Estimation of Nutraceutical Astaxanthin by RP-HPLC Method: Development and Validation

Abstract

Background: Nutraceuticals have always been considered natural and safe supplements that may prevent disease, may substitute prescription drugs, compensate for a poor diet or promote health. Several nutraceutical products in the market today play roles as antioxidants and probiotics. One such nutraceutical is Astaxanthin, a red-pigmented keto-carotenoid, which offers a plethora of uses and health benefits. Astaxanthin, like many carotenoids, is a lipid-soluble pigment which faces solubility and bioavailability issues owing to its lipophilicity, due to which its estimation is challenging. And the analysis of a simple solvent extract of Astaxanthin would produce many ester-astaxanthin “peaks” and utilize costly organic solvents. Additionally, the literature review indicates the use of very complex mobile phases and tedious solvent extraction procedures involved in the estimation of Astaxanthin.  Objective: The manuscript describes the development and validation of a rapid, sensitive and specific Reverse Phase-High Performance Liquid Chromatographic (RP-HPLC) method involving Ultraviolet (UV) detection for the determination and quantification of nutraceutical Astaxanthin in bulk and capsule dosage forms.   Methods: Chromatography was carried out on an Inertsil ODS C18 (250 x 4.6 mm), 5µ, using a filtered and degassed mixture of Water: Acetonitrile (30:70% v/v) as the mobile phase at a flow rate of 1 mL /min with isocratic elution method and effluent was monitored at 478 nm.  Results: The method was linear in the range of 5 – 15 μg mL-1 of Astaxanthin. The method was validated with respect to precision, accuracy, specificity and robustness. The run time of the analyte was 8.45 min.  Conclusion: The developed method is simple, specific, precise, accurate, and robust and shows no interference with diluents which proves the adaptability of the method for routine analysis of the nutraceutical Astaxanthin in bulk and marketed formulations.