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Lecture 3B- Chromatography Drug Analysis and Systems.docx

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Biochemistry 3386B
L.Graham Smith

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Lecture 3 Drug Analysis SystemsInstruments Part II Chromatographic AssaysChromatographic Assays done in Specialty Labs 1 High Performance Liquid Chromatography HPLC 2 GasLiquid Chromatography GC 3 Thin Layer Chromatography TLC 4 Gas ChromatographyMass Spectrometry GCMS 5 HPLCMS LCMSDefine A method used primarily for the separation of components of a sample in which the components are distributed between two phases one of which is stationary and the other is mobile ie Moving The stationary phase can be o Solid o Liquid or gum o Supported by solid or gel o Can be packed in a column spread as a layer or distributed as a filmThe mobile phase can be liquid or gaseousGas Chromatography GCThe support for the stationary phase is a hollow capillary glass quartz o 025 mm diameter o 30m lengthcolumn is lined with stationary phasea rubbery silicone polymer o polymerization within tube molecules stick and interactmobile phase is the carrier gas ie Heliumthere is a separation of components on the basis of their ability to vaporize and the presence of polar functional groups o smaller more in vaporgas phase o larger more in condensedstationary phaseapplicable for the analysis of drugs which can be vaporized o Max MW 800900o Thermally stable o ie NOT cyclosporine MW1100 or phenobarbital thermally unstableresolution is superior to HPLC and far superior to TLCID drugs by their retention timeSome FYI FactsCan be labor intensive requires some special trainingLow sample throughputSample is extracted and concentrated before analysisNitrogenphosphate NP detector is the most common detector for drug analysis amino groups
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