Practical Protein Chemistry - A. Darbre 1989
Analytical Methods
Determination of protein attached to a solid support
Fluorometric determination of carbodiimide
Mix 100 µL of the test solution with 100 µL of a 20 mM trans-aconitic acid solution in anhydrous dioxane (dried and distilled over metallic sodium) in a Glass-stoppered test tube. Heat for 10 min at 50 °C. Then, add 50 µL of 0.2 M pyridine in anhydrous dioxane and dilute with 2.5 mL of methanol. Measure the fluorescence immediately at an excitation wavelength of 400 nm and an emission wavelength of 490 nm against a control solution. The fluorescence intensity is linearly dependent on the carbodiimide amount up to 50 nmol.
trans-Aconitic acid can be used as a spraying reagent for plates in Thin-Layer Chromatography to detect carbodiimides.
Last update: 06/08/2026
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