Analysis of Glycated Hemoglobin by HPLC Technology Shalini Kini Product Specialist Diabetic EpidemiologyWhere Do we Stand!?! World Scenario * IDF Diabetes Atlas 5th Edition 2012 update. http://www.idf.org/diabetesatlas/5e/Update2012 Did U Know!!??! One Adult in Ten will have diabetes by 2030!!! Year 2012 Year 2030 63.01m India 101.2m India • India Ranked Second in the world in Diabetes Prevalence‚ behind China. *IDF Diabetes Atlas- Fifth Edition http://www.idf
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purification and lysing methods: Brad Ford Assay‚ SDS-PAGE‚ affinity purification‚ lysozyme treatment‚ nickel column‚ sonication‚ and flavin reeducate activity assay. His-tagged protein is expressed in E. coli from pGhis‚ with the T7 RNA lac repressor induction system.5 Sonication is used‚ it’s why bacteria are lysed. The tag allows purification for ion affinity chromatography‚ following a gel filtration column‚ defined as removing ions and other small molecules. Where then‚ the Bradford assay was made use of
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C2.1.1 Structure and bonding a) Compounds are substances in which atoms of two or more elements are chemically combined. b) Chemical bonding involves either transferring or sharing electrons in the highest occupied energy levels (shells) of atoms in order to achieve the electronic structure of a noble gas. c) When atoms form chemical bonds by transferring electrons‚ they form ions. Atoms that lose electrons become positively charged ions. Atoms that gain electrons become negatively charged
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Backround: Gas Chromatography separates organic samples much in the same way as column chromatography. The only differences are that it uses a moving gas phase and a stationary liquid phase‚ and that the temperature of the gas system can be controlled. In a gas chromatograph the sample is shot in with a syringe and is immediately vaporized in a heated injection chamber. It is then introduced to a moving stream of gas called the carrier gas which sweeps the vaporized sample into a column filled with
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| 10-15 | Beer | S7 | 10 | 15 | 0 | 2-5 | Table 1.1 Showing the solutions made for analysis and the expected percentage of ethanol in each sample. After creating the solutions we would run the solutions through the Gas Chromatography instrument. The Gas Chromatography instrument was a Focus GC and the experimental
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Experiment 1: Preparation of 1-Bromobutane and Reactivity of Alkyl Halides Objective: The purpose of this lab is to prepare 1-bromobutane from 1-butanol in an acid-catalyzed substitution reaction. While the reaction would be expected to occur as SN2 due to the primary nature of the substrate‚ because H2SO4 is used as a solvent‚ the conditions are very polar and the reaction can proceed via an SN1 reaction. The main objective is to obtain test results to determine the mechanism of the reaction
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A rotovap was then used to concentrate the solution to dryness. When packing the chromatography column‚ cotton is added to prevent the
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Chapter 1: Measuring the amount of substance Analytical chemistry: science of chemical measurement. Its object is the generation‚ treatment and evaluation of signals from which information is obtained on the composition and structure of matter Measurement: process of obtaining the magnitude of a quantity Example: The amount of saturated fat in the sample is 3 g/serving. Quantity: attribute of a phenomenon that may be distinguished qualitatively and determined quantitatively Value: magnitude
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The resultant solution diluted up to the mark with diluents to obtain 1000 μg/ml. Further stock solution was used to prepare plasma standards to construct the calibration curve. C. Pre-chromatographic isolation of baclofen from plasma Prior to chromatography‚ baclofen from plasma constituents was isolation by adding 0.25 ml of acetonitrile and approximately 25 mg of zinc sulphate crystals to 0.50 ml of plasma. The aged plasma was stored at - 250C. This plasma was thawed at room temperature and 0.50
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phosphate pathway. The homogenate was made by adding the buffer to the liver of the quail after one hour centrifuge at 12300 RPM materializing at 4°C. Precipitation protein was working by ammonium sulfate; it loaded directly to column 2’‚ 5’ ADP Sepharose 4B affinity chromatography‚ which has been a high affinity to NADP+ substrate. The flow rate of elution was 23 ml/hr. Taking 18 fractions‚ checking activity for each fraction at 340 nm. The activity found in fraction tube six to fourteen. This process
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