TLC Identification of Yao Medicine Pileostegia tomentellal and Extraction Technology and Content Determination of Umbelliferone

[Objectives]To establish a TLC and content determination method of Pileostegia tomentellal,with umbelliferone as the indicator component.[Methods]TLC identification was performed by silica gel G thin layer plate with n-hexane-ethyl acetate(4:3)as the developing agent,and the plate was examined by UV lamp(365 nm).The umbelliferone content was determined by HPLC:Inertsil ODS-3 C 18 column(4.60 mm×250 mm,5μm);mobile phase acetonitrile-0.2%phosphoric acid gradient elution;detection wavelength 320 nm,flow rate 1.0 mL/min,column temperature 30℃,injection volume 10μL.[Results]The chromatogram of P.tomentellal showed the same color spot in the same position as that of reference medicinal material,and the spot was clear with good specificity.Umbelliferone showed a good linear relationship when the injection volume was 2.63-131.27μg/mL(R^(2)=0.9997).The average recovery of umbelliferone in the low,middle and high adding groups of P.tomentellal was 99.57%and the RSD was 2.15%.[Conclusions]The method can effectively identify Yao medicine P.tomentellal and accurately determine the content of umbelliferone in medicinal materials,which will provide a scientific basis for the development and utilization of medicinal resources of Yao medicine P.tomentellal.

Purification Process,Content Determination,Pharmacological Activity and Molecular Mechanism of Neogambogic Acid

Neogambogic acid is characterized by broad antitumor spectrum,good antitumor effect and low toxicity and side effects.This paper reviews the purification process,content determination and pharmacologic activity of neogambogic acid,in order to provide a theoretical reference for the research and application of neogambogic acid.

Research Progress on Purification Process Optimization and Content Determination of Zeaxanthin

At present,the purification process of zeaxanthin mainly includes organic solvent extraction,ultrasonic-assisted extraction and enzyme extraction,and the content determination technology mainly includes ultraviolet-spectrophotometry and high performance liquid chromatography.In this paper,the purification process and content determination technology of zeaxanthin in recent years are reviewed in order to provide ideas and theoretical basis for further research and application of zeaxanthin.

Extraction Process and Content Determination of Caffeic Acid in Laggera alata from Different Production Areas of Guangxi

[Objectives] To establish a method for determining the content of Laggera alata( D. Don) Sch. Bip. Ex Oliv. using caffeic acid the target component,and to compare the content of caffeic acid in the medicinal materials of L. alata in different production areas of Guangxi.[Methods]The content was determined by Inertsil~ODS-3 chromatographic column C_(18)( 4. 60 mm × 250 mm,5 μm,mobile phase: acetonitrile-0. 1% phosphoric acid( 22∶ 78),detection wavelength: 320 nm,flow rate: 1. 0 m L/min,column temperature: 30℃,and injection volume: 10 μL. [Results] The caffeic acid showed a good linear relationship in the range of injection volume of 0. 025 92-0. 259 2 μg( R =0. 999 5). The average recovery rate was 98. 33%( RSD = 1. 85%). L. alata in different production areas of Guangxi contained the caffeic acid,and there was a great difference in the caffeic acid. L. alata in Baise had the highest content of caffeic acid,while that in Guilin had the lowest content of caffeic acid. [Conclusions]This method can accurately determine the content of caffeic acid and is expected provide a scientific basis for the development and utilization of herbal medicine L. alata.

Thin-layer Identification,Extraction Process and Content Determination of Chlorogenic Acid in Laggera alata(D.Don)Sch.Bip.ex Oliv.

