Study on HPLC characteristic fingerprint of substance benchmark of classical famous prescription of Jichuan Decoction / 中草药

Objective: To establish an HPLC characteristic fingerprint of substance benchmark (standard decoction) of classical famous prescription of Jichuan Decoction (JD), and provide reference for the quality study of substance benchmark of JD. Methods: JD standard decoction was prepared according to the ancient method, 15 batches JD standard decoction were determined by HPLC. The similarity analysis and characteristic peak analysis of 15 batches JD were carried out by the "Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica 2012 version". Results: A total of 18 common characteristic peaks were screened by automatic matching method, peaks 1 and 3 were from Angelicae Sinensis Radix and Cimicifugae Rhizoma, peaks 2, 5, 6, 7, 9, 11 and 13 were from Cistanches Herba, peaks 4, 12, 14, 15 and 17 from were Cimicifugae Rhizoma, peaks 8, 10 and 18 from Aurantii Fructus, and peak 16 was from Angelicae Sinensis Radix. Seven characteristic components were identified by the reference substance, including caffeic acid (peak 1), echinacoside (peak 2), ferulic acid (peak 3), isoferulic acid (peak 4), mullein glycoside (peak 6), naringin (peak 8) and neohesperidin (peak 10). The similarities of 15 batches substance benchmark of JD were greater than 0.9. Conclusion: The HPLC method established for substance benchmark of JD is simple, accurate, stable and sensitive. It can be used for the quality study for JD substance benchmark, and provides a reference for the transformation and development of JD for modern preparations.

Comprehensive Quality Evaluation of Lysimachia christinae Based on the Entropy Weight TOPSIS Method Combined with Multiple Indicator Components / 中国药房

OBJECTIVE：To e stablish UPLC characteristics fingerprints of Lysimachia christinae ，and to simultaneously determine 3 effective components and to comprehensively evaluate the quality of L. christinae from different production areas. METHODS：UPLC method was adopted to establish characteristics fingerprint of the whole plant ，stem and leaves of 10 batches of L. christinae ，and determine the contents of kaemperfol- 3-O-rutinoside，quercetin，kaemperfol. The determination was performed on Waters CORTECS UPLC T 3 column with mobile phase consisted of acetonitrile- 0.2％ phosphoric acid aqueous solution （gradient elution ）at the flow rate of 0.2 mL/min. The detection wavelength was set at 364 nm，and column temperature was 30 ℃. The sample size was 1 µL. Similarity Evaluation System for TCM Chromatographic Fingerprint （2012 edition）was adopted to evaluate its similarity ， and common peaks were confirmed. Using the contents of kaemperfol- 3-O-rutinoside， quercetin， kaemperfol，total ash ，acid-insoluble ash and sulfur dioxide residue ，the ethanol-soluble extract as index ，entropy weight TOPSIS was used to evaluate the overall quality of L. christinae comprehensively. RESULTS ：There were 7 common peaks in the whole plant，stem and leaves of 10 batches of L. christinae ，among which 3 peaks were identified as kaemperfol- 3-O-rutinoside， quercetin and kaemperfol. The similarity of same part in the whole plant of L. christinae from different batches were not lower than 0.830. The similarity between stem and leaves of L. christinae in same batch was 0.504-0.859； the similarity between whole plant and stem was 0.593-0.904；the similarity between whole plant and leaves was 0.885-0.995. The linear ranges were 0.392 0-39.197 0 μg/mL for kaempferol-3-O-rutinoside， 0.397 0- 39.703 4 μg/mL for quercetin，0.380 9-38.093 0 μg/mL for kaempferol（r＞0.999 0）. RSDs of precision ，stability and repeatability tests were all lower than 2％. The recoveries were 96.43％（RSD＝0.63％，n＝9），100.32％（RSD＝0.46％，n＝9）， 101.80％（RSD＝0.32％，n＝9），respectively. The content range of above components in L. christinae were 0.006 3％-0.041 1％， 0.002 9％-0.008 6％，0.004 4％-0.017 5％（stem）；0.024 8％-0.290 5％，0.000 9％-0.009 0％，0.001 3％-0.012 4％（leaves）； 0.007 9％-0.118 0％，0.001 5％-0.008 8％，0.002 8％-0.012 5％（whole plant ）. There was no significant difference in the contents of 3 components in L. christinae among different producing areas （P＞0.05）. The order of the contents of kaempferol- 3-O- rutinoside in different parts of L. christinae was leaves ＞whole plant ＞stem. The contents of quercetin and kaempferol were high relatively in the stem. Results of entropy weight TOPSIS method showed that mean values of Ci for L. christinae from Zhongjiang county and Shuangliu county of Sichuan province ，Shizhu county of Chongqing city were 0.446，0.512，0.287. CONCLUSIONS ： Established fingerprint and content determination method are stable and feasible ，and multi-index evaluation model constructed by characteristic chromatogram combined with entropy weight TOPSIS analysis method can be used for comprehensive quality evaluation of L. christinae . The quality of L. christinae from Sichuan province is better.

