Simultaneous Determination of 7 Active Components in Mori Australis Cortex and Mori Cortex from Different Sources by HPLC / 中国药房

OBJECTIVE：To establish a meth od for the simultaneous determination of 7 active components in Mori Australis Cortex and Mori Cortex from different sources in Chongqing area ，so as to provide reference for improving the quality control standards of Mori Australis Cortex and Mori Cortex and comparing the equivalence of their quality. METHODS ：HPLC method was used to determine the contents of neochlorogenic acid ，mulberroside A ，chlorogenic acid ，astragalin，kaempferol，morusin and isoquercetin in 58 batches of Mori Australis Cortex and Mori Cortex. The chromatographic column was Diamonsil C 18 with mobile phase consisted of 0.1％ formic acid solution-acetonitrile （gradient elution ） at the flow rate of 1.0 mL/min. The detection wavelength was 280 nm，column temperature was 30 ℃，and the injection volume was 10 μL. Using SPSS 22.0 software， independent sample t-test，principal component analysis and cluster analysis were used to analyze the content difference of the above-mentioned 7 active components in Mori Australis Cortex and Mori Cortex. RESULTS ：There was a good linear relationship between the peak area and the concentration of the above 7 active components （r≥0.999 0）. The RSDs of precision ，stability（24 h），repeatability，durability and recovery were less than 3％. The average contents of neochlorogenic acid ，mulberroside A ， chlorogenic acid ， astragalin， kaempferol， morusin and 023-58576130。E-mail：1025473978@qq.com isoquercetin in Mori Australis Cortex were 0.304，22.462， 1.730，1.308，1.593，2.842 and 0.657 mg/g，respectively. Those of Mori Cortex were 0.305，22.995，2.486，2.438， 2.916，4.158 and 1.264 mg/g，respectively. The results of independent sample t-test showed that only the content of kaempferol in the above 7 active components of Mori Australis Cortex and Mori Cortex had significant difference （P＜0.05）. The results of principal component analysis and cluster analysis showed that there was no significant difference in the contents of above 7 active components between Mori Australis Cortex and Mori Cortex. CONCLUSIONS：The established HPLC method is simple ，sensitive and accurate ，which can provide a reference for improving the quality control standard of Mori Australis Cortex and Mori Cortex. Mori Australis Cortex and Mori Cortex have certain quality equivalence in main active components ，and the Mori Australis Cortex from M. australis and M. cathayana can be used as a substitute for the Mori Cortex.

Establishment of UPLC Fingerprint of Pyrrosia petiolosa from Southwest China and Content Determination of Phenolic Acids Component / 中国药房

OBJECTIVE：To esta blish a UPLC fingerprint of Pyrrosia petiolosa from southwest China ，and to determine the contents of 4 kinds of phenolic acids （neochlorogenic acid ，caffeic acid ，chlorogenic acid and cryptochlorogenic acid ）. METHODS：The determination was performed on Waters Cortecs T 3 C18 column（100 mm×2.1 mm，1.6 μm）with mobile phase consisted of methanol- 0.1％ phosphoric acid （gradient elution ）at the flow rate of 0.35 mL/min. The detection wavelength was set at 326 nm. The column temperature was 30 ℃，and injection volume was 1 μL. UPLC method was used to establish the UPLC fingerprint of P. petiolosa in combination with the Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 edition）. Cluster analysis and principle component analysis （PCA）were performed by using SPSS 20.0 software. The contents of 4 kinds of phenolic acids in 20 batches of P. petiolosa were determined by external standard method. RESULTS ：There were 9 common peaks for the UPLC fingerprint of P. petiolosa . Peaks 1，3，4，5 and 9 were identified as neochlorogenic acid ，caffeic acid，chlorogenic acid ，cryptochlorogenic acid and isochlorogenic acid C ，respectively. RSDs of the relative retention time of each peak in different batches of P. petiolosa were 0-0.68％，and the RSDs of the relative peak area were 0-62.35％. The similarities between the fingerprint of 20 batches of medicinal materials and the control chromatogram were not less than 0.990. The result of cluster analysis showed that P. petiolosa from different regions could be sorted into three species. Results of PCA showed the differences among P. petiolosa from different regions. The linear range of neochlorogenic acid ，caffeic acid ，chlorogenic acid and cryptochlorogenic acid were 0.61-61.41，0.18-17.60，2.00-200.11，0.62-61.51 μ g/mL （R2＞0.999 9）. RSDs of precision ， reproducibility and stability tests were all lower than 2.00％. The recoveries were 96.23％-98.17％（RSD＝0.96％-2.28％， n＝6）. Among 20 batches of samples ，the contents of above 4 kinds of phenolic acids were 0.385 3-1.891 9，0.018 0-0.129 5，2.569 5-10.676 0，0.563.5-1.860 5 mg/g. CONCLUSIONS ： The established UPL C fingerprint could reflect the main chemical constituents of P. pedunculata . Phenolic acids could be used as the main evaluation indexes for the quality of P. petiolosa . The quality order of P. petiolosa from southwest China was Chongqing product＞Sichuan product ＞Guizhou product.

