Study on HPLC fingerprint of Leonurus japonicus granules and the contents of 4 index components / 中国药房

OBJECTIVE To establish HPLC finger print of Leonurus japonicus granules，and to determine the contents of 4 index components such as leonurine hydrochloride ，ferulic acid ，rutin，hyperoside. METHODS The determination was performed on Inertsil TM ODS-3 column with mobile phase consisted of acetonitrile （A）-0.1％ formic acid solution （B）in the form of gradient elution；the flow rate was 1.0 mL/min，the detection wavelength was 280 nm，the column temperature was 25 ℃，and the sample size was 5 µL. Similarity Evaluation System of Chromatogram Fingerprint of TCM （2012 edition）was used for establishing the HPLC fingerprints of 10 batches of L. japonicus granules and analyzing their similarities. By comparing with HPLC fingerprints of reference substance ，the common peaks were identified. SPSS 25.0 and SIMCA 13.0 software were used for cluster analysis and principal component analysis ；the above HPLC method was used for the content determination of 4 index components in L. japonicus granules such as leonurine hydrochloride ，ferulic acid ，rutin，hyperoside. RESULTS HPLC fingerprints of 10 batches of L. japonicus granules were established ，and 16 common peaks were matched ，and 4 peaks identified were leonurine hydrochloride （peak 6），ferulic acid （peak 13），rutin（peak 14），hyperoside（peak 16）；the similarities of 10 batches of samples were all higher than 0.970. The 10 batches of samples could be divided into four categories by cluster analysis and principal component analysis；the classification results were consistent. The contents of leonurine hydrochloride ，ferulic acid ，rutin and hyperoside were 122.10-138.82 μ g/g，9.33-10.45 μ g/g，14.12-18.95 μ g/g，5.87-8.06 μ g/g，respectively. CONCLUSIONS Established HPLC fingerprint of L. japonicus granules and the method for the content determination of 4 index components are simple and easy to operate，and have high precision and good repeatability ，which provide reference for the quality evaluation of L. japonicus granules.

Fingerprint Establishment ，Content Determination and Chemical Pattern Recognition Analysis of Tibetan Medicine Ershiwuwei Luxue Pills / 中国药房

OBJECTIVE：To establish fingerprint of Tibetan medicine Ershi wuwei luxue pills ，and determine the contents of 5 components，and to conduct chemical pattern recognition. METHODS ：HPLC method was adopted. Using aesculetin as reference ， HPLC fingerprint of 10 batches of Tibetan medicine Ershi wuwei luxue pills were drawn. The similarity evaluation was conducted by using Evaluation System of TCM Chromatogram Fingerprint Similarity （2012 edition），and the common peaks were determined. Same HPLC method was adopted to determine the contents of 5 components in Tibetan medicine Ershi wuwei luxue pills. The cluster analysis and principle component analysis were performed by using SPSS 19.0 software. RESULTS ：Totally 11 common peaks were calibrated ，and the similarity was higher than 0.98. Five common peaks were identified ，as aesculetin ，orientin， isovitexin，isoscoparin and ellagic acid. The linear range of aesculetin ，orientin，isovitexin，isoscoparin and ellagic acid were 1.232-11.092 μg/mL（r＝0.999 6），2.766-24.893 μg/mL（r＝0.999 5），1.400-12.600 μg/mL（r＝0.999 8），0.600-5.400 μg/mL（r＝ 0.999 5），49.447-445.025 μg/mL（r＝0.999 4），respectively. RSDs of precision ，stability（24 h）and reproducibility tests were all lower than 2％. The average recoveries were 101.29％（RSD＝2.33％，n＝3），91.39％（RSD＝1.22％，n＝3），90.28％（RSD＝ 1.88％，n＝3），98.76％（RSD＝2.53％，n＝3），101.45％（RSD＝2.84％，n＝3），100.44％（RSD＝1.38％，n＝3），100.91％ （RSD＝1.73％，n＝3），97.78％（RSD＝2.07％，n＝3），99.15％（RSD＝1.28％，n＝3），100.27％（RSD＝1.81％，n＝3），98.38％ （RSD＝1.89％ ，n＝3），101.92％（RSD＝1.17％ ，n＝3），95.50％（RSD＝0.67％ ，n＝3），99.89％（RSD＝0.38％ ，n＝3）， 100.10％（RSD＝0.65％，n＝3），respectively. Their contents were 0.175-0.310， 0.351-0.632， 0.274-0.395， 0.186-0.278， 61932600 6.956-8.636 mg/g，respectively. Cluster analysis showed that 10 batches of Tibetan medicine Ershi wuwei luxue pills were clustered into two category ，with S1-S4 as one category and S5-S10 as one category. Principal component analysis showed that accumulative contribution rate of two principle components was 89.178％. CONCLUSIONS ：Established fingerprint is stable and feasible ，and the method of content determination is simple ， accurate and reproducible. They combined with chemical pattern recognition can be used for the quality control of the Tibetan medicine Ershiwuwei luxue pills.

