Role of lab in tox
- 1. العظي هللا صدقم االسراء سورة(85)
- 2. Role of laboratory in Toxicological Studies Prepared by Reda M. Elsayed 29/4/2015
- 4. Types of Biological samples: 1- BODY FLUIDS: •Blood (plasma, serum and whole blood) •Urine, semen, saliva, gastric contents..etc. 2- Tissue samples: liver, kidney, muscles….etc and Hair.
- 9. a) Biochemical and toxicological study Homogenized by Tissue Homogenizer/ Grinder
- 12. b) Histopathology exam.: Organs Preserved as it is in different preservative according to type of tissue then processed by the specialist.
- 13. a- Biochemical Tests. b- Toxicological Tests. c- Histopathological Exam.
- 14. 1. Spectrophotometery: Used for liver and kidney functions, glucose, pseudocholine estrase...etc 2. Immunoassay ( ELISA and RIA): Hormones and some enzymes….etc.
- 15. 1. Spectrophotometery 2. Immunoassay ( ELISA and RIA): 3. Chromatography: TLC HPLC, LC- MS GC, GC-MS
- 18. •“Immuno” refers to an immune response that causes the body to generate antibodies. •“Assay” refers to a test. Immunoassays are chemical tests used to QUALITATIVE or SEMIQUANTITATIVE to determine a specific substance, the analyte, in a blood or body fluid sample, using an immunological reaction.
- 19. RIA In which, there is a competition between two antigens: the labeled antigen reagent (Ag*) and the unlabeled specimen (Ag) (or test sample analyte) Both compete for a limited amount of antibody.
- 20. Tramadol PROCEDURE The assay is based on the competition of Tramadol labeled enzyme (G6PDH) and the free drug in the urine sample for the fixed amount of antibody binding sites. In presence of the drug, The enzyme (G6PDH) is free so activity is determined by the conversion of NAD to NADH…. Colour change …positive test. In the absence of the free drug in the sample, the antibody binds the drug enzyme conjugate and enzyme activity is inhibited.
- 22. ELISA An enzyme-linked immunosorbent assay (ELISA) used to detect the presence and/or amount of a target protein of interest within an experimental sample. Detection of the target protein is made by antibodies, which make the ELISA an immunoassay. Through a series of incubation and washing steps.
- 24. Architect plus I 1000 Immuno- analyser
- 27. Liquid-liquid extraction is a useful method to separate components (compounds) of a mixture
- 28. For example: Suppose that you have a mixture of sugar in vegetable oil (it tastes sweet!) and you want to separate the sugar from the oil. You observe that the sugar particles are too tiny to filter and you suspect that the sugar is partially dissolved in the vegetable oil.
- 29. How about add water to the mixture Will it separate the sugar from the oil? Sugar is much more soluble in water than in vegetable oil, and, as you know, water is immiscible (=not soluble) with oil. هليختلطالزيت بالماء The water phase is the bottom layer and the oil phase is the top layer, because water is denser than oil.
- 30. By shaking the layers (phases) well, you increase the contact area between the two phases. The sugar will move to the phase in which it is most soluble (the water layer)
- 32. In this example: water was the extraction solvent. the original oil-sugar mixture was the solution to be extracted. sugar was the compound (analyte) extracted from one phase to another.
- 33. the concept of liquid-liquid extraction: Liquid-liquid extraction is based on the transfer of a solute substance from one liquid phase into another liquid phase according to the solubility. You can use extraction to separate a substance selectively from a mixture, or to remove unwanted impurities from a solution.
- 34. In the practical use, usually one phase is a water or water-based (aqueous) solution and the other an (organic solvent) which is immiscible with water. The success of this method depends upon the difference in solubility of a compound in various solvents. And the choose of a suitable extraction solvent.
- 35. 1. Thin layer chromatography 2. Liquid Chromatography 3. Gas Chromatography (GC)
- 36. Chromatographic separation process is based on the difference in the surface interaction of the analyte (sample) and eluent molecules (mobile phase) Analyte molecules while moving through the porous packing materials tend to interact with the surface adsorption .
- 39. C)GC-MS:
- 40. HPLC is a form of liquid chromatography used to separate compounds that are dissolved in solution. Compounds are separated by injecting a sample mixture onto the column. The different components in the mixture pass through the column at differentiates due to differences in their partition behavior between the mobile phase and the stationary phase.
- 41. Solvent (mobile phase) Solvent Delivery System (Pump) Sample Injector Column Detectors (Diode Array) Data Collection
- 42. The column is one of the most important components of the HPLC chromatograph. It is made out of stainless steel tubes with a diameter of 3 to 5mm and a length ranging from 10 to 30cm. Normally, columns are filled with silica gel. Silica is inert to most compounds and has high surface activity which can be modified easily with water and other agents. Silica can be used to separate a wide variety of chemical compounds.
- 44. Photo-Diode Array UV detector (200-400nm ) Fluorescence Electrochemical Mass- Spectrometric IR Absorbance
- 46. Retention time (Rt): It is the characteristic time it takes for a particular analyte to pass through the system (from the column inlet to the detector) under set conditions.