Simultaneous Determination of Seven Constituents in Qinggan Lidan Mixture by HPLC / 中国实验方剂学杂志

Objective: To establish an HPLC method for simultaneous determination of geniposide,chlorogenic acid,neochlorogenic acid,cryptochlorogenic acid,isochlorogenic acid A,B and C in Qinggan Lidan mixture,in order to provide references for its quality control. Method: The analysis of methanol extract of this drug was performed on a 35℃ Luna C18 column (4.6 mm×250 mm,5μm),with the mobile phase comprised of acetonitrile-0.4% phosphoric acid flowing at 1.0 mL·min-1 in a gradient elution mode (0-10 min,8%-12%A;10-30 min,12%A;30-60 min,12%-35%A),and the detection wavelengths were set at 238 and 327 nm. Result:Geniposide,chlorogenic acid,neochlorogenic acid,cryptochlorogenic acid,isochlorogenic acid A,B and C were completely separated,and well separated from other constituents. The linear ranges of geniposide,chlorogenic acid,neochlorogenic acid,cryptochlorogenic acid,isochlorogenic acid A,B and C were 0.188-2.355,0.083-1.040,0.074-0.920,0.075-0.940,0.064-0.800,0.076-0.955,0.071-0.888 μg (r ≥ 0.999 0),respectively. The average recoveries were 99.45%,98.45%,99.06%,98.50%,98.16%,101.01%,96.93%,with the RSDs of 0.5%,1.8%,1.3%,2.4%,2.3%,1.6%,1.6%,respectively.The contents of geniposide,chlorogenic acid,neochlorogenic acid,cryptochlorogenic acid,isochlorogenic acid A,B and C were 3.420-3.794,0.835-0.890,1.222-1.275,1.064-1.210,0.377-0.398,0.419-0.464 and 0.362-0.405 g·L-1,respectively. Conclusion:This method can be used for simultaneous determination of muti-ingredients in Qinggan Lidan mixture,and the established method is simple and accurate,with a good reproducibility and high sensitivity. It can be used for the quality control of Qinggan Lidan mixture.

Study on Frosting Quality Markers of Mori Folium Based on Component Characteristic / 中国实验方剂学杂志

Objective: To screen the frosting qualitymarkers (Q-marker) of Mori Foliumand identify Mori Folium after frost. Method: The HPLC-DAD-MSn fingerprints of Mori Folium before and after frost were established,the common peaks were markedand the characteristic components were selected by orthogonal partial least squares discrimination analysis (OPLS-DA). The components with a large content change were used as marker components,and the relatively stable component was used as an internal reference component. The ratio of the marker components to the peak area of the internal reference component was used as component characteristic to identify the frost of Mori Folium,andqualitative study of frosting markerswere performed by MSn and authentic standards. Result: The fingerprints were established and 33 common peaks were marked. Through OPLS-DA,peaks 1,23,14 were determined as marker components,peak 12 was determined as internal reference component. The frosting quality markers of Mori Folium were characterized as citric acid derivatives,saponin F,tryptophan and neochlorogenic acid by MSn and standards. The ratios of the peak areas of citric acid derivatives,saponin F,tryptophan and neochlorogenic acid before and after frost were 0.15±0.054,1.0±0.48; 0.14±0.073,0.98±0.48,0.13±0.088,0.89±0.49,respectively. Conclusion: In the fingerprints established in this study,the peak area ratios of citric acid derivatives,saponin F,tryptophan to new chlorogenic acid had specificity,which can be used as frosting quality markers for identification of Mori Folium. This study expanded the connotation of the Q-marker specificity of traditional Chinese medicine. The results can provide experimental data for the quality evaluation and drug-forming properties of Mori Folium,and provide reference for similar research.

