Involvement of a Novel Peptide in the Heat Shock Response of Australian Wheats

1990 ◽  
Vol 17 (4) ◽  
pp. 441 ◽  
Author(s):  
CS Blumenthal ◽  
IL Batey ◽  
CW Wrigley ◽  
EWR Barlow
Keyword(s):  
Heat Shock ◽  
High Performance ◽  
Storage Proteins ◽  
Seed Proteins ◽  
Reversed Phase ◽  
Terminal Sequence ◽  
Chinese Spring Wheat ◽  
End Use ◽  
Peptide Gene

A low molecular weight peptide, induced by exposure of coleoptiles to heat stress at 41°C, has been detected by reversed-phase high performance liquid chromatography of extracts from coleoptiles of five wheat (Triticum aestivum) cultivars. This component is detected within 1 h of a 41°C heat shock, is not detected 48 h after cessation of the heat shock, and remains present during a continuous 24 h heat treatment. The appearance of this component is also induced by exposure of the coleoptile to 0.1 M sodium arsenite or 10% ethanol. When other species such as barley (Hordeum vulgare), soybean (Glycine max), sorghum (Sorghum bicolor), maize (Zea mays), mungbean (Vigna radiata), or rice (Oryza sativa) were examined for the presence of a component eluting in the same position, it was only detected in maize. The amino acid sequence for the heat-induced peptide from wheat was determined to be: V-L-V-P-V-P-Q-L-Q-P-Q-N-Q-P/Q. The sequence of 12 of these amino acids is the same as the N-terminal sequence of α- and β gliadins (wheat endosperm storage proteins). The production of this heat-induced peptide in aneuploids of Chinese spring wheat indicated that the peptide gene was located on the same chromosome arm as one of the gliadin genes. The presence of this gliadin-like peptide in heat-stressed coleoptiles may be due to the presence of five heat shock elements in the gene sequence of gliadins. The potential heat inducibility of the gliadin gene has important implications for end-use quality of wheat. The results also imply that seed proteins may have a function other than storage of nitrogen.

SELECTION OF COLUMN AND OPERATING CONDITIONS FOR REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY OF PROTEINS IN CANADIAN WHEAT

1985 ◽  
Vol 65 (2) ◽  
pp. 285-298 ◽  
Author(s):  
J. E. KRUGER ◽  
B. A. MARCHYLO
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Storage Proteins ◽  
Sodium Dodecyl ◽  
Reversed Phase ◽  
Operating Conditions ◽  
Protein Patterns ◽  
Optimal Separation ◽  
Rp Hplc

Chromatographic conditions were optimized and three commercially available columns were evaluated for separation of alcohol-soluble storage proteins of Neepawa wheat using reversed-phase high-performance liquid chromatography (RP-HPLC). Optimal separation was achieved using an extracting solution of 50% 1-propanol, 1% acetic acid, and 4% dithiothreitol and an HPLC elution time of 105 min at a flow rate of 1.0 mL/min. HPLC columns evaluated (SynChropak RP-P, Ultrapore RPSC and Aquapore RP-300) varied in selectivity and resolution. The column providing the greatest versatility was Aquapore RP-300 available in cartridge form. Sodium dodecyl sulfate gradient-gel electrophoresis analysis of protein peaks resolved by RP-HPLC indicated that many of the eluted peaks contained more than one protein species. Chromatographic protein patterns obtained for Neepawa wheat grown at different locations and in different years were qualitatively the same.Key words: Protein, high-performance liquid chromatography, wheat

ON THE IDENTITY OF PLATELET FIBRINOGEN WITH PLASMA FIBRINOGEN

1987 ◽  
Author(s):  
H Kaudewitz ◽  
A Henschen ◽  
R E Zimmermann
Keyword(s):  
High Performance ◽  
Reversed Phase ◽  
Plasma Fibrinogen ◽  
Side Chain ◽  
Terminal Sequence ◽  
Hplc Fingerprint ◽  
Amino Terminal ◽  
Chain Composition ◽  
Established Fact ◽  
Amino Terminal Sequence

