scholarly journals High-Performance Liquid Chromatography Under Partially Denaturing Conditions (dHPLC) is a Fast and Cost-Effective Method for Screening Molecular Defects: Four Novel Mutations Found in X-Linked Chronic Granulomatous Disease

2012 ◽  
Vol 76 (2) ◽  
pp. 158-166 ◽  
Author(s):  
E. B. de Oliveira-Junior ◽  
C. Prando ◽  
J. A. Lopez ◽  
J. C. Arango ◽  
M. Buzolin ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Chronic Granulomatous Disease ◽  
High Performance ◽  
Cost Effective ◽  
Granulomatous Disease ◽  
Novel Mutations ◽  
Molecular Defects ◽  
Cost Effective Method

scholarly journals Flavonoid Quantification in Onion by Spectrophotometric and High Performance Liquid Chromatography Analysis

HortScience ◽  
2002 ◽  
Vol 37 (4) ◽  
pp. 682-685 ◽  
Author(s):  
Kevin A. Lombard ◽  
Emmanuel Geoffriau ◽  
Ellen Peffley
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Hplc Analysis ◽  
Correlation Coefficients ◽  
Cost Effective ◽  
Chromatography Analysis ◽  
Cost Effective Method ◽  
Performance Liquid Chromatography Analysis

Direct spectrophotometric determination of quercetin content in onions (Allium cepa L.) was investigated as a possible alternative to high-performance liquid chromatography (HPLC) analysis. Quercetin content in five onion varieties was monitored at 362 nm and quantified using simple spectrophotometric and HPLC methods. HPLC revealed that 3,4'-Qdg and 4'-Qmg comprised up to 93% of total flavonol content detected in the studied varieties. These major quercetin conjugates combined (3,4'-Qdg + 4'-Qmg) and total flavonol conjugates quantified by HPLC correlated closely with spectrophotometer values. Correlation coefficients were 0.96 (P < 0.0001) for 3,4'-Qdg + 4'-Qmg and 0.97 (P < 0.0001) for total flavonol conjugates in onion. Simple spectrophotometric procedure proved to be a valid, efficient, and cost-effective method for the quantification of total quercetin in onion. Chemical names used: quercetin-3,4'-O-diglucoside (3,4'-Qdg); quercetin-4'-O-glucoside (4'-Qmg).

scholarly journals Rapid and Accurate Approach for Screening of Microsatellite Unstable Tumours Using Quasimonomorphic Mononucleotide Repeats and Denaturating High Performance Liquid Chromatography (DHPLC)

2009 ◽  
Vol 26 (1) ◽  
pp. 19-26 ◽  
Author(s):  
Gašper Berginc ◽  
Damjan Glavač
Keyword(s):  
High Performance Liquid Chromatography ◽  
Capillary Electrophoresis ◽  
Liquid Chromatography ◽  
Microsatellite Markers ◽  
High Performance ◽  
Cost Effective ◽  
Slovenian Population ◽  
Cost Effective Approach ◽  
Msi Analysis ◽  
Colorectal Tumours

MSI analysis is becoming increasingly important for the detection of both hereditary non-polyposis colorectal cancer and sporadic primary colorectal tumours with MSI high phenotype. The Bethesda panel of five microsatellite markers has been proposed to provide uniform criteria for MSI analysis. Here we report on an MSI analysis approach using quasimonomorphic mononucleotide repeats and denaturating high performance liquid chromatography (DHPLC). We analysed 595 newly diagnosed colorectal tumours and 145 normal samples. Microsatellite markers BAT-25, BAT-26, NR-21, NR-22, and NR-27 were amplified in multiplex reaction and analysed using DHPLC and capillary electrophoresis (CE). DHPLC conditions for analysis of MSI multiplex assay were evaluated and tested. Analysis and cross-examination of the results obtained from 96 samples using DHPLC and capillary electrophoresis showed the same sensitivity and specificity of the two approaches for detecting MSI-H tumours. Using our new approach we showed that the tested markers are quasimonomorphic in a Slovenian population, with frequencies of polymorphisms 0.07%, 1.4%, 2.1%, 1.4%, and 1.4% for BAT-25, BAT-26, NR-21, NR-22, and NR-27, respectively. Forty-three (7.2%) new MSI-H tumours were identified, of which 84% showed instability in all 5 tested markers. Overall, we developed a high-throughput, robust, accurate and cost-effective approach for the detection of MSI-H tumours.

scholarly journals The role and impact of high throughput biomimetic measurements in drug discovery

