Research Progress on Purification Process, Content Determination and Pharmacological Action of Atractylodin

Atractylodis Rhizoma comes from the dry rhizome of Atractylis lancea or Atractylodes chinensis in the Compositae family,and it is suitable for preventing and treating diseases such as cold,edema,night blindness and rheumatic arthralgia.Atractylodin is the main active component extracted and isolated from Atractylodis Rhizoma.A large number of studies have found that atractylodin has excellent drug activity in improving gastrointestinal emptying,anti-inflammation,inhibiting malignant tumor and reducing blood lipid.In this paper,the purification process and pharmacological activity of Atractylodin were summarized to provide a theoretical basis for basic research,clinical application and further development and utilization of atractylodin.

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.

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 Content of the Magnolol from Magnolia officinalia Leaves by HPLC

[Objective] This study aimed to extract magnolol from wild Magnolia offici- nalia leaves by ultrasonic-assisted extraction. [Method] Magnolol was qualitatively id- entified by coloration method and thin layer chromatography, and magnolol content in Magnolia officinalia leaves was measured by high performance liquid chromatog- raphy （HPLC）. The HPLC was conducted with C18 as the stationary phase while different mobile phases were selected. The measurement wavelength, flow velocity and sample size adopted in the HPLC were 294 nm, 1 ml/min and 20 μl, respec- tively. [Result] Magnolol content in Magnolia officinalia leaves was 0.75%. When methyl alcohol and water with a volume ratio of 78：22 was used as the mobile phase, the retention time of magnolol was 4.528 min and the separation effect was good. In addition, it was easy to operate with good reproducibility and sensitivity. [Conclusion] This method is suitable for the measurement of magnolol content in Maonolia officinalia leaves.

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 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 Benzoylaconitine Compounds in the Decoctions of Aconiti Lateralis Radix Praeparata(Heishun Pieces),Trichosanthis Fructus and Their Combination

[Objectives]This study was conducted to determine the contents of benzoylmesaconine,benzoylaconitine and benzoylhypacoitine in the decoctions of Heishun pieces,Trichosanthis Fructus and their combination.[Methods]Heishun pieces,Trichosanthis Fructus and their combination were extracted for different time periods,and then grouped.HPLC was performed using an Agilent ZORBAX SB-C 18 chromatographic column(4.6 mm×250 mm,5μm)and acetonitrile-0.02 mol/L sodium dihydrogen phosphate as the mobile phase at a flow rate of 1 mL/min and a column temperature of 30℃,and the sample volume was 20μL.The detection wavelength was 230 nm.[Results]The total amounts of benzoylmesaconine,benzoylaconitine and benzoylhypacoitine in the single decoction group of Heishun pieces were all significantly different from those in the combined decoction group at corresponding time.[Conclusions]The total content of the benzoylaconitine type increased significantly after the combined decoction of Heishun pieces and Fructus Trichosanthis,which proves the scientificity of"eighteen incompatible medicaments,19 counteraction"in traditional Chinese medicine to some extent.

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.

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 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.

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.

Determination of Naringin Content in Peel of Guangxi Citrus maxima (Burm.) Merr by HPLC

[Objectives]To establish a method for determining the naringin content in the peel of Guangxi Citrus maxima(Burm.)Merr.[Methods]The high performance liquid chromatography(HPLC)method was applied.The chromatographic column was Agilent HC-C18(4.6 mm×250 mm,5μm);the mobile phase was acetonitrile-0.1%phosphoric acid(18∶82);the flow rate was 1.0 mL/min;the column temperature was 25℃;the detection wavelength was 283 nm.[Results]Naringin showed a good linear relationship in the range of 0.164-3.27μg,r=0.9999.The average recovery rate was 98.66%,and RSD=1.80%(n=6).[Conclusions]This method is simple,feasible,reproducible,and accurate,so it can be used for the determination of naringin content in the peel of Guangxi C.maxima.

