scholarly journals Fast identification of wheat 1BL.1RS translocation by reversed-phase ultra-performance liquid chromatography (RP-UPLC)

2013 ◽  
Vol 64 (9) ◽  
pp. 865 ◽  
Author(s):  
Jianwen Zhou ◽  
Caixia Han ◽  
Hui Cao ◽  
Shoumin Zhen ◽  
Zitong Yu ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Large Scale ◽  
Polyacrylamide Gel Electrophoresis ◽  
Ultra Performance Liquid Chromatography ◽  
Reversed Phase ◽  
Wheat Breeding ◽  
Parental Line ◽  
Wheat Cultivars ◽  
Protein Markers ◽  
Fast Identification

The 1BL.1RS chromosomal translocation in wheat is the result of replacement of the short arm of chromosome 1B of wheat by the short arm of chromosome 1R of rye, which had been widely used as a parental line in worldwide wheat breeding, resulting in a high percentage of wheat cultivars containing this translocation. A fast and reliable approach to identify this translocation is highly desirable in modern wheat breeding. This study compared reversed-phase ultra-performance liquid chromatography (RP-UPLC), acidic polyacrylamide gel electrophoresis (A-PAGE), liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), allelic-specific PCR, and reversed-phase high-performance liquid chromatography (RP-HPLC) approaches to identify the 1BL.1RS translocation in 76 bread wheat cultivars. Two gliadin bands in the Gli-B1 region of A-PAGE separation were confirmed by LC-MS/MS to be omega secalins from the 1BL.1RS translocation, and they can be used as reliable protein markers for identifying the translocation. A few specific minor peaks eluted at 12–13 min on the RP-UPLC patterns can readily differentiate the 1BL.1RS translocation. Of the 76 wheat cultivars tested, 40 were identified as carrying the 1BL.1RS translocation by RP-UPLC, which was consistent with the results of A-PAGE, HPLC, and PCR. Compared with other established methods, RP-UPLC showed a clear advantage in fast identification of the 1BL.1RS translocation with higher reliability and lower costs, and it is therefore ideal for large-scale screening of the 1BL.1RS translocation in wheat breeding.

scholarly journals A Rapid, Reliable RP-UPLC Method for Large-Scale Analysis of Wheat HMW-GS Alleles

Molecules ◽  
2021 ◽  
Vol 26 (20) ◽  
pp. 6174
Author(s):  
Su-Bin Lee ◽  
Yu-Jeong Yang ◽  
Sun-Hyung Lim ◽  
Yong Q. Gu ◽  
Jong-Yeol Lee
Keyword(s):  
Large Scale ◽  
Storage Proteins ◽  
Ultra Performance Liquid Chromatography ◽  
Reversed Phase ◽  
Wheat Breeding ◽  
Pcr Analysis ◽  
Large Scale Analysis ◽  
End Use ◽  
Acquity Uplc ◽  
Wheat Storage Proteins

High-molecular-weight glutenin subunits (HMW-GS) account for only 10% of total wheat storage proteins, but play an important role in the processing quality of wheat flour. Therefore, identifying HMW-GS alleles associated with good end-use quality provides important information for wheat breeders. To rapidly, accurately and reproducibly identify HMW-GS, we established an optimized reversed-phase ultra-performance liquid chromatography (RP-UPLC) method. Separation parameters were optimized using an ACQUITY UPLC Protein BEH C4 column and stepwise ACN gradient, and the separation patterns and retention times (RTs) of 22 subunits were comparatively analyzed in 16 standard wheat cultivars. All HMW-GS proteins were well separated within about 5.5 min, and all analyses were complete within 12 min. We distinguished the 16 subunits based on RT, although three subunits in 1Bx (1Bx7/1Bx7OE and 1Bx17) and three subunits in 1By (1By8*, 1By9 and 1By15) had overlapping RTs; these were differentiated by SDS-PAGE. To distinguish 1Bx7 and 1Bx7OE, which differ in protein abundance, RP-UPLC was combined with PCR analysis of DNA junction markers. The optimized method was successfully applied to determine HMW-GS alleles in a large collection of bread wheat germplasm (1787 lines). This protocol is an appropriate option for selecting lines harboring favorable HMW-GS alleles in wheat breeding.

scholarly journals Structural elucidation of triacylglycerol using online acetone Paternò–Büchi reaction coupled with reversed-phase liquid chromatography mass spectrometry

The Analyst ◽  
2020 ◽  
Vol 145 (20) ◽  
pp. 6532-6540 ◽  
Author(s):  
Elissia T. Franklin ◽  
Yu Xia
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Large Scale ◽  
Reversed Phase ◽  
Structural Elucidation ◽  
Scale Analysis ◽  
Liquid Chromatography Mass Spectrometry ◽  
Reversed Phase Liquid Chromatography ◽  
Large Scale Analysis ◽  
Phase Liquid

The developed online RPLC-PB-MS/MS system allows large scale analysis of isomeric triacylglycerol lipids differing in CC locations.

