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.

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 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.

Determination of the Content of Kaempferol from Fermented Ginkgo( Ginkgo biloba L.) Leaves

[Objectives] This study was conducted to determine kaempferol content in ginkgo( Ginkgo biloba L.) leaves subjected to microbial fermentation.[Methods]Bacillus licheniformis was selected for solid-state fermentation of ginkgo leaves,and the content of kaempferol in ginkgo leaves was determined by RPHPLC method. At first,methanol was used to extract flavonoid glycosides,which were then hydrolyzed by hydrochloric acid solution. HPLC was performed with Platisil ODS column C18( 150 mm ×4. 6 mm,5 μm) using mobile phase Vmethanol∶ Vwater( 0. 4% phosphoric acid solution) = 55∶45 at a flow rate of 1 ml/min,and the eluate was detected with a shimadzu HPLC ultraviolet detector at 360 nm. [Results]With kaempferol as the reference substance,the correlation coefficient was0. 999 2 in the range of 0. 001 06-0. 016 96 g/L. The content in the fermented product was less than that in the non-fermented product by 28%. [Conclusions]The method is simple,accurate,and is suitable for determination of kaempferol. This study will provide an experimental basis for the development and utilization of ginkgo.

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.

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.

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 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.

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.

TLC Identification and Extraction Process of Rubiasin-1-methyl Ether from Yao Medicine Chuanlianzhu

[Objectives]To establish a thin-layer chromatography(TLC)method for the determination of rubiadin-1-methyl ether in Yao Medicine Chuanlianzhu(Damnacanthus giganteus).[Methods]A silica gel G thin-layer plate was adopted for TLC.Petroleum ether(60-90℃)-chloroform-methanol-water(7:15:3:1)was used as the developing solvent and inspected under ultraviolet lamp(365 nm).The content was determined by Inertsil ODS-3 C 18 column(4.60 mm×250 mm,5μm),mobile phase:acetonitrile-0.2%phosphoric acid gradient elution,detection wavelength 277 nm,flow rate 1.0 mL/min,column temperature 30℃,injection volume 10μL.[Results]The spots of 10 Chuanlianzhu samples from different origins showed the same color at the same position as the control,and the spots were clear and specific.The injection volume of rubiadin-1-methyl ether showed a good linear relationship in the range of 2.90-145μg(R=0.9996).The average recovery rate of rubiadin-1-methyl ether in the low,medium and high dose groups of Yao Medicine Chuanlianzhu was 98.72%,and RSD=1.78%.[Conclusions]This method can effectively identify Yao Medicine Chuanlianzhu medicinal materials and accurately determine the content of rubiadin-1-methyl ether in the medicinal materials.It provides a scientific basis for the development and utilization of Yao Medicine Chuanlianzhu medicinal resources.

Oil Content Conversion between Ether-based and Ethanol-based Methods and Establishment of Admissible Error for Imported Wool

Due to various reasons, the inspection methods often need to be changed, and the detection reagents often need to be replaced. In this study, a comparative experiment was conducted between the ethanol-based and ether-based determination methods for oil content in imported wool. The determination results obtained from the two methods were treated as abscissa and ordinate respectively,and their linear relationship was analyzed. According to the linear regression analysis, the conversion equation of determination result between the two methods was obtained. In addition, the repeatability admissible error and reproducibility admissible error were established through analyzing the comparative experimental results by scientific software. This study will bring new ideas for further researches in this field, and provide reference for solving the similar problems in actual inspection work.

A rapid and accurate direct measurement method of underground coal seam gas content based on dynamic diffusion theory

Coal seam gas content is frequently measured in quantity during underground coal mining operation and coalbed methane(CBM)exploration as a significant basic parameter.Due to the calculation error of lost gas and residual gas in the direct method,the efficiency and accuracy of the current methods are not inadequate to the large area multi-point measurement of coal seam gas content.This paper firstly deduces a simplified theoretical dynamic model for calculating lost gas based on gas dynamic diffusion theory.Secondly,the effects of various factors on gas dynamic diffusion from coal particle are experimentally studied.And sampling procedure of representative coal particle is improved.Thirdly,a new estimation method of residual gas content based on excess adsorption and competitive adsorption theory is proposed.The results showed that the maximum error of calculating the losing gas content by using the new simplified model is only 4%.Considering the influence of particle size on gas diffusion law,the particle size of the collected coal sample is below 0.25 mm,which improves the measurement speed and reflects the safety representativeness of the sample.The determination time of gas content reduced from 36 to 3 h/piece.Moreover,the absolute error is 0.15–0.50 m^3/t,and the relative error is within 5%.A new engineering method for determining the coal seam gas content is developed according to the above research.

Application of near infrared spectroscopy in monitoring the moisture content in freeze-drying process of human coagulation factor VIII

As an important process analysis tool,near infrared spectroscopy(NIRS)has been widely used in process monitoring.In the present work,the feasibility of NIRS for monitoring the moisture content of human coagulation factor VIII(FVIII)in freeze-drying process was investigated.A partial least squares regression(PLS-R)model for moisture content determination was built with 88 samples.Different pre-processing methods were explored,and the best method found was standard normal variate(SNV)transformation combined with 1st derivation with Savitzky–Golay(SG)15 point smoothing.Then,four different variable selection methods,including uninformative variable elimination(UVE),interval partial least squares regression(iPLS),competitive adaptive reweighted sampling(CARS)and manual method,were compared for eliminating irrelevant variables,and iPLS was chosen as the best variable selection method.The correlation coe±cient(R),correlation coe±cient of calibration set(Rcal),correlation coefficient of validation set(Rval),root mean square errors of cross-validation(RMSECV)and root mean square errors of prediction(RMSEP)of PLS model were 0.9284,0.9463,0.8890,0.4986% and 0.4514%,respectively.The results showed that the model for moisture content determination has a wide range,good linearity,accuracy and precision.The developed approach was demonstrated to be a potential for monitoring the moisture content of FVIII in freeze-drying process.