[Objectives]This paper aims to establish thin-layer identification and content determination method for Laggera alata(D.Don)Sch.Bip.ex Oliv.with chlorogenic acid as the index component and compare the content of chlorogenic acid in L.alata from different places in Guangxi.[Methods]Silica gel GF254 thin-layer plate was used for identification under an ultraviolet lamp(365 nm),with butyl acetate-formic acid-water(V∶V∶V=7∶2.5∶2.5)as a developing agent.The content of chlorogenic acid was determined under the following chromatographic conditions:column,Inertsil ODS-3 C18 column(4.60 mm×250 mm,5μm);mobile phase,methanol-0.1%phosphoric acid(28∶72);detection wavelength,329 nm;flow rate,1.0 mL/min;column temperature,25℃;and injection volume,10μL.[Results]Chlorogenic acid can be detected by thin layer chromatography with clear spot and good specificity.Chlorogenic acid showed a good linear relationship in the injection amount range of 0.099-0.99μg(R^(2)=0.9999).The content of chlorogenic acid in L.alata varied greatly among the 10 different producing areas in Guangxi.L.alata produced in Dee Township,Longlin,Baise,Guangxi showed the highest chlorogenic acid content,and that produced in Shangsi County and Pingle County showed the lowest chlorogenic acid content.[Conclusions]This method can effectively identify L.alata and accurately determine the content of chlorogenic acid,thereby providing a scientific basis for the development and utilization of L.alata resources.

Content Determination of Total Flavonoids from Paulownia fortunei Flower

[Objectives]The research aimed to explore the determination method for content of total flavonoids from Paulownia fortunei flower.[Methods]Rutin was taken as control.By comparing absorption characteristics of total flavonoids in NaNO_(2)-Al(NO_(3))_(3)-NaOH and AlCl_(3) chromogenic system,an appropriate method for the determination of total flavonoids from P.fortunei flower was screened,and the method was verified by methodology.On the basis of single factor experiment,the coloration time of the method was optimized by orthogonal experiment.[Results]NaNO_(2)-Al(NO_(3))_(3)-NaOH chromogenic system was more suitable for the determination of total flavonoids from P.fortunei flower,and the adding standard recovery was between 99.2%and 105.5%,and RSD was 2.18%,with better repeatability,precision,stability and accuracy.The optimized coloration time of NaNO_(2),Al(NO_(3))_(3) and NaOH was 6,6,and 10 min.Under the condition,average content of total flavonoids from P.fortunei flower was 3.78%,and RSD was 2.05%.[Conclusions]The optimized NaNO_(2)-Al(NO_(3))_(3)-NaOH chromogenic method is simple,rapid,and accurate,and could be used as a method for the determination of total flavonoids from P.fortunei flower.

Optimization of Extraction Process for Total Flavonoids from Penthorum chinense Pursh and Comparison of Their Contents from Different Parts

[Objectives]This study was conducted to optimize the extraction process of total flavonoids from Penthorum chinense Pursh and compare their contents from different parts.[Methods]Single factor and orthogonal experiments were designed to optimize the extraction process of total flavonoids from P.chinense Pursh with the volume fraction of ethanol,the ratio of material to liquid,heating reflux extraction time and extraction times as factors,and the content of total flavonoids as the index.A verification test was carried out.The optimized extraction process was adopted to compare the contents of total flavonoids from different parts of P.chinense Pursh.[Results]The best extraction process was extracting the powder of P.chinense Pursh for 2.0 h with 20 times of 55%ethanol by reflux twice.Under this condition,the contents of total flavonoids were 3.63%,8.90%,11.28%,and 4.36%from stems,leaves,flowers and whole grass of P.chinense Pursh,respectively.[Conclusions]The process is reasonable,feasible and stable,and can effectively extract total flavonoids from P.chinense Pursh.The contents of total flavonoids from different parts of P.chinense Pursh were quite different,and the value was higher in the leaves and flowers,so the proportions of leaves and flowers should be paid attention to in the industrial processing of P.chinense Pursh.

