Difference between HPLC and HPTLC and Applications.

Difference between HPLC And HPTLC comparative method

1. DIFFERENCE BETWEEN HPLC AND HPTLC CHOTHISH JAYARAM 3 rd PHARM D 1

2. What is HPLC ? HPLC ( high-pressure liquid chromatography or high-performance liquid chromatography ) is an enhanced form of liquid chromatography. Therefore, it is a type of column chromatography, which involves the high-pressure pumping of a mixture dissolved in the liquid mobile phase through a column filled with the solid stationary phase. Moreover, a carrier gas such as helium or nitrogen is responsible for the movement of the sample through the column. Furthermore, HPLC is a versatile technique which allows the separation and identification of compounds dissolved in the sample with low concentrations such as parts per trillion. The separation of components is due to the differential interactions of each component with stationary and mobile phases. Generally, these differential interactions occur due to the different polarities of components.  What is HPTLC ? HPTLC ( high-performance thin-layer chromatography ) is a sophisticated form of TLC or thin-layer chromatography. Therefore, it provides a high-efficiency separation. Generally, the main difference between HPTLC and TLC is the characteristics of the separation plate. The plate used for HPTLC contains a significantly smaller particle size. It produces a higher packing density as well as a smoother surface. Therefore, with the reduction of the sample diffusion, compact bands or spots can be obtained in HPTLC. On the other hand, the thinner layer in HPTLC increases the detection sensitivity and analysis speed. Moreover, the other benefits of HPTLC include higher reproducibility, sharp bands for quantitative analysis, usability in the mass spectroscopy, etc.     2

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6. 6 APPLICATIONS OF HPLC & HPTLC APPLICATIONS OF HPLC The information that can be obtained using HPLC [1-8] includes identification, quantification, and resolution of a compound. Preparative HPLC refers to the process of isolation and purification of compounds. This differs from analytical HPLC, where the focus is to obtain information about the sample compound. Chemical Separations It is based on the fact that certain compounds have different migration rates given a particular column and mobile phase, the extent or degree of separation is mostly determined by the choice of stationary phase and mobile phase. Purification Purification is defined as the process of separating or extracting the target compound from a mixture of compounds or contaminants. Each compound showed a characteristic peak under certain chromatographic conditions. The migration of the compounds and contaminants through the column need to differ enough so that the pure desired compound can be collected or extracted without incurring any other undesired compound. Identification Generally assay of compounds are carried using HPLC. The parameters of this assay should be such that a clean peak of the known sample is observed from the chromatograph. The identifying peak should have a reasonable retention time and should be well separated from extraneous peaks at the detection levels which the assay will be performed.

7. 7 Other applications of HPLC Other applications of HPLC includes Pharmaceutical applications Tablet dissolution study of pharmaceutical dosages form. Shelf-life determinations of pharmaceutical products Identification of active ingredients of dosage forms Pharmaceutical quality control Environmental applications Detection of phenolic compounds in Drinking Water Identification of diphenhydramine in sedimented samples Bio-monitoring of pollutant Forensics Quantification of the drug in biological samples. Identification of anabolic steroids in serum, urine, sweat, and hair Forensic analysis of textile dyes. • Determination of cocaine and metabolites in blood

8. 8 Clinical Quantification of ions in human urine Analysis of antibiotics in blood plasma. Estimation of bilirubin and bilivirdin in blood plasma in case of hepatic disorders. Detection of endogenous neuro-peptides in extracellular fluids of brain. Application of HPTLC HPTLC Technique can be used for Fingerprint analysis and the determination of identity and quality of Herbal drugs effectively. Applications of HPTLC for phyto -chemical analysis, biomedical analysis, herbal drug quantification, finger print analysis are in the routine practice. HPTLC-MS, HPTLC-FTIR and HPTLC-Scanning Diode Laser mode are powerful analytical tools in the field of analysis. HPTLC method deals with qualitative and quantitative analytical applications such as herbal and dietary supplements, nutraceuticals, and various types of medicines. It is used in quality control, purity check; clinical applications: metabolism studies, drug screening etc.; forensic : poisoning investigations, assaying radiochemical impurities of radiopharmaceuticals, detection and identification of pharmaceutical raw materials, drugs and their metabolites in biological media.

9. 9 CONCLUSION It can be concluded from the entire review that HPLC is a versatile, reproducible chromatographic technique for the estimation of drug products. It has wide applications in different fields in term of quantitative and qualitative estimation of active molecules. HPTLC is an ideal tool for identification of herbal materials. In addition, it is used for semi-quantitative comparison to provide quantitative results. HPTLC use for screening pharmaceutical compounds for the antimicrobial activities is emerging. The uses in validation of new incoming products and its introduction into the regulatory systems are of much importance towards the future of HPTLC.

10. Conclusion   HPLC is an enhanced chromatographic technique of liquid chromatography. Therefore, it contains a solid stationary phase filled into a column. Moreover, it produces results with high efficiency and resolution. On the other hand, HPTLC is an enhanced form of thin-layer chromatography. Therefore, it contains a solid stationary phase fixed on to a plate. However, it produces moderately resolved but, high-efficiency results. Both HPLC and HPTLC are two types of automated chromatographic techniques used to separate components of a mixture. However, the main difference between HPLC and HPTLC is the type of chromatographic technique.   References: “High Performance Liquid Chromatography (HPLC).” HiQ, The Linde Group 2. “HPTLC.” Merck, Merck KGaA, 10

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