Content Determination of Nine Constituents in Different Medicinal Parts of Pimpinella thellungiana / 中国实验方剂学杂志

Objective:To develop a method to quantify nine constituents in different medicinal parts of Pimpinella thellungiana, in order to compare the content difference of the nine constituents, namely protocatechuic acid,gallic acid,neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,luteolin-7-O-β-D-glucuronide,isochlorogenic acid A,apigenin-7-O-β-D-glucuronide and isochlorogenic acid C. Method:The analysis was performed on an Agilent TC-C18 column (4.6 mm×250 mm,5 μm) with acetonitrile and mixed acid solution (0.1% phosphoric acid-0.1% glacial acetic acid) as mobile phase for gradient elution. The handover detection wavelengths were at 265 and 325 nm. The column temperature was 20℃, and the flow rate was 1.0 mL·min-1. The experiment data was analyzed using the software of Markerlynx XS. Result:The nine constituents of protocatechuic acid,gallic acid,neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid, luteolin-7-O-β-D-glucuronide,isochlorogenic acid A,apigenin-7-O-β-D-glucuronide and isochlorogenic acid C had a good degree of separation and a good linearity in their respective linear ranges(r>0.999 8). The average recoveries ranged from 99.11% to 100.76%,and the RSD ranged from 0.9% to 2.0% 。The results showed that the contents of the nine constituents had significant differences in different medicinal parts of P. thellungiana. The average contents of the nine constituents were the highest in leaves,which was followed by stem,and the lowest was in root. Conclusion:The study could provide evidence for the quality control,clinical application,and scientific resources utilization of P. thellungiana.

Study on HPLC Fingerprints of Nauclea officinalis Extract Syrup and Content Determination of 9 Components / 中国药房

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.

Content Determination of 6 Active Components in Morus alba by HPLC / 中国药房

OBJECTIVE:To establish the method for the simultaneous determination of content of 6 active components as neochlorogenic acid,mulberroside A,chlorogenic acid,astragalin,sanggenon C and morusin in Morus alba,and to provide reference for improving quality control standard of M. alba. METHODS:HPLC method was adopted. The determination was performed on Agilent 5 TC-C18with mobile phase consisted of acetonitrile-0.1% formic acid(gradient elution)at the flow rate of 1 mL/min. The detection wavelength of 280 nm. RESULTS:The mass concentration linear range of neochlorogenic acid, mulberroside A,chlorogenic acid,astragalin,sanggenon C and morusin were 0.001 06-0.042 4,0.001 67-0.066 8,0.007 95-0.318, 0.001 65-0.066 0,0.005 00-0.200 and 0.001 24-0.049 6 mg/mL,respectively(all r≥0.999 6);the limits of quantitation were 0.11, 0.14,0.81,0.17,0.45 and 0.12 μg/mL,respectively;the limits of detection were 0.04,0.05,0.41,0.07,0.18 and 0.04 μg/mL, respectively;RSDs of precision test were 0.26%,0.31%,0.24%,0.27%,0.36% and 0.44%(n=6),respectively;RSDs of stability test were 0.68%,0.54%,0.62%,0.53%,0.41% and 0.73%(n=6),respectively;average method recovery rates were 99.1%,98.8%,98.8%,98.4%,98.5% and 99.9%(RSDs were 0.5%-1.5%,n=9),respectively. CONCLUSIONS:The method is simple,accurate,and can be used for simultaneous determination of 6 active components in M.alba.

