Validation of an Innovative Chromatographic Method for Hypericin Quantification in Nanostructured Lipid Carriers.

BACKGROUND: Hypericin (HYP) is a natural compound widely used as a food supplement. The encapsulation of HYP into nanosystems, such as nanostructured lipid carriers (NLC), is a promising strategy for delivering this lipophilic molecule and protecting it from degradation. OBJECTIVE: This study aims to develop and validate an analytical method to quantify the encapsulation efficiency of HYP in NLC. METHOD: A reverse-phase high-performance liquid chromatography (HPLC) method was developed and validated according to the International Conference on Harmonization (ICH) guide Q2 (R1). NLC was prepared through the ultrasonication method, and HYP encapsulation efficiency was evaluated using the validated method. RESULTS: Separation was achieved using an isocratic mobile phase composed of acetonitrile, methanol, and ammonium acetate buffer (10 mM, pH 5.0) (54:36:10, v/v/v) and a reverse stationary phase. The specificity, linearity, precision, accuracy, and robustness of the method were assessed and confirmed during the validation. Furthermore, the validated method was able to determine the encapsulation efficiency of HYP in NLC. CONCLUSIONS: The HPLC method was validated, and the results indicated the ability of NLC to deliver HYP compounds for further application as a food supplement. HIGHLIGHTS: HYP is used as a food supplement and for photodynamic therapy (PDT). The developed method was specific, linear, precise, accurate, and robust. NLCs showed a high ability to encapsulate HYP.

In vivo study of hypericin-loaded poloxamer-based mucoadhesive in situ gelling liquid crystalline precursor system in a mice model of vulvovaginal candidiasis.

The present study reports the performance of the pigment hypericin (HYP)-loaded poloxamer-based mucoadhesive in situ gelling liquid crystalline precursor system (LCPS) for the treatment of vulvovaginal candidiasis (VVC) in mice. LCPS composed of 40% of ethoxylated and propoxylated cetyl alcohol, 30% of oleic acid and cholesterol (7:1), 30% of a dispersion of 16% poloxamer 407 and 0.05% of HYP (HYP-LCPS) was prepared and characterized by polarized light microscopy (PLM), small-angle X-ray scattering (SAXS) and ex vivo permeation and retention studies across vaginal porcine mucosa were performed. In addition, the antifungal properties of the HYP-LCPS were evaluated in a murine in vivo model; for this, infected C57BL female mice groups were treated with both HYP in solution and HYP-LCPS, and after 6 days colony forming unit (CFU)/ml count was performed. PLM and SAXS confirmed that HYP-LCPS is a microemulsion situated in boundary transition region confirming its action as an LCPS. When in contact with simulated vaginal fluid, HYP-LCPS became rigid and exhibited maltase crosses and bragg peaks characteristics of lamellar phase. Ex vivo permeation and retention studies showed that HYP-LCPS provides a localized treatment on the superficial layers of porcine vaginal mucosa. HYP-LCPS induced a significant reduction in the number of CFU/ml in the mice; thus this formulation indicated it is as effective as a commercial dosage form. It was concluded that LCPS maintains the biological activity of HYP and provides an adequate drug delivery system for this lipophilic molecule at the vaginal mucosa, being a promising option in cases of VVC.

A Review of Analytical Methods for the Determination of Hypericin in Foods, Herbal, Biological and Pharmaceutical Matrices.

AIMS: A review of analytical methods for the determination of hypericin in foods, herbal, biological and pharmaceutical matrices. BACKGROUND: Hypericin (HYP) is a naturally-occurring pigment obtained from some plants of the genus Hypericum. Although HYP has been known for many years, it has recently attracted attention due to its varied biological properties, such as anti-inflammatory and antidepressant activity and it is also an efficient photosensitizer. OBJECTIVE: The objective of this review is to provide insights into the physicochemical properties of HYP, as well as to report the analytical methods existing in the literature and official compendia for different matrices. METHODS: The survey data were collected by Google Scholar® and Scopus® using keys terms. RESULT: Analytical methods involving HYP are mainly concerned with the quality control of pharmaceutical preparations, foods, beverages, biological samples and drug delivery systems using different types of analysis methods. Some difficulties have also been identified due to the physicochemical properties of HYP. It presents great solubility in alkaline solutions, organic bases and common polar organic solvents. CONCLUSION: It can be analyzed by thin layer chromatography, spectrophotometry in the ultraviolet region, but the most commonly used method is by HPLC. HYP presents monographs in the American, British and European Pharmacopoeias, however, the methods of analysis are not yet harmonized.

