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High-Performance Liquid Chromatography
Types of Techniques
- Liquid Chromatography Mass Spectrometry (LC-MS)
- Gas Chromatography-Mass Spectrometry (GC-MS)
- High-Performance Liquid Chromatography (HPLC)
- Inductively Coupled Plasma Mass Spectrometry (ICP-MS)
- Combustion Ion Chromatography (CIC)
- Ion Chromatography
- Gel Permeation Chromatography (GPC) / Size Exclusion Chromatography (SEC)
- Multi-Column Chromatography (MCC)
- LC-Orbitrap
- Accelerator Mass Spectrometry (AMS)
- Nano scale Secondary Ion Mass Spectroscopy
- Secondary-ion mass spectrometry (SIMS)
- SEC-MALS
High-Performance Liquid Chromatography (HPLC)
HPLC is mainly used for the separation, identification, and quantification of components in the sample matrix. This analytical technique can study the non-volatile, thermally unstable, and polar components. The high-pressure chromatography column has been used to separate the components and liquids followed by quantification and detection. The sample is dissolved using the suitable solvent and is passed through the stationary phase (HPLC column). In HPLC, 44 the components are separated based on the difference in the affinity between the mobile phase and the stationary phase. The components with the greater affinity exit from the column after a longer duration as compared to the components with less affinity. All the components were detected using the detector and peak intensity is employed to determine the components and its quantity. This technique works based on various separation mechanisms such as reverse phase, normal phase, ion pair, ion exchange, and size exclusion chromatography. The determination of active ingredients in the product, determination of molecular weight, separation and purification, and polymer formulation can be accomplished using this technique.
The specific intermolecular inter- actions between the molecules of a sample and the packing material define their time “on-column”. Hence, different constituents of a sample are eluted at different times. Thereby, the separation of the sample ingredients is achieved.
HPLC is crucial in the pharmaceutical industry for analyzing drug purity, identifying impurities, and ensuring the consistency of formulations. It plays a key role in quality control by quantifying active pharmaceutical ingredients (APIs) and monitoring degradation products. HPLC methods are essential for regulatory compliance, supporting the development, validation, and routine testing of pharmaceutical products to meet stringent quality standards and ensure patient safety.
Food Science and Safety:HPLC plays a critical role in food science and safety by detecting and quantifying contaminants such as pesticides, mycotoxins, and heavy metals in food samples. It ensures compliance with regulatory limits, safeguarding public health and enhancing food safety. Additionally, HPLC analyzes nutrients, additives, and flavors, supporting quality control measures and ensuring accurate labeling of food products.
Environmental Monitoring:HPLC is utilized in environmental monitoring to analyze organic pollutants such as pesticides, herbicides, and industrial chemicals in water, soil, and air samples. It provides precise quantification of contaminants, aiding in regulatory compliance and assessing environmental impact. HPLC’s sensitivity and versatility make it indispensable for monitoring pollution levels and ensuring environmental health and safety.
Forensics:In forensics, HPLC is instrumental for analyzing drugs, toxins, and poisons in biological fluids and tissues obtained from crime scenes or post-mortem examinations. It facilitates the identification and quantification of substances such as opioids, cocaine metabolites, and pharmaceuticals, aiding in criminal investigations and toxicological assessments. HPLC’s high sensitivity and ability to separate complex mixtures make it a valuable tool in forensic laboratories for accurate and reliable chemical analysis.
Chemical Industry:In the chemical industry, HPLC is widely used for quality control, process optimization, and product development. It enables the analysis of raw materials, intermediates, and final products to ensure purity, identify impurities, and monitor reaction kinetics. HPLC’s capability to separate and quantify components in complex mixtures makes it essential for maintaining high standards of product quality and ensuring regulatory compliance.
Other Industries:- Cosmetics and personal care product analysis: Identifies and quantifies ingredients in cosmetics and personal care products.
- Consumer product testing: Analyzes the composition of consumer products to ensure safety and identify potential hazards.
- High Resolution: HPLC can separate complex mixtures with high resolution, allowing for the identification of closely related components.
- Versatility: HPLC can analyze a wide range of samples, including non-volatile, heat-sensitive, and ionic compounds.
- Sensitivity: Modern HPLC detectors offer high sensitivity, allowing for the detection of trace levels of analytes.
- Quantification: HPLC provides quantitative information about the amount of each component present in the sample.
- Solid Sample – 1 gm.
- Liquid Sample – 1 ml.