Secondary metabolite credentials ofEvolvulus alsinoides by high performance thin layer chromatography(HPTLC)

Plants and plant-based products are the bases of many modern pharmaceuticals that are current in use today for various diseases.The aim of the study was to investigate the biochemical constituents and high performance thin layer chromatography（HPTLC） finger printing of the ethanolic extract of Evolvulus alsinoides.Phytochemical screening was done by standard procedures and HPTLC method was also established to analyze alkaloids,flavonoids and phenolic compounds from the ethanolic extract of Evolvulus alsinoides.Preliminary phytochemical screening showed that ethanol extracted more secondary metabolites than other solvents.HPTLC fingerprinting analysis showed the presence of various alkaloids,flavonoids and phenols（quercetin） in the ethanolic extract.It can be concluded that Evolvulus alsinoides may serve as a source of potent antioxidants that may be used in the prevention of various diseases such as cancer,diabetes and cardiovascular diseases due to the presence of phenolic compounds.HPTLC finger print of Evolvulus alsinoides may be useful in the differentiation of the species from adulterants and act as a biochemical marker for this medicinally important plant in the pharmaceutical industry and plant systematic studies.

Comparison of liquid-liquid extraction-thin layer chromatography with solid-phase extraction-high-performance thin layer chromatography in detection of urinary morphine

Liquid-liquid extraction-thin layer chromatography （LLE-TLC） has been a common and routine combined method for detection of drugs in biological materials. Solid-phase extraction （SPE） is gradually replacing the tra- ditional LLE method. High performance thin layer chromatography （HPTLC） has several advantages over TLC. The present work studied the higher efficiency of a new SPE-HPTLC method over that of a routine LLE-TLC method, in extraction and detection of urinary morphine. Fifty-eight urine samples, primarily identified as mor- phine-positive samples by a strip test, ＇were re-screened by LLE-TLC and SPE-HPTLC. The results of LLE-TLC and SPE-HPTLC were then compared with each other. The results showed that the SPE-HPTLC detected 74% of total samples as morphine-positive samples whereas the LLE-TLC detected 48% of the same samples. We further discussed the effect of codeine abuse on TLC analysis of urinary morphine. Regarding the importance of morphine detection in urine, the present combined SPE-HPTLC method is suggested as a replacement method for detection of urinary morphine by many reference laboratories.

Evaluation of quantitative variation of secondary metabolites in Bergenia ciliata(Haw.)using high performance thin layer chromatography

Dear Editor： Therapeutically active metabolite contents in a med- icinal plant vary in nature, which may impact on its therapeutic efficacy. Bergenia （Saxifragaceae） is an evergreen perennial herb widely distributed in Central and East Asia with about 30 species reported worldwide. It grows at a range of altitudes from the Khasia hills at 400 feet to the temperate Himalayas from Kashmir to Bhutan at 7,000-10,000 feet. Its distribution over a wide range of altitudinal zones makes it a good candidate for studying variations in its metabolic profiles under different climatic conditions. Bergenin （C-glycoside of 4-O-methyl gallic acid） has been identified as a potent active secondary metabolite in Bergenia and other therapeutically active constituents including, among others, gallic acid （3,4,5 trihydroxybenzoic acid）, （＋） catechin, and gallicin （Fig. 1）.

Study of Cuscuta chinensis Lam. in Nuhuang Fuzheng Oral Liquid by Thin Layer Chromatography

[Objectives] To better control the quality of Nuhuang Fuzheng Oral Liquid and study the main component Cuscuta chinensis Lam.by Thin Layer Chromatography. [Methods] Through changing the treatment methods of the test sample solution,proportion of the developing solvent and sample application volume,taking the spot resolution,definition,and Rf value,optimal Thin Layer Chromatography conditions were screened for Cuscuta chinensis Lam. [Results] After the test sample solution passing the neutral alumina column,it was extracted two times using the ethyl acetate. Methanol was added to dissolve. Benzene-ethyl acetate-formic acid( 5∶5∶2.5) was used as developing solvent.And ammonia fumigation was carried out to develop color. In the thin layer chromatograph obtained through these conditions,Nuhuang Fuzheng Oral Liquid test sample solution showed the same stripe in the same position of the control drug chromatogram and there was no obvious tailing phenomenon and the spot was clear. [Conclusions] The thin layer chromatography identification conditions can be used as the method for quality control of Cuscuta chinensis Lam. in Nuhuang Fuzheng Oral Liquid.