Simultaneous Determination of 6 Flavonoids in Zhuang Medicine Engelhardia roxburghiana by HPLC-QAMS and Multivariate Statistical Analysis / 中国药房

OBJECTIVE：To establish a m ethod for simultaneous determination of neoastilbin ，astilbin，neoisoastilbin，isoastilbin， quercitrin and engeletin in Engelhardia roxburghiana，and conduct multivariate statistical analysis. METHODS ：HPLC-QAMS method was adopted. The determination was performed on Phenomenex SuperLu C 18 column with mobile phase consisted of acetonitrile-0.1％ formic acid （19 ∶ 81，V/V）at the flow rate of 1.0 mL/min. The detection wavelengths were set at 254 nm （neoastilbin，astilbin，neoisoastilbin，isoastilbin，engeletin）and 291 nm（quercitrin）. The column temperature was 30 ℃，and sample size was 10 μL. Using astilbin as internal substance，and the relative correction factors of other 5 factors were calculated. The contents of each component were calculated according to relative correction factor ，and were compared with the results of external standard method. SPSS 22.0 software was used for cluster analysis and principal component analysis. RESULTS ：The linear range of neoastilbin ，astilbin，neoisoastilbin，isoastilbin，quercitrin and engeletin were 0.007-0.311，0.871-18.184，0.002-0.119， 0.052-1.251，0.105-2.202，0.020-2.319 μg（r＞0.999），respectively. RSDs of precision ，reproducibility and stability （24 h）tests were all lower than 3％. The average recoveries were 97.32％，94.89％，97.15％，96.90％，97.52％ and 97.53％（RSDs were 1.09％ -2.60％ ，n＝6），respectively. The relative correction factors of neoastilbin ，neoisoastilbin，isoastilbin，quercitrin and engeletin were 1.252 6，1.198 3，0.958 6，0.807 1 and 1.138 1， respectively. The contents of neoastilbin ， neoisoastilbin， qq.com isoastilbin，quercitrin and engeletin measured by QAMS were 0.394 2-2.067 2，0.139 1-0.804 7，2.864 8-8.554 8，4.581 2- 11.371 1，1.028 9-13.401 5 mg/g；the contents of neoastilbin ， astilbin，neoisoastilbin，isoastilbin，quercitrin and engeletin were 0.367 2-1.925 3，46.361 1-126.342 1，0.138 1-0.798 8，2.966 2-8.857 8， 4.642 5-11.523 3，0.970 6-12.641 9 mg/g，respectively. Relative errors of two methods was lower than or equal to 3.55％. The results of cluster analysis showed that 9 batches of samples could be clustered into two categories ；S8 sample was one category and others were one category. The results of principal component analysis showed that accumulative contribution rate of former 2 principle components was 84.745％，and the results of sample classification were consistent with those of cluster analysis. CONCLUSIONS ： The established HPLC-QAMS method is accurate ，feasible and repeatable ，and can be used for simultaneous determination of 6 flavonoids in E. roxburghiana ，and it can provide reference for quality control.

