Introduction to Analytical Science Report - Science Assignment Help

Assignment Task

Question 1 Describe the issues involved in sampling and analysis of a blood sample.

Question 2 Ibuprofen is a propionic acid derivate and nonsteroidal anti-inflammatory drug (NSAID) with antiinflammatory, analgesic, and antipyretic effects. The chemical structure of ibuprofen is reported below.

Each aliquot of 0.5 ml plasma was spiked with 25 µL of I.S. solution (final concentration 4 µg mL-1), mixed by vortex-mixing for 30 s, then equilibrated for 10 min at room temperature. Acetonitrile (1 ml) was then added and the sample was vortex-mixed for 30 s. Following centrifugation in a clinical centrifuge at 1200 g, the supernatant was removed into a screw-top tube and mixed with 8.5 ml of 10 mM phosphoric acid, pH 2. Extraction of drug and metabolite from plasma was done by solid-phase extraction. The extraction cartridges (Bond-Elut, C18, size 100 mg) were prewetted sequentially with 4 ml each of methanol, water, then 10 mM phosphoric acid, before loading the diluted supernatant from the plasma samples. The sample was loaded with vacuum gravity flow onto a Bond-Elut cartridge, then the column was washed with 2 ml of acetonitrile-10 mM phosphoric acid (20:80, v/v). IBP was eluted by gravity with 1 ml of acetonitrile-10 mM phosphoric acid (50:50, v/v), with a final centrifugation for 3 min at 1800 g to collect all the eluent. The eluent was evaporated to dryness under a stream of nitrogen gas at 37°C, the residue was reconstituted into 400 µL of acetonitrile-methanol-1% acetic acid (pH 3) (10:15:75, v/v), and the sample was dissolved by ultrasonic bath for 3 min and vortex-mixing for 1min. Finally, 100 µL were injected onto the HPLC system with the chromatography conditions as described above for the analytical HPLC system.

Please address the following questions:

• Were proteins removed from the sample and how? (max 150 words)
• Why was the sample mixed with 10 mM phosphoric acid, pH 2? (max 100 words)
• How was the conditioning accomplished? and the equilibration? (max 100 words)
• Was the sample concentrated and if so by how many times? What are the implications of this in the analysis (max 200 words)?
• The human serum #14 was run in liquid chromatography and a peak at rt of IBP provided a signal equal to 20000 mAu. Quantify the true mass of IBP which was present in the original volume by using the calibration curve below. Explain your reasoning (max 200 words).

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