Determination of Lucidin in Rubia tinctorum Aglycones by an HPLC Method with Isocratic Elution

An HPLC method has been developed for the separation of anthraquinones with particular attention to the determination of lucidin in commercially available sources of Rubia tinctorum aglycones. Variations of chromatographic conditions have been studied in order to provide suitable resolution of anthraquinones under isocratic conditions on an end‐capped RP C18‐column.     

High-performance liquid chromatographic determination of some anthraquinone and naphthoquinone dyes occurring in historical textiles

A reversed-phase HPLC method has been developed for identification and quantitation of nine natural quinone dyes and applied to historical textile fibres. A Purospher RP18e column was used with a convex gradient of methanol in a mobile phase of 0.1 M aqueous citrate buffer (pH 2.5) and spectrophotometric diode-array detection at 270 nm. For identification of alizarin, purpurin and xanthopurpurin, occurring together in the madder plant, an isocratic method was used with a methanol-0.2 M acetate buffer (pH 4.3) (75:25) as the mobile phase. After an acid extraction of textile fibres and the analysis of the extracts, alizarin and purpurin were identified and quantitated in three fibres.     

HPLC Analysis of Alizarin and Purpurin Produced by Rubia tinctorum L. Hairy Root Cultures

We have determined the quantities of the anthraquinones alizarin and purpurin, with especial regard to their effective antigenotoxic activity, in genetically transformed hairy root cultures of Rubia tinctorum L. Hairy roots were cultured on solid and in liquid Gamborg B5 and 1/2 NMS media in a shaking cabinet and in a bioreactor. Methanolic extracts of lyophilized hairy roots were hydrolysed, and then purified by solid phase extraction with good recovery. A new HPLC method was developed for the determination of alizarin and purpurin. The analysis was performed on a Luna C8 RP column using a 45:55 (v/v) mixture of acetonitrile:20 mM ammonium formate-formic acid buffer (pH 3.00) as eluent. Peaks were identified by addition of standards and by diode-array detection. External standardization allowed the determination of alizarin and purpurin with good sensitivity and reliability. The maximum purpurin content was observed in cultures cultivated on solid B5 medium (5.94 mg g⁻¹). The highest alizarin content was measured in cultures cultivated on solid 1/2 NMS medium (2.14 mg g⁻¹).

     

Identification of photoproducts from imazosulfuron by HPLC

Photolysis of imazosulfuron was studied in aqueous solution under UV light. The reaction followed a pseudo-first-order kinetic with significant correlation coefficient. The major photodegradation products of imazosulfuron after irradiation under UV light were separated and tentatively identified by HPLC-MS analysis as (4,6-dimethoxypyrimidine-2-yl)aminocarbonylsulfamic acid, 4,6-dimethoxy-2-ureidopyrimidine and 2,2'-dichloro-[3,3'] bi [imidazo[1,2-a] pyridinyl]. The results indicate that different reaction pathways are followed: (1) cleavage of the sulfonylurea bridge; (2) desulfonylation, which can proceed either by a carbon-sulfur cleavage or a nitrogen-sulfur cleavage. A mechanism for the formation of the photoproducts is proposed.     

Identification of natural dyes in archeological Coptic textiles by liquid chromatography with diode array detection

Reversed-phase HPLC with diode-array UV-Vis spectrophotometric detection has been used for identification of natural dyes in extracts from wool and silk fibres from archeological textiles. The examined objects originate from 4th to 12th Century Egypt and belong to the collection of Early Christian Art of the National Museum in Warsaw. Extraction from fibres was carried out with HCl solution containing ethanol or with warm pyridine. As the main individual chemical components of natural dyes, anthraquinone, indigoid and flavonoid dyes including alizarin, purpurin, luteolin, apigenin, carminic acid, ellagic acid, gallic acid, laccaic acids A and B and indigotin were found. For pyridine extracts another mobile phase with an optimized gradient of organic modifier concentration was used. With such an eluent the appearance of double peaks for indigotin and indirubin was eliminated. For acidic extraction of dyes from fibres, ethanol was used. Due to its higher boiling point than methanol it evaporates slower from the extraction solution enabling a more efficient extraction of dyes.     

Identification of Antiangiogenesis Protein by HPLC and ESIMS

The bio-mass spectrometry has been become an important analytical tool in Life Science such as in searching for a functional protein in disease treatments, for example, the proteins with a function of antiangiogenesis. If an unknown functional protein can be identified and characterized, it is then possible to mass-produce this protein through bio-techniques and the technique of genetic engineering.     