Development and validation of a UPLC method for screening potentially counterfeit anti-hypertensive drugs using design of experiment

2014 ◽  
Vol 6 (13) ◽  
pp. 4610-4616 ◽  
Author(s):  
Chiguru Vishnuvardhan ◽  
R. Srinivas ◽  
N. Satheeshkumar
Keyword(s):  
Liquid Chromatography ◽  
Design Of Experiment ◽  
Ultra Performance Liquid Chromatography ◽  
Reversed Phase ◽  
Uv Detection ◽  
Counterfeit Medicines ◽  
Development And Validation

An isocratic reversed phase ultra performance liquid chromatography (RP-UPLC) method was developed for screening counterfeit medicines with UV detection at 210 nm.

Development and Analytical Validation of the Methodology for Vitamins in Tablets by Ultra-Performance Liquid Chromatography

2019 ◽  
Vol 57 (10) ◽  
pp. 881-891
Author(s):  
Giulio D C d’Oliveira ◽  
Andréa R Chaves ◽  
Caridad N Pérez
Keyword(s):  
Liquid Chromatography ◽  
Ultra Performance Liquid Chromatography ◽  
Reversed Phase ◽  
Analysis Time ◽  
Photodiode Array Detection ◽  
Total Analysis Time ◽  
Quality Control Laboratory ◽  
Photodiode Array ◽  
Array Detection ◽  
Total Analysis

Abstract In the present study, we developed a reliable and robust chromatographic method for the quantification of multivitamins in tablet samples by ultra-performance liquid chromatography (UPLC) with photodiode array detection. The vitamins nicotinamide, pyridoxine, riboflavin, and thiamin were analyzed and quantified in a total analysis time of 2.5 minutes, using hydrophilic interaction liquid chromatography stationary phase. Tocopherol acetate and cyanocobalamin were analyzed and quantified in a total analysis time of 2.5 minutes, using reversed-phase (RP)-UPLC. The analysis time reported here is lower than that of similar methods reported in the literature for single vitamin determination. The method linearity exhibits a good correlation coefficient (R2 = 0.998) with the relative residual standard deviation in the acceptable limit of 2.0%. The developed methods were validated, and the results demonstrated that the proposed analytical method showed to be selective, sensitive, accurate, and robust for the quantification of evaluated vitamins in multivitamin tablets. The work was fully developed in the quality control laboratory of a pharmaceutical industry in the Agroindustrial District of Anápolis (DAIA, Goiás, Brazil), where the product is manufactured.

Characterization by high-performance liquid chromatography of circulating growth hormone-releasing factors in normal subjects

1986 ◽  
Vol 111 (3) ◽  
pp. 507-511 ◽  
Author(s):  
E. S. Penny ◽  
A. M. Sopwith ◽  
R. L. Patience ◽  
J. A. H. Wass ◽  
G. M. Besser ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Large Scale ◽  
Molecular Form ◽  
Normal Subjects ◽  
Reversed Phase ◽  
Percentage Increase ◽  
Mixed Meal ◽  
Analytical Separation

ABSTRACT Four forms of circulating immunoreactive human GH-releasing factor (ir-hGRF) have been identified in each of four normal subjects, with a mean increase in total ir-hGRF of twofold over basal levels following a mixed meal. Plasma samples (200 ml) from each individual were subjected to large-scale Vycor extraction with initial purification by high-performance liquid chromatography on a reversed-phase C18 column, followed by analytical separation of the ir-hGRF components using a C3 wide-pore reversed-phase column, and subsequent radioimmunoassay of the fractions. The mean recovery of total ir-hGRF from the plasma (fasted and non-fasted) was 76±16% (2×s.e.m.). Analytical separation of the ir-hGRF revealed four components which co-eluted with synthetic hGRF-37, hGRF-40 and hGRF-44, and a peak eluting between hGRF-40 and -44 which may represent hGRF-42. The hGRF-40 was shown to be the predominant circulating molecular form in the fasted state in each subject, and in three out of four subjects following a mixed meal. The hGRF-44 showed the greatest percentage increase over basal in all four individuals. J. Endocr. (1986) 111, 507–511

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