Application of Thin Layer Chromatography in Preparation of Drug Containing Serum of Eucalyptus Oil

[Objectives] To explore whether the thin layer chromatography( TLC) can be used to guide the preparation of drug containing serum of eucalyptus oil. [Methods]Eucalyptus oil samples with different dilution ratio were detected by TLC. Eucalyptus oil was intragastrically administered to mice,serum samples of different eucalyptus oil doses,different intragastric administration methods and different blood sampling times were collected. The above samples were detected by TLC,and the above results were verified by gas chromatography. [Results] TLC can detect concentration difference of eucalyptus oil samples with different dilution ratio. TLC can provide qualitative and semi-quantitative detection of eucalyptus oil in serum. High,medium and low dose of eucalyptus oil in serum had different effects on the growth of MCF-7 cells in the SD rats( P < 0. 05). [Conclusions] TLC can be used to guide the preparation of drug containing serum of eucalyptus oil.

DETERMINING THE CONTENT OF JASMINODIN IN MEI RONG CAPSULE WITH THIN LAYER CHROMATOGRAPHY

Objective Jasminodin in Mei Rong capsule was determined by the method or thin layer chromatog- raphy. Mothods The sample Jasminodin was extracted with methanol and then the extract was chromatographed on silica gel GF254 plate with chloroform-methanol-water(10:3:1) as mobile phase and the chromatograms were scanned by using Shimadzu CS-930 dual wavelength TLCS in zegzag scanning mode at λ_s 370nm and λR 250nm. Results Callbration graph was rectilinear between 2.7μg^16.2μg per spot for the jasminodin. Conclusion Satisfa- tory results were obtained with the average recovery ratio of 99.76%,RSD 2.27%.

Identification of Ethnic Medicine Polygonum capitatum by Thin Layer Chromatography

[Objectives]To establish a thin-layer chromatography identification method for the ethnic medicinal materials of Polygonum capitatum.[Methods]The thin layer chromatography(TLC)method was used to identify materials.[Results]The TLC identification showed that in the chromatogram of the test sample,there were spots of the same color in the corresponding position of the chromatogram of the reference medicinal material,and the reproducibility was good.[Conclusions]The TLC identification and the determination results of the inspection items can provide a basis for the quality control of the medicinal materials of P.capitatum.

Preparation Process and Thin Layer Chromatography Identification of Wufang Babu Poultice

[Objectives] To establish the process flow of preparation of Wufang Babu Poultice and the identification method of thin layer chromatography (TLC). [Methods] For the forming process of Wufang Babu Poultice, the preparation method of mixed pharmaceutical powder with suitable pharmaceutical excipients was adopted. Qualitative identification of medicinal materials in Wufang Babu Poultice (Strychni Semen, Rhei Radix Et Rhizoma, Dipsaci Radix, and Angelicae Sinensis Radix) was carried out by TLC. [Results] Mixed pharmaceutical powder mixed with glycerol, gelatin and other pharmaceutical excipients can be prepared for forming. The test solution chromatography of each medicinal material (Strychni Semen, Rhei Radix et Rhizoma, Dipsaci Radix, Angelicae Sinensis Radix) showed pigment spots of the same color at the same position as its corresponding control medicinal materials and reference chromatography, and the display was clear. [Conclusions] The preparation process is simple and feasible, and can be used as the forming process of Wufang Babu Poultice. The TLC determination method is simple to operate, has good specificity, and has no effect on negative results, and can be used for identification of Wufang Babu Poultice.

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.

Algerian mint species:high performance thin layer chromatography quantitative determination of rosmarinic acid and in vitro inhibitory effects on linoleic acid peroxidation

Objective:To determine the quantitative paremeters of rosmarinic acid in Algerian mints,Mentha spicata L.(M.spicata),Mentha pulegium L.and Mentha rotundifolia(L.)Huds by high performance thin layer chromatography(HPTLC)-densitometric method and screen the effects of these plant extracts on linoleic acid peroxidation.Methods:The analyses were performed on HPTLC silica gel 60 F254 plates with chloroform:acetone:formic acid(75:16.5:8.5,v/v)as the mobile phase.Rosmarinic acid was determined in UV at 365 nm and fluorescence at λ_(exc)325 nm with a 550 nm filter,respectively.The effects of plants extracts on linoleic acid peroxidation were measured by an indirect in vitro colorimetric method.Results:Chromatographic resolution permitted reliable quantification in both measurement modes and calibration curves were linear in a range of 150-1000 ng/spot.M.spicata was found to contain significantly higher concentrations of rosmarinic acid.The densitometric quantification allowed the analysis of many samples in a short time with reasonable precision(total precision for Mentha spp extracts,5.1%and 5.8%for UV and fluorescence detection,respectively).The HPTLC data,allied to assays of linoleic acid peroxidation prevention,suggested the potential of M.spicata(52%Trolox®equivalents)as a natural source for inhibitors of lipid peroxidation.Conclusions:Densitometry can be used for routine determination and quality control of rosmarinic acid in herbal and formulations containing Mentha species.

