Development and validation of a HPLC-method for the quantitative determination of coumarin in tonka beans

Introduction. The tonka beans (seeds of Dipteryx odorata, family Fabaceae) is a valuable source of coumarin used in the food, perfume and cosmetic industries thanks to its rich aroma. Despite its pharmacological value (anticoagulant, antioxidant activity), coumarin has a proven hepatotoxicity and haematotoxicity, which limits its use in a number of countries. Despite the control of synthetic coumarin in aromatics, the natural analog of tunica beans does not have clear rules, although it has identical activity. Growing demand for organic products has increased the popularity of tonka beans, but lack of scientific data on their safety and methods of analysis makes quality control difficult. The development of validated techniques, such as HVACR, is an urgent task to ensure the safety of its application in industry.

Aim. Development of a HPLC-method for the quantitative determination of coumarin in beans and its validation with further testing on several batches of the studied object.

Materials and methods. Five batches of Tonka beans from different producing countries were purchased as research objects, for sample preparation was used US-bath. Chromatography conditions: Luna 5 μm C18(100) Å LC Column 250 × 4.6 mm, thermostat temperature – 40 °C, detection wavelength – 275 nm. Mobile phase A – acetonitrile, mobile phase B – water. Elution mode: isocratic mobile phase B – 50 % for 12 minutes.

Results and discussion. The method meets the criteria of suitability of the chromatographic system, specificity, linearity (range 0.001–0.1 mg/ml) and precision. Its advantages are expressiveness and economy, which simplifies the quality control of raw materials. The quantitative content of coumarin in the studied object ranges from 1.25 ± 0.07 % to 4.29 ± 0.12 %.

Conclusion. Developed and validated by HPLC-methodology for the quantitative determination of coumarin in tonka beans. The developed methodology can be used to standardize this promising raw material in various industries, ensuring the necessary level of safety in its application.

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