The solution homogeneity expelled, by centrifugation for 10 min. The sample was centrifuged and separated into two layers, and took the top of the sample is injected for HPLC (11,12). Measured concentration in total lipid: The total fats balanced concentration of the pesticide getting by dividing the measured pesticide residue concentration in the overalll tissue sample by the decimal fraction of the sample that consisted of ether-extractable lipid. The total lipid content of each specimen was estimated from its total cholesterol & triglycerides levels by using a summation method. Analytical results for organochlorine pesticides were reported on a lipid-adjusted basis (nanograms per gram or parts per billion) (14).
known as cation-exchange or anion-exchange chromatography, depending on whether the solutes to be exchanged are positively or negatively charged. Size Exclusion Chromatography: Here the molecules are separated according to their molecular weight and it is suitable for molecules having molecular weight of 2000 Daltons or more. Largest molecules are eluted first and the smallest molecules last. Affinity Chromatography: Here the stationary phase contains specific groups of molecules which can absorb the sample if stearic and charge related conditions are satisfied d. This technique is used to isolate prooteins, enzymes as well as antibodies from m mixtures. Partition Chromatography: Here the stationary phase is a thin liquid film either adsorbed or
After that the seeds were crushed to a fine paste with the 10ml of buffer added as the seeds were crushed, the paste is then subsequently filtered into a measuring cylinder. The solution collected after filtration is the amount of amylase extracted solution (AE), with its volume being recorded. The amylase extracted solution(5ml) is then transferred into a beaker, then a five-fold dilution of the amylase extract using 20ml of buffer. Resulting in 25ml of diluted amylase extract (DAE). A control extract is prepared (5ml of DAE) to a test tube, which is then placed in boiling waterbath for 10minutes, after 10minutes remove the control extract and leave it to cool at room temperature.
After which the digestions were examined by gel electrophoresis. The samples were run on a 50 mL 0.9% (w/v) agarose gel in 1X TAE buffer at 100 V until the leading track dye traveled 2/3 the distance of the gel. The gel was then soaked in GelRed for 20 minutes and examined under UV light. To prepare the digestions 10 μL of each digestion was mixed with 2 μL of 6X track dye in a micro centrifuge tube. 12 μL of 1 kb DNA ladder and each digestion was run on the
Rose Bengal-(bis(aminoethyl)ethylene glycol) (2) from Rose Bengal disodium salt (1) The synthesis was done following procedure from . Rose Bengal Na+ salt (915 mg, 0.90 mmol) was dissolved in DMF (2ml) and DIPEA (0.312 ml, 1.80 mmol), HATU (308 mg, 0.81 mmol) were added. After activation for 15 min, the mixture was added to O-Bis-(aminoethyl)ethylene glycol trityl resin (309 mg, 0.31 mmol) preswollen in DMF for 2 hours. The coupling reaction wrapped in aluminum foil was allowed to proceed overnight on a nitrogen bubbler apparatus. The resulting red-burgundy coloured resin was filtered and washed well with DMF.
The developing chamber for the TLC plates was prepared by adding ethyl acetate that contained 0.5% acetic acid to the glass jar. The TLC plates were prepared by drawing a horizontal line from bottom margin (0.5cm) where the four spots were placed. Separate small capillary tubes were used to spot solutions of Tylenol, Anacin, acetaminophen and acetylsalicylic acid respectively on the drawn line. The spotted TLC plate was placed in the developing chamber (glass jar) ensuring the solution in the glass jar is below the drawn line, followed by covering the top of the glass jar(Qui, Haixing, and Yusheng
The volumetric flask was then filled up to its 100 mL mark with deionized water. The buret was washed out with dionized water and then with the strong base NaOH before being filled up with NaOH. About 20 mL of the unknown weak acid was pipetted into a beaker. The starting volume of the NaOH in the buret was recorded before about 4 mL of the strong base was titrated into the weak acid solution. The final volume was recorded.
The final volume was adjusted with the same solvent to get concentration of 100 µg/ml. the solution was further diluted to get the concentration of 10 µg/ml, filtered through 0.45 µm filter tips , and aliquots of 20 µl from this solution was injected into HPLC by using an
(2004) with some modifications. Briefly, 10 µL of liver homogenate were mixed with 90 µL of low melting point agarose (0.7% in PBS) at 37 ºC and loaded on a fully frosted slide coated with 110 µl of normal melting point agarose (1% in PBS). Then this slide was coverslipped and the agarose layer was left 10 min at 4 °C to solidify. The cover slip was removed and the previous step was repeated for 5 min at 4 ºC. The slides were placed for 2 hrs at 4 °C in a lysis buffer containing (2.5 mol/L NaCl, 100 mmol/L Na2EDTA, 10 mmol/L Tris, [pH 10] and a freshly prepared 1% Triton X-100 and 10% Dimethyl sulfoxide were added to the buffer just before use).
The ratio of the absorbances of the glycosylated hemoglobin & the total hemoglobin fraction of the control and the test is used to calculate the percent glycosylated hemoglobin of the sample . Reagents: 10 Tests 25 Tests Ion Exchange Resin (Predispensed Tubes) 10 × 3ml 25 × 3ml Lysing reagent 5 ml 12.5 ml Resin separators 10 pieces 25 pieces Procedure: Wavelength: 415 nm (Hg 405 nm) Temperature: R.T Light Path: 1cm A.Hemolysate preparation : 1-Dispense 0.5 ml lysing reagent in to tube as labeled as test (T) 2-Add 0.1 ml of the reconstituted well mixed blood sample into the appropriatelylabeledtubes. Mix unit completes lysis is evident. 3-Allow to stand for 5 minutes . B.Glycosylated haemoglobin (GHb) separation : 1-Remove cap from the ion – exchange resin tubes and label as test .