Separation and Detection of Ceramides by HPLC Followed by Evaporative Light-Scattering Detection and Thin Layer Chromatography

Ceramides are important signaling molecules involved in a variety of cellular processes, including cell growth, differentiation, and apoptosis. Currently, different methods are used for ceramide analysis, some of which are insensitive or cumbersome. This paper described methods utilizing thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) followed by evaporative light scattering detection (ELSD) to detect ceramide directly in cell extracts without derivatization, which was proved to be efficient and reproducible. Five kinds of ceramides were used as standards. Both TLC and normal-phase HPLC/ELSD results indicate that yeast contains several kinds of ceramides.

Thin Layer Chromatography and Methodological Investigation of Matrine in Zhukuqin Granule

[Objectives] To establish a sensitive and simple thin layer chromatography method of matrine in Zhukuqin granule. [Methods]With Zhukuqin granule as control medicinal materials,matrine as reference substance,we chose orthogonal test method,systematically investigated various factors affecting the thin layer chromatography,and selected the best thin layer chromatography conditions. [Results] We screened out the best thin layer chromatography of matrine in the Zhukuqin granule with chloroform as extraction solvent,solvent extraction and ultrasonic extraction as extraction method,toluene-acetone-ethyl acetate-ammonia solution( 2:3:4:0. 5) as developing agent,improvement of bismuth potassium iodide solution as color developing agent. [Conclusions] The separating degree of this method was high,it had good repeatability,and it could be used to control quality of Zhukuqin granule.

Thin Layer Chromatography Screening for Glycyrrhiza uralensis

[Objectives] To establish a simple and rapid,specific and reproducible thin layer chromatography( TLC) method,and accordingly establish a thin layer identification method for quality control of Glycyrrhiza uralensis. [Methods] According to the physical and chemical properties,components of Glycyrrhiza uralensis were identified by thin layer chromatography. According to Veterinary Pharmacopoeia of the People's Republic of China 2010( Volume Ⅱ) and related literature report,the thin layer chromatography identification method was studied and improved for Glycyrrhiza uralensis.[Results] When the preparation method for test sample solution was 6g Glycyrrhiza uralensis added with 30 mL saturated n-butanol,ultrasonic processing 20 min,filtered,filtrate dried,added 3 mL methanol and obtained Glycyrrhiza uralensis test sample solution. Then,chloroform-methanol-formic acid-ethyl acetate( 10∶ 3∶ 2∶ 3) as developing solvent,clear spot with high degree of separation and reproducibility could be obtained,and can be used for thin layer chromatography identification of ammonium glycyrrhetate in Glycyrrhiza uralensis.[Conclusions] The modified method omitted the step of ether treatment of samples,and directly adopted saturated n-butanol to extract components. It does not influence the detection of chromatographic spots,and solves the issue of sample gelatinizing sticking and filtration,simplifies the test process,and reduces the harm to human body.

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.

Optimizing the nickel boride layer thickness in a spectroelectrochemical ATR-FTIR thin-film flow cell applied in glycerol oxidation

The influence of the drop-casted nickel boride catalyst loading on glassy carbon electrodes was investigated in a spectroelectrochemical ATR-FTIR thin-film flow cell applied in alkaline glycerol electrooxidation.The continuously operated radial flow cell consisted of a borehole electrode positioned 50μm above an internal reflection element enabling operando FTIR spectroscopy.It is identified as a suitable tool for facile and reproducible screening of electrocatalysts under well-defined conditions,additionally providing access to the selectivities in complex reaction networks such as glycerol oxidation.The fast product identification by ATR-IR spectroscopy was validated by the more time-consuming quantitative HPLC analysis of the pumped electrolyte.High degrees of glycerol conversion were achieved under the applied laminar flow conditions using 0.1 M glycerol and 1 M KOH in water and a flow rate of 5μL min^(–1).Conversion and selectivity were found to depend on the catalyst loading,which determined the catalyst layer thickness and roughness.The highest loading of 210μg cm^(–2)resulted in 73%conversion and a higher formate selectivity of almost 80%,which is ascribed to longer residence times in rougher films favoring readsorption and C–C bond scission.The lowest loading of 13μg cm^(–2)was sufficient to reach 63%conversion,a lower formate selectivity of 60%,and,correspondingly,higher selectivities of C_(2)species such as glycolate amounting to 8%.Thus,only low catalyst loadings resulting in very thin films in the fewμm thickness range are suitable for reliable catalyst screening.

