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.

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.

Quality Evaluation of Leontopodium franchetii Beauv. from Different Localities Based on Moisture,Ash and Extract Contents and Thin-layer Chromatography

[Objectives] The aim was to determine the moisture,ash and extract contents in Leontopodium franchetii Beauv. from different localities. [Methods] The contents of moisture,total ash,acid-insoluble ash and extract in L. franchetii from different localities were determined according to the methods described in Chinese Pharmacopoeia( 2015 Edition). [Results] In the 10 samples,the contents of moisture were all less than or equal to 15. 0%; the contents of total ash were all below 12. 0%; and the contents of acid-insoluble ash were all less than 3. 0%. The contents of water-soluble extract( by cold-soaked extraction method) in the samples were all below 12. 0% except those from Hongyuan Prairie of Sichuan Province and Xinglong Mountain in Lanzhou City,Gansu Province. [Conclusions]The contents of the four indicators measured in this experiment varied by small margins,indicating that the quality was relatively stable. This study provides theoretical data for the revision of the quality standards for L. franchetii.

Separation of Amino Acids Based on Thin-Layer Chromatography by a Novel Quinazoline Based Anti-Microbial Agent

A newly designed quinazoline based compound, 6-pyridin-2-yl-5,6-dihydro-benzo[4,5]imidazo[1,2-c] quinazoline (PDBIQ) has shown the ability for the easy detection of nineteen amino acids on thin-layer chromatography plates as a spray reagent. This new reagent enabled to produce various distinguishable colors with amino acids with different RF values. The detection limits and the binding ability of PDBIQ with amino acids have been calculated. PDBIQ is also able to detect aminoacids from hydrolised seed protein. The title compound also exhibited profound inhibitory action against some gm (+ve) and gm (-ve) bacterial organisms. This paper deals with synthesis, spectroscopic application and biological evaluation of the organic moity.

ANALYSIS OF HUMAN PLACENTA BY ^(31)P NUCLEAR MAGNETIC RESONANCE AND THIN-LAYER CHROMATOGRAPHY SCANNING COMBINED WITH THE CORRECTIVE METHOD OF ABSORBANCE PROPORTIONAL COEFFICIENT

Five phospholipids in human placenta were determined by phosphorus 31 nuclear magnetic resonance(^(31)P NMR)spectroscopy and thin-layer chromatography(TLC) scanning combined with the corrective method of absorbance proportional coefficient. The NMR spectrometer used this investigation was a Bruker AM-500 spectrometer operating at 202.4 MHz for ^(31)P chemical shifts are relative to 85% phosphoric acid. TIC was carried out by silica gel H plate developed in chloroform-methanol-glacial acetic acid-ethanol-water(25:4:6:2:0.5),with Vaskovsky reagent as colour -developing agent of phospholipids.

Identification of Citric Acid in Wuhuang Oral Liquid by Thin Layer Chromatography

[Objectives] To screen the identification conditions of citric acid in Wuhuang Oral Liquid by thin layer chromatography,and establish the quality control method for citric acid in Wuhuang Oral Liquid.[Methods] Different treatment methods were adopted for test sample,developing agent,and drying time,thin layer chromatography separation condition and spot definition were taken as indicators to conduct experiment,to select optimal thin layer identification method. [Results] Methanol was used as the extraction solvent,ultrasonic treatment,ether extraction,dissolution by anhydrous ethanol as treatment conditions of test sample; upper solution of butyl acetate-formic acid-water(4 ∶ 2 ∶ 2) after placing one hour was taken as developing agent; 0. 1% bromocresol green(BCG) as the developer; when developing the color in 3 hours after development,in thin layer chromatograph,there appeared the same strip in the same position of test sample of Wuhuang ORAL Liquid and control substance,no obvious trailing phenomenon,and the color was uniform and clear.[Conclusions]The thin layer chromatography identification conditions can be used as the method for quality control of Wuhuang Oral Liquid.