Fast measurement method based on near-infrared spectroscopy in concentration of alcohol precipitation proee for Artemisiae Annuae Herba and Lonicerae Flos in Reduning Injection / 中草药

Objective: The calibration models were developed in the concentration of alcohol precipitation proee for Artemisiae Annuae Herba (AAH) and Lonicerae Flos (LF) in Reduning Injection (RI) to realize the on-line monitoring of production process. Methods: Based on the near infrared reflectance spectroscopy (NIRS), partial least regression (PLS) models were developed to fast measure the contents of neochlorogenic acid, chlorogenic acid, and 4-O-caffeoylquinic acid in the concentration of the alcohol precipitation proee for AAH and LF. Results: In the quantitative models of neochlorogenic acid, chlorogenic acid, and 4-O-caffeoylquinic acid, the coefficient of determination (R2) of cross validation sets were 0.954 5, 0.975 2, and 0.969 1; The root mean square errors of calibration (RMSEC) were 0.213, 0.676, and 0.225; The root mean square errors of cross-validation (RMSECV) were 0.233, 0.692, and 0.258. When the established models were applied to on-line monitoring, the coefficient of determination of neochlorogenic acid, chlorogenic acid, and 4-O-caffeoylquinic acid were 0.984 2, 0.983 7, and 0.987 0, the residual predictive deviation (RPD) were 4.77, 5.29, and 4.37; The relative standard errors of prediction (RSEP) were 3.519%, 3.778%, and 3.895%. Conclusion: The models above are proved to fast measure the contents of neochlorogenic acid, chlorogenic acid, and 4-O-caffeoylquinic acid in the concentration of alcohol precipitation proee for AAH and LF in RI.

Optimization on Transformation from Chlorogenic Acid to Neochlorogenic Acid by Box-Behnken Design-response Surface Methodology / 世界科学技术-中医药现代化

Box-Behnken design-response surface methodology was used to optimize the transformation from chlorogenic acid to neochlorogenic acid.Based on single factor experiments,the experiment design were developed by box-benhnhen central composite design with causal factors of the reaction temperature,time and PH to neochlorogenic acid.The optimum transformation conditions were as follow:reaction temperature at 107℃,reaction time of 60 min,the PH of 4.72.Under the optimum extraction technology conditions,the productivity of neochlorogenic acid was 64.20％.Neochlorogenic acid was isolated and purified.The determination and characterization of neochlorogenic acid was detected by HPLC,1H-NMR,13C-NMR and ESI-MS.The results showed that the content of neochlorogenic acid reached to 98.78％ and the yield of 87.37％.

Study on HPLC fingerprint of Yinhuang Drop Pills / 中草药

Objective: To establish the specific chromatogram of Yinhuang Drop Pills (YDP) by High performance liquid chromatography. Methods: The chromatographic column Kromasil 100-5C18 (250 mm × 4.6 mm, 5 μm) was used, acetonitrile-0.4% phosphoric acid as mobile phase with gradient elution, and the detection wavelength was 372 nm. Through similarity evaluation software, the similarity of specific chromatogram of 10 batches YDP were calculated. Results: The HPLC specific chromatogram of YDP showed 7 common peaks, of which 6 peaks (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, 3,4-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, and 4,5-O-dicaffeoylquinic acid) from Lonicerae Flos, 1 common peaks (baicalin) from Scutellariae Radix; By comparing with the reference substances, identified 7 components, respectively, the neochlorogenic acid (peak 1), chlorogenic acid (peak 2), cryptochlorogenic acid (peak 3), 3,4-O-dicaffeoylquinic acid (peak 4), 3,5-O-dicaffeoylquinic acid (peak 5), 4,5-O-dicaffeoylquinic acid (peak 6), baicalin (peak 7). By the methodology validation, verified that this method has good precision, repeatability and stability, 10 batches of sample similarity is greater than 0.9. Conclusion: This method was available for the quality control of YDP.

