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Analytical technique
Clinical Chemistry Ch. 5
| Term | Definition |
|---|---|
| Chromatography | technique used to separate complex mixtures based on different physical interactions between the compounds and the instrument thin-layer chromatography, gas chromatography |
| Ion-Exchange Chromatography | separates ions by charge |
| Thin-Layer Chromatography | used to: monitor the progress of a reaction identify compounds present in a given substance determine the purity of a substance |
| High Performance Liquid Chromatography (HPLC) | uses pressure to increase the speed of separation |
| Gas Chromatography | used to separate mixtures of volatile or substances that can be made volatile |
| Gas Chromatography-Mass Spectrometry (GC-MS) | completed method that is used to identify components of a test substance applications: drug detection fire investigation environmental analysis explosive investigation identification of unknown substances |
| Spectrophotometry | instrument used to measure the light transmitted by a solution to determine the concentration of the light-absorbing substances in the solution |
| Electrophoresis | the migration of charged solutes or particles in an electrical field |
| capillary electrophoresis: | performed in a capillary tube |
| Electrophoresis samples | serum (should be diluted before testing due to high amounts of proteins) CSF (should be concentrated before testing) urine (should be concentrated before testing) |
| Electrophoresis steps | sample is placed at the cathode (-) charged pole and migrates to the anode (+) charged pole cellulose acetate-one of the most common support media used in clinical labs |
| Iontophoresis | migration of small ions |
| Osmometry | measures concentration of solid particles in a solution |
| vapor pressure: | pressure at which the liquid solvent is in equilibrium with the water vapor |
| freezing point: | temperature at which the vapor pressures of the solid and liquid phases are the same |
| boiling point: | temp at which the vapor pressure of the solvent reaches on atmosphere |
| osmotic pressure: | pressure that allows solvent to flow through a semipermeable membrane |
| Fluorometry | use fluorescent molecules for measuring emission is a more specific and sensitive method than spectrophotometry is very sensitive to environmental changes such as pH, temp, or UV light |
| Chemiluminescence | light produced by a chemical reaction |
| Beer's Law | relationship between absorption of light and concentration of a substance is directly proportional to the amount of light absorbed or inversely proportional to the logarithm of the transmitted light |
| Anion | negative charged ion migrates toward the anode (positive pole) |
| Cation | positive charged ion migrates toward the cathode (negative pole) |
| Anode | positive pole |
| Cathode | negative pole |
| Electrophoresis Support Media | paper, cellulose acetate, agarose gel film, starch gel, polyacrylamide gel |
| Atomic Absorption Spectrophotometer | measures concentrations by detecting absorption of electromagnetic radiation by atoms rather than molecules usual light source is a hollow-cathode lamp |
| Spectrophotometer Components | light source: incandescent tungsten or tungsten iodide lamp deuterium-discharge and mercury arc lamp are used in the ultraviolet UV radiation monochromator: colored glass filters, interference, filters, prisms, and diffraction gratings |
| Electromagnetic Radiation | short wavelengths correspond to high energy and long wavelengths correspond to lower energy visible light: 400 nm (violet) - 700 nm (red) |
| Spectrophotometer QA | check wavelength accuracy, stray light, and linearity |
| Stray Light | wavelength of light outside the measured range; can be caused from scratches on optical surface, or dust particles |
| 4 Chromatography | ion-exchange thin-layer high performance liquid gas |
| Turbidity | measurement of transmitted light (from one source to another) measurements are based on size of particles as well as concentration measurements made using a spectrophotometer for amount of particles in specimen |
| Nephelometry | a measurement of scattered light based on wavelength and particle size measured by a spectrophotometer differs from turbidity in that only small particles are measured instead of all particles |
| LASER | Light Amplified by Stimulated Emission of Radiation |
| Visible Light | 400nm (violet)-700nm (red) |
| Fluorometry | uses fluorescent molecules for measuring emission more specific and sensitive than spectrophotometry very sensitive to environmental changes such as pH, temp, or UV light |
| Chemiluminescence | light produced by a chemical reaction differs from fluorescence in that np excitation radiation or monochromators are needed |
| Ion Selective Electrodes | charged particles move towards the opposite charged electrode |
| Universal ISE | pH electrode |
| Zone Electrophoresis | migration of charged macromolecules in porous support medium (paper, cellulose acetate, agarose gel film) |
| Thin-Layer Chromatography Use | monitor the progress of a reaction identify compounds present in a given substance determine the purity of a substance |
| HPLC Benefits | uses pressure to increase the speed of separation detection methods are very sensitive and highly automated allows smaller particle size to be used |
| Gas Chromatography | used to separate mixtures f volatile or substances that can be made volatile tests substances purity, identifies a compound, or separate different components of a mixture |
| Gas Chromatography-Mass Spectrometry Application | drug detection fire investigation environmental analysis explosive investigation identification of unknown substance |
| Serum Osmolality Formula | (2 x (Na + K)) + (BUN / 2.8) + (glucose / 18) |
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