HPLC Method for Determination of Content of Total Flavonoids from Ginkgo (Ginkgo biloba L.) Leaves

[Objectives]This study was conducted to determine the content of total flavonoids in ginkgo ( Ginkgo biloba L.) leaves.[Methods]The content of total flavonoids from ginkgo leaves was determined by reversed phase-high performance liquid chromatography (RP-HPLC).The flavonoid glycosides were first extracted with methanol,and hydrolyzed with hydrochloric acid solution to prepare a test solution.Platisil ODS C18 column (150 mm×4.6 mm,5 μm) and the mobile phase V methanol ∶ V water (0.4% phosphoric acid solution)=85∶ 15 were selected for HPLC separation.The HPLC separation was performed with the column at a column temperature of 25℃ using the mobile phase at a flow rate of 1 ml/min.The sample size was 10 μl,and detection was performed with an Agilent HPLC ultraviolet detector at 360 nm.[Results]The reference substance,quercetin,had good linearity in the range of 0.002 6-0.052 0 g/L,with a correlation coefficient of 0.999 7;and the RSD was 1.26%.[Conclusions]The determination method has rapid and simple operation with accurate results and is good in repeatability.This method is suitable for the determination of content of total flavonoids in ginkgo leaves.

Determination of Isoflavone from Soybean Lines Cultivated in Jilin Province and Correlation Analysis between Isoflavone Content and Soybean Quality

[Objective]The aim of this study was to set up a high performance liquid chromatography for rapid determination of isoflavones from soybean and analyze the correlation between isofalvone content and protein or fat content. [Method]The isoflavones were firstly extracted by 80% methanol and then hydrolyzed at 100 ℃. The chromatographic separation adopted a reversed-phase C18 analytical column with binary high-pressure gradient elution,while its analysis time was 25 min and column temperature was 40 ℃. The diode array detector was used for monitoring with wavelength of 260 nm. The correlation between isofalvone content and protein or fat content was analyzed by data processing system Origin 6.0. [Result]The high performance liquid chromatograph for determination of isoflavones from soybean was verified to be accurate and reliable by methodology. The isoflavones of 85 soybean lines cultivated in Jilin Province were determined,and the results primarily showed the characters and ranges of isoflavones from soybean lines cultivated in Jilin Province,while the isoflavone content of soybeans ranged from 2.29 to 4.89 mg/g,and the average content was 3.36 mg/g. The isoflavone content of 5 soybean lines exceeded 4 mg/g,while there was a remarkably negative correlation between isoflavone content and protein content,and there was no significant positive correlation between isoflavone content and fat content. [Conclusion]The isoflavone content of soybean lines cultivated in Jilin Province is higher,so it is feasible for breeding the soybean lines with high isoflavone content and fat contetnt.

Simultaneous Determination of Four Pesticide Residues in Fruit Juice by HPLC

[Objective] This study was conducted to develop a system for simultaneous determination of imidacloprid, diflubenzuron, thiabendazole and carbendazim in fruit juice by HPLC. [Method] Using acetonitrile as the extraction solvent, the pesticides in fruit juice were purified through a NH2 solid phase extraction （SPE） cartridge, then detected by HPLC. [Result] There was a good linear relationship between the peak area and the concentrations of imidacloprid, diflubenzuron, thiabendazole and carbendazim in a range of 0.05-5.0 μg/ml, and the linear correlation coefficient varied in a range of 0.999 0-0.999 8; the limit of detection for imidacloprid, diflubenzuron, thiabendazole and carbendazim was 0.003, 0.005, 0.003 and 0.007 mg/kg, respectively. The recovery rate of imidacloprid, diflubenzuron, thiabendazole and carbendazim standards added at three levels （0.1, 0.5 and 1.0 mg/kg） ranged from 82% to 107%, with RSD less than 4.5%. [Conclusion] The sensitivity, accuracy and precision of this method were able to meet the requirements for pesticide residue analysis.

