Application Note Categories
HPLC
High Performance Liquid Chromatography (HPLC)is a technique for separating components in a sample on the basis of interactions between the component, a liquid mobile phase and a solid stationary phase.
HPLC or high performance liquid chromatography is a technique for separating components in a sample on the basis of interactions between the component, a liquid mobile phase and a solid stationary phase. Both individual compounds and more complex fractions of a sample can be separated in this manner with detection possible using UV absorption, refractive index measurement, evaporative light scattering or mass spectrometry. The sample is introduced into a metal column coated with a stationary phase through which the mobile phase flows under pressure. The stronger the interaction with the stationary phase, the longer the component will be retained on the column and the stronger the interaction with the mobile phase the shorter will be its residence time on the column and vice versa. A typical separation may require 5 minutes to an hour. HPLC has found its greatest applicability in the pharmaceutical and biotechnology areas in the last 20 years due to its ability to separate complex molecules like proteins and drugs.
Application Notes - HPLC
Three steroid conjugates, estrone 3-sulphate, b-estradiol 17-(b-D glucaronide) and 5b-pregnane-3a, 20a diol glucaronide were separated by reversed phase HPLC/ELSD.
A mixture of four muscle relaxant drugs, rocuronium, pancuromium, pipecuronium and vecuronium bromide was separated by reversed phase HPLC and the drugs detected by ELSD, UV and refractive index.
Ginseng root extracts were separated by reversed phase HPLC and detected by ELSD.
A mixture of twelve underivatized amino acids was separated by reversed phase HPLC and the amino acids detected by ELSD.
Examples of the use of reversed phase HPLC with ELSD detection are given for combinatorial drug analysis. Separations of an aspartate antagonist in synthesis solution and mixture of catecholamines are shown.
Polyethylene glycol MW 600 was separated by reversed phase HPLC and detected by ELSD.
To detect adulteration of olive oil with vegetable oil by measuring triglycerides, a reversed phase HPLC procedure with ELSD detection was developed.
The determination of fatty acids by reversed phase HPLC with ELSD detection was found to be superior to that by UV detection. Analytical-bore columns with flow splitters were found to produce equivalent results when narrow-bore columns were not available
The ELSD and UV detection of five fatty acid methyl esters separated by reversed phase HPLC was compared.
Oligosaccharides found in beer were separated by reversed phase HPLC and detected by evaporative light scattering.
Simple sugars and sugar alcohols found in fruit juices were separated by reversed phase HPLC and detected by evaporative light scattering.
Adulteration of orange juice with high fructose corn syrup detected by reversed phase HPLC with evaporative light scattering.
The use of acetone in the mobile phase to speed up run times for carbohydrates in beer by reversed phase HPLC and evaporative light scattering detection is described.
Carbohydrate sugars in soy infant formula were determined by reversed phase HPLC with evaporative light scattering detection.
Six organic acids in a standard were separated using a GROM-RESIN AC organic acids HPLC column and detected by conductivity.