It is a well established fact that fibrinogen occurs not only in blood plasma but also in the α-granules of the platelets. Recently, it has been shown that fibrinogen is synthesised in the megakaryocytes as well as in the liver. Plasma fibrinogen is derived from the liver, but platelet fibrinogen either exclusively or partially from the megakaryocytes. Conclusive, proteinche-mical evidence for the identity of the fibrinogens from the two biosynthetic sources has previously not been produced. However, the two fibrinogen preparations have been shown to have the same overall peptide chain composition, to be thrombin-clottable and release fibrinopeptides of A- and B-type, and to react with antibodies against plasma fibrinogen. The two preparations differ in the way that platelet fibrinogen lacks the higher-molecular-mass γ-chain variant.The aim of the present investigation was to conduct a detailed proteinchemical comparison between human plasma and platelet fibrinogen. For this purpose, fibrin(ogen)s from the two sources were mercaptolysed, alkylated and the three peptide chains isolated by reversed-phase high-performance liquid chromatography (HPLC). The peptide chains were then analysed directly for amino-terminal sequence and for carboxyterminal sequence by isolation of a terminal fragment. The HPLC-fingerprint patterns of the cyanogen bromide-cleaved chains were compared. The native fibrinogens were also treated with thrombin and the fibrinopeptide type distribution determined by reversed-phase HPLC. The carbohydrate side chain compositions were established by ion-exchange-chromatographic methods after acid hydrolysis. So far no previously unrecognized differences have been observed.

Bioactive Compounds from the Flower Part of Polygonum maritimum L. Collected from Algerian Coast

2020 ◽  
Vol 16 (4) ◽  
pp. 543-545 ◽  
Author(s):  
Imad Abdelhamid El Haci ◽  
Wissame Mazari ◽  
Fawzia Atik-Bekkara ◽  
Fatma Mouttas-Bendimerad ◽  
Fayçal Hassani
Keyword(s):  
Phenolic Compounds ◽  
Bioactive Compounds ◽  
High Performance ◽  
Reversed Phase ◽  
Uv Spectra ◽  
Local Database ◽  
Rp Hplc ◽  
Algerian Coast ◽  
First Time

Background: Polygonum maritimum is one of the spontaneous halophyte plants of the Algerian coast. Many studies were carried out to evaluate the contents and the quality of phenolic compounds of this plant around the Mediterranean region. Objective: This paper intends to identify, for the first time, the phenolic compounds from the flower part of P. maritimum. Methods: RP-HPLC-PDA (Reversed Phase-High Performance Liquid Chromatography-Photo Diode Array) material was used for this purpose. Many standards were used and their retention times were stored in a local database. Identification was made on the basis of retention times of retained compounds and those found in the literature, and UV spectra of each peak. Results: This study intends to identify five phenolic acids (gallic, ferulic, sinapic, caffeic and syringic acids), one flavonol (rutin) and one flavanone (naringenin). Conclusion: P. maritimum is an important source of natural bioactive compounds that can be exploited for the benefit of many fields.

Reversed Phase High Performance Liquid Chromatography of Essential Oils

1980 ◽  
Vol 35 (9-10) ◽  
pp. 675-681 ◽  
Author(s):  
Dieter Strack ◽  
Peter Proksch ◽  
Paul-Gerhard Gülz
Keyword(s):  
Essential Oils ◽  
Chromatographic Analysis ◽  
High Performance ◽  
High Performance Liquid Chromatographic ◽  
Reversed Phase ◽  
Complex Mixtures ◽  
Naturally Occurring ◽  
Chromatographic Procedure ◽  
Liquid Chromatographic

Abstract A reversed phase high performance liquid chromatographic procedure for the separation of essential oils is described. Applying water-acetonitrile elution systems on octyl and octadecylsilane- bonded silica, complex mixtures of sequiterpenes and oxygenated volatile constituents can be resolved, comparably to the quality of gas chromatographic analysis. Photometrical detection at different wavelengths can be an important parameter in optimizing the separation of essential oil constituents. The application of the described method is demonstrated with naturally occurring mixtures of terpenes from leaves of Cistus ladanifer.

