English (United Kingdom)

https://curated-unify.zendy.io/wp-json/zendy-region/v1/featured_content/oa?rat=en

https://curated-unify.zendy.io/wp-json/zendy-region/v1/highlighted_journal/

Global: Zendy Tools annual

Presents the keyphrase highlighting as premium feature

Keyphrase Highlighting

Presents the summarisation as premium feature

Summarisation

Insights

Presents the pdf analysis as premium feature

PDF Analysis

Presents the zaia usage as premium feature

ZAIA

Global: Zendy Tools monthly

Global: Zendy Plus monthly

Presents the access of premium content as premium feature

Premium Content

Global: Zendy Plus annual

Global: Zendy Free Plan

HPLC is an analytical technique widely used for identification, separation, detection and quantification of various drugs and its related degradants. Appropriate mobile phase, stationary phase, column, column size, temperature, wavelength and gradient selection is important for the suitable compatibility and stability of drug as well as impurities and degradants. Most of the drugs as well as other compounds can be analyzed by HPLC method because of the several advantages like rapidity, specificity, accuracy, precision and ease of automation in this method. This article has been prepared with an aim to review different aspects of HPLC, such as principle, types, mode of separation, characteristics, instrumentation, important parameters and various applications of HPLC in different fields. Keyword: HPLC, Analysis, Method development and Validation, Detection, Identification, Quantitation. Introduction High-performance liquid chromatography or High pressure liquid chromatography (HPLC) is a specific form of column chromatography generally used in biochemistry and analysis to separate, identify, and quantify the active compounds. 1 HPLC is the method of choice for checking peak purity of new chemical entities, monitoring reaction changes in synthetic procedures or scale up, evaluating new formulations and carrying out quality control / assurance of the final drug products. 2,3 HPLC is now one of the most powerful tools in analytical chemistry. It has the ability to separate, identify, and quantify the compounds that are present in any sample that can be dissolved in a liquid. HPLC is the most accurate analytical methods widely used for the quantitative as well as qualitative analysis of drug product and used for determining drug product stability. 4,5 High pressure liquid chromatography is the full form for HPLC and as given in the name, there is use of high pressure in the principle of its operation. Also due to its efficiency in analysis of compounds it is regarded as High performance liquid chromatography. Some have even gone to the extent of calling it as High patience liquid chromatography based on the long human time requirement and patience needed in its operation. HPLC is basically a highly improved form of column chromatography. Instead of allowing the solvent to drip through a column under just the force of gravity, it is externally forced through the column under high pressures of up to 400 atm. This makes the chromatographic process a lot faster. It also allows the use of very small particle size for the column packing material which gives a much greater surface area for interactions between the stationary phase and the molecules flowing through it. Thus, it allows a much better separation of the components of the mixture. 6,7 Now a days, highperformance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) is one of the most versatile analytical tools available due to its rapid and diverse growth in applications in the analysis of drugs and metabolites from biological fluids. In HPLC, the essential equipment consists of an eluent, reservoir, a high-pressure pump, and an injector for introducing the sample, a column containing the stationary phase, a detector and recorder. 8 The development of highly efficient micro particulate bonded phases has increased the versatility of the technique and has greatly improved the analysis of multi component mixtures. 9 Principle HPLC principle is that solution of sample is injected into a column of porous material (stationary phase) and liquid phase (mobile phase) is pumped at higher pressure through the column. The principle of separation followed is the adsorption of solute on stationary phase based on its affinity towards stationary phase. 2 HPLC is a special branch of column chromatography in which the mobile phase is forced through the column at high speed. As a result the analysis time is reduced by 1-2 orders of magnitude relative to classical column chromatography and the use of much smaller particles of the adsorbent or support becomes possible increasing the column efficiency substantially. 10

A review of HPLC technique covering its pharmaceutical, environmental, forensic, clinical and other applications