Validation of spectrophotometric and colorimetric methods for determination of efavirenz in various biological matrices: Application to the development of dissolvable microneedle-based efavirenz complex inclusion for vaginal delivery

• Published in: Microchemical journal Vol. 200; p. 110361
• Main Authors: Hidayat, Muh. Taufik, Fitrayani, Nurul, Alik Samma, Abigael, Anggriani, A., Permana, Andi Dian
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.05.2024
• Subjects: Dissolving microneedle; Efavirenz; Inclusion complex; Method validation; UV–Vis spectrophotometer; Vaginal; Dissolving microneedle; UV–Vis spectrophotometer; Method validation; Efavirenz; Vaginal; Inclusion complex
• ISSN: 0026-265X; 1095-9149
• DOI: 10.1016/j.microc.2024.110361

[Display omitted] •A spectrophotometric and colorimetric methods to quantify efavirenz was developed.•The analytical method was validated according to the ICH and FDA guidelines.•Inclusion complex formulations incorporated into microneedle dissolutions were developed.•The validated method was applied in ex vivo and in vivo vaginal delivery studies. One of the antiretroviral treatments (ART) used to treat Human Immunodeficiency Virus (HIV) disease is efavirenz (EFV). EFV itself has bioavailability issues during oral therapy due to its poor solubility. When injected, administration issues arise. Hence, creating medication delivery methods was crucial to creating EFV delivery. HIV therapy benefits from the vaginal medication delivery system's ease of use, higher bioavailability, and quick systemic absorption in the surface folds. In this study, an inclusion complex (IC) was added to the EFV formulation to increase solubility, and dissolving microneedles (DMN) was used for delivery to increase penetration. Here, in support of this formulation creation process, we carried out validation utilizing spectrophotometric and colorimetric techniques. The amount of EFV was determined by validating ethanol and artificial vaginal fluid (AVF) using UV–Vis spectrophotometry. Vaginal tissue and plasma were also validated using colorimetric with the β-naphthol reagent. The International Council for Harmonization (ICH) and US Food and Drug Administration (FDA) requirements were adhered to in verifying the creation of analytical procedures. The outcomes demonstrated a linear correlation value of ≥0.9998. The LLOQ levels were 0.99 g/mL, 0.97 g/mL, 0.66 g/mL, 0.74 g/mL, and 0.74 g/mL in ethanol, AVF, porcine vaginal tissue, rat vaginal tissue, and vaginal plasma, respectively. The developed precision and accuracy were confirmed. The approach was successfully used to determine the amount of EFV in inclusion complex integration and DMN platform. The outcomes demonstrated high IC-EFV incorporation release and penetration profiles, permeation and high drug retention andex vivokinetics in the vagina and enhanced bioavailability fromin vivostudy pharmacokinetic profiles of DMN-IC-EFV.