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.

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.

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.

Identification of Xunxi Shujin Decoction by TLC

[Objectives]To establish a TLC qualitative method for the determination of Radix Aconiti,Radix Aconiti Kusnezoffii,Caesalpinia sappan L.and Radix zanthoxyli.[Methods]TLC was used to study the quality of Radix Aconiti,Radix Aconiti Kusnezoffii and C.sappan L.in Xunxi Shujin Decoction,and the influencing factors were investigated to optimize the TLC conditions.[Results]In the TLC identification test of Radix Aconiti,Radix Aconiti Kusnezoffii,C.sappan L.and R.zanthoxyli in Xunxi Shujin Decoction,the spots were clear,the resolution was good,the specific value was moderate,the negative control was not disturbed,and the specificity was strong.[Conclusions]The established TLC method was simple,specific and reproducible,and could be used for the quality control of Xunxi Shujin Decoction.

TLC联合UPLC-QQQ-MS分析天王补心丸中4种外源性污染成分

目的对天王补心丸质量分析中发现的异常成分进行研究,建立薄层色谱(TLC)、超高效液相色谱—三重四极杆质谱(UPLC-QQQ-MS)同时测定天王补心丸中盐酸小檗碱、表小檗碱、盐酸黄连碱、盐酸巴马汀等4种外源性污染成分的分析方法。方法采用TLC法,通过优化展开系统,对158批样品进行定性鉴别;针对异常斑点,采用HPLC-Q/TOF-MS进行成分鉴定确认;采用UPLC-QQQ-MS进行定量分析。结果158批天王补心丸样品中有9批检出盐酸小檗碱、表小檗碱、盐酸黄连碱、盐酸巴马汀等4种外源性污染成分,分别在0.97~483.34、0.98~98.49、1.00~100.50、0.88~87.75ng·mL^(-1)内呈线性,回收率为99.30%~103.99%,平均标准偏差(RSD)为0.90%~2.62%(n=6),9批样品中上述4种成分的含量分别为20.3~104.1、1.0~17.6、0.4~30.6、3.2~24.8μg·g^(-1)。结论该方法简便、准确、专属性强,可为天王补心丸的质量控制提供参考。

TLC法鉴别复方氨酚烷胺片主成分

用薄层色谱法来鉴别复方氨酚烷胺片中的5种主成分,对原检验标准进行优化。以对乙酰氨基酚、马来酸氯苯那敏、咖啡因、盐酸金刚烷胺为对照品,采用硅胶GF_(254)薄层板,以三氯甲烷-甲醇-丙酮-氨水(9∶1.5∶1∶0.05)为展开剂,先在紫外光灯(254 nm)下检视,再喷以稀碘化铋钾显色剂;以胆酸、猪去氧胆酸为对照品,采用硅胶G薄层板,以乙酸乙酯-正己烷-甲醇-醋酸(25∶20∶3∶2)为展开剂,喷以10%硫酸乙醇溶液,105℃加热显色。改进的薄层色谱方法能有效分离4种主成分,显3个荧光斑点和两个色点;人工牛黄的薄层鉴别方法能定性胆酸和猪去氧胆酸。本方法结果准确,专属性强,薄层斑点清晰,分离度好,可替代原标准鉴别方法。

TLC-FID法测定延迟焦化原料油族组成

用三种不同极性展开剂在棒状薄层色谱棒上较好地将延迟焦化原料油分离为饱和烃、芳烃、胶质和沥青质四个族组分,建立了用棒状薄层色谱法-氢焰检测器(TLC-FID)快速测定延迟焦化原料油族组成的方法。通过重复性试验考察本方法的精密度,四组分相对标准偏差均小于3.00%,极差值均小于3.50%。

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）.

中兽药苦虎益母口服液TLC与HPLC检测方法的建立

为更好地控制苦虎益母口服液的品质和安全性,研究并制定苦虎益母口服液质量标准,采用薄层色谱法(TLC)对方中苦参、虎杖、益母草、黄芪进行定性鉴别,采用高效液相色谱法(HPLC)对虎杖苷进行定量检测。结果显示,苦参、虎杖、益母草、黄芪薄层鉴别斑点显色清晰,分离效果较好,阴性无干扰,能很好地鉴别目标成分。虎杖苷质量浓度在0.035~0.282 mg/mL范围内与峰面积有良好的线性关系,回归方程为y=29086x-173.16,相关系数r=0.9993,平均回收率为98.82%,RSD值为0.73%。苦虎益母口服液的质量控制中定性鉴别分离度好,含量测定结果重现性好。

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.

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.

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%.

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.

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 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.

动物食品中硫酸沙丁胺醇残留物的TLC-SERRS分析

目的:建立薄层色谱与表面增强共振拉曼光谱联用技术检测苯乙胺类拟肾上腺素硫酸沙丁胺醇药物残留的方法。方法:使用微波加热法用硝酸银和柠檬酸钠制备表面增强剂银溶胶;对所制备的银溶胶进行参数测定,选择表面增强效果较好的银溶胶;应用表面增强共振拉曼光谱法检测硫酸沙丁胺醇时,在TLC板分离出的目标斑点上滴加芳香族重氮盐和表面增强剂银溶胶后再检测其拉曼光谱,并对市售猪肝进行样品检测。结果:该方法可显著增强拉曼光谱的检测信号,增强因子可达到1.65×107。结论:本方法专属性强、耐用性好,为苯乙胺类拟肾上腺素药物残留物的鉴别技术提供了一种快速、高效的新方法。

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.