High Performance Liquid Chromatography HPLC originally refered to:

High Performance Liquid Chromatography HPLC  originally refered to:

High Performance Liquid Chromatography HPLC

originally refered to: High Pressure Liquid Chromatography currently refers to: High Precision Liquid Chromatography

high pressure to be able to use small particle size to allow proper separation at reasonable flow rates Applications of Liquid

Chromatography Partition chromatography Adsorption, or liquid-solid chromatography

Ion exchange chromatography Size exclusion, or gel, chromatography Components

Solvent Reservoir and Degassing System Pumps Precolumns Sample Injection System

Columns Temperature Control Detectors Schematic

of an apparatus for HPLC

Components Solvent Reservoir and Degassing System isocratic elution - single solvent separation technique

gradiant elution - 2 or more solvents, varied during separation Isocratic mixture of A (aqueous buffer)

and B (acetontrile) Isocratic elution: performed with a single solvent (or

constant solvent mixture) Gradient Elution

Gradient elution: continuous change of solvent composition to increase eluent

strength Improvement in separation efficiency by gradient elution. Column: 1m x

2.1mm id, precision-bore stainless; packing: 1% PermaphaseR ODS. Sample: 5L of chlorinated benzenes in isopropanol.

Detector: UV photometer (254 nm). Conditions: temperature 60oC, pressure, 1200 psi.

Components Pumps 1000 - 6000 psi at rate of @ 3 mL/min reproducible to 2% screw driven syringe

reciprocating pump A reciprocating pump for HPLC.

Components Precolumns remove impurities from solvent saturates mobile phase with liquid of stationary phase before the analytical column

Components Sample Injection System sample valve syringe

A sampling loop for liquid chromatography Components http://www.youtube.com/watch?v=mPO7Tv2mIJU

Columns straight, 15 to 150 cm in length; 2 to 3 mm i.d. packing - silica gel, alumina, Celite

Normal phase Polar Stationary Phase and Nonpolar Solvent Reverse phase Nonpolar Stationary Phase and Polar Solvent Size exclusion (gel permeation) - Small molecules penetrate into the pores within the packing while larger molecules only partially penetrate the pores. The large molecules elute before

the smaller molecules. Effect of Particle Size Columns Cont.

Components Temperature Control Components

Detectors single wavelength UV tuneable UV diode-array refractive index

MS Performances of LC Detectors Ultraviolet

detector cell for HPLC Diode-Array Absorption

Spectra Absorption spectra of the eluent from a mixture of

three steroids taken at 5-second intervals.

Recently Viewed Presentations

  • "Mirror Image" Lena Coakley - Weebly

    "Mirror Image" Lena Coakley - Weebly

    Plot "Mirror Image" by Lena Coakley is a scientific allegorical tale that depicts the internal quest to find ones true self.The story takes the reader through the experience of a teenager named Alice who underwent a brain transplant into a...
  • Community Language Learning (Cll) - Drrz

    Community Language Learning (Cll) - Drrz

    COMMUNITY LANGUAGE LEARNING (CLL) " " " " Background developed by Charles Curran in the 1970s primary insights from Rogerian counseling theory counselor client (knower) teacher (learner) student Maskowitz's Humanistic Techniques which engages the whole person: blend what students feel,...
  • lubrication.narod.ru

    lubrication.narod.ru

    ОАО ММК ЛПЦ-5 2-х клетевой реверсивный стан холодной прокатки ф. SMS-Demag Общее описание установки Осн
  • Integration with Workforce Solutions and Career Services

    Integration with Workforce Solutions and Career Services

    System focus, not just co-location Required-partner programs under WIOA and state law are "system partners" Focus is on providing customers with access to partner programs whether they are co-located in a Workforce Solutions Office or not
  • Tardigrada - Minnesota State University Moorhead

    Tardigrada - Minnesota State University Moorhead

    Tardigrada Tardigrada: "water bears" First discovered in 1773 800 spp 0.1-0.5mm in length Widespread Live in semi aquatic habitats, water film Cryptobiotic, some are cyclomorphic Temp range from +149C to -272C, withstand high vacuum, high UV, long periods without O2...
  • Breast is Best - Yola

    Breast is Best - Yola

    Use a balloon whisk to whisk eggs and yolks in a bowl. while whisking, gradually add cream mixture until well combined. Pour evenly among four 1-cup capacity ovenproof dishes. Place in a roasting pan. Pour enough boiling water into pan...
  • Home Usability - RTC:Rural

    Home Usability - RTC:Rural

    Look at home usability across three urban communities using a random sample of approx. 2,500 address from zip codes surrounding local CILs in: Atlanta, Georgia (33 Zips) Fresno, California (25 Zips) Indianapolis, Indiana (30 Zips) Total Design Survey Method (Dillman)
  • World Cities - SLC Geog A Level Blog

    World Cities - SLC Geog A Level Blog

    Urban Processes Many developing cities are continuing to grow though a number of processes: Urban sprawl: Urban areas growing outwards in an uncontrollable way. In the developing world this results in slums Suburbanisation: The wealthy choosing to live on the...