ON THE FACTOR XIIIa-INDUCED CROSSLINKING OF HUMAN FIBRIN α-CHAINS

1987 ◽  
Author(s):  
A Henschen ◽  
E Müller
Keyword(s):  
High Performance ◽  
Cyanogen Bromide ◽  
Control Sample ◽  
Reversed Phase ◽  
Factor Xiiia ◽  
Total Amino Acid ◽  
Terminal Sequence ◽  
Linkage Pattern ◽  
Early Protein ◽  
Α Chain

Factor XIIIa catalysis the formation of isopeptide bonds Between γ-carbamoyl groups of peptide-bound glutamines and ε-amino groups of lysines or lysine analogues. During fibrin crosslinking two such bonds are rapidly formed between the C-termini of two γ-chains in adjacent molecules and then several bonds are more slowly formed between several α-chains. The crosslinking sites in the γ-chain were identified already 15 years ago, those in the α-chain are still only tentatively or partially identified,, However, by determining the incorporation of lysine analogues in the α-chain it could be shown that the glutamines in positions 328, 366 and possibly also 237 may participate in crosslinking reactions. Analyses of cyanogen bromide fragments isolated from crosslinked fibrin indicated the segments 271-776 and 518-587 to contain the primary crosslinking sites.In the present study factor XHI-containing fibrinogen was incubated over night with thrombin in presence of calcium ions and cysteine or, as a control, in presence of EDTA. The fibrin material was cleaved with cyanogen bromide, mercaptolysed, pyri-dylethylated and then subjected to Sephacryl S-300 chromatography. The early protein fractions were tested by reversed-phase high-performance liquid chromatography (HPLC) using fibrinogen fragments as reference. In the control sample Aa-chain fragment 271-776 eluted first but in the crosslinked sample it was missing and instead a heterogeneous mixture of higher-molecular weight components was observed. N-Terminal sequence analysis showed the mixture to contain not only the expected fragments 241-476 and 518-584,but in fact all glutamine- or lysine-containing Aα-chain fragments between positions 208 and 611. In the corresponding 6 fragments a total of 6 glutamines and 21 lysines as potential crosslinking sites are present. Two fragments contain only one each of these residues which therefore must be true crosslinking sites. Remaining sites and the actual linkages were identified after reversed-phase HPLC of the tryptic peptide mixture by N-terminal sequence and total amino acid analyses.The linkage pattern will provide information about the localisation and conformation of the C-terminal part of the α-chain and its contribution to the fibrin polymer structure.

scholarly journals Combined Determination of Succinic Acid and Cetylpyridinium Chloride in Medicinal Films by Gradient High Performance Liquid Chromatography

Medicina ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 75-88
Author(s):  
O. N. Dvorskaya ◽  
◽  
N. N. Nozhkina ◽  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Succinic Acid ◽  
High Performance ◽  
Cetylpyridinium Chloride ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Gradient Elution Mode

A technique has been developed based on reversed-phase high-performance liquid chromatography with diode-matrix detection for the joint determination of succinic acid and cetylpyridinium chloride in complex action medicinal films. Efficient chromatographic separation of active drug components was achieved in a gradient elution mode on a Luna C18 (2) 100A column (4.6 × 250 mm, 5 µm) using a mobile phase consisting of a 0.1% solution of phosphoric acid and acetonitrile. The detection wavelength was 210 nm for both compounds. The developed method is validated in terms of specificity, linearity, precision, accuracy and can be used to determine the authenticity and quantitative content of succinic acid and cetylpyridinium chloride in the joint presence in assessing the quality of medicinal films.

scholarly journals Quick Detection of Aldehydes and Ketones in Automotive Textiles

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Yanxue Ma ◽  
Wenliang Xue ◽  
Mengyuan Wei ◽  
Jingfang Qian
Keyword(s):  
High Performance ◽  
Development Process ◽  
Linear Models ◽  
Determination Limit ◽  
Aldehydes And Ketones ◽  
Actual Environment ◽  
End Use ◽  
Fabric Production ◽  
Automotive Textiles