HPLC Fingerprint Establishment ，Chemistry Pattern Recognition Analysis and Content Determination of Arnebia euchroma / 中国药房

OBJECTIVE：To es tablish the HPLC fingerprint of Arnebia euchroma ，analyze them with chemical pattern recognition technology ， and determine the contents of 3 components. METHODS ： HPLC method was adopted. Using acetylshikonin as reference ，HPLC fingerprint of 34 batches of A. euchroma from different sources were drawn. Similarity Evaluation System for TCM Chromatographic Fingerprint （2012A edition ）was used to evaluate the similarity of the samples ，and common peaks were determined. SPSS 19.0 and SIMCA 14.1 statistical software was used for cluster analysis ，principle component analysis and orthogonal partial least squares-discriminate analysis. According to the standard of the variable importance in the project greater than 1，the differential markers affecting the quality difference of A. euchroma were screened. Meanwhile ，the contents of 3 components were determined by the same HPLC method. RESULTS ：There were 12 common peaks in HPLC fingerprints for 34 batches of A. euchroma . The similarity of other samples were more than 0.86，except that t he three （No.2016A3005-5） batches of medicinal herbs on the market were less than 0.72；3 common peaks were identified ， such as shikonin ，acetylshikonin， β ，β ′-dimethylacrylic acanine. These 34 batches of samples could be classified into two categories . S 1， qq.com S4-S6，S13，S15-S20，S22，S26-S34 were clustered into one category，and others clustered into the other category. By principal component analysis ，the contribution rates of three principle components were 52.834％ ，18.600％ and 8.387％ . Accumulative contribution rate was 79.821％ . Six constituents，such as shikonin，acetylshikonin and β，β'-dimethylacrylic acanine were screened as differential markers，representing the major differences of A. euchroma . The linear range of above three components were 0.72-90，2.05-410，2.50-500 µg/mL（r all more than 0.999）， respectively. The limits of quantification were 0.132，0.135，0.118 µg/mL，respectively. The limits of detection were 0.040，0.041， 0.036 µg/mL，respectively. RSDs of precision ，stability（24 h），reproducibility and durability tests were all lower than 3％. Recoveries were 95.959％-100.201％（RSD＝1.669％，n＝6），97.818％-102.698％（RSD＝1.788％，n＝6），95.831％-99.344％ （RSD＝1.600％，n＝6）. The contents of above three components were 0.002％-0.134％，0.025％-1.388％，0.022％-0.881％. CONCLUSIONS：Established HPLC fingerprint and content determination method are simple and stable ，can be used for quality evaluation and quantitative analysis of A. euchroma . Shikonin ，acetylshikonin and β，β'-dimethylacrylic acanine are different in the content and are differential markers of A. euchroma from different source.

Evaluation of the Process Control Capability of Lorazepam Tablets by Near-Infrared Spectroscopy / 中国药学杂志

OBJECTIVE: To evaluate the process control capability of lorazepam tablets produced in China. METHODS: Near-infrared spectroscopy combined with cluster analysis(CLA) and principal component analysis(PCA) were used to characterize the different processes and process control space of lorazepam tablets produced in China. Universal quantitative model was built to obtain the content predictions of individual units(tablets), on base of which process mean value,intra-batch and inter-batch differences and distribution status were calculated by univariate statistics analysis methods. RESULTS: Three different manufacturing processes of lorazepam tablets were characterized by both CLA and PCA. The process control spaces reconstructed by the second and third principal components indicated that the process of manufacturer B had smaller variation than that of manufacturer A. The universal quantitative model had a principal component number of 5, r2 square value of 93.89% and bias of -0.008 56. The statistic distribution of API contents showed that 9 batches out of the total 27 batches had relative lager intra-batch differences and manufacturer B had better inter-batch differences than manufacturer A. CONCLUSION: The method this study established can reveal the control levels of different processes of lorazepam tablets, which provide an efficient quality consistency evaluation means for generic drug consistency assessment.

Content Determination and Principle Components Analysis of 4 Kinds of Flavones in Scutellaria barbata Decoction Pieces by HPLC / 中国药房

