PMID- 35214021 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20220301 IS - 1999-4923 (Print) IS - 1999-4923 (Electronic) IS - 1999-4923 (Linking) VI - 14 IP - 2 DP - 2022 Jan 26 TI - Validation of a Rapid and Easy-to-Apply Method to Simultaneously Quantify Co-Loaded Dexamethasone and Melatonin PLGA Microspheres by HPLC-UV: Encapsulation Efficiency and In Vitro Release. LID - 10.3390/pharmaceutics14020288 [doi] LID - 288 AB - 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((R)) C18 HPLC Column (25 cm x 4.6 mm, 5 microm) 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. FAU - Brugnera, Marco AU - Brugnera M AUID- ORCID: 0000-0003-2600-6446 AD - Innovation, Therapy and Pharmaceutical Development in Ophthalmology (InnOftal) Research Group, UCM 920415, Department of Pharmaceutics and Food Technology, Faculty of Pharmacy, Complutense University of Madrid, Plaza Ramon y Cajal, s/n, 28040 Madrid, Spain. AD - Sanitary Research Institute of the San Carlos Clinical Hospital (IdISSC), San Carlos Clinical Hospital, Calle Profesor Martin Lagos, s/n, 28040 Madrid, Spain. FAU - Vicario-de-la-Torre, Marta AU - Vicario-de-la-Torre M AUID- ORCID: 0000-0002-0048-1794 AD - Innovation, Therapy and Pharmaceutical Development in Ophthalmology (InnOftal) Research Group, UCM 920415, Department of Pharmaceutics and Food Technology, Faculty of Pharmacy, Complutense University of Madrid, Plaza Ramon y Cajal, s/n, 28040 Madrid, Spain. AD - Sanitary Research Institute of the San Carlos Clinical Hospital (IdISSC), San Carlos Clinical Hospital, Calle Profesor Martin Lagos, s/n, 28040 Madrid, Spain. FAU - Andres-Guerrero, Vanessa AU - Andres-Guerrero V AUID- ORCID: 0000-0003-0157-1932 AD - Innovation, Therapy and Pharmaceutical Development in Ophthalmology (InnOftal) Research Group, UCM 920415, Department of Pharmaceutics and Food Technology, Faculty of Pharmacy, Complutense University of Madrid, Plaza Ramon y Cajal, s/n, 28040 Madrid, Spain. AD - Sanitary Research Institute of the San Carlos Clinical Hospital (IdISSC), San Carlos Clinical Hospital, Calle Profesor Martin Lagos, s/n, 28040 Madrid, Spain. FAU - Bravo-Osuna, Irene AU - Bravo-Osuna I AUID- ORCID: 0000-0003-3133-7872 AD - Innovation, Therapy and Pharmaceutical Development in Ophthalmology (InnOftal) Research Group, UCM 920415, Department of Pharmaceutics and Food Technology, Faculty of Pharmacy, Complutense University of Madrid, Plaza Ramon y Cajal, s/n, 28040 Madrid, Spain. AD - Sanitary Research Institute of the San Carlos Clinical Hospital (IdISSC), San Carlos Clinical Hospital, Calle Profesor Martin Lagos, s/n, 28040 Madrid, Spain. FAU - Molina-Martinez, Irene Teresa AU - Molina-Martinez IT AUID- ORCID: 0000-0002-9157-571X AD - Innovation, Therapy and Pharmaceutical Development in Ophthalmology (InnOftal) Research Group, UCM 920415, Department of Pharmaceutics and Food Technology, Faculty of Pharmacy, Complutense University of Madrid, Plaza Ramon y Cajal, s/n, 28040 Madrid, Spain. AD - Sanitary Research Institute of the San Carlos Clinical Hospital (IdISSC), San Carlos Clinical Hospital, Calle Profesor Martin Lagos, s/n, 28040 Madrid, Spain. FAU - Herrero-Vanrell, Rocio AU - Herrero-Vanrell R AD - Innovation, Therapy and Pharmaceutical Development in Ophthalmology (InnOftal) Research Group, UCM 920415, Department of Pharmaceutics and Food Technology, Faculty of Pharmacy, Complutense University of Madrid, Plaza Ramon y Cajal, s/n, 28040 Madrid, Spain. AD - Sanitary Research Institute of the San Carlos Clinical Hospital (IdISSC), San Carlos Clinical Hospital, Calle Profesor Martin Lagos, s/n, 28040 Madrid, Spain. LA - eng GR - 813440/ORBITAL-ITN/ PT - Journal Article DEP - 20220126 PL - Switzerland TA - Pharmaceutics JT - Pharmaceutics JID - 101534003 PMC - PMC8878730 OTO - NOTNLM OT - HPLC-UV OT - PLGA OT - co-delivery OT - dexamethasone OT - encapsulation efficiency OT - glaucoma OT - in vitro release OT - melatonin OT - microspheres OT - validation COIS- The authors declare no conflict of interest. EDAT- 2022/02/27 06:00 MHDA- 2022/02/27 06:01 PMCR- 2022/01/26 CRDT- 2022/02/26 01:02 PHST- 2021/09/27 00:00 [received] PHST- 2021/10/28 00:00 [revised] PHST- 2022/01/25 00:00 [accepted] PHST- 2022/02/26 01:02 [entrez] PHST- 2022/02/27 06:00 [pubmed] PHST- 2022/02/27 06:01 [medline] PHST- 2022/01/26 00:00 [pmc-release] AID - pharmaceutics14020288 [pii] AID - pharmaceutics-14-00288 [pii] AID - 10.3390/pharmaceutics14020288 [doi] PST - epublish SO - Pharmaceutics. 2022 Jan 26;14(2):288. doi: 10.3390/pharmaceutics14020288.