Determination of Chlorogenic Acid,Geniposide,Total Flavonoids and Total Triterpenes in Wulan-13

[Objectives]This study was conducted to establish a method for the determination of chlorogenic acid,geniposide,total flavonoids and total triterpenes in Wulan-13.[Methods]The contents of chlorogenic acid and geniposide were determined by HPLC,and the contents of total flavonoids and total triterpenes were determined by an ultraviolet spectrophotometer.[Results]There was a good linear relation between the mass of chlorogenic acid reference substance and the peak area in the range of 0.05-0.45μg,and the regression equation was Y=2524.1X+3.1943,(r=0.9998).A good linear relationship was found between the mass of gardenoside reference substance and the peak area in the range of 0.776-6.984μg,and the regression equation was Y=1670.5X+64.804,(r=0.9998).There was also a good linear relation between the mass of rutin reference substance and its absorbance in the range of 0.00808-0.04848 mg,and the regression equation was Y=12.916X+0.014,(r=0.999).The mass of oleanolic acid reference substance had a good linear relation with its absorbance in the range of 0.00418-0.0209 mg,and the regression equation was Y=51.89X-0.0839,(r=0.9991).[Conclusions]The content determination method is simple,reliable and reproducible,and suitable for controlling the contents of chlorogenic acid,geniposide,total flavonoids and total triterpenes in Wulan-13.

Determination of Salvianolic Acid B in Yiqi Huayu Prescription by HPLC

[Objectives]To determine the content of salvianolic acid B in Yiqi Huayu Prescription by HPLC.[Methods]The chromatographic column was ZORBAX Eclipse Plus C 18(4.6 nm×250 nm,5μm);the mobile phase was acetonitrile-0.1%phosphoric acid(21:79),the detection wavelength was 286 nm,the column temperature was 30℃,and the flow rate was 1.0 mL/min.A method for determination of salvianolic acid B in Yiqi Huayu Prescription was established.[Results]The linear relationship of salvianolic acid B was good in the range of 0.0214-0.4064 mg/mL.The regression equation was Y=5995.98984 X-0.07332,r=0.9999.The average recovery rate was 98.88%(RSD=1.6%).[Conclusions]The method is reliable,accurate and specific,and can be used for the determination of salvianolic acid B in Yiqi Huayu Prescription.

Determination of the Content of Five Active Components in Toad Skin

[Objectives] To establish a method for the determination of active components in toad skin. [Methods] HPLC method was used to determine the content of five active components (bufotalin, cinobufotalin, bufalin, cinobufagin and resibufogenin) in toad skin. [Results] Chromatographic conditions are as follows: Agilent ZORBAX SB-C 18 chromatographic column was used;acetonitrile (A)-0.3% glacial acetic acid (B) gradient elution (0-15 min, 28%A-54%A;15-35 min, 54%A-54%A) was conducted;the flow rate was 0.6 mL/min;the detection wavelength was 296 nm;the column temperature was 30 ℃;the sample size was 10 μL. Under the above conditions, the determination method of the five components can be established at one time. [Conclusions] The method was stable and reliable, and can provide experimental basis for the development and utilization of active ingredients in toad skin.

Determination of Phytoene Content in Tomato Ketchup by HPLC

[Objective] The aim was to establish an HPLC method for determination of phytoene content in tomato ketchup. [Method] The chromatographic conditions were as follows： column, Agilent AT C18 （250 mm×4.6 mm, 5 μm）; mobile phase, methanol-THF （75：25）; detection wavelength, 287 nm; flow rate, 1.0 ml/min; column temperature, 30 ℃; sample size, 50 μl. [Result] There was a good linear relation- ship in the phytoene content range of 0.186-1.116μg. The average recovery rate was 103.8% with RSD of 1.47%. The phytoene content in the tomato ketchup sample was determined as 10.7 rag/100 g simple, accurate, rapid and reliable, and phytoene content in tomato ketchup. [Conclusion] The established method is it can be used for the determination of

Determination of Coenzyme Q_(10) Content by RP-HPLC and Daily Change of Transmittance of Co-Q_(10)

[Objective] The aim of this study was to establish a reversed phase high-performance liquid chromatography（RP-HPLC）method for the content determination of Co-Q10.[Method] The RP-HPLC method was used to detect the Co-Q10,and the ultraviolet spectro-photometry was used to analyze the daily change of transmittance of Co-Q10.[Result] By using RP-HPLC,Co-Q10 had a good linear relationship between 40-300 μg/ml（r=0.999 9）.The limit of detection was 0.4 ng and the average recovery was 97.44%（n=3）.The system suitability of HP-HPLC was good,and the average recovery and precision results could meet the needs of assay.[Conclusion] This method was convenient,accurate and reproducible and could be used in quality control of Co-Q10.However,when it operates,light should be evaded.