Characteristic fingerprint and multi-components quantitative determination for Fuke Qianjin Capsules by HPLC / 中国中药杂志

To establish an HPLC characteristic fingerprint method of Fuke Qianjin Capsules,and determine the contents of its main components. The analysis was carried out on a Kromasil 100-5-C18 analytical column(4. 6 mm ×250 mm,5 μm) with gradient elution by acetonitrile(A)-0. 1% phosphoric acid aqueous solution(B),a flow rate at 1. 0 m L·min-1 and the detection wavelength of 254 nm.The column temperature was 30 ℃,and the injection volume was 10 μL. The determination method of genistin,jatrorrhizine,andrographolide and 14-deoxy-11,12-didehydroandrographolide index components were studied methodologically. The common mode of the characteristic fingerprint of Fuke Qianjin Capsules was set up with 8 common peaks,which were identified as genistin,jatrorrhizine,palmatine,berberine,andrographolide,14-deoxy-11,12-didehydroandrographolide,Z-ligustilide,and Z-3-butylidenephthalide,respectively,in comparison with the references. The similarities of 20 batches of Fuke Qianjin Capsules samples were above 0. 95. All of the above-mentioned 4 analytes could be well separated under the optimized chromatographic conditions. RSD of precision and repeatability experiment were both less than 1. 5%,and the sample solution was stable during 72 h. All of the compounds had a good linearity and linear range. The contents of genistin,jatrorrhizine,andrographolide,and 14-deoxy-11,12-didehydroandrographolide in 20 batches of Fuke Qianjin Capsules samples were 28. 66-56. 04,94. 77-197. 92,1 705. 33-4 148. 93 and 462. 16-1 225. 96 μg in each capsule,respectively. The developed HPLC characteristic fingerprint and quantitative analysis methods were reliable,accurate and sensitive,and could be used effectively evaluate the quality of Fuke Qianjin Capsules samples.

Comparation of chemical composition of Cyathula offinalis between different growth years by HPLC characteristic fingerprint combined with chemometrics / 中草药

Objective HPLC characteristic fingerprint and chemometrics method were used to compare the chemical characteristics difference between 1-6 years old Cyathula offinalis. Methods The peak area data matrix of 6 × 54 and the difference of characteristic peak of 1-6 years old C. offinalis were analyzed by HPLC. Similarity analysis the samples was investigated with “Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica 2004A”. Hierarchical clustering analysis (HCA) and principal component analysis (PCA) were carried out with SPSS 22.0 software. Results According to the analysis of characteristic peak difference, 1 and 6 years old C. offinalis showed exclusive difference in four chromatographic peaks and five chromatographic peaks, respectively. The peak area of 1 year old sample was significantly higher than other years in four chromatographic peaks, which showed obvious differences in components cumulant. The similarity degree analysis showed that the similarity degree of the 3, 4, and 5 years old C. offinalis were more than 0.98, but the 1, 2 and 6 years old were less than 0.85. Dendrogram of HCA and 3D scatter plot of PCA showed that the 3, 4, and 5 years old C. offinalis were clustered into one class. Conclusion There are certain differences in the chemical components among different growth years of C. offinalis. The chemical components of the 3, 4, and 5 years old C. offinalis tend to be uniform, which provides a scientific reference for the harvest time of C. offinalis.