OBJECTIVE： To establish the method for simultaneous determination of 4 kinds of flavones such as sutellarin， sutellarein， luteolin and apigenin in Scutellaria barbata decoction pieces， and to conduct principle component analysis. METHODS： HPLC method was adopted. The determination was performed on Agilent ZOXDB-C18 column with mobile phase consisted of methanol-acetonitrile （80 ∶ 20，V/V）-1％ acetic acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 335 nm， and column temperature was 30 ℃. The sample size was 10 μL. Principal component analysis was carried out by SPSS 20.0 and SIMCA-P 13.0 software. RESULTS： The linear ranges of sutellarin， sutellarein， luteolin and apigenin were 0.131-1.446 μg（r＝0.999 0）， 0.031-0.345 μg（r＝0.999 7）， 0.005-0.055 μg（r＝0.999 2）， 0.024-0.268 μg（r＝0.999 2）， respectively. The limits of quantitation were 1.178 8， 0.602 9， 0.744 1， 1.079 1 ng； the limits of detection were 0.353 6， 0.106 1， 0.223 2， 0.323 7 ng；RSDs of precision， stability and reproducibility tests were all lower than 2％. The recoveries were 99.38％-100.56％（RSD＝0.44％，n＝6）， 91.01％-96.81％（RSD＝2.43％， n＝6）， 91.44％-97.34％（RSD＝2.59％， n＝6）， 96.21％- 99.26％（RSD＝1.23％，n＝6）， respectively. By principal component analysis， principal component 1 and prinicipal component 2 were main influential factors of sample， quality accumulative variance contribution rate of them was 92.573％（＞80％）. The comprehensive score of sample S14-3 was the highest， and the overall quality was relatively good； samples S14-2， S14-3 were the second. These 3 batches of sample were processed and produced in S. barbata planting base with stable quality. CONCLUSIONS： Established method is simple and rapid， and can be used for simultaneous determination of 4 kinds of flavones in S. barbata decoction pieces. Principle component analysis can provide reference for the quality control of S. barbata decoction pieces.

Establishment of UPLC Fingerprint of Ficus tikoua and Its Cluster Analysis and Principle Component Analysis / 中国药房

OBJECTIVE： To establish a UPLC fingerprint of Ficus tikoua. METHODS： UPLC method was adopted. The determination was performed on Waters ACQUITY UPLC BEF C18 column with mobile phase consisted of 0.2％ aqueous acetic acid-acetonitrile （gradient elution）； the detection wavelength was 254 nm； the flow rate was 0.1 mL/min； the column temperature was 25 ℃， and sample size was 2 μL. UPLC fingerprints of 10 batches of samples and 2 batches of adulterants were determined by using No. 14 peak as reference. The similarity evaluation was carried out by using the TCM Chromatographic Fingerprint Similarity Evaluation System （2012 edition） so as to determine common peak. The cluster analysis was performed by using SPSS 20.0 software. SIMCA 13.1 software was used to conduct the principal component analysis and orthogonal partial least squares discriminant analysis （OPLS-DA）. RESULTS： There were 28 common peaks in UPLC fingerprint of 10 batches of F. tikoua. The similarity of 10 batches of F. tikoua was between 0.839 and 0.935， and the similarities of the 2 batches of adulterants were 0.503 and 0.173 respectively， which indicated that F. tikoua could be distinguished from adulterants. 10 batches of F. tikoua could be divided into 2 categories by cluster analysis and principle component analysis， and S3-S5， S9 and S10 were grouped into one category， and the remaining batches were grouped into one category. 7 components with a variable importance in projection （VIP） value ＞1 were screened by OPLS-DA analysis. These 7 components may be the main components that caused the quality difference of 10 batches of F. tikoua samples. CONCLUSIONS： Established fingerprint， cluster analysis， principle component analysis and OPLS-DA can be used for the identification and quality control of F. tikoua.

Comprehensive evaluation of Yiqi Fumai Injection quality by UPLC-Q-TOF- MSE fingerprint and chemical pattern recognition / 中草药

