Hplc test

1 Overview of HPLC HPLC is an abbreviation for High Performance Liquid Chromatography. "Chromatography" is a technique for separation, "chromatogram" is the result of chromatography, and "chromatograph" is the instrument used to conduct chromatography. Among the various technologies developed for chromatography, devices dedicated for molecular separation called columns and high-performance pumps for delivering solvent at a stable flow rate are some of the key components of chromatographs. As related technologies became more sophisticated, the system commonly referred to as High Performance Liquid Chromatography, simply became referred to as "LC". Nowadays, Ultra High Performance Liquid Chromatography (UHPLC), capable of high-speed analysis, has also become more wide-spread. Only compounds dissolved in solvents can be analyzed with HPLC. HPLC separates compounds dissolved in a liquid sample and allows qualitative and quantitative analysis of what components and how much of each component are contained in the sample. Fig.1 shows a basic overview of the HPLC process. The solvent used to separate components in a liquid sample for HPLC analysis is called the mobile phase. The mobile phase is delivered to a separation column, otherwise known as the stationary phase, and then to the detector at a stable flow rate controlled by the solvent delivery pump. A certain amount of sample is injected into the column and the compounds contained in the sample are separated. The compounds se...

HPLC Basics What is HPLC? HPLC is an analytical chemistry technique applied to separate, identify, and quantify semi- and non-volatile compounds in liquid samples.Industries like pharma and biopharma, environmental, and food and beverage use HPLC for the characterization, purification, and quantitation of compounds. Contact us Product tour Blog The mobile phase carries a liquid sample through the column to the detector, and compounds or analytes separate due to varying degrees of interaction with the stationary phase. A detector measures the analytes after elution from the column, and a chromatography data system (CDS) translates the detected signal. The translated data output of an HPLC analysis is called a chromatogram, where the x-axis is a measure of time and the y-axis measures a specific signal generated by the detector. Analyte – target compound(s) of interest for detection in an HPLC analysis Mobile phase – phase in motion and composed of solvent or eluents flowing from injection to detection Stationary phase – the still phase where physical separation of analytes occurs Flow rate – how fast the mobile phase flows with respect to time Retention time – time between sample injection and the maximum peak signal of the analyte in a chromatogram Efficiency – given as the number of theoretical plates, a key metric for quantifying performance of a separation Resolution – ability to distinguish between peaks and the primary concern for any separation Selectiv...

HPLC Method Development Steps How to develop an HPLC method Developing an HPLC method involves four basic steps: scouting, optimization, robustness testing, and validation. Learn best practices for sample preparation plus expert tips and tricks for developing an HPLC method. Contact us Product tour Blog Preparation method Analytical principle Application(s) Dilution Decrease analyte, solvent, or matrix concentration in sample Preventing column/detector overloading, reducing sample solvent elution strength Centrifugation Sedimentation based on density Removing large cellular components from solution Filtration Remove particulates from sample Extending column lifetime, preventing clogging of fluidics Protein precipitation Desolubilize proteins by adding salt, solvent, or altering pH Removal of protein from solution Liquid-liquid extraction Isolate sample components based on solubility differences in two miscible solvents Purifying compounds based on polarity/charge Solid phase extraction Selective separation/purification of target analytes using a sorbent stationary phase Isolating small molecules from biological matrices, desalting large biomolecules Immunoaffinity capture Selective purification of analyte using an antibody Isolating small molecules from biological/environmental/food and beverage matrices Protein digestion Enzymatic cleavage of protein into peptides Generating peptides for bottom-up proteomics/peptide mapping Derivatization Chemical reaction to alter the ph...

Only orderable as part of a profile. For more information see: -HAEV1 / Hemolytic Anemia Evaluation , Blood -HBEL1 / Hemoglobin Electrophoresis Evaluation , Blood -MEV1 / Methemoglobinemia Evaluation , Blood -REVE1 / Erythrocytosis Evaluation , Whole Blood -THEV1 / Thalassemia and Hemoglobinopathy Evaluation , Blood Cation Exchange/High-Performance Liquid Chromatography (HPLC) Only orderable as part of a profile. For more information see: -HAEV1 / Hemolytic Anemia Evaluation , Blood -HBEL1 / Hemoglobin Electrophoresis Evaluation , Blood -MEV1 / Methemoglobinemia Evaluation , Blood -REVE1 / Erythrocytosis Evaluation , Whole Blood -THEV1 / Thalassemia and Hemoglobinopathy Evaluation , Blood A large number of variants of hemoglobin (Hb) have been recognized. Although many do not result in clinical or hematologic effects, clinical symptoms that can be associated with hemoglobin disorders include microcytosis, sickling disorders, hemolysis, erythrocytosis/polycythemia, cyanosis/hypoxia, anemia (chronic, compensated or episodic), and increased methemoglobin or sulfhemoglobin results (M-hemoglobins). For common, and many of the uncommon, hemoglobin variants, protein studies will be sufficient for definitive identification. High-performance liquid chromatography is a method that provides useful and supplementary information on most hemoglobin variants. Some hemoglobin disorders and variants, including common alpha thalassemia conditions, are not detected by screening methods and r...