UPLC characteristic fingerprint of leaves of Moringa oleifera / 中国中药杂志

This study aims to establish the characteristic fingerprint of the leaves of Moringa oleifera by Ultra High Performance Liquid Chromatography (UPLC) for its quality control. The method was developed on a column of Agilent Eclipse XDB-C₁₈ with acetonitrile-0.01% TFA solution as the mobile phase by gradient elution at a flow rate of 0.5 mL·min⁻¹. The detective wavelength was 210 nm, and the column temperature was 35 °C. The 14 batches of the leaves of M. oleifera were compared for the similarity by using Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2004A). The UPLC characteristic fingerprint was established, and twelve common peaks were identified by comparison with the references and UPLC-MS. The relative retention times were 0.08 (No. 1, adenosine), 0.14 (No. 2, L-phenylalanine), 0.22 (No. 3, 5-caffeoylquinic acid), 0.28 (No. 4, L-tryptophane), 0.42 (No. 5, 4-caffeoylquinic acid), 0.65 (No. 6, vicenin-2), 0.94 (No. 7, vitexin), 0.96 (No. 8, isovitexin), 1.00 (No. 9, isoquercitrin), 1.11 [No. 10, quercetin 3-O-β-D-(6"-malonyl)-glucopyranoside], 1.21 (No. 11, astragalin) and 1.37 [No. 12, kaempferol 3-O-β-D-(6"-malonyl)-glucopyranoside]. It is the first time to establish the UPLC characteristic fingerprint of the leaves of M. oleifera. The method is simple, quick and reproducible with high precision, which can provide a scientific basis for the quality control of the leaves of M. oleifera.

HPLC characteristic fingerprints of Gentiana scabra Bge. and Gentiana rigescens Franch. based on the polarity orientation technology / 中国药学杂志

OBJECTIVE: To establish the HPLC characteristic fingerprints of Gentiana scabra Bge. and G. rigescens Franch. METHODS: The characteristic fingerprints were established based on polarity orientation technology. Similarity analysis, cluster analysis, and principal component analysis were used to evaluate the fingerprints. RESULTS: The HPLC fingerprints of ethyl acetate fraction from the water extract of G. scabra and G. rigescens showed obvious characteristics. The overall similarity of 21 batches of samples was 0.29-0.95. The overall similarity of G. scabra was 0.82-0.99 except S4. And that of G. rigescens was 0.75-0.99 among which 10 batches were greater than 0.90. In the cluster analysis, the samples were divided into three clusters: S4, G. scabra, and G. rigescens. In the principal component analysis, 21 batches of G. scabra. and G. rigescens could be clearly divided into two categories, while S4 was far away from the other samples of G. scabra. CONCLUSION: G scabra and G. rigescens can be identified effectively by the characteristic fingerprints. The method can be used for the quality control of Radix Gentianae.

A system research on HPLC characteristic fingerprint of Pinelliae Rhizoma and Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine / 中草药

Objective: To study the characteristic fingerprint of different extract parts from Pinelliae Rhizoma (PR) and Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine (PRPZA) by HPLC, to elucidate the major material foundations before and after processing, and to provide reliable method and scientific basis for the quality control of PR and PRPZA. Methods: The HPLC fingerprints of water, 75% ethanol and 95% ethanol extracts from PR and PRPZA were established. The similarity was analyzed with "Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica". The principal component analysis (PCA) was performed by SPSS software 17.0. Results: Six common modes of the HPLC characteristic fingerprint of different extract parts from PR and PRPZA have been established. Six specific peaks were identified as inosine, guanosine, adenosine, succinic acid, ephedrine hydrochloride, and 6-gingerol, respectively. Compared with the characteristic fingerprint of PR, there were two more peaks existed in retention time 18.3 (peak 10) and 73.5 min (peak 19, 6-gingerol) of PRPZA. Conclusion: It is the first time to establish the HPLC characteristic fingerprint of different extract parts from PR and PRPZA. The method is stable, time-saving, and reliable, and could provide an efficient basis for the quality control of PR and PRPZA.