Objective To establish a UPLC-Q-TOF-MSE fingerprint of Yiqi Fumai Injection (YQFM) for providing reference for visual, easy and overall control of its quality. Methods The chromatographic separation was performed on a Waters Acquity UPLC HSS T3 (100 mm × 2.1 mm, 1.8 μm) column with the mobile phase consisting of acetonitrile and 0.1% formic acid for gradient elution. The flow rate was 0.3 mL/min, and the column temperature was 30 ℃. The capillary voltage was set at 2.5 kV. The nebulization gas was set to 800 L/h at 400 ℃, and the source temperature was 100 ℃. The BPC obtained with negative ion ESI mass spectra were selected for the fingerprint analysis. Similarity evaluation was used to evaluate the quality of YQFM from different batches. Based on the intensities of the ions for common peaks, HCA and PCA were performed using SPSS 19.0 and Simca-P software. Results The UPLC-Q-TOF-MSE fingerprint of YQFM was established by using 28 batches of sample and 18 common peaks were found, of which 15 mutual peaks from Ginseng Radix et Rhizoma rubra, three mutual peaks from Ophiopogonis Radix. Compared with the reference substances and references, 16 of the common peaks were identified and the similarity of 28 batches samples were over 0.970. 28 batches of YQFM could be divided into four grades when the sum of squared Euclidean distance is 5-10 in the result of HCA; PCA got seven principal components through dimension reduction and accumulative contribution rate reached 84.989%. By fitting the load factor model of the first principal component, ten markers greatly impacting on the quality were found. The comprehensive evaluation function of YQFM in different batches was constructed according to the principal component score. Among 28 batches of YQFM, the comprehensive score of S28 was the best, closely followed by S22, S11 and S9, while S14 and S13 was the worst. Conclusion The utilization of UPLC-Q-TOF-MSE fingerprint coupled with chemical pattern recognition could objectively and effectively assess the quality of YQFM, can provide a more comprehensive reference for the quality control of YQFM.

The analysis of multivariate image and chemometrics in TLC fingerprinting of artificial cow-bezoar / 中国中药杂志

A method of thin-layer fingerprinting chromatogram of artificial cow-bezoar was established with the developing solvent consisting of cyclohexane, ethyl acetate, acetic acid and methanol (2∶7∶1∶2), and 10% sulfuric acid ethanol solution sprayed as colour-developing agent. After heated at 105 ℃, TLC was recorded as an image in ultraviolet light at 366 nm which was converted into grayscale. By the gray value extracted from the grayscale, the multivariate data obtained from TLC of samples could be analyzed by chemometric method. The results indicated that samples from different manufacturers could be distinguished by this method and some specific bands were found out. All in one, this simple and practical method was suitable for the evaluation of quality difference.

Diagnostic Value of Serum CA72-4,CA242,CA19-9 and CEA for Gastric Cancer Based on PCA-Decision Tree Model / 现代检验医学杂志

Objective To evaluate the diagnostic value of serum tumors CA72-4,CA242,CA19-9 and carcino-embryonic anti-gen (CEA)for patients with gastric cancer based on principle component analysis (PCA)-decision tree analysis.Methods Serum levels of CA72-4,CA242,CA19-9 and CEA in 193 patients with gastric cancer,106 patients with benign gastric disea-ses and 86 nornal controls were measured by electrochemiluminescence assay,and data were analyzed by the receiver operat-ing characteristic (ROC)curve,PCA and PCA-decision tree analysis.Results The area under the ROC curve of CA72-4, CA242,CA19-9 and CEA was 0.741[95% confidence interval (95%CI),0.692~0.791],0.863 (95%CI,0.827~0.898), 0.783 (95%CI,0.737~0.828)and 0.827 (95%CI,0.785~0.869),respectively.The combined four serum tumor markers in the PCA-AUC model was 0.935 (95%CI,0.912~0.958)at the cutoff value (PC score)of 44.13 with 78.2% of sensi-tivity and 94.8% of specificity.The accuracy of serum CA72-4,CA242,CA19-9 and CEA for the diagnosis of gastric cancer group and nongastric cancer group (benign gastric diseases and nornal controls)in the decision tree model were 76.2% and 94.8%,56.5% and 96.5% for prediction,respectively.The combined four serum tumors for the diagnosis of gastric cancer group and nongastric cancer group in PCA-decision tree model were 90.3% and 100%,72.4% and 92.2% for prediction,re-spectively.Conclusion The PCA-decision tree model based on serum CA72-4,CA242,CA19-9 and CEA were helpful for the diagnosis of gastric cancer.

Study on fingerprint analysis of Xihuang Pills by HPLC-ELSD / 中草药

Objective: To establish the quality evaluation of fingerprints of Xihuang Pills by HPLC-ELSD. Methods: The chromatography conditions were defined as waters C18 column (250 mm × 4.6 mm, 5 μm); Mobile phase was methanol-0.5% acetic acid, gradient elution; temperature of column was set at 30℃. The ELSD conditions were as follows: the temperature of drift tube was 45℃, the gas speed was 1.5 L/min. Ten batches of Xihuang Pills samples were analyzed for similarity analysis (SA), hierarchical clustering analysis (HCA) and principle component analysis (PCA). Results: The chromatographic fingerprint was completed with 25 recognizable peaks, and the samples with great differences and the compounds with greater impact on the quality were obtained through HCA and PCA. Conclusion: HPLC fingerprint combining with pattern recognition could reflect the intrinsic quality to provide a scientific basis for the quality control of Xihuang Pills.