AbstractThis study was aimed to develop a quick detection method to test aldehydes and ketones in textiles in order to control the quality of automotive textiles in the development process from fabric production to end-use in vehicles. In this study, a pretreatment of samples was applied to simulate the actual environment of textiles used in vehicles. Collected volatiles were reacted with 2,4-dinitrophenylhydrazine and then eluted with acetonitrile tetrahydrofuran. The eluent was analyzed with high-performance liquid chromatography. Findings showed more than 90% volatiles could be detected in the established method; the lowest determination limit was 0.0297 mg/mL; and the lowest quantification limit was 0.0991 mg/mL, which meant sensitivity and capability of the method were high. Regression coefficients of linear models between volatile concentrations and chromatographic peak characteristics were >0.995, indicating that the method could effectively and efficiently determine the contents of volatiles in automotive textiles.

scholarly journals High Performance Liquid Chromatography versus Stacking-Micellar Electrokinetic Chromatography for the Determination of Potentially Toxic Alkenylbenzenes in Food Flavouring Ingredients

Molecules ◽  
2021 ◽  
Vol 27 (1) ◽  
pp. 13
Author(s):  
Huynh N. P. Dang ◽  
Joselito P. Quirino
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Micellar Electrokinetic Chromatography ◽  
Reversed Phase ◽  
Food Samples ◽  
Electrokinetic Chromatography ◽  
Analytical Methodology

Alkenylbenzenes, including eugenol, methyleugenol, myristicin, safrole, and estragole, are potentially toxic phytochemicals, which are commonly found in foods. Occurrence data in foods depends on the quality of the analytical methodologies available. Here, we developed and compared modern reversed-phase high performance liquid chromatography (HPLC) and stacking-micellar electrokinetic chromatography (MEKC) methods for the determination of the above alkenylbenzenes in food flavouring ingredients. The analytical performance of HPLC was found better than the stacking-MEKC method. Compared to other HPLC methods found in the literature, our method was faster (total run time with conditioning of 15 min) and able to separate more alkenylbenzenes. In addition, the analytical methodology combining an optimized methanol extraction and proposed HPLC was then applied to actual food flavouring ingredients. This methodology should be applicable to actual food samples, and thus will be vital to future studies in the determination of alkenylbenzenes in food.

scholarly journals Stereospecific Assay of (R)- and (S)-Goitrin in Commercial Formulation of Radix Isatidis by Reversed Phase High-Performance Liquid Chromatography

2017 ◽  
Vol 2017 ◽  
pp. 1-6 ◽  
Author(s):  
Lixing Nie ◽  
Zhong Dai ◽  
Shuangcheng Ma
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Reversed Phase ◽  
Commercial Formulation ◽  
Radix Isatidis ◽  
Enantioselective Resolution ◽  
Pharmacological Studies ◽  
Optimum Separation

Radix isatidis (Banlangen) is a widely used traditional Chinese medicine for treating fever and removing toxic heat. Pharmacological studies have indicated that (R)-goitrin (epigoitrin) is one of the main constituents accounting for its antiviral activity, while (S)-goitrin (goitrin) is known as an antithyroid factor. To better control the quality of radix isatidis and its formulations, it is imperative to enantiomerically determine the contents of R- and S-goitrin. In this study, an enantioselective method based on reversed phase chromatography was developed for the assay of (R, S)-goitrin enantiomers. Optimum separation was obtained on an S-Chiral A column (4.6 mm × 250 mm, 5 μm) using methanol/water (30 : 70, v/v) as the mobile phase. After validation, the method was applied to quantify the enantiomers in Banlangen granules, which is the most prescribed commercial formulation of radix isatidis. Compared to enantioselective resolution approaches based on normal phase chromatography, the new method could be conveniently performed using regular reversed phase high-performance liquid chromatography (RP-HPLC) equipment and was found to be more suitable for the enantioselective quality control of water-extracted and soluble medicines.

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