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.

Extraction Process and Content Determination of Total Flavonoids in Ginkgo( Ginkgo biloba L. ) Leaves

[Objectives]This study was conducted to study the extraction process and the content determination of flavonoids in ginkgo( Ginkgo biloba L.) leaves.[Methods]Ethanol extraction and methanol extraction of total flavonoids in ginkgo leaves were studied,and the optimal extraction conditions for flavonoids were determined by orthogonal test; and with quercetin as reference substance,total flavonoid content in ginkgo leaves was determined by UV spectrophotometry.[Results]The optimal extraction process was 4 h of Soxhlet extraction with methanol; and the total flavonoid contents had a good linear relation in the range of 0. 006 5-0. 039 mg/ml( R^2= 0. 999 9),the average content was stabilized at 1. 135%,and the average recovery of the method was 102. 0%. [Conclusions]This study selected the optimal extraction process for total flavonoids in ginkgo leaves. The test method is simple with high accuracy and precision,and is suitable for the extraction and determination of total flavonoids in ginkgo leaves.

Content Determination of Astragaloside Ⅳ in Astragalus from Three Different Regions by HPLC

[Objectives] The content of astragaloside in Astragalus membranaceus(Fisch.) Bge.var.mongholicus(Bge.) Hisao from three different regions was determined.[Methods] Referring to the method recorded in the Chinese Pharmacopoeia(2015 edition),the content of astragaloside IV in A.membranaceus was determined by HPLC.[Results] There were great differences in the astragaloside IV content of A.membranaceus among different regions.The content of astragaloside IV in A.membranaceus cultivated in Inner Mongolia was highest(0.155%),followed by that(0.143%) in A.membranaceus cultivated in Gansu,and the content of astragaloside IV in A.membranaceus cultivated in Shanxi was lowest(0.080%).The contents of astragaloside IV in A.membranaceus from different regions were all in line with the standard(not less than 0.040%) of Chinese Pharmacopoeia(2015 edition).[Conclusions]The content of astragaloside IV in A.membranaceus cultivated in three different regions met the medicinal standards.

Content Determination of Quercetin in Ferment of Ginkgo biloba L. Leaves by HPLC

[Objectives] This study aimed to determine the content of quercetin in ferment of Ginkgo biloba L.leaves.[Methods]Bacillus licheniformis was selected for solid-state fermentation of G.biloba leaf powder,and the content of quercetin in ferment of G.biloba leaves was determined by reversed-phase high-performance liquid chromatography.First,the flavonoid glycosides were extracted with methanol.Then,the flavonoid glycosides were hydrolyzed with hydrochloric acid to prepare the test solution.The chromatographic conditions were as follows:Platisil ODS C_(18) column(150 mm × 4.6 mm,5 μm);V_(methonal)∶V_(water)(0.4% phosphoric acid solution) =55∶45;flow rate of 1 m L/min;Shimadzu UV detector;detection wavelength of 360 nm.[Results] Quercetin was used as a reference substance.In the range of 0.002 6-0.036 0 g/L,there was a good linear relationship,with correlation coefficient of 0.999 8 and RSD of 1.26%.[Conclusions] This method is simple,easy to operate,accurate,and reproducible.It is suitable for the determination of quercetin content in G.biloba leaves.

Extraction Technology of Total Flavonoids from Pueraria lobata (Willd.) Ohwi and Content Determination of Pueraria lobata (Willd.) Ohwi at Different Altitudes

[Objectives]To optimize the extraction technology of total flavonoids from Pueraria lobata( Willd.) Ohwi. [Methods]The ultraviolet-visible spectrophotometry was used to determine the content of total flavonoids in Pueraria lobata( Willd.) Ohwi. With the puerarin as index,the reflux extraction and single factor test were employed to investigate the effects of temperature,time,ethanol concentration and solid-liquid ratio on the content of total flavonoids in Pueraria lobata( Willd.) Ohwi,respectively. Under the optimal extraction technology,the content of total flavonoids in Pueraria lobata( Willd.) Ohwi at different altitudes was determined.[Results] The optimum extraction process was as follows: 70%ethanol; solid-liquid ratio of 1∶ 30; 1 h reflux extraction. Under these conditions,the extraction rate of flavonoids in Pueraria lobata( Willd.) Ohwi was 11. 48%,the total flavonoids content of different kudzu parts was in the order of roots > stems > leaves,and the total flavonoids content of the sample at about an altitude of 1000 m was significantly higher than at the altitudes of 1400 m and 1700 m.[Conclusions]It was suggested that the Pueraria lobata( Willd.) Ohwi should not be cultivated as medicinal plant in too high mountains,and the stems and leaves of Pueraria lobata( Willd.) Ohwi could be used as raw materials for extracting total flavonoids.

Study on Colorimetric Method and Content Determination of Total Phenol in Homemade Plum Wine

[Objectives] To study on colorimetric method and content determination of total phenol in homemade plum wine. [Methods]The optimal determination condition of total polyphenols content in plum wine( commercially available and homemade) by Folin-Ciocalteu method was inspected,and commercially available and homemade plum wine was evaluated by the method. [Results]The optimal determination conditions of total polyphenols content: sample dose of 1. 0 m L,Folin-Ciocalteu reagent of 1 m L,4% Na_2CO_3 solution of 4. 0 m L,reaction temperature of 50 ℃,reaction time of 1. 5 h,and determination wavelength of 756 nm. Absorbance showed good linear relationship with total polyphenols content within the range of 17. 73-59. 12 μg/m L( y = 14. 878 x + 0. 0739,R^2= 0. 9998). Recovery rate of adding standard sample was between 98. 8% and 103. 5%,and relative standard deviation was 2. 0%( n = 5). [Conclusions]The method had high precision degree and good stability,which was suitable for measuring total polyphenols content in plum wine( commercially available and homemade).

