How to Use Rothera’s Mixture Powder to Detect Ketone Bodies in Urine Sample

Ketone bodies refer to three molecules which are water soluble and are produced by the liver in human body. Ketone bodies are typically composed of 20% acetoacetic acid, 78% beta-hydroxybutyric acid and 2% acetone. Concentration of ketone bodies in blood usually remains below 1mg/dl but if its concentration in urine and subsequently in blood increases, a sample of urine is tested for the presence and determination of the amount of ketone bodies. Rothera’s mixture powder is used to test for the presence of ketone bodies or acetone in urine. There are several methods for detection of ketone bodies out of which the most commonly used ones are Rothera’s test, ferric chloride test, ace-test tablet test and strip reagent test.

Rothera’s nitroprusside test and its derivative methods detect acetone and acetoacetic acid, which are two of the components that ketone bodies are made up of. Beta-hydroxybutyric acid cannot be detected through any screening tests.

Rothera’s test using classic nitroprusside technique to detect acetone and acetoacetic acids

Rothera’s mixture powder manufacturers mix ammonium sulphate, sodium nitroprusside and liquid ammonia solution to form the powder. The acetone or acetoacetic acid in the sample reacts with nitroprusside component of Rothera’s mixture in an alkali solution to form a purple mixture with a calomel red colour ring at the rim. Rothera’s test gives the most accurate results with acetoacetate concentration of 1-5 mg/dl and acetone of 10-25 mg/dl. The test is performed in the following manner:

• 5ml of test sample is transferred to a dish or test tube.
• The sample is then saturated with sulphate of ammonia.
• One sodium nitroprusside crystal is added
• The liquid ammonia solution is layered on the side of the tube
• A purple or calomel red coloured ring appears at the rim or junction of the two layers

The clinical significance of the test is most pronounced in cases of increased:

• Propanol poisoning
• Diabetes mellitus
• Severe starvation
• Fasting
• Anorexia
• Fever
• Lactic acidosis
• Prolonged vomiting
• Salicyclate toxicity
• Severe level of carbohydrate restriction

The Rothera mixture suppliers and where do they usually supply their stock

Chemical compounds used for testing the presence of some or the other important factor related to the human body are usually supplied to pathological or research laboratories. Chemists perform experiments using these compounds on the test samples they are provided with and are able to determine the severity of a condition or disease by the test results. Rothera mixture powder suppliers have to ensure that the chemical compounds are carried and preserved according to mentioned guidelines in order to prevent them from spoiling. The manufacturers and suppliers are responsible for producing and supplying the best quality chemical compounds.

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