Principle Component Analysis of Health Expenditure Per Capital in China / 中国卫生经济

Objective:To analyze the influencing factors for health expenditure per capital in China,solved the correlation problem occurred in linear regression.Methods:Principle Component Analysis was generated by using data of health expenditure per capital as well as GDP per capital,disposable income per capital,urbanization,aging of population,infant mortality rate,number of medical practitioners and beds per 1 000 people from year 1990 to 2014.Results:Explanatory variables all significantly affected health expenditure per capita.GDP per capita,disposable income per capita,number of medical practitioners and beds per 1 000 people positively influenced health expenditure per capita.While infant mortality rate,urbanization and aging of population negatively affected health expenditure per capita.Conclusion:Single factor had no impact on health expenditure per capital.The rise of health expenditure per capital was resulted by multiple factors.

The diagnostic value of four serum tumor markers for gastric cancer based on pattern recognition techniques* / 重庆医学

Objective To evaluate the diagnostic value of serum tumors CA72-4,CA242,CA19-9 and carcino-embryonic antigen(CEA)in patients with gastric cancer based on pattern recognition techniques.Methods Data of serum concentrations of CA72-4,CA242,CA19-9 and CEA of 212 patients with gastric cancer,116 patients with benign gastric disease and 117 healthy subjects were retrospectively analyzed;and the diagnostic performance of each tumor marker,four tumor markers based principle component analysis(PCA),decision tree,PCA-decision tree and the fisher discriminant analysis models were established.Results CA242 had the best diagnostic effect on gastric cancer,and the area under the ROC curve(AUC)was 0.841(95％CI:0.804-0.877).PCA model showed that the serum levels of four tumor markers in patients with gastric cancer were significantly different from those in benign and healthy patients,and obvious metabolic disorders of serum with four tumor markers were found among the patients with gastric cancer.The diagnosis accuracy of the decision tree,PCA-decision tree and the Fisher discriminant analysis models for gastric cancer patients was 58.6％,65.5％and 58.6％respectively,and for non-gastric cancer patients(benign gastric diseases and healthy controls)was 94.7％,99.4％and 97.6％.And the prediction accuracy of the decision tree,PCA-decision tree and the fisher discriminant analysis models for gastric cancer patients was 65.7％,77.6％and 73.1％,and for non-gastric cancer patients was 87.5％,96.9％and 96.9％,respectively.Conclusion The PCA-decision tree model of serum CA72-4,CA242,CA19-9 and CEA might be helpful for the diagnosis and prediction of patients with gastric cancer.

Role of Static Posturography in Fatigue Assessment due to Flight TasksLoad / 现代生物医学进展

Objective:To explore the role of static posturography in the assessment of fatigue due to flight tasks.Methods:Thirtymale college students were asked to perform simulated flight tasks consecutively forfour hours.Meanwhile their statie posturography and tasks performance would be repeatedly measured during the task-load at end of every hour.Based on the changed significantly parameters,the static balance index would be built by principle component analysis.Then its correlation with task-load level would be further analyzed by curve estimation.Results:Static postural control declined significantly under effect of simulated flight tasks.With task load sustaining,static balance index increased significantly and correlated linearly with duration of task load (R2=0.949).Besides,there was quadratic relationship between the change of multi-tasks performance and duration of task load (R2=0.968).And correlation of multi-tasks performance with static standing balance level also had been proved to be quadratic (R2=0.976).Conclusions:Static posturography correlated linearly with flight task-load level,which could reflect fatigue level caused by task load.