Degradation kinetics of chlorogenic acid, cryptochlorogenic acid, and neochlorogenic acid at neutral and alkaline pH values / 药学学报

The degradation kinetics of chlorogenic acid (5-CQA), cryptochlorogenic acid (4-CQA), and neochlorogenic acid (3-CQA) in aqueous solution at 37 ℃ and different pH values (7.05, 7.96, 9.25) were investigated in the present work. The results indicated that 3-, 4- and 5-CQA tended to remain stable in acidic pH circumstance, and unstable in neutral and alkaline pH circumstance. With the increase of the alkalinity, the degradation of 3-, 4- and 5-CQA was increased leading to a less amount of total CQA and was satisfactorily described by the Weibull equation. Meanwhile, caffeic acid was not detected after the degradation of CQA. Moreover, the degradation of 3-CQA and 5-CQA tended to be converted to 4-CQA, and the degradation of 4-CQA tended to be converted to 3-CQA rather than 5-CQA. The comparison of the degradation kinetics parameters of 3-, 4- and 5-CQA at neutral and alkaline pH values showed that the orders of the rate constant (k) values were 4-CQA > 3-CQA > 5-CQA, while the orders of the degradation half life (t1/2) values were 4-CQA < 3-CQA < 5-CQA, indicating the orders of the stabilities of 3-, 4- and 5-CQA at 37 ℃ and neutral and alkaline pH values were 4-CQA < 3-CQA <5-CQA.

Comparative Analysis on Contents of Phenolic Acids in the Decoction of Xanthii Fructus before and after Frying / 中国药师

Objective:To examine the content changes of neochlorogenic acid, chlorogenic acid, 1, 5- dicaffeoylquinic acid and total phenolic acids in the water extract from raw and fried Xanthii Fructus. Methods:The stir-frying method was used to process fried Xanthii Fructus from different habitats. The contents of neochlorogenic acid, chlorogenic acid and 1,5-dicaffeoylquinic acid in the wa-ter extract were determined by HPLC, the total phenolic acids content was determined by UV. Results:The contents of neochlorogenic acid, chlorogenic acid, 1,5-dicaffeoylquinic acid and total phenolic acids in the water extract from fried Xanthii Fructus were all in-creased. Conclusion:Fried Xanthii Fructus can increase the contents of effective ingredients in the decoction resulting in the enhanc-ment of clinical curative effect.

Optimization on alcohol precipitation process for Lonicerae Flos and Artemisiae Annuae Herba in Reduning Injection by central composite design and response surface methodology / 中草药

To optimize the alcohol precipitation process for Lonicerae Flos and Artemisiae Annuae Herba (LA) in Reduning Injection and obtain the relationship equation of the key process parameters and quality attributes, which could provided a theoretical basis of the control automization in the alcohol precipitation process. With the transfer rates of neochlorogenic acid, chlorogenic acid, cryptochlorogenin acid, caffeic acid, secoxyloganin and solid content in precipitation liquid as evaluation indexes, the effect of the seven factors, such as relative density and the temperature of LA in liquid phase before alcohol precipitation, alcohol concentration in the end of alcohol precipitation, stirring speed in alcohol precipitation, adding alcohol speed, standing temperature, and standing time, on the alcohol precipitation process was investigated by the single factor experiment. By analysis of variance, the key factors that could influence the alcohol precipitation process were determined. Then the range of parameters of key factors was further studied and explored by Box-Behnken response surface methodology. The optimum preparation conditions of the alcohol precipitation process of LA were as follows: stirring speed was 550 r/min, adding alcohol speed was 4.0 mL/s, standing temperature was 30℃, standing time was 24 h, alcohol concentration was 75%, the relative density of LA was 1.10, and the temperature of LA was 25℃. Under these conditions, the transfer rates of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, and secoxyloganin were 94.8%, 97.6%, 97.4%, 97.2%, and 96.1%, and the solid content was 4.2%. The alcohol precipitation process of LA have been optimized by the response surface method, which could help to enhance the stability of the process.

