Method development and validation on RP-HPLC method for estimation of xanthohumol in nanostructured lipid carriers...

Xanthohumol(Xn) is isolated from female inflorescences of Humulus lupulus. It has been discovered that Xn and its formulation are useful in the treatment of cancer. As this bioactive compound has medicinal importance, hence, a novel, precise, and sensitive HPLC method should be developed. In the present study, an RP-HPLC method has been developed and validated as per ICH Q2(R1) guidelines using column C18 having particle size 5 µm and dimension 250 × 4.6 mm. The detection wavelength (λ) used was 370 nm. The mobile phase consisted of a combination of HPLC grade Methanol and HPLC grade water buffer at a ratio of 95:5 v/v, with a flow rate of 1.0 mL/min. The total run time was 10 min with Xn retention time (Rt) at 4.5 min. The calibration plot was found linear in the range 2-10 μg/mL. The recovery of 97.65 % indicated good accuracy of method. The method's precision is within 2 % of acceptable limits. LOD and LOQ values of 0.85 and 2.6 μg/mL indicated good sensitivity of method. The uniqueness of the research work is relying on achieving peak of Xn at lesser retention time as compared to existing methods. Further the results of specificity studies revealed absence of any interference of Xn peak with the excipients used in the nanostructured lipid carriers (NLCs). Overall, the study provided an accurate, precise, sensitive and specific method to quantify Xn in bulk and NLCs.