Discrimination of Rhodiola crenulata by HPLC fingerprinting and similarity analysis / 中国药学杂志

OBJECTIVE: To establish the HPLC fingerprint of Rhodiola crenulata herbs from Sichuan plateau, and compare them with commercially available samples. METHODS: RP-HPLC analysis was applied using Agilent Zorbax C18 chromatographic column (4.6 mm×250 mm, 5 μm). The mobile phase A was 0.1% formic acid aqueous solution and mobile phase B was 0.1% formic acid in acetonitrile. The flow rate was 1 mL·min-1 and the column temperature was maintained at 35℃. The detection wavelength was set at 245 nm and injection of sample was 20 μL. The traditional Chinese medicine fingerprint chromatogram similarity evaluation system (Version 2004A), principal component analysis, and cluster analysis were used to compare 30 Rhodiola crenulata samples from various locations based on their HPLC chromatograms. RESULTS: The established HPLC fingerprint of Rhodiola crenulata was able to analyze Rhodiola crenulata from different sources. CONCLUSION: The method has good repeatability and stability, and can be used for the quality management standard of Rhodiola crenulata.

Comparative study on HPLC specific chromatogram of Oldenlandia corymbosa Linn. and Oldenlan diadiffusa(Willd.) Roxb / 中国药学杂志

OBJECTIVE: To establish the HPLC specific chromatogram of Oldenlandia corymbosa Linn. and Oldenlan diadiffusa (Willd.) Roxb. in different regions so as to distinguish these two traditional Chinese medicinal materials. METHODS: HPLC was performed on an Agilent Eclipse XDB-C18(4.6 mm×250 mm,5 μm) column with mobile phase consisting of methanol-water at a flow rate of 1 mL·min-1, column temperature at 30℃ and detection wavelength at 254 nm. Similarity evaluation system for chromatographic fingerprint of TCM was used to conduct similarity evaluation and Matlab7.0 software was used for principal component and cluster analysis. RESULTS: Oldenlandia corymbosa Linn. and Oldenlan diadiffusa (Willd.) Roxb. had eight common peaks, namely, a-h; the characteristic peaks of O. corymbosa were 7 to 12, while the characteristic peaks of O. diffusa were 1 to 6. The similarity values of O. corymbosa and O. diffusa collected from different sources were 0.733-0.984 and 0.873-0.951, respectively. According to the principal components analysis and cluster analysis, the tested samples were classified into three categories: O. corymbosa collected from wild, O. diffusa collected from wild and purchased O. diffusa. CONCLUSION: The established method is reliable and rapid to distinguish the two kinds of easily confused traditional Chinese medicinal materials and also can offer reference for their quality control and clinical use.

Topological analysis of carbon flux during multi-stress adaptation in Halomonas sp. AAD12

Background Osmolytes with their effective stabilizing properties are accumulated as protectants not only against salinity but also against denaturing harsh environmental stresses such as freezing, drying, high temperatures, oxygen radicals and radiation. The present work seeks to understand how Halomonas sp. AAD12 cells redirect carbon flux specifically to replenish reactions for biomass and osmolyte synthesis under changing salinity and temperature. To accomplish this goal, a combined FBA-PCA approach has been utilized. Results Experimental data were collected to supply model constraints for FBA and for the verification of the model predictions, which were satisfactory. With restrictions on the various combinations of selected anaplerotic paths (reactions catalyzed by phosphoenolpyruvate carboxylase, pyruvate carboxylase or glyoxylate shunt), two major phenotypes were found. Moreover, under high salt concentrations, when the glucose uptake rate was over 1.1 mmoL DCW- 1 h- 1, an overflow metabolism that led to the synthesis of ethanol caused a slight change in both phenotypes. Conclusions The operation of the glyoxylate shunt as the major anaplerotic pathway and the degradation of 6-phosphogluconate through the Entner-Doudoroff Pathway were the major factors in causing a distinction between the observed phenotypes.

Quality evaluation of Rabdosiae Rubescentis Herba from different areas by HPLC-PDA fingerprint combined with principle component analysis / 中草药

Objective: To establish HPLC-PDA fingerprint for study on Rabdosiae Rubescentis Herba (RRH) from different areas and to scientifically evaluate and effectively control the quality of RRH from different areas by HPLC fingerprint combined with principal component analysis (PCA). Methods: The fingerprint of RRH was established by HPLC-PDA and PCA and semblance analysis methods were applied to treating the experimental data in order to fine the similarity and difference among 18 batches of RRH from four different areas. The chromatographic procedure was carried out with DiamonsilTM C18 (250 mm × 4.6 mm, 5 μm) as an analytic column and a mixture consisting of acetonitrile-0.05% phosphoric acid in gradient as mobile phase, and the temperature of column was 30 ℃. The detection wavelength was set at 238 nm and the flow rate was 0.8 mL/min. Results: The common mode of HPLC characteristic chromatographic profile was set up with 19 common peaks, and four of them were identified as oridonin, ponicidin, lasiodonin, and rosmarinic acid. The similarities of 18 batches of RRH were between 0.421- 0.984. The result of PCA showed that the accumulative contribution rate of the first three factors was up to 87.3%, which were chosen to comprehensively evaluate the quality of RRH. Conclusion: The application of HPLC combined with PCA could quickly and objectively evaluate the quality difference of RRH in different batches.