Research on the HPLC characteristic fingerprints of Rhizoma Pinelliae and processed products / 中国药学杂志

OBJECTIVE: To establish and compare the HPLC fingerprints of different parts of Rhizoma Pinelliae (RPI), Rhizoma Pinelliae Praeparatum Cum Alumine (RPA), Rhizoma Pinelliae Praeparatum Cum Zingibere Et Alumine (RPZ), Rhizoma Pinelliae Praeparatum (RPP) and Jing Pinelliae (JPI) and provide reliable method and scientific basis for their quality control. METHODS: The HPLC fingerprints of water, 75% ethanol and 95% ethanol extracts of Rhizoma Pinelliae and processed products were established and analyzed with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(Version 2004A). RESULTS: The chromatograms of water, 75% ethanol and 95% ethanol extracts were generated as the representative standard fingerprints. Thirteen common peaks were obtained in the fingerprints of Rhizoma Pinelliae and Rhizoma Pinelliae Praeparatum Cum Alumine; 15 common peaks were obtained in the fingerprints of Rhizoma Pinelliae Praeparatum Cum Zingibere Et Alumine; 17 common peaks were obtained in the fingerprints of Rhizoma Pinelliae Praeparatum and Jing Pinelliae. Among the common peaks, 8 characterized peaks were identified as inosine, guanosine, adenosine, succinic acid, ephedrine hydrochloride, liquiritin, glycyrrhizic acid, and 6-Gingerol. Guanosine, adenosine, succinic acid, and ephedrine hydrochloride existed in Rhizoma Pinelliae and processed products. 6-gingerol was only detected in Rhizoma Pinelliae Praeparatum Cum Zingibere Et Alumine. Liquiritin and glycyrrhizic acid were detected in Rhizoma Pinelliae Praeparatum and Jing Pinelliae. A new peak (peak 8) appeared in the chromatograms of Rhizoma Pinelliae Praeparatum Cum Alumine and Rhizoma Pinelliae Praeparatum Cum Zingibere Et Alumine; Rhizoma Pinelliae Praeparatum and Jing Pinelliae did not show inosine and had a new peak (peak 11) compared to Rhizoma Pinelliae. CONCLUSION: This study established the HPLC characteristic fingerprints of different parts of Rhizoma Pinelliae and processed products. The method is stable, time-saving, reliable and can identify Rhizoma Pinelliae and its four different processed products, which provides a scientific basis for the quality control of Rhizoma Pinelliae.

Research on quality specification of the seeds of Peganum harmala L. of a uygur traditional medicine / 中国药学杂志

