Abstract
Objective: The aim of the present study is to develop and validation of a ultra-high performance liquid chromatography (UHPLC) method to determine the ursolic acid content and its encapsulation efficiency (EE) in lipid-core nanocapsules prepared from poly (L-lactic acid).
 Methods: A simple UHPLC-PDA method was developed and validated for the quantitative determination of ursolic acid in poly(L-lactic acid) nanocapsules. The chromatographic conditions used were: RP-C18 column, isocratic mobile phase containing acetonitrile:water (92:8, v/v), flow rate of 0.8 ml/min, column temperature of 50°C, and detection at 203 nm. The following parameters were evaluated: Specificity, linearity, limits of detection and quantification, precision, accuracy, and robustness.
 Results: The method was specific to the ursolic acid and linear (r=0.9998) in the range of 10–100 μg/ml. The limits of detection and quantification were 1.35 and 4.10 μg/ml, respectively. The precision was demonstrated by a relative standard deviation less than 2%. Adequate accuracy (98.35%±0.82) was obtained. Changes in flow rate, mobile phase, and column temperature did not significantly alter the peak area and the retention time of the ursolic acid. The mean EE was 99.89%.
 Conclusion: The method proved to be fast, sensitive, and simple for quantifying ursolic acid in nanocapsules and was successfully used for determining the EE.
Highlights
Pentacyclic triterpenoids are phytochemicals widely distributed in nature and ursolic acid (3β-hydroxy-urs-12-en-28-oic) (UA) is one of their main representatives [1]
The present paper describes the development and the validation of a ultra-high performance liquid chromatography (UHPLC)-PDA method for determining the Ursolic acid (UA) content and its EE in lipid-core nanocapsules prepared from poly(L-lactic acid) (PLA)
The total analysis time and the retention time were 2.1 and 1.78 min, respectively. This fast response is very important for the analysis routine. These conditions provided a lower tail size and a more symmetrical peak for the UA quantification when compared to other methods that used HPLC [22,23]
Summary
Pentacyclic triterpenoids are phytochemicals widely distributed in nature and ursolic acid (3β-hydroxy-urs-12-en-28-oic) (UA) is one of their main representatives [1]. UA is usually found in human diet [3] and has a wide range of biological activities, such as the antioxidant effect [4], the anti-obesity and muscle synthesis properties [5], the hepatoprotective potential [6], the blood sugar-lowering effect [7], the neuroprotective potential [8], the antiinflammatory activity [9], and the antitumor properties [10,11] In spite of this pharmacotherapeutic potential, some limitations can be related to the UA use due to its low aqueous solubility and its reduced permeability through biological membranes, which decrease its absolute bioavailability to about 8% [12] and lead to a classification as a class IV drug by the biopharmaceutical classification system. A validated method that is able of quantifying the drug loading in nanoformulation is essential during the research and development process
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