Presentation HPLC-MSfor biomedical bio enginnering
- 1. HPLC-MS High-Performance Liquid Chromatography-Mass Spectrometry Technique & Applications A Presentation By Mr. MD SUMON ALI
- 2. INTRODUCTION High-Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS) is a powerful analytical technique that combines the separation capabilities of HPLC with the detection and identification capabilities of MS. This technique has become a crucial tool in various fields, including Biomedicine, pharmaceuticals, biotechnology, environmental monitoring, and food safety.
- 3. What is HPLC? - A highly sophisticated analytical technique for separating complex biological mixtures. • - Operates under high pressure for precise resolution of components.
- 4. What is MS? • A method that measures the mass-to-charge ratio of ions, providing detailed molecular and isotopic data.
- 5. How does HPLC-MS work In HPLC-MS, the HPLC system separates the components of a mixture based on their interactions with the stationary phase and the mobile phase. The separated components are then introduced into the MS system, where they are ionized and detected based on their mass-to-charge ratio.
- 6. COMPONENET HPLC Component: 1. Solvent Reservoir: Holds the mobile phase solvents, which are used to separate the analytes. 2. Pump: Delivers the mobile phase solvents to the column at a precise flow rate. 3. Injector: Introduces the sample into the mobile phase stream. 4. Column: Separates the analytes based on their interactions with the stationary phase and mobile phase. 5. Detector: Measures the absorbance or fluorescence of the analytes as they elute from the column. MS Component: 6. Ion Source: Ionizes the analytes, converting them into charged particles. 7. Mass Analyzer: Separates the ions based on their mass-to-charge ratio (m/z). 8. Detector: Measures the intensity of the ions as they reach the detector.
- 7. HPLC-MS Technique: Sample Preparation: Prepare the sample by filtering, diluting, or extracting the analytes of interest. HPLC Separation: Separate the analytes using HPLC, with the column and mobile phase conditions optimized for the specific analysis. Ionization: Ionize the analytes using the chosen interface (e.g., ESI or APCI). Mass Analysis: Separate the ions based on their m/z using the mass analyzer (e.g., quadrupole, time-of-flight, or orbitrap). Detection: Measure the intensity of the ions as they reach the detector. Data Analysis: Analyze the resulting mass spectra to identify and quantify the analytes.
- 8. APPLICATIONS IN BIOMEDICINE Pharmacokinetics and Pharmacodynamics: HPLC-MS is used to study the absorption, distribution, metabolism, and excretion of drugs and their metabolites in the body. Biomarker Discovery: HPLC-MS is used to identify and quantify biomarkers for diseases, such as cancer, diabetes, and cardiovascular disease.
- 9. • Toxicology: HPLC-MS is used to study the toxicity of drugs, chemicals, and environmental pollutants, including the identification and quantification of toxic metabolites. • Infectious Disease Diagnosis: HPLC-MS is used to identify and quantify microorganisms, such as bacteria, viruses, and fungi, in clinical samples. • Clinical Chemistry: HPLC-MS is used to measure the levels of various analytes, such as hormones, vitamins, and minerals, in biological fluids.
- 10. Cancer Research: HPLC-MS is used to study the molecular mechanisms of cancer, including the identification and quantification of cancer biomarkers.
- 11. Neurodegenerative Disease Research: HPLC-MS is used to study the molecular mechanisms of neurodegenerative diseases, such as Alzheimer's and Parkinson's. Parkinson's disease and Alzheimer's disease are progressive brain diseases caused by gradual damage to brain cells. Each of these conditions has its own set of symptoms, stages, and treatments.
- 12. In conclusion, HPLC-MS is a powerful analytical technique that has become a crucial tool in various fields. Its high sensitivity, specificity, and quantitative capabilities make it an ideal technique for the analysis of complex mixtures. THANK YOU
- 13. Reference • "Introduction to Modern Liquid Chromatography" by Lloyd R. Snyder, Joseph J. Kirkland, and John W. Dolan.“ • Mass Spectrometry: Principles and Applications" by Edmond de Hoffmann and Vincent Stroobant. • "High-Performance Liquid Chromatography" by Sandie Lindsay. • Articles and application notes from analytical instrument manufacturers like Agilent, Waters, Thermo Fisher Scientific, and Shimadzu. • Review papers and open-access journals discussing HPLC-MS principles.