Development and validation of a reversed-phase high-performance liquid chromatographic method with solid-phase extraction for the quantification of hydrochlorothiazide in ex vivo permeation studies

ONNAINTY RENEE; SCHENFELD ESTEBAN; QUEVEDO MARIO ALFREDO; GRANERO G.

BIOMEDICAL CHROMATOGRAPHY

JOHN WILEY & SONS LTD

Lugar: LOndres; Año: 2017

0269-3879

ydrochlorothiazide (HCT) is a diuretic used to treat hypertension. In order to study its intestinalpermeation behavior applying an ex vivo methodology, a rapid, sensitive and selective reversed‐phase liquid chromatography (RP‐HPLC) method coupled with UV detection (RP‐HPLC UV)was developed for the analysis of HCT in TC199 culture medium used as mucosal and serosalsolutions in the everted rat intestinal sac model. Also, analytical procedures for the quantificationof HCT by RP‐HPLC with UV detection required a sample preparation step by solid‐phase extraction.The method was validated in the concentration range of 8.05 × 10−7 to 3.22 × 10−5 M forHCT. Chromatographic parameters, namely carry‐over, lower limit of quantification(1.4491 × 10−7 M), limit of detection (3.8325 × 10−8 M), selectivity, inter‐ and intraday precisionand extraction recovery, were determined and found to be adequate for the intended purposes.The valida