Development and validation of analytical method by HPLC-DAD for determination of vasodilator active in pharmaceutical ophthalmic forms

Abstract

The substance 4-Aminobenzamidine dihydrochloride (4-AD) is one of the degradation products of diminazene aceturate and has demonstrated antiglaucomatous potential. Glaucoma is the second leading cause of blindness worldwide; thus, new therapeutic alternatives must be studied, for example, the molecule 4-AD vehiculated into polymeric inserts for prolonged release. The present work aims to develop and validate an analytical method to quantify 4-AD in pharmaceutical ophthalmic forms. A HPLC was used with UV-Vis detector, at 290 ƞm and ACE ® C18 column (125 × 4.6 mm, 5 μm), in which the mobile phase consists of phosphate buffer (pH 7.4) and triethylamine (30 mmol/L), under an isocratic flow of 1.0 mL/min. The retention time of 3.2 minutes was observed. The method was developed and validated in accordance with ANVISA recommendations and ICH guides. The linearity range was established between the concentrations 5 and 25 μg/mL (correlation coefficient r = 0.993). The accuracy, repeatability, and intermediate precision tests obtained a relative standard deviation less than or equal to 5%. In addition, the method was considered selective, exact. and robust, with pH being its critical factor. Therefore, the HPLC analysis method is robust and can be used to quantify 4-AD in pharmaceutical forms for ocular application.

Etnobotânica de plantas medicinais aromáticas: preparações e usos da flora local em cinco comunidades rurais localizadas na região do Baixo Tocantins, Pará, Brasil

“…A comunidade Mamangal situa-se as margens do Rio Mamangal, no município de Igarapé-Miri (1º58'37" Sul e 48º57'34" Oeste) (Pinto et al, 2014). A Comunidade de Nossa Senhora do Livramento, está localizada na Rodovia PA 252, Km 05, município de Acará (1º57'37" Sul e 48º11'47" Oeste).…”