Rapid determination of retinoic acid and its main isomers in plasma by second order high-performance liquid chromatography data modeling

Abstract

This paper reports the development of a method based on high-performance liquid chromatography (HPLC) coupled to second-order data modeling with multivariate curve resolution-alternating least-squares (MCR–ALS) for quantification of retinoic acid and its main isomers in plasma in only 5.5 min. The compounds retinoic acid (RA), 13-cisretinoic acid, 9-cis-retinoic acid, and 9,13-di-cis-retinoic acid were partially separated by use of a Poroshell 120 EC–C18 (3.0 mm×30 mm, 2.7 μm particle size) column. Overlapping not only among the target analytes but also with the plasma interferents was resolved by exploiting the second-order advantage of the multi-way calibration. A validation study led to the following results: trueness with recoveries of 98.5– 105.9 % for RA, 95.7–110.1 % for 13-cis-RA, 97.1– 110.8 % for 9-cis-RA, and 99.5–110.9 % for 9,13-di-cis-RA; repeatability with RSD of 3.5–3.1 % for RA, 3.5–1.5 % for 13-cis-RA, 4.6–2.7 % for 9-cis-RA, and 5.2–2.7 % for 9,13- di-cis-RA (low and high levels); and intermediate precision (inter-day precision) with RSD of 3.8–3.0 % for RA, 2.9– 2.4 % for 13-cis-RA, 3.6–3.2 % for 9,13-di-cis-RA, and 3.2– 2.9 % for 9-cis-RA (low and high levels). In addition, a robustness study revealed the method was suitable for monitoring patients with dermatological diseases treated with pharmaceutical products containing RA and 13-cis-RA.