Preparation of Neochlorogenic Acid Reference Substances by Medium-low-pressure Preparative Chromatography / 世界科学技术-中医药现代化

This study was aimed to establish a separation method for neochlorogenic acid reference substances from Lonicera japonica. Refined neochlorogenic acid inL. japonica water extract was separated and concentrated by HPD200A macroporous resin, which was isolated and purified by medium-low-pressure preparative chromatography and determined by HPLC. The structure was identified by various spectroscopic data including ESI-MS,1H-NMR and13C-NMR. The results showed that the optimal purification technology conditions were as follows: washed with 5BV of water, collected elution, concentration, drying; neochlorogenic acid crude products were eluted with acetonitrile-0.5% formic acid solution (10:90) with the flow rate of 20 mL·min-1; and the detection wavelength was 326 nm. The contents of the prepared neochlorogenic acid reached to 98.86% and the yield was 89.1%. It was concluded that the method was effective for the preparation of neochlorogenic acid with high purity. It can be used to prepare the reference substances for quantitative analysis and content determination of Chinese materia medica.

Determination of eight components in Lonicerae Japonicae Flos by HPLC / 中草药

Objective: To establish an HPLC method for simultaneously determining eight components such as neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, and secoxyloganin in Lonicerae Japonicae Flos. Methods: A RP-HPLC method was established with Luna C18 column (250 mm × 4.6 mm, 5 μm) by a gradient elution using methanol (A) and 0.1% phosphoric acid (B): 0-20 min, 12%-30% A; 20-60 min, 30%-50% A; The flow rate was 1.0 mL/min with double wavelength (237 and 324 nm), and the column temperature was 30°C. Results: The eight components was separated to baseline; Each component had a wide linear range and a good linear relationship (r > 0.9998); The recoveries were 98.72% to 102.50%. Conclusion: The method is accurate, sensitive, reliable, and has a good reproducibility. It could be used for the quality control of Lonicerae Japonicae Flos.

Simultaneous determination of the contents and fingerprint of multi-components in honeysuckle extract by HPLC / 中国药学杂志

OBJECTIVE: To establish a HPLC method for the simultaneous determination of the contents and fingerprint of 7 active constituents (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C) in honeysuckle extract. METHODS: The chromatographic separation was achieved on a AkzoNobel Kromasil C18 column(4.6 mm × 250 mm, 5 μm), the mobile phase was acetonitrile-0.1% phosphoric acid aqueous solution with gradient elution at a flow rate of 1.0 mL · min-1, the detection wavelength was set at 326 nm, and the column temperature was 30°C. RESULTS: All the 7 compounds showed good linearity in the ranges of the test concentrations. The RSDs of the precision, stability and reproducibility tests were less than 3%. The average recoveries were in the range of 97.98%-99.29%. CONCLUSION: This method is simple, sensitive, accurate, and can be used for quality control of honeysuckle extract. Copyright 2012 by the Chinese Pharmaceutical Association.

Determination of Six Phendic Acids in Mailuoning Injection by RP-HPLC / 中国中医药信息杂志

Objective To establish a method for determination the contents of six phendic acids in Mailuoning injection by HPLC at same time. Methods Separation was performed on a Kromasil-C18 (4.6 mm? 250 mm, 5 ?m) column with mobile phase consisting of 0.1% phosphoric acid-water and Acetonitrile with gradient elute at the flow rate of 1 mL/min. The UV detection wavelength was set at 327 nm and the column temperature was set at 25 ℃. The sample size was 20 ?L. Results The standard cures of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, 3,5-Dicaffeoylquinic acid, 3,4- Dicaffeoylquinic acid were linear within the contentration range of 0.010 90～0.544 8 ?g (r=0.999 9), 0.024 06～1.203 2 ?g (r =1), 0.011 06～0.552 8 ?g (r =1), 0.015 94～0.796 8 ?g (r =1), 0.009 104～0.455 2 ?g (r =1), 0.010 37～0.518 4 ?g (r=1) respectively. And the recovery rates of them were 101.06%, 100.22%, 101.54%, 99.42%, 100.21%, 101.65% respectively. Conclusion The method appeared to be simple, reliable, accurate and can be used to determine the contents of six phendic acids in Mailuoning injection.