ADMET & DMPK ◽  
2018 ◽  
Vol 6 (2) ◽  
pp. 74-84 ◽  
Author(s):  
Shenaz Bunally ◽  
Robert J Young
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Drug Discovery ◽  
Physicochemical Property ◽  
Physicochemical Properties ◽  
High Throughput ◽  
High Performance ◽  
Cost Effective ◽  
High Quality ◽  
The Right

During the early phase of drug discovery, it is becoming increasingly important to acquire the full physicochemical profile of molecules. For this purpose, there is a strong interest in developing efficient and cost-effective platforms for fast and reliable measurements of physicochemical properties. We have developed an automated physchem platform which ensures that consistent, comprehensive, and high-quality physicochemical property measurements and derived property information for 100's of compounds per week are available alongside potency data at the right time to guide compound progression decisions. We discuss the routine assessments of biomimetic properties using high throughput automated high-performance liquid chromatography (HPLC) platforms, with details of the methods and hardware employed, also with illustrations of the quality and impact of the data generated.

scholarly journals In House Validated UHPLC Protocol for the Determination of the Total Hydroxytyrosol and Tyrosol Content in Virgin Olive Oil Fit for the Purpose of the Health Claim Introduced by the EC Regulation 432/2012 for “Olive Oil Polyphenols”

Molecules ◽  
2019 ◽  
Vol 24 (6) ◽  
pp. 1044 ◽  
Author(s):  
Maria Tsimidou ◽  
Michaela Sotiroglou ◽  
Aspasia Mastralexi ◽  
Nikolaos Nenadis ◽  
Diego García-González ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Olive Oil ◽  
High Performance ◽  
Virgin Olive Oil ◽  
Cost Effective ◽  
Health Claim ◽  
Relative Standard ◽  
Oil Sector ◽  
Hydrolysis Conditions

An ongoing challenge in olive oil analytics is the development of a reliable procedure that can draw the consensus of all interested parties regarding the quantification of concentrations above the required minimum value of 5 mg of bioactive “olive oil polyphenols” per 20 g of the oil, to fulfill the health claim introduced by the European Commission (EC) Regulation 432/2012. An in-house validated ultra-high performance liquid chromatography (UHPLC) protocol fit for this purpose is proposed. It relies on quantification of the total hydroxytyrsol (Htyr) and tyrosol (Tyr) content in the virgin olive oil (VOO) polar fraction (PF) before and after acidic hydrolysis of their bound forms. PF extraction and hydrolysis conditions were as previously reported. The chromatographic run lasts ~1/3 of the time needed under high performance liquid chromatography (HPLC) conditions, this was also examined. Eluent consumption for the same piece of information was 6-fold less. Apart from being cost effective, a larger number of samples can be analyzed daily with less environmental impact. Two external curves, detection at 280 nm and correction factors for molecular weight difference are proposed. The method, which is fit for purpose, is selective, robust with satisfactory precision (percentage relative standard deviation (%RSD) values < 11%) and recoveries higher than 87.6% for the target analytes (Htyr, Tyr). Standard operational procedures are easy to apply in the olive oil sector.

scholarly journals Rapid Genotyping of CTX-M Extended-Spectrum β-Lactamases by Denaturing High-Performance Liquid Chromatography

2007 ◽  
Vol 51 (4) ◽  
pp. 1446-1454 ◽  
Author(s):  
Li Xu ◽  
Jason Evans ◽  
Thomas Ling ◽  
Kathy Nye ◽  
Peter Hawkey
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Cost Effective ◽  
Epidemiological Studies ◽  
Survey Study ◽  
The United Kingdom ◽  
Extended Spectrum ◽  
Surveillance Programs ◽  
Clinically Significant

ABSTRACT Denaturing high-performance liquid chromatography (dHPLC) is a powerful technique which has been used extensively to detect genetic variation. This is the first report of the application of dHPLC for rapid genotyping of bacterial β-lactamase genes. The technique was specifically developed to genotype members of all bla CTX-M DNA homology groups. Thirteen well-defined bla CTX-M extended-spectrum β-lactamase (ESBL)-producing strains were used to develop and optimize the dHPLC genotyping assay. Further evaluation was carried out with a blinded panel of 62 clinical isolates. The results of bla CTX-M genotyping achieved by dHPLC were comparable to the typing results obtained by DNA sequencing. Applying the newly developed dHPLC-based genotyping method, we successfully genotyped all 73 bla CTX-M ESBL-producing strains from the 4-month survey study. Furthermore, we found the first reported cases in the United Kingdom of clinically significant disease caused by CTX-M-14- and CTX-M-1-producing Escherichia coli strains. We conclude that the novel dHPLC assay is highly accurate, rapid, and cost-effective for the genotyping of bla CTX-M-producing ESBLs and has great potential for determining the clinical relevance of different and new bla CTX-M genotypes, as well as for epidemiological studies and surveillance programs.

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