Optimization of Extraction Process and Content Determination of Total Flavonoids from Sanguisorbae Radix and Carbonized Sanguisorba Root by Orthogonal Experimental Design

[Objectives] To optimize the extraction process of total flavonoids from Sanguisorbae Radix and carbonized Sanguisorba root,compare quality of different batches of Sanguisorbae Radix,study the effects of processing on the content of flavonoids,and provide scientific basis for reasonable utilization of Sanguisorbae Radix. [Methods] Test samples were prepared by heating,refluxing,and extraction,the extraction process was optimized by orthogonal experiment design,color was developed by NaNO_2-Al( NO_3)3-NaOH,and total flavonoids were measured by UV method at the wavelength of 510 nm. [Results] The linear relationship of rutin was excellent in the concentration range of 0. 1248 mg/mL-0. 5712 mg/mL,R^2= 0. 9997; the average recovery was 99. 67% and the RSD was 0. 70%. The optimum extraction conditions were as follows: the volume fraction of ethanol was 50%,the extraction temperature was 90℃,the extraction time was 90 min,and the solid-to-liquid ratio was 1∶ 20( g/mL). [Conclusions] After optimization of the extraction process,the extraction rate of total flavonoids in samples of Sanguisorbae Radix was significantly increased; there was certain difference in the content of total flavonoids between different batches of Sanguisorbae Radix and processed products; the total flavonoids significantly declines in carbonized sanguisorba root,and the influence of processing on its curative effect was to be further studied.

Extraction Process and Content Determination of Caffeic Acid in Zhuang Medicine Cryptolepis buchananii

[Objectives]To establish a method for determining the content of caffeic acid in Zhuang Medicine Cryptolepis buchananii.[Methods]The content of caffeic acid in C.buchananii leaves was determined by high performance liquid chromatography(HPLC).[Results]The results of HPLC determination showed that the RSD values were less than 3%,and the content of caffeic acid in 10 batches of C.buchananii leaves was in the range of 0.0965%-0.4772%.[Conclusions]This method is accurate and reliable,has good linear relationship and high precision,and can be used for quality evaluation of C.buchananii.

Technical Study on Extraction of Procyanidins in Lycium ruthenicum Murr. Using Sub-critical Fluid 1,1,1,2-Tetrafluoroethane( R134a)and Content Determination from Different Producing Areas

[Objectives] To study the optimal conditions for extracting procyanidins fromLycium ruthenicum Murr. with sub-critical fluid R134 a( 1,1,1,2-tetrafluoroethane) in 1 L extraction kettle. [Methods]Taking the extraction rate of procyanidins as an indicator,the influence of pressure,temperature,and extraction time on extraction rate of procyanidins fromL. Ruthenicum Murr. was studied by single factor experimental methods and orthogonal array design. [Results]The order of factors affecting extraction rate of procyanidins was extraction temperature > extraction pressure > extraction time. The optimum extraction conditions were as follows: the extraction rate of procyanidins fromL. ruthenicum Murr. was the highest with extraction pressure of 1. 2 MPa,extraction temperature of 50℃ and extraction time of 90 min. The content of procyanidins in L. ruthenicum Murr. from different producing areas was determined by vanillin-HCl method under the optimal conditions. [Conclusions] The method has the advantages of easy operation,good selectivity,low extraction temperature and high extraction efficiency,which is suitable for extraction of procyanidins in L. ruthenicum Murr.

Content Determination of Chlorogenic Acid and Luteoloside in Flos Lonicerae

The contents of chlorogenic acid and luteoloside in Flos Lonicerae from Binhai New Area and Jinnan District of Tianjin were determined to provide basis for the quality identification of this medicinal material. The content of chlorogenic acid was determined by HPLC. In Flos Lonicerae from Binhai New Area and in Flos Lonicerae harvested at different stages from Jinnan District,the contents of chlorogenic acid were 3. 804%,5. 507%( three green stage),4. 855%( silver flower stage) and 4. 220%( golden flower stage),respectively,and the contents of luteoloside were 5. 53%,12. 405%( three green stage),14. 370%( silver flower stage) and 0. 917%( golden flower stage),respectively. The contents of chlorogenic acid in Flos Lonicerae from Jinnan District were higher than that from Binhai New Area. Among different stages,the content of chlorogenic acid was highest in three green stage,followed by that in silver flower stage. As the flowers bloomed,the content of chlorogenic acid in the medicinal material showed a significant downward trend. In Flos Lonicerae from Jinnan District,the content of luteoloside was highest in silver flower stage and lowest in golden flower stage.

Content Determination of Total Flavonoids in Tetracera asiatica from 10 Different Production Areas in Guangxi

[Objectives] To determine the content of total flavonoids in Tetracera asiatica from different production areas in Guangxi and provide a basis for improving the quality standards of crude drug. [Methods] Using 50% ethanol as extractor,NaNO_2-Al(NO_3)_3-NaOH as chromogenic system and rutin as the reference substance,the content of total flavonoids from the crude drug was determined. [Results] A method for determining the content of total flavonoids was established,the content of total flavonoids was 11.35-21.35 mg·g^(-1),and the average content was 17.64 mg·g^(-1). [Conclusions] The method had high repeatability and stability,and the method was simple,rapid and sensitive and suitable for quantitative analysis.

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.