PilonM-Experimental

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School

University of Minnesota-Twin Cities *

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Course

2111

Subject

Chemistry

Date

Dec 6, 2023

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docx

Pages

2

Uploaded by ChancellorIron12716

Materials: Five solutions were made through the mixture of five liquid antihistamines. The antihistamines that were added were Amitriptyline, Triprolidine, Promethazine, Pheniramine, and Pyrilamine which were all purchased from Sigma Aldrich. The dead time marker and the internal standard, Uracil and Hexanophenone, were purchased from Acros. Method: The five antihistamines were added to 10 mL volumetric flasks in varying concentrations through the use of micropipettes to create different solutions. The amounts of each antihistamine added to the solutions is outlined in Table 1. A dead time marker and an internal standard, Uracil and Hexanophenone respectively, were also added to each solution with consistent volumes. Each flask was diluted to the line with eluent A. The flasks were inverted to create a homogenous solution. Each solution was drawn up into a syringe and then pushed out through a nylon tip to filter the samples. These samples were put into labeled 1 mL Eppendorf tubes. 500 μL of the unknown were added to a 10 mL volumetric flask along with the dead time marker and internal standard added in the same amounts as the other samples. This solution was diluted with eluent A to the line and inverted several times. Table 1. Volume in μL Solute Solution 1 Solution 2 Solution 3 Solution 4 Solution 5 Amitriptyline 40 50 60 70 80 Triprolidine 50 60 70 80 40 Promethazine 60 70 80 40 50 Pheniramine 70 80 40 50 60 Pyrilamine 80 40 50 60 70
Uracil 50 50 50 50 50 Hexanophenone 90 90 90 90 90 A blank was run in the HPLC before any of the solutions were analyzed. The syringe was prepared for injection by rinsing with eluent A in triplicate. 50 μL of eluent A were drawn into the syringe without the presence of air bubbles. The solution was injected into the injector valve in the load position. To start the method, the injector valve was moved to the downward position. After the method was completed, the blank run was analyzed for peaks. Once no peaks were observed, the samples could be run. This process was repeated for each of the five samples as well as the unknown which was run in triplicate. After each of the solutions were analyzed by the instrument, the seven largest peaks were selected and manually integrated. Instrumentation: A micropipette with a range from 10 μL to 100 μL was used to prepare the solutions, while a micropipette with a range from 100 μL to 1000 μL was used to prepare the unknown. Plastic syringes with nylon tips were used to filter the solutions and the unknown. A 50 μL syringe was used to inject the solutions into the HPLC instrument. The HPLC used for this experiment was purchased from Agilent Technologies and was the 1200 Infinity Series. The stationary phase of the HPLC used was silica based, namely C18. The injection port was 3.5 μm while the diameter and length of the column was 4.6×50 mm.
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