Diversity Analysis of Some Traditional Rice Genotypes in Sri Lanka.

Aims: To study the diversity of traditional rice genotypes in Sri Lanka using cluster analysis and principle component analysis. Study Design: The experiment was carried out using one hundred rice genotypes with six modern rice cultivars and ninety four traditional rice cultivars. Rice genotypes were planted according to a randomized complete block design with four replications and 20 plants per plot with 15 cm X 20 cm spacing. Place and Duration of Study: A field experiment was carried out during 2011/2012 Maha season and 2012 Yala season at Faculty of Agriculture, University of Ruhuna, Sri Lanka. Methodology: Plant height (cm), number of tillers/plant and number of productive tillers/plant were measured before harvesting. Panicle length (cm), panicle weight (g), number of spikelets/panicle, number of fertile spikelets/panicle, 100 grain weight (g) and yield/plant (g) were measured after harvesting and drying of grains for 14% moisture content. Principal component analysis (PCA) and cluster analysis were performed using SPSS statistical software. Results: Among nine studied variables three principal components exhibited more than one Eigen value and showed about 89.6 % variability. The principal components (PC) 1, 2 and 3 had 51.07%, 22.08% and 16.46% variability among the genotypes for the evaluated traits respectively. The first PC was more related to panicle weight, number of spikelets/panicle, number of fertile spikelets/panicle, spikelet fertility percentage and yield (g/plant). Number of tillers/plant, number of productive tillers/plant and yield (g/plant) were more related traits in the second principal component. The highest contribution in third principal component was from the panicle weight, 100 grain weight and yield (g/plant). Based on the nine yield and yield attributing characters, the genotypes were grouped in to seven clusters in cluster analysis. The genotypes under cluster V recorded the highest divergence among them as it exhibited the highest intra-cluster distance. The lowest intracluster distance was recorded in the cluster VI. The modern rice cultivar BG 379/2 was fallen in to the cluster VI with 3 traditional rice cultivars namely Karayal I, Bathkiri el and Hondarawala. Conclusion: One hundred rice genotypes were grouped in to divergent groups by principle component analysis and cluster analysis. This clustering pattern can be used for the selection of parental materials with diverse characters.

HPTLC fingerprint analysis of Aconitum herbals in Xinjiang / 中国药学杂志

OBJECTIVE: To establish the HPTLC fingerprint of Aconitum plants in Xinjiang and to carry out similarity comparison, cluster analysis and principal component analysis. METHODS: Using high performance thin layer silica gel G plate, n-hexane-ethyl acetate-methanol (6.4:3.6:1) as developer, diluted bismuth potassium iodide solution as colour-developing agent, of the HPTLC fingerprints were obtained. The common pattern was explored by chemical fingerprint analysis system software. Similarity analysis, cluster analysis and principal component analysis were carried out. RESULTS: The conditions for thin layer chromatography were optimized, the spots were clear, and the separation was good; similarity analysis showed that Xinjiang Aconitum had some differences in chemical composition between different plant species, which can be broadly divided into three categories. Aconitum leucostomum Wo-rosch. and Aconitum apetalum (Huth) B. Fedtsch. had high similarity with Aconitum carmichaelii Debx. and Aconitum kusnezojfii Reichb. CONCLUSION: The established thin-layer chromatographic fingerprints have laid the methodological foundation for the study of the fingerprint of Xinjiang Aconitum plants. Aconitum leucostomum Worosch, which is a widely distributed and abundantly reserved Aconitum species in Xinjiang, has potential medicinal value and similarity with Aconitum carmichaelii Debx. and Aconitum kus-nezoffii Reichb. Which have national quality standard, and is worthy of further studies.