Difference analysis of different parts of chicory based on HPLC fingerprint and multi-component content determination

Objective:To establish HPLC fingerprints of different parts of chicory stems,leaves,roots,flowers and seeds,and compare the similarities and differences of chemical components in different parts,so as to provide a scientific basis for the comprehensive utilization of chicory.Methods:To establish the HPLC fingerprint of chicory,the chromatographic column was chosen with Agilent ZORBAX Eclipse XDB-C_(18),the mobile phase was methanol(A)-0.2%formic acid(B),the flow rate was 1 mL/min,the column temperature was 30℃,and the detection wavelength was 254 nm.The Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese Medicine(2012 Edition)was used to evaluate the similarity of different parts of decoction pieces,and the determination method of multi-component content was established based on fingerprint identification chromatographic peaks,and the determination results were analyzed.Results:The HPLC fingerprinting method of chicory was established.Sixteen chromatographic peaks were identified and 10 of them were identified as:caftaric acid(1),esculin(2),chlorogenic acid(3),esculetin(4),caffeic acid(5),cichoric acid(8),hyperoside(11),rutin(12),isochlorogenic acid C(14)and luteolin(16).The similarity of different parts was 0.084–0.701.At the same time,the total content of detected chemical components was ranked as flower>leaf>stem>root>seed.Roots did not contain caftaric acid,rutin,and luteolin,flowers did not contain luteolin,and seeds did not contain caftaric acid,cichoric acid,and luteolin.The content of cichoric acid in leaves was the most,and esculin in flowers was the most.Conclusion:The results of HPLC fingerprint and multi-component content determination revealed the similarity and difference of different parts of chicory from chemical composition,indicating that there were certain differences in different parts of chicory.The established HPLC fingerprinting method can provide a reference for quality control and evaluation of different parts of the chicory.

Determination of the Contents of Seven Chemical Components in Vidal Grape by Quantitative Analysis of Multi-components by Single Marker(QAMS)

[Objectives]This study was conducted to establish a method of quantitative analysis of multi-components by single marker(QAMS)for the simultaneous determination of such seven chemical components as gallic acid,epicatechin,catechin,ferulic acid,chlorogenic acid,rutin and caffeic acid in Vidal grape.[Methods]The high performance liquid chromatography was carried out using a COSMOSIL C18-MS-II column(4.6 mm×250 mm,5μm)with the mobile phase acetonitrile-2%acetic acid aqueous solution(gradient elution)at a flow rate of 1.0 ml/min.The detection wavelength was 280 nm,and the column temperature was 25℃.Using caffeic acid as an internal reference,the relative correction factors between it and other six to-be-detected components,and the contents of the seven components were calculated using the correction factors.The established was compared the results with the external standard method to verify the feasibility and accuracy of the method.[Results]The seven components had a good linear relationship in the ranges of 1.060-10.60,1.419-14.19,1.062-10.62,0.2950-2.950,0.1019-1.019,0.2014-2.014,and 0.1498-1.498μg,respectively,and the relative correction factors of gallic acid,epicatechin,catechin,ferulic acid,chlorogenic acid and rutin were 0.9760,0.7806,0.3277,1.640,1.161,2.778,respectively.There was no significant difference between the results of the QAMS method and the external standard method.[Conclusions]The QAMS method using caffeic acid as an internal reference is accurate and feasible,and provides a reliable method for the quality evaluation of Vidal ice grape.

Identification of variety of rice and determination of content of each rice in mixtures of two rices by nonlinear chemical analysis

Fingerprints of two varieties of rice and their mixtures were investigated by a nonlinear chemical reaction system consisting of rice components,sodium bromate,manganese sulfate,sulfuric acid and acetone.The variety of rice was identified by the visual characteristic of fingerprint and system similarity pattern recognition,and the content of each variety of rice in the mixture was determined by the quantitative information of fingerprint.The results show that nonlinear chemical analysis may be used to exactly identify the variety of pure rice and to accurately determine the content of each variety of rice in the mixture,indicating the method is simple and convenient.

