Development and Validation of an HPLC–MS/MS Method for Pioglitazone from Nanocarriers Quantitation in Ex Vivo and In Vivo Ocular Tissues

Abstract

Pioglitazone (PGZ) is an oral anti-hyperglycemic agent, belongs to the class of thiazolidinediones, and is used for the treatment of diabetes mellitus type 2. In recent years, its anti-inflammatory activity has also been demonstrated in the literature for different diseases, including ocular inflammatory processes. Additionally, this drug belongs to Class II of the Biopharmaceutical Classification System, i.e., slightly soluble and highly permeable. The main objective of this study was to validate a new analytical HPLC-MS/MS method to quantify free-PGZ and PGZ from polymeric NPs to conduct nanoparticle application studies loaded with this active ingredient to transport it within ocular tissues. An accurate, sensitive, selective, reproducible and high throughput HPLC-MS/MS method was validated to quantify PGZ in cornea, sclera, lens, aqueous humor, and vitreous humor. The chromatographic separation was achieved in 10 min on a Kinetex C18 column. Linear response of PGZ was observed over the range of 5–100 ng/mL. The recovery of free-PGZ or PGZ from NPs was in the range of 85–110% in all tissues and levels tested. The intra-day and inter-day precision were <5% and <10%, respectively. The extracts were shown to be stable in various experimental conditions in all matrices studied. The range of concentrations covered by this validation is 80–1600 µg/kg of PGZ in ocular tissues. It is concluded that this method can be applied to quantify PGZ for in vivo and ex vivo biodistribution studies related to the ocular administration of free-PGZ and PGZ from nanoparticles.

Assessment of Efficacy and Safety Using PPAR-γ Agonist-Loaded Nanocarriers for Inflammatory Eye Diseases

“…In our previous studies, PZ-NCs of PLGA-PEG were optimized and characterized physiochemically and biopharmaceutically as a new delivery system suitable for the ocular route [ 36 , 37 ]. PZ-NCs were previously synthesized using the solvent displacement technique, showing a particle size of 247.22 ± 2.77 nm with polydispersity index (PI) values lower than 0.2 (0.17 ± 0.03) and a negative charge with zeta potential (ZP) values of −3.34 ± 0.42 mV.…”

“…After that, the NCs were resuspended in water, and the values found were a particle size of 271.31 ± 2.78 nm, PI of approximately 0.24 ± 0.00, ZP of −7.57 ± 1.53 mV, and EE of 89.13% ± 2.21%. These physiochemical parameters before and after the spray-drying process were considered suitable for ocular delivery [ 36 , 37 ]. These parameters are in concordance with other authors for the use of polymeric systems in ocular therapies [ 18 , 28 , 38 , 39 ].…”

Development of an Improved Method for the Determination of Iodine/β-Cyclodextrin by Means of HPLC-UV: Validation and the Thyroid-Stimulating Activity Revealed by In Vivo Studies

“…Accuracy is the closeness of agreement between the accepted true value, or a reference value, and the measured result obtained. Accuracy studies are usually evaluated by determining the recovery of a spiked sample of the analyte into the matrix of the sample to be analyzed [ 34 ]. Three different samples of the working solution of KI (low, medium and high, i.e., 80%, 100% and 120%, respectively) were analyzed ( Table 4 ).…”

Stabilization by Nano Spray Dryer of Pioglitazone Polymeric Nanosystems: Development, In Vivo, Ex Vivo and Synchrotron Analysis

“…Rabbits were euthanized after 10 d of topical administration with PGZ-NPs and the Spray 2 formulation (0.05 mL/eye/day). The ocular tissues were isolated from the eyes and kept at −80 • C until quantification by liquid chromatography-mass spectrometry (LC-MS) [62]. The values are described as the average ± SD (n = 3/group).…”