中药材黄芪与红芪的色谱鉴别法

目的：鉴别中药材黄芪和红芪．方法：高效液相色谱法,数据分析用waters公司．Patten Mattch软件．结果：黄芪和红芪药材的区别是在高效液相色谱指纹图谱中黄芪药材有26个特征峰,红芪药材有23个特征峰;红芪中毛蕊异黄酮的含量低于芒炳花素,黄芪中毛蕊异黄酮的含量高于芒炳花素;红芪中不含黄芪甲苷,黄芪中含有黄芪甲苷．结论：可以用色谱法准确、可靠地鉴别黄芪和红芪药材．     

Identification of anthraquinone coloring matters in natural red dyes by electrospray mass spectrometry coupled to capillary electrophoresis

Capillary electrophoresis with UV/visible diode-array detection (DAD) and electrospray mass spectrometric (ESI-MS) detection were used for the identification of anthraquinone color components of cochineal, lac-dye and madder, natural red dyestuffs often used by ancient painters. For the purpose of such analysis, ESI-MS was found to be a much more appropriate detection technique than DAD one owing to its higher sensitivity (detection limits in the range 0.1-0.5 micro g ml(-1)) and selectivity. The method developed made it possible to identify unequivocally carminic acid and laccaic acids A, B and E as coloring matters in the examined preparations of cochineal and lac-dye, respectively. In madder, European Rubia tinctorum, alizarin and purpurin were found. The method allows the rapid, direct and straightforward identification and quantification of components of natural products used in art and could be very helpful in restoration and conservation procedures.     

Identification and determination of corticosteroids in adrenal gland extracts for pharmaceutical use by HPLC

Summary Some HPLC procedures with isocratic or gradient elution are reported for the identification and determination of most of the characteristic components of cortical extracts. The proposed solvent systems were: A) for normal phase chromatography, mixtures of chloroform-methanol-water on silica columns. B) For reversed phase chromatography, mixtures of methanol-water or acetonitrile-water or tetrahydrofuran-water on octadecyl silica columns of different brands. With these systems it was possible to identify and determine, in addition to the principal corticosteroids, some minor components of the cortical extracts as the 20β-dihydroderivatives of compounds F, E, A, B, the 17-ketosteroids adrenosterone, 11β-hydroxyandrostendione and androstendione and finally, progesterone and 17-OH progesterone. In reversed phase chromatography it was also possible, by monitoring the effluent at 205 nm, to reveal the 5α- and 5β-tetrahydroderivatives of the main corticosteroids and to separate them from most of the steroidal components of the adrenal extracts; in these conditions it was also possible to reveal some characteristic, unknown components of the cortical extracts. Some results of quantitative analysis of cortical extracts are also reported, comparing different analytical procedures. Presented at the 15th International Symposium on Chromatography, Nürnberg, October 1984     

Fingerprint analysis of Shankhpushpi for species discrimination by HPLC coupled with chemometric methods

Abstract

Shankhpushpi, traded in the Indian market is having different controversial botanical sources, that is, Convolvulus pluricaulis Choisy (Convulvulaceae), Clitoria ternatea Linn. (Papilionaceae), and Evolvulus alsinoides Linn. (Convulvulaceae). Simple, accurate and precise RP-HPLC-PDA method was established for quality assessment and discrimination of Shankhpushpi samples using chromatographic profiling method. In distinction to the routine method of quality assessment which uses single or dual marker peaks, all chromatographic data (retention times/variables) were used. Fifteen shankhpushpi samples were purchased from the market including authenticated samples of all three C. pluricaulis, C. ternatea, E. alsinoides. A total of 18 samples were analyzed by HPLC and the dataset was then treated with multivariate analyses like PCA and HCA by using MINITAB software. Thus, the developed method was useful in discriminating the Shankhpushpi samples and for the perseverance of quality control.     

鲨鱼软骨血管抑制因子的高效液相色谱与电喷雾质谱研究

电喷雾质谱 ( ESIMS)具有快速、灵敏等特点 ,近年来已成为鉴定和分析多肽、蛋白质和核酸的有力工具 .将 ESIMS与高效液相色谱 ( HPLC)联用 ,在蛋白质分子量、结构及活性位点等方面的研究已取得较大的进展 [1 ] .鲨鱼软骨血管抑制因子 ( SCAIF- I)是作者之一从鲨鱼软骨中提取的一种未知蛋白质[2 ] ,研究表明 ,SCAIF- I对肿瘤、癌症的抑制及治疗具有明显的疗效[3] .因此 ,对其结构的研究具有重要的意义 .本文报道了用 HPL C与 ESIMS对未知蛋白质 SCAIF- I的分子量及肽段部分序列的测定结果 .1　实验部分1 .1　样品制备　 SC…     