Determination of the Content of Gastrodin in Jingtianshenma Tablets by High Performance Liquid Chromatography

[Objectives]The research aimed to establish a high performance liquid chromatography method for the content determination of gastrodin in Jingtianshenma Tablets.[Methods]A phenomenex Luna C18(250 mm×4.6 mm,5μm)column was used;the mobile phase was acetonitrile-0.05%phosphoric acid solution(3∶97);the detection wavelength was 220 nm,and the column temperature was set at 25℃.[Results]Gastrodin showed a good linear relationship in the range of 0.0984-0.5904μg with the peak area,and regression equation was Y=2000000X-51999(r=0.9999).The limits of detection and quantification for gastrodin were 2.50 and 4.20 ng respectively,and the average recovery rate was 95.95%.[Conclusions]This method is sensitive,accurate and reproducible,with good linearity,and it is suitable for the content determination of gastrodin in health food Jingtianshenma Tablets.

Determination of the Content of Astragaloside IV in Yikangshu Granules by High Performance Liquid Chromatography

[Objectives]The research aimed to establish a high performance liquid chromatography method for the content determination of astragaloside IV in Yikangshu Granules.[Methods]Kromasil 5μm C18(2),100 A,250 mm×4.6 mm was used;the mobile phase was acetonitrile-water(32∶68);flow velocity was 1.0 mL/min;the temperature of evaporator and sprayer was 80 and 30℃;the column temperature was set at 30℃,and injection volume was 20μL.[Results]Astragaloside IV showed a good linear relationship in the range of 1.01-10.14μg with the peak area,and regression equation was lgY=1.7728lgX+1.597(r=0.9999).The limit of detection for astragaloside IV was 1.96 ng,and the average recovery rate was 95.31%.[Conclusions]This method is sensitive,accurate and reproducible,with good linearity,and it is suitable for the content determination of astragaloside IV in health food Yikangshu Granules.

Determination of Salbutamol Sulfate Content by Molecular Spectroscopy with Cu2+ as a Medium

Salbutamol sulfate was coordinated with Cu2+ when pH was 10, and the characteristic absorption peak of the complex was generated at 334 nm. The absorption intensity of the complex was in linear relation with the concentration of salbutamol sulfate. The regression equation was A=0.011 ρsalbutamol sulfate-0.123 0, correlation coefficient r=0.999 5, and the detection limit was 4.6 μg/ml. It was used for the determination of salbutamol sulfate samples successfully. The recovery rate was 100.5%-103.0%, and the RSD was 1.5%.

Study on Method for Determination of Artemether Content in New Artemether Injection

This study was conducted to establish a high performance liquid chromatography（ HPLC） method for the determination of artemether in the artemether injection using Hypersil ODS C18 chromatographic column（ 5 μm,4. 6 mm × 150 mm）. Mobile phase,column temperature,flow rate and detection wavelength were optimized. Acetonitrile-water-tetrahydrofuran（ 62∶ 37∶ 1,V/V） was selected as the mobile phase,and the HPLC was performed with column temperature at30 ℃ and the flow rate at 1. 0 ml/min; and the detection wavelength was set at 216 nm. The HPLC detection system of artemether had good suitability. The linearity was good in 100-800 μg/ml concentration range,and the regression equation was y = 302. 36 x-682. 02,R2= 0. 999 8. The overall average recovery was97. 58%,and the RSD was 1. 58%. Three batches of artemether injection samples were determined by the method,showing RSD of 1. 42%. The method could be used for the detection of artemether content in artemether injection.

CONTENT DETERMINATION OF HYPERIN IN RHODODENDRON BY HPLC

As one of the cultural heritages of the nation,Mongolian Medicine was formed and developed in the long-term medical practice of the Mongolian people,and has become a major constituent part in the Traditional Chinese Medicine.In the past,Mongolian medicine made great contributions to the living and breeding of generations of the Mongolian people and had a positive impact on the formation of