Thin Layer Chromatographic Study of Bauxite and Quantitative Estimation of Co-Existing Al^(3+), Fe^(2+) and Ti^(4+)

Thin layer chromatography in combination with spectrophotometry and titrimetry has been used to evaluate chromatographic characteristics of bauxite constituents. The retention behaviors of four major constituents (Al 3+ , Fe 2+ , Ti 4+ , Si 4+ ) in bauxite mineral have been examined on plain and modified layers of silica gel G, silica gel H and cellulose with mobile phases containing aqueous sodium chloride, formic acid and hydrochloric acid. Ternary separation of Al-Fe-Ti was achieved on chromatographic plates made of silica gel H. The pH effect and presence of impurity elements in samples, nature of stationary phases on the ternary separation and detection limits of bauxite constituents were studied. Silicon in bauxite was detected on cellulose plates. Quantitative determinations of Al 3+ , Fe 2+ and Ti 4+ on silica gel H impregnated with sodium formate layers were achieved by titrimetry and spectrophotometry.

Novelties Filtration Theory of Liquid Chromatography-Mass Spectrometry in Volume Nanotube of Cotton Filament of Layers Woven Fabrics

Objectives of the research to present a modern theory of water purification for multiple purposes entitled “a novelties filtration theory of liquid chromatography-mass spectrometry” is an exceedingly sensitive and specific analytical technique in volume layers woven fabrics that can precisely determine the identities and quantities of compounds within volume Nanotube of cotton filament of layers woven fabrics. The problems are that the filters in the local and international markets have increased complications in configuration, installation and cost without reaching the efficiency that humanity hopes. Throw materials and methods the chromatography-mass spectrometry in layers woven fabrics, and throw the nanotube of cotton filament for purification of water dyes and smells. Industry, in which mass spectrometry is a convenient, versatile method for characterization and identification of process throw the Nanotube of cotton filament for purification of water dyes and smells. Results came up with a theme “innovations in textiles”, and also, for characterization of fibers and contaminants of the fabrics. Additive manufacturing in layers woven fabrics, are the processes used to synthesize a volume object under computer control with successive material layers that have been used and highlighted. The conclusions has included chromatography-mass spectrometry drop, physico-chemical, biological, combined physical-biological and chemical-biological treatment processes recently being developed to meet Jet-filtration, the strict discharging limits set by ASTM standards. Some important aspects of both qualitative and quantitative data analysis have been described and the power of using mass profiles to enhance selectivity and sensitivity has been demonstrated.

基于Box-Behnken设计-响应面法优化调肝活血稳压颗粒提取工艺及其质量标准的研究

目的优化调肝活血稳压颗粒的提取工艺,并对其质量标准进行研究。方法本研究以浸膏得率及丹酚酸B的转移率的综合评分为指标,采用单因素考察结合Box-Behnken设计-响应面法优化调肝活血稳压颗粒的提取工艺,采用薄层色谱法对方中药材进行鉴别。结果确定最终提取工艺为:第一次加入10倍量水煎煮90 min,第二次加入6倍量水煎煮60 min。测定方法结果准确可靠。薄层色谱法检测出天麻、栀子、丹参等三味中药的特征性斑点。结论优化后的提取工艺操作简单,提取效果良好,结果稳定可靠。

基于评价抽检的市售钩藤质量评价

目的全面评价市售钩藤的质量。方法对29批钩藤进行法定标准(性状、鉴别、检查、浸出物)检验;使用薄层色谱(TLC)法对钩藤对照药材、毛钩藤碱、去氢毛钩藤碱、钩藤碱、异钩藤碱进行鉴别;使用高效液相色谱(HPLC)法建立钩藤特征图谱,采用聚类分析、主成分分析(PCA)-X及正交偏最小二乘法判别分析(OPLS-DA)法对共有峰峰面积进行统计分析,筛选出钩藤的关键质量控制成分,同时测定钩藤中喜果苷、异去氢钩藤碱、去氢钩藤碱、异钩藤碱、钩藤碱、去氢毛钩藤碱、毛钩藤碱的含量,整体评价钩藤质量。结果29批样品中,合格样品23批,合格率79.31%;TLC法鉴别发现:不同基源钩藤化学成分差异较大,大叶钩藤未检出毛钩藤碱,华钩藤未检出去氢毛钩藤碱,毛钩藤未检出4种成分,仅基源为钩藤的样品在与对照药材和对照品色谱相应的位置上显相应的斑点;HPLC法测定结果表明:不同基源钩藤的化学成分及含量差异较大,大叶钩藤未检出去氢钩藤碱和毛钩藤碱,华钩藤未检出异去氢钩藤碱、去氢钩藤碱和去氢毛钩藤碱,毛钩藤仅检出喜果苷,且喜果苷含量较低。基于统计分析,不同批次钩藤间存在差异,并发现可能导致钩藤质量差异的3个已知关键质控成分:异去氢钩藤碱、去氢钩藤碱、异钩藤碱。结论钩藤整体质量不理想,现行炮制规范和检验标准无法确保产品质量,影响临床用药的安全性和有效性,不同基源钩藤的化学成分类别及含量差异较大,增加了制定统一质量标准的难度。