SYSTEM IDENTIFICATION METHOD OF LINEAR AND NON-LINEAR LIQUID PREPARATIVECHROMATOGRAPHIC SEPARATION

The paper proposes the identification method of linear and non-linear chromatographic system. Isotherms and lumped mass transfer coefficients of chromatography separating isomer sorbitol and mannitol on D80 adsorbent are determined. The analyses of root mean square error of chromatography elution curves are carried out. The results show that the system studied is non-linear and the theoretical elution curves of the non-linear chromatographic model are more accurate than those of the linear chromatographic model. Results of parameter sensitivity analyses show that elution curves of non-linear chromatographic separation are more sensitive to the variation of parameter "ai" in the adsorption isotherms than to the variation of parameter "bi" as well as lumped mass transfer coefficients.

Pharmacognosy Identification of Xiaohuangcao (Dendrobium loddigesi Rolfe)

[Objectives]Pharmacognosy identification was performed on Xiaohuangcao ( Dendrobium loddigesi Rolfe).[Methods]The medicinal materials were identified through original plants,characters,microscopic structure and thin-layer identification characteristics.[Results]D.loddigesi has obvious plant morphology,characters,microscopic structure and thin-layer identification characteristics.The stem of Xiaohuangcao is slender and cylindrical,and the surface is golden yellow;and the fiber bundles outside the vascular bundles are crescent-shaped or semi-circular in the stem transection.For the powder,crystal fiber can be observed;the vascular bundles are embedded with siliceous block cells;and there are more starch grains.In the thin layer chromatography,petroleum ether-ethyl acetate-butanone-glacial acetic acid (8.5∶ 3.5∶ 1.5∶ 5 d) was used as a developing solvent,and 10% sulfuric acid ethanol solution was used as a color developing agent.[Conclusions]The research results provide reference for the application of the medicinal material and the formulation of its related quality standards.

Isolation and Identification of Flavone Type of Two Compounds from Salix matsudana

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Rapid Laboratory Comparison Method for Identification of Hazardous Wastes from Illegal Oil Refining

[Objective] This paper aimed to study the rapid identification method of hazardous wastes from illegal oil refining,and provide a new theoretical basis and practical basis for shortening the appraisal cycle and reducing the cost of identification.[Method] The identified substance and the qualified oil products were diluted by carbon disulfide with a ratio of 1∶100 for the gas chromatographic analysis.According to the corresponding national standard,the oil change indexes,such as the moisture content,viscosity,acid value and mechanical impurities of the identified substance,were measured.[Result] By the comparison between the gas chromatogram spectrograms of the identified substance and the qualified oil,it is found that the main component of the identified substance was the mineral oils.And the test results of oil change index showed that the mineral oil was unqualified.[Conclusion] This method was of great significance to save the cost of appraisal and improve the efficiency of case handling.

Qualitative Identification and Content Determination of Psoralen in Ficus Pandurata Hance

[Objectives]This study aimed to establish a qualitative identification and content determination method for psoralen in Ficus pandurata Hance and compare the psoralen contents in roots,stems and leaves of F.pandurate Hance and Ficus pandurata Hance var.holophylla Migo.[Methods]Thin-layer chromatography(TLC)was used for qualitative identification,and the content of psoralen was determined by high-performance liquid chromatography.The chromatographic conditions were as follows:column,Agilent ZORBAX Eclipse Plus C18(250 mm×4.6 mm,5μm);mobile phase,methanol-water(55∶45);flow rate,1 mL/min;detection wavelength,246 nm;column temperature,30℃;and injection volume,10μL.[Results]The TLC chromatogram of F.pandurate Hance showed clear spots and good separation.The concentration of psoralen detected in the range of 2.06-41.20μg/mL had a good linear relationship with the peak area(r=0.9999).The RSD values of the precision,stability and reproducibility tests were all less than 2%.The average recovery rate was 100.2%(RSD=1.13%,n=6).[Conclusions]The established method is simple,easy,accurate and reproducible.It can be used for quality control of F.pandurate Hance.