Numerical Experiment of Combined Infrared and Ultraviolet Radiation Remote Sensing to Determine the Profile and Total Content of Atmospheric Ozone

A new remote sensing method is described to determine the vertical distribution and total content of atmospheric ozone. The method combines surface infrared, satellite infrared and ultraviolet channels. The width of the infrared channels is 0.01 cm-1, less than Lorentz half-width at the earth's surface, rather than the present width, because these channels can obtain information about variations in the ozone profile below the profile main-peak. The numerical experiments show that the method has a satisfactory precision in determining total ozone content, just about I percent error, and vertical distribution from the earth to 65 km space. In addition, some semi-analysis functions lor calculating backscattered ultraviolet and a relaxation equation are described in this paper.

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.

Determination of Oleanolic Acid in Achyranthes aspera L. of Different Production Areas by HPLC

[Objectives] To determine the content of oleanolic acid in 10 batches of Achyranthes aspera L. from different production areas,and to establish a high performance liquid chromatography( HPLC) method for the determination of oleanolic acid in A. aspera. [Methods]Agilent C18 liquid chromatography column( 250 mm × 4. 6 mm,5 μm) was used for gradient elution with acetonitrile∶ water = 61∶ 39 as the mobile phase. The flow rate was 1 mL/min,the detection wavelength was 210 nm,and the column temperature was 35℃. [Results] The oleanolic acid injection volume showed a good linear relationship with the peak area in the range of 0. 382-7. 640 μg. The linear equation of oleanolic acid was: A = 530. 76 C,R = 1. 000 0; the range of sample recovery rate was 96. 03%-102. 73%,and the RSD value was 2. 16%( n =9). The content of the oleanolic acid was the highest in A. aspera produced in Guilin City of Guangxi( 0. 92%),and the lowest oleanolic acid content was in sample of Sitang in Nanning City of Guangxi( 0. 27%). It is recommended that the oleanolic acid content of A. aspera should not be lower than 0. 20%. [Conclusions]The HPLC method is accurate,reliable,easy to operate,and has good resolution. It is suitable for the determination of the content of oleic acid in A. aspera. It can provide reference for the quality control and standard drafting of A. aspera.

Establishment of a Method for the Determination of Emodin in Wudajiangjun Liquor

[Objectives]To establish a method for the determination of emodin in Wudajiangjun liquor,a hospital preparation.[Methods]A high-performance liquid chromatography(HPLC)method for the determination of emodin in Rhizoma Polygontum Cuspidatum in hospital preparation Wudajiangjun liquor was established.Using HPLC method,octadecylsilane chemically bonded silica gel was used as filler for the chromatographic column[Inertsil-C 18 chromatographic column(5μm,4.6 mm×250 mm)].Methanol-0.1%phosphoric acid water(76∶24)was used as mobile phase.The flow rate was 1.0 mL/min,the column temperature was 30℃,the detection wavelength was 254 nm,and the injection volume was 10μL.[Results]The content of emodin in Wudajiangjun liquor was determined by reversed-phase high performance liquid chromatography(RP-HPLC).When the injection volume of emodin was in the range of 5.45-54.5μg/mL,there was a good linear relationship between the injection volume and the peak area.The regression equation is Y=42.952-15.068(r=0.9998),the average recovery rate is 98.23%,and RSD=1.45%.[Conclusions]A method for the determination of emodin in Wudajiangjun liquor by HPLC was established.This method can be used as a quality control method for Wudajiangjun liquor with high accuracy and good repeatability,which lays a foundation for the quality control of the medicinal liquor.

Effect of Different Processing Methods on Content ofβ-Asarone in the Zhuang Medicine Rhizoma Acori Tatarinowii

[Objectives]This paper aims to establish a method to simultaneously determine the content ofβ-asarone in Rhizoma Acori Tatarinowii dried by three different methods.[Methods]Reversed-phase high-performance liquid chromatography was used,and the chromatographic conditions were as follows:column,Thermo SCIENTIFIC Hypersil GOLD Dim.(mm);mobile phase,methanol-0.1%phosphoric acid(63∶37);column temperature,30℃;flow rate,1mL/min;detection wavelength,257 nm;sample size,10μL.[Results]The linear range of the injection volume ofβ-asarone was 49.28-246.40μg/mL(R=0.9993);the limit of quantification was 0.85 ng and the detection limit was 0.34 ng;the RSD values of precision,stability and reproducibility tests were all less than 3%;and the sample recovery rate was 98.53%-98.97%(RSD<3.00).The results show that the content ofβ-asarone was highest in shade-dried Rhizoma Acori Tatarinowii.The order ofβ-asarone content was as follows:Rhizoma Acori Tatarinowii dried in shade>Rhizoma Acori Tatarinowii dried at 55℃>Rhizoma Acori Tatarinowii dried at 60℃.[Conclusions]This method is sensitive,reliable,and reproducible.It can be used to simultaneously determine the content ofβ-asarone in Rhizoma Acori Tatarinowii dried by three different methods.