OBJECTIVE: To develop the quality specification of seeds of Peganum harmala. METHODS: According to the Chinese Pharmacopoeia (2010 Version, Volume 1) and its appendix method, the water, total ash, acid insoluble ash, 50% ethanol extractives, and heavy metal were analyzed for seeds of P. harmala. TLC method was used to separate harmaline (HAL), harmine (HAR) and vasicine (VAS) in seed samples using mixture of ethyl acetate-methanol-ammonia water (10:1.5:0.5) as a developing solvent on high performance silica G pre-coated plate with 254 nm fluorescent (GF254) and to identify them inspected under UV 366 nm, 254 nm, visualized by spraying with both Dragendorff reagent and by bioautographic assay. In the HPLC method, HAL and HAR were separated on a C18 column with acetonitrile-ammonium acetate water (19:81) as the mobile phase and detected at 330 nm. The HPLC fingerprints were performed on the same C18 column and eluted by using a linear gradient of acetonitrile (A) and 0.1 mmol · L-1 ammonium acetate buffer under the flow rate at 0.7 mL · min-1 and detected at 280 nm. RESULTS: In the TLC procedures, 254 and 366 nm fluorescent, Dragendorff reagent, and bioautographic assay for the detection of acetylcholinesterase inhibitor can be used for qualitative identification of the active ingredients. For the HPLC quantitative method, the calibration curve of HAR displayed ideal linearity over the range of 1.97-198.68 μg · mL-1 with average recovery of 99.69% (RSD of 1.89%). HAL displayed ideal linearity over the range of 1.70-345.30 μg · mL-1 with average recovery of 100.66% (RSD of 1.78%). The contents of HAL and HAR in 11 batches of seeds of P. harmala were 3.234% and 3.755%. In the characteristic fingerprints of seeds of P. harmala, four common peaks were identified. CONCLUSION: The results indicated that the water, total ash, acid insoluble ash, and 50% ethanol extractives were not more than 9.0%, 8.0%, 1.0%, and 22.0%, respectively. The heavy metal of plumbum, cadmium, arsenic, mercury, and copper were not more than 5 × 10-6, 3 × 10-6, 2 × 10-6, 2 × 10-6 and 20 × 10-6, respectively. The content limit of the sum of HAL and HAR was not lower than 5.5%. With the peak of HAL as reference peak, the variance of relative retention time of the four common peaks, in the characteristic fingerprints of seeds off. harmala, should be fluctuated in the range of 5% of the specified value. The qualitative and quantitative method established was suitable for the quality evaluation and assessment of seeds of P. harmala.

Identification of active components in Syngnathus acus by HILIC-ESI-TOF/MS and study on their specific fingerprint chromatograms / 中草药

Objective: To develop a new method based on hydrophilic interaction chromatography-electrospray ionization-time of flight-mass spectrometry (HILIC-DAD-ESI-TOF/MS) for the rapid identification of the active components in Syngnathus acus and the development of their specific fingerprint chromatograms. Methods: Samples were extracted by accelerated solvent extraction, and the extraction conditions were optimized. The developed HILIC-DAD-ESI-TOF/MS method was used to identify the components in water extract from S. acus, and a chromatographic fingerprint based on HILIC analysis was established. Results: Ten compounds in S. acus extract could be primarily identified by HILIC-DAD-ESI-TOF/MS on-line detection, in which seven nucleosides were determined. The HPLC characteristic fingerprint was established on the basis of analysis on the multi batches of S. acus, which could be used to evaluate the quality of S. acus combined with similarity calculation. Conclusion: This method is simple and rapid, and is a powerful tool for the identification of S. acus.

Verification of Digitized Information of GC-MS Characteristic Fingerprint of Amomum villosum by GC / 中药新药与临床药理

Objective To investigate the applicability of the digitized standard of GC-MS characteristic fingerprint in GC. Methods The GC-MS and GC were used to analyze the essential components of thirteen batches of Amomum villosum samples. The digitized standard of characteristic fingerprint of Amomum villosum essential oil was compared with the result of the samples detected by GC-MS and GC. Results Ten essential characteristic components have been identified in thirteen batches of Amomum villosum samples,and their relative content was (88.15?2.97)%,which are the representative components. The main components were ?-pinene,camphene,?-pinene,?-myrcene,limonene,linalol,camphor,isoborneol,borneol and borneol acetate. With the ten components as indexes,the sample similarity calculated by cosin method was 0.994～1.000 when analyzed by GC-MS and GC,0.978～0.999 when analyzed by GC-MS and the digitized standard of GC-MS characteristic fingerprint,and 0.986～0.998 when analyzed by GC and the digitized standard of GC-MS characteristic fingerprint. Conclusion The digitized standard of GC-MS characteristic fingerprint of Amomum villosum can be used in GC,which will ensure the comparison of results obtaining at different time by different types of machines,on different chromatographic columns and under different conditions. The method can be used for quality evaluation of Amomum villosum.