Metabonomic analysis of hepatitis B virus-induced liver failure:identification of potential diagnostic biomarkers by fuzzy support vector machine

Hepatitis B virus(HBV)-induced liver failure is an emergent liver disease leading to high mortality.The severity of liver failure may be reflected by the profile of some metabolites.This study assessed the potential of using metabolites as bio- markers for liver failure by identifying metabolites with good discriminative performance for its phenotype.The serum samples from 24 HBV-induced liver failure patients and 23 healthy volunteers were collected and analyzed by gas chromatography-mass spectrometry(GC-MS)to generate metabolite profiles.The 24 patients were further grouped into two classes according to the severity of liver failure.Twenty-five commensal peaks in all metabolite profiles were extracted,and the relative area values of these peaks were used as features for each sample.Three algorithms,F-test,k-nearest neighbor(KNN)and fuzzy support vector machine(FSVM)combined with exhaustive search(ES),were employed to identify a subset of metabolites(biomarkers)that best predict liver failure.Based on the achieved experimental dataset,93.62%predictive accuracy by 6 features was selected with FSVM-ES and three key metabolites,glyceric acid,cis-aconitic acid and citric acid,are identified as potential diagnostic bio- markers.

Systematically structural identification of nitric compounds in crude oil with chemometric resolution

Aimed at the problem of classification of non-hydrocarbons of crude oil, the theoretical standpoint that the polarity of a compound depends on the whole structure and composition of molecule instead of a kind of heteroatom or its functional group was presented. A method was established for the systematically structural identification of nitric compounds in crude oil. The pre-fractionation of a crude oil sample into 7 fractions was performed by di- adsorption column chromatography with neutral aluminum oxide and silica gel. Subsequently, the individual components were obtained by using capillary column gas chromatography, and the types of compounds were detected by a mass spectrometer. In combination with a chemometric resolution, the compounds of fraction were further identified. This method can relieve the difficulty of classical analysis in identifying those species with very low contents or without being completely separated. The structures of 168 nitric compounds in a crude oil sample were determined by this method.

改进和优化黑果枸杞及其制品中花青素测定的pH示差法

建立一种基于美国官方分析化学师协会(Association of Official Analytical Chemists,AOAC)方法检测黑果枸杞及其制品中花青素含量的改进pH示差法。考察了黑果枸杞及其制品中花青素的最佳提取和检测条件,通过液相色谱-三重四级杆串联质谱法鉴别出黑果枸杞中花青素的具体化学结构,并计算出混合花青素的平均摩尔质量。通过分光光度法测得混合花青素的平均摩尔消光系数,对改进后的pH示差法进行方法学验证和花青素的含量测定。结果显示,最佳提取和检测条件如下:黑果枸杞花青素提取溶剂为盐酸-80%(体积分数)乙醇(3∶97,体积比),料液比为1∶100(g∶mL),提取温度为50℃,提取时间为30 min,缓冲溶液稀释5倍后静置平衡20 min。液相色谱-三重四级杆串联质谱法鉴别黑果枸杞中主要以矮牵牛素类花青素为主(占97.96%),黑果枸杞特有的混合花青素平均摩尔质量为912.7 g/mol,平均摩尔消光系数为29591 L/(mol·cm)。pH示差法改进后能够满足方法学验证要求,固体样品和液体样品最低检出限分别为28.2 mg/100 g、0.282 mg/100 mL。方法改进后花青素提取增长率均大于20%,静置平衡20 min后单次检测结果精密度小于0.3%。以矮牵牛素类花青素代替矢车菊素-3-O-葡萄糖苷计算花青素含量平均提高了2.41倍,能真实地反映黑果枸杞及其制品中花青素的含量。

白肋烟中糖酯类化合物的分离与鉴定

烟草中糖酯类化合物是重要的致香前体物,对烟叶的品质具有重要影响。但目前还未见关于白肋烟烟叶中糖酯类化合物的分离分析研究。本实验利用高分辨液相色谱-串联质谱(LC-MSn)技术,从白肋烟叶提取物中共分离鉴定出4类蔗糖酯和2类葡萄糖酯,每类均包含多种同系物。其中,4种蔗糖酯为首次发现,8种葡萄糖酯的结构鉴定在栽培烟草中为首次报道。本研究丰富了烟草中糖酯类化合物的种类,对烟草中不同类型蔗糖酯和葡萄糖酯的分析鉴定和代谢研究具有参考意义。

