Diacetyl monoxime, chemically known as CH3C(=NOH)COCH3, is a white to off-white powder that's slightly soluble in water. It's used as a reagent in a number of reactions in organic chemistry, especially for the detection of urea. The Blood Urea Nitrogen (BUN) test is used primarily to detect problems with the kidneys and uses diacetyl monoxime as its primary reagent. This test is routinely performed in many medical laboratories.
Instructions
1. Label the three test tubes as "Blank," "Test" and "Standard." Fill the "Blank" test tube with 0.05 ml of distilled water, the "Test" test tube with 0.05 ml of the blood serum sample and the "Standard" test tube with 0.05 ml of a standard solution. The standard solution represents a reference BUN concentration, usually about 15 mg of urea per deciliter of blood (mg/dl).
2. Add 3.0 ml of diacetyl monoxime to all three test tubes and mix it thoroughly by lateral tapping (tapping on the side of the test tube). Add 3 ml of ferrithiocyanate to each test tube and mix thoroughly again with lateral tapping.
3. Fill one of the water baths with water and heat to a boil. While the water in the first water bath is boiling, fill the second water bath with ice water. Place all three tubes in the boiling water for 15 minutes and then place them in the ice water for two minutes. Let the test tubes stand at room temperature for one minute.
4. Calibrate the spectrophotometer. Load the "Blank" test tube into the spectrophotometer and subject it to light with a wavelength of 520 nanometers (nm). Adjust the spectrophotometer to show an absorbance of zero.
5. Record the absorbance of both the standard and test samples for 520 nm light. Calculate the BUN level of the test sample as (test absorbance/standard absorbance) multiplied by the reference concentration.
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