Abstract
This paper discusses the development and validation of a rapid method for the reversed phase HPLC-UV quantification of biodegradable poly(D,L-lactic-co-glycolic) acid (PLGA) microspheres co-loaded with two neuroprotective agents (dexamethasone and melatonin) (DX-MEL-MSs) to be intravitreally administered as a promising glaucoma treatment. The study was performed to validate two procedures that quantify the content of the two active substances entrapped into the polymer matrix during an encapsulation efficiency assay and the amount of drugs liberated over time during the in vitro release assay. The reversed-phase method allowed for the simultaneous determination of dexamethasone and melatonin, which were respectively detected at 240.5 and 222.7 nm. Chromatographic separation was performed using an Ascentis® C18 HPLC Column (25 cm × 4.6 mm, 5 µm) with an isocratic mobile phase composed of methanol-water (70:30, v/v) with 1.0 mL min−1 flow rate. The two procedures were validated analytically in terms of system suitability testing, specificity, linearity, precision, accuracy, sensitivity, and robustness. Both the validated procedures were applied to characterize DX-MEL-MSs and were found appropriate to quantify the drug quantities encapsulated and estimate their release profile over 10 days. The validation study designed in this work can be helpful for planning any other protocols that refer to the quantification of PLGA based drug delivery systems.
Highlights
Glaucoma, diabetic retinopathy, age-related macular degeneration, and retinitis pigmentosa are some of the optic neuropathies that have a major impact worldwide and can lead to progressive and permanent blindness [1]
The two compounds were loaded in biodegradable PLGA microspheres for intravitreal administration as a neuroprotective treatment for glaucoma
One procedure (A) was proposed to quantify the content of the two active substances entrapped into the polymer matrix, the second one (B) was intended for the quantification of DX and MEL liberated over time during in vitro release assay
Summary
Diabetic retinopathy, age-related macular degeneration, and retinitis pigmentosa are some of the optic neuropathies that have a major impact worldwide and can lead to progressive and permanent blindness [1]. Neurodegenerative processes lead to RGC death and involve several molecular mechanisms, such as neuroinflammation, excitotoxicity, axonal transport damage, and oxidative stress [6,7]. As all these mentioned events have demonstrated effects that can interact and be triggered by different pathways, co-delivery and administration of different agents can represent an effective clinical approach to treat glaucoma [8,9]. For this reason, neuroprotective strategies, such as the use of antioxidant or anti-inflammatory agents in combination, can represent a promising therapy for this chronic disease
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