Analysis of 25-hydroxyvitamins D2 and D3 in Human Plasma by High-Performance Liquid Chromatography (HPLC) (CAT#: STEM-CT-3112-CJ)

High performance liquid chromatography separates mixtures based on a variety of interaction forces. This interaction force is usually a non-covalent property between the analytes and the analytical tube column. When using HPLC, liquid analytes to be detected are injected into the column at different times and moved through the stationary phase by pressure. As different substances in the analytes interact with the stationary phase differently, different substances leave the column in sequence, and different peak signals are obtained through the detector, with each peak top representing an additional compound species, and finally the analyzed signals are compared to determine the substances contained in the analytes to be detected. The final analysis of these signals is used to determine the substances contained in the material to be measured.

Biochemistry; Biomedical

1. Sample preparation: Sample preparation is central to successful HPLC and UHPLC analyses.
2. Method scouting. Involves screening various column and eluent conditions. The purpose of this phase is to select the best combinations for a successful HPLC separation.
3. Method optimization. Includes iterative testing of various separation conditions of the HPLC method and is performed to achieve the best possible resolution, speed, and reproducibility. This step is the most time-consuming part of method development and often requires expert knowledge to perfect.
4. Robustness testing. Done to determine the impact of changing parameters of the separation method. Optimizing robustness is important for many method development and validation processes.
5. Method validation.

• Sample: Urine; Bodily fluids; Blood; Saliva; Serum; Plasma; Drugs; Food; Metabolites of illicit and synthetic drugs; Environmental pollutants; Liquid & More
• Equipment: High-Performance Liquid Chromatography (HPLC) System; Fluorescence Detection; UV Detection
• (Optional): High-Performance Liquid Chromatography (HPLC) Columns

1. The chromatographic analysis instrument designed with liquid as mobile phase is called liquid chromatograph, and the liquid chromatograph with high pressure infusion pump, high efficiency fixed phase and high pressure sensitive detector is called high performance liquid chromatograph.
2. There are many kinds of high-performance liquid chromatography, but no matter what type of high-performance liquid chromatography, it is basically divided into four parts: high-pressure infusion device, sampling system, separation system and detection system.
3. Volatile substances are better separated by gas chromatography.