HPLC法测定TALICA缓释胶囊中3种主成分和葡甲胺的含量

目的基于高效液相色谱法(HPLC)建立TALICA缓释胶囊中阿莫西林、奥美拉唑镁、利福布汀3种主成分和葡甲胺辅料的含量.方法应用HPLC法,色谱柱为Agilent Zorbax SB-C18柱(150 mm×4.5 mm,5μm);流动相分别为乙腈(A)-磷酸盐缓冲液(B)和乙腈(A)-庚烷磺酸钠缓冲液(B),按时间程序梯度洗脱;VWD检测器,检测波长分别为254 nm和195 nm;进样量分别为10μL和20μL;流速均为1 mL·min^(-1);柱温分别为26℃和35℃;通过外标法计算阿莫西林、奥美拉唑镁、利福布汀3种主成分和葡甲胺辅料的含量.结果在波长254 nm下,阿莫西林、奥美拉唑镁、利福布汀分别在质量浓度0.02~2.12、0.02~2.16、0.02~2.16 mg·mL^(-1)内线性关系良好(r均大于0.9997),平均回收率分别为99.5%、98.9%、99.6%,RSD分别为0.5%、1.1%和0.6%,测得3种主成分的标示百分含量范围为98.8~101.1%.在波长195 nm下,葡甲胺在质量浓度1.00~50.00 mg·mL^(-1)内线性关系良好(r为0.9999),平均回收率及RSD为100.8%、0.9%,测得葡甲胺的平均含量为6.6%.结论HPLC法建立的TALICA缓释胶囊中阿莫西林、奥美拉唑镁、利福布汀3种主成分和葡甲胺辅料的含量测定分析方法稳定、简单、可靠,可用于TALICA缓释胶囊的质控研究.

引火汤冻干粉HPLC指纹图谱建立及8种成分含量测定

目的建立引火汤冻干粉HPLC指纹图谱,并测定地黄苷D、麦冬甲基黄烷酮A、毛蕊花糖苷、原儿茶酸、梓醇、五味子醇甲、五味子乙素、五味子甲素的含量。方法指纹图谱建立采用Supersil AQ18色谱柱(4.6 mm×250 mm,5μm);流动相乙腈-0.1%磷酸,梯度洗脱;体积流量1.0 mL/min;柱温30℃;检测波长230 nm。UPLC-MS/MS含量测定采用Alphasil VC-C_(18)色谱柱(2.1 mm×100 mm,2.5μm);流动相乙腈-0.1%甲酸,梯度洗脱;体积流量0.3 mL/min;柱温30℃;电喷雾离子源;正负离子扫描;多反应监测模式。结果10批冻干粉指纹图谱中有17个共有峰,相似度大于0.90,指认出毛蕊花糖苷、水晶兰苷、去乙酰车叶草苷酸、五味子醇甲。8种成分在各自范围内线性关系良好(R^(2)>0.9906),平均加样回收率98.7%~101.1%,RSD 2.0%~5.2%。结论指纹图谱结合含量测定能完整表征引火汤基准样品质量,从而为其关键化学属性质量评价提供参考。

HPLC法同时测定脑脉泰胶囊中12种成分的含量

目的建立HPLC法同时测定脑脉泰胶囊中3′-羟基葛根素、2-羟基红花黄色素A、葛根素、葛根素芹菜糖苷、3′-甲氧基葛根素、大豆苷、二苯乙烯苷、木犀草苷、染料木苷、木犀草素、丹酚酸B、迷迭香酸的含量。方法分析采用KromasilC_(18)色谱柱(250mm×4.6mm,5μm);流动相乙腈-水(含1.0g/L磷酸),梯度洗脱;体积流量0.8mL/min;柱温30℃;检测波长280nm。再进行聚类分析、主成分分析。结果12种成分在各自范围内线性关系良好(r>0.9990),平均加样回收率97.07%~98.30%,RSD0.86%~1.82%。10批样品聚为4类,4种主成分累积方差贡献率达86.262%。结论该方法简便稳定,可靠准确,可为脑脉泰胶囊的质量控制提供科学依据。