Mucoadhesive In Situ Gelling Liquid Crystalline Precursor System to Improve the Vaginal Administration of Drugs.

The vaginal mucosa is a very promising route for drug administration due to its high permeability and the possibility to bypass first pass metabolism; however, current vaginal dosage forms present low retention times due to their dilution in vaginal fluids, which hampers the efficacy of many pharmacological treatments. In order to overcome these problems, this study proposes to develop a mucoadhesive in situ gelling liquid crystalline precursor system composed of 30% of oleic acid and cholesterol (7:1), 40% of ethoxylated and propoxylated cetyl alcohol, and 30% of a dispersion of 16% Poloxamer 407. The effect of the dilution with simulated vaginal fluid (SVF) on this system was evaluated by polarized light microscopy (PLM), small-angle X-ray scattering (SAXS), rheological studies, texture profile analysis (TPA), mucoadhesion study, in vitro drug release test using hypericin (HYP) as drug model, and cytotoxicity assay. PLM and SAXS confirmed the formation of an isotropic system. After the addition of three different concentrations of SVF (30, 50, and 100%), the resultant formulations presented anisotropy and characteristics of viscous lamellar phases. Rheology shows that formulations with SVF behaved as a non-Newtonian fluid with suitable shear thinning for vaginal application. TPA and mucoadhesion assays indicated the formation of long-range ordered systems as the amount of SVF increases which may assist in the fixation of the formulation on the vaginal mucosa. The formulations were able to control about 75% of the released HYP demonstrating a sustained release profile. Finally, all formulations acted as safe vaginal drug delivery systems.

Recent Advances in the Use of Metallic Nanoparticles with Antitumoral Action - Review.

The term cancer represents a set of more than 100 diseases that are caused due to an uncontrolled growth of cells; and their subsequent spread to the other tissues and organs of the body by a phenomenon, called 'metastasis'. According to the estimates provided by the World Health Organization (WHO), cancer is expected to account for about 10 million deaths per year by 2020 and 21 million cancer cases, which may lead to 13 million deaths by 2030, making cancer as the cause of highest mortality in contrast to other diseases. The search for potential therapeutics against cancer, which can reduce the side-effects that occur due to the difficulty of recognition between cancerous and normal cells, has ever been increased. In this view, nanotechnology, especially metallic nanoparticles (MNPs), comes to aid in the development of novel therapeutic agents, which may be synthesized or modified with the most diverse functional chemical groups; this property makes the metallic nanoparticles suitable for conjugation with already known drugs or prospective drug candidates. The biocompatibility, relatively simple synthesis, size flexibility and easy chemical modification of its surface, all make the metallic nanoparticles highly advantageous for opportune diagnosis and therapy of cancer. The present article analyzes and reports the anti-tumor activities of 78 papers of various metallic nanoparticles, particularly the ones containing copper, gold, iron, silver and titanium in their composition.

Characteristics, Biological Properties and Analytical Methods of Ursolic Acid: A Review.

Ursolic acid (UA) is a naturally occurring triterpenoid which is a promising candidate for the development of new therapeutic approaches and for the prevention and treatment of several diseases owing to its pharmacological importance. However, its low solubility in aqueous medium affects its therapeutic application. Several strategies have been used to overcome this obstacle. In this study, the incorporation of UA in to different drug delivery systems was found to be highly efficient. In addition, important investigations were performed about methods for qualitative and quantitative analyses of UA in various raw materials, including plants, biological fluids, and drug delivery systems, were investigated. Most recently high performance liquid chromatography coupled with various detectors, gas chromatography-mass spectrometry and capillary electrophoresis were used for this purpose. Thus, this review was performed to evaluate the biological effects of UA demonstrated thus far as well as the currently used, delivery systems and analytical methods.