流动注射-化学发光法测定决明子蒽醌类化合物

基于在碱性条件下,蒽醌对ClO- Luminol化学发光体系的显著抑制作用,结合反向流动注射技术,提出了蒽醌的流动注射 化学发光分析新方法。蒽醌的质量浓度在0.5～100μg mL范围内与化学发光的抑制强度呈良好的线性关系。检出限为0.05μg mL,RSD=1.6%,采样频率为150次 h,回收率为105%～107.5%。     

Simultaneous Identification and Determination of Several Barbiturates in Plasma by Reverse-Phase HPLC

Abstract

A Reverse-Phase HPLC method is described for the simultaneous identification and determination of 5 barbiturates: Allobarbital, Phencbarbital, Butabarbi -tal, Butalbital and Pentobarbital. Barbital is used as internal standard. The mobile phase is Milli Q Water/ Acetonitrile/ 1.75 M Phosphoric Acid (738:200:2, v/v/v) at a flow rate of 0.8 mL/min and UV detection is carried out at 195 nm. The single extraction procedure requires only small samples and analysis is quick.     

Identification of the individual molecular species of ceramides derived from human erythrocytes using HPLC/MS and HPLC/MS/MS

A preparative separation of total ceramide fraction from the crude extract of the erythrocytary lipids was done by means of normal-phase column chromatography. This was followed by comprehensive profiling of the molecular species in the obtained ceramide by means of HPLC/MS and HPLC/MS/MS. The MS/MS analysis displayed that human erythrocytes contain 19 molecular species of the ceramide of which 12 can be unambiguously identified; erythrocytary ceramides may contain not only sphingosine but also sphinganine as their building blocks; one of the species (namely Cer 24:2/S18) previously has managed to escape identification. We also obtained a quantitative profile of major ceramide species showing the prevalence of Cer 22:0/S18, 24:0/S18 and 24:1/S18.     

Identification of flavonoids in Dendrobium huoshanense and comparison with those in allied species of Dendrobium by TLC,HPLC and HPLC coupled with electrospray ionization multi‐stage tandem MS analyses

Dendrobium huoshanense, a unique species in the genus Orchidaceae, is only found in China and is known as “mihu”. Due to the lack of quality control, the use of D. huoshanense in the herbal market has been limited. In this study, methods based on thin‐layer chromatography, high‐performance liquid chromatography and high‐performance liquid chromatography coupled with electrospray ionization multi‐stage tandem mass spectrometry were used to identify the flavonoids in D. huoshanense and distinguish this species from other Dendrobium species. Using thin‐layer chromatography, a characteristic band was observed for D. huoshanense, and this band was absent from the thin‐layer chromatography plates of other Dendrobium species. Then, using high‐performance liquid chromatography, nine peaks of flavonoids were observed in the chromatograms of ten batches of D. huoshanense. Ultimately, 22 flavonoids in D. huoshanense were identified by multi‐stage tandem mass spectrometry, and 11 of these compounds are being reported from D. huoshanense for the first time. In addition, two compounds both with molecular weights of 710, were identified as being unique to D. huoshanense; one of these compounds, apigenin‐6‐C‐α‐L‐rhamnosyl‐(1→2)‐β‐D‐glucoside‐8‐C‐α‐L‐arabinoside, was proven to be responsible for the characteristic thin‐layer chromatography band of D. huoshanense. These analysis methods can be applied for the identification and quality control of D. Huoshanense.     

Identification of gasolines with hydrocarbon markers

A procedure for the identification of gasoline samples containing hidden chemical markers based on the results of fuel analysis by gas-liquid chromatography with sample preevaporation is proposed. Tetracosane (C₂₄H₅₀) in a concentration of 2.0 × 10⁻³ wt % was used as a marker. The procedure allowed us to detect the presence of the marker in gasoline samples subjected to uncontrollable evaporation in storage or on fire.     

Resolution and Identification of Steroid Oxime Syn and Antiisomers by HPLC

Abstract

Oxime derivatives of a number of steroidal 4-en-3-one 17-alcohols and 17-esters have been separated into syn and anti isomers by HPLC under normal or reverse-phase conditions. The elution order of the isomeric pairs was found to vary with the nature of the steroid and the type of stationary phase: a peak ratio method at two wavelengths provides a very convenient method for unambiguously identifying the geometries of the isomeric pairs.