination was performed on YMC Triart C 18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid water solution （gradient elution ）at the flow rate of 0.3 mL/min. The detection wavelength was set at 254 nm，and column temperature was 30 ℃. The sample size was 1 μL. Using limonin as reference，the characteristic chromatograms of E. rutaecarpa ， processd E. rutaecarpa decoction piece ，water decoction and formula granules （each 10 batches，totally 60 batches）were drawn. The similarity was evaluated with TCM Chromatographic Fingerprint Similarity Evaluation System （2012 edition），and to determine the common characteristic peak. The differences of ratio of common characteristic peak area were evalucoted according to variance analysis. Meanwhile ，the cluster analysis and principal component analysis （PCA） were performed to research the differences of E. rutaecarpa ，processed E. rutaecarpa decoction piece ，water decoction and formula granules by using SPSS 20.0 software. RESULTS ：Totally 16 and 17 common peaks were respectively confirmed in characteristic chromatograms of E. rutaecarpa samples and processed E. rutaecarpa samples（decoction piece ，water decoction and formula granules ）. No. 8，9，11， 17 peaks were identified as limonin ，evodiamine，rutaecarpine and glycyrrhizic acid. Compared with decoction piece ，the similarities of characteristic peak between water decoction and formula granules were lower than 0.55，while those between water decoction and formula granule were higher than 0.95. Cluster analysis and PCA results showed that E. rutaecarpa decoction piece and processed E. rutaecarpa decoction piece could be clustered into one category ；E. rutaecarpa water decoction and formula granules could be clustered into one category ；processed E. rutaecarpa water decoction and formula granules could be clustered into one category. CONCLUSIONS ：Compared with E. rutaecarpa ，processed E. rutaecarpa additionally contain glycyrrhizic acid ； main che mical components of decoction piece ，water decoction and formula granules are basically the same ，but the contents of the components between decoction piece to water decoction and formula granules are different.
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OBJECTIVE：To establish HP LC ch aracteristic ch romatogram of different medicinal parts of Cirsium japonicum ， and to compare the difference of chemical components in different medicinal parts of C. japonicum according to chemical identification method ，and to provide reference for quality control and evaluation of C. japonicum . METHODS ：Medicinal material （overground part ），leaves，flower，main stem and lateral stem of C. japonicum were determined by HPLC. According to the TCM Chromatographic Fingerprint Similarity Evaluation System （2012A edition ），the chromatograms were matched to generate the HPLC characteristic chromatogram of each medicinal part. The differences of common characteristic peak area were analyzed according to variance analysis of single factor. The chromatographic peaks were identified by comparison of reference substance. Meanwhile，the chemical pattern recognition was performed to research the different medicinal parts of C. japonicum according to principal component analysis （PCA）and cluster analysis. RESULTS ：HPLC characteristic chromatograms of medicinal material ， leaves，flower，main stem and lateral stem from C. japonicum were established respectively ，and 15 common peaks were confirmed for medicinal material ，leaves and flower of C. japonicum ；11 common peaks were confirmed in chromatograms of main stem and lateral stem from C. japonicum （absence of No. 7，9，12，13 peak）. The contents of chemical components were different greatly among different medicinal parts. No. 1，2，3，10，11 peaks were identified as neochlorogenic acid ，chlorogenic acid ， cryptochlorogenic acid ，linarin and pectolinarin. Results of PCA and cluster analysis showed that chemical pattern recognition and clustering of the flower and stem of C. japonicum were distinct and can be clustered into one category respectively. However ，the leaves distribution of C. japonicum was relatively scattered ，so it was difficult to cluster . CONCLUSIONS ：Established HPLC characteristic chromatogram-chemical pattern recognition can reflect the differences of different medicinal parts of C. japonicum integrally， comprehensively and truly ， which has vital significance for origin indentification ， quality control and overall evaluation of C. japonicum .
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Objective To compare the effects of two crushing methods on dissolution rate and decocting rate of water-soluble protein in Plastrum Testudinis.Methods The dissolution rate of water-soluble protein was compared in Plastrum Testudinis processed by the traditional crushing method(passing 100-eyes screen,fine powder)and ultra-fine crushing method(passing 300-eye screen,ultra-fine powder);besides,orthogonal design was applied to study the process technical condition of decocting rate in Plastrum Testudinis.Results Water-soluble protein in fine powder of Plastrum Testudinis was hardly detected while the dissolution rate of water-soluble protein in ultra-fine powder was 51.2 %in 20 minutes and up to 70.5 %at 2 hours.Three influencing factors of fineness,decocting frequencies and decocting time were measured with orthogonal test.The results of variance analysis showed that fineness significantly influenced the decocting rate(P .05).Conclusion The ultra-fine powder technique is propitious to the increase of dissolution rate and decocting rate of water-soluble protein in Plastrum Testudinis.