基于液相色谱-串联质谱肽组学技术鉴定地龙制剂中的多肽及含量检测

建立了液相色谱-串联质谱(LC-MS/MS)肽组学鉴定多肽地龙蛋白压片糖果中多肽及高效液相色谱(HPLC)检测多肽含量新方法。本文联用LC-MS/MS与De novo分离初步鉴定多肽地龙蛋白压片糖果中多肽成分,采用化学合成法制备多肽,进一步利用HPLC验证并测定其多肽含量。总共鉴定出482个多肽序列,对相对强度和评分较高的10条多肽进行化学合成,成功在多肽地龙蛋白压片糖果实际样品中验证到氨基酸序列为LAPVNKPGRF(LF-10)的多肽,实验测得LF-10的含量为0.00182%。LC-MS/MS与De novo的联用为地龙多肽类成分的鉴定分析提供有力的支持,建立的LF-10多肽HPLC定量分析方法简便易行、准确可靠,有助于多肽地龙蛋白压片糖果的质量控制及地龙多肽的进一步研究。

制备高效液相色谱用于纽莫康定杂质B1和B5的纯化

目的开展低含量成分的富集和纯化研究,从Glarea lozoyensis菌体的发酵产物(即纽莫康定B0粗品)中制备出杂质B1和B5,并对其进行结构鉴定。方法建立了基于制备高效液相色谱(prep-HPLC)技术的两步纯化法,第一步使用亲水(HILIC)模式固定相对目标杂质进行富集,第二步采用反相(RPLC)模式固定相对杂质进行制备。得到的两个杂质采用质谱(MS)、核磁(NMR)进行鉴定,确定化合物结构。结果两个杂质为B1和B5,相对分子质量均为1049 Da,色谱纯度分别为97.83%和98.13%。经核磁鉴定,B1为B0的高酪氨酸类似物,B5为B0的鸟氨酸类似物。结论本研究发展的基于prep-HPLC的两步纯化方法为低含量成分制备提供参考,第一步分离实现目标成分富集,第二步利用正交性的色谱柱提高分离选择性,成功制备出杂质B1与B5,有助于B0杂质谱的建立和进一步的质量控制研究。

广西特色药材横经席鉴别方法的研究

目的:建立横经席专属性鉴别方法。方法:采用性状、显微及薄层色谱相结合的方法对横经席及其同属品种绣毛红厚壳、红厚壳进行对比研究,找到横经席的鉴别特征。结果:横经席性状鉴别特征为叶呈长圆状披针形,两面无毛;锈毛红厚壳叶呈矩圆形至椭圆形,两面主脉及小枝均密被锈红色柔毛;红厚壳叶呈宽椭圆形或倒卵状椭圆形,两面无毛。横经席粉末显微特征可观察到叶表皮细胞、纤维等,无非腺毛;锈毛红厚壳粉末可见较多非腺毛;红厚壳叶下表皮细胞垂周壁波浪状弯曲并呈连珠状增厚。薄层色谱鉴别结果显示,横经席比同属其他品种多1个特征斑点。结论:研究建立的横经席鉴别方法具有专属性,可区别横经席正品及其混伪品。

米糠油中呈色物质的分离和鉴定

米糠油的色泽是影响其品质的因素之一。该研究对米糠油中呈色物质的提取方法进行优化,并利用高效液相色谱串联质谱法对呈色物质进行分离和鉴定。以甲醇∶水(8∶2,体积比)为萃取剂,用液液萃取法提取米糠油中的强极性呈色物质,然后将样品皂化后利用正己烷提取弱极性呈色物质。强极性和弱极性提取液经质谱分别鉴定出9类物质,共37种,其中强极性提取液中二氢异黄酮类含量最多,弱极性提取液中二氢黄酮醇类为主要化合物。

白薇及其伪品的理化鉴定

目的:鉴定白薇与其常见伪品(大理白前、合掌消、徐长卿、老瓜头和潮风草),为白薇的理化鉴定提供理论依据,确保用药安全。方法:通过化学方法和薄层色谱法,根据呈色反应和荧光斑点的颜色、位置,鉴定白薇及其伪品。结果:蔓生白薇和直立白薇加入试剂后即显深棕红色和紫红色,随着溶剂挥干,颜色变深,最后变成深棕色和墨绿色,其他伪品与之大有不同;不同样品薄层色谱的斑点位置及颜色有明显的差异,白薇主要在Rf=0.56和Rf=0.75有紫红色的荧光斑点,而混淆品斑点大都在起始位置有明显的荧光斑点。结论:理化鉴定法可用于鉴定白薇的真伪,确保白薇质量,并且为白薇药用的真实性、有效性、安全性提供了科学理论依据。