Application of solid-phase extraction in the method development for determination of SEPA, an acronym for soft enhancement of percutaneous absorption, in human, rat, and rabbit serum using GC-FID method.
A new nonaqueous topical minoxidil formulation containing SEPA (2-n-nonyl-1,3-dioxolane) for enhancement of percutaneous absorption was under evaluation. SEPA does not have chromophore for either ultraviolet or fluorescence detection using liquid chromatography and has no functional groups for derivatization. Therefore, a direct gas-chromatographic method with flame-ionization detection (GC-FID) was developed. Owing to the limited detection response of the FID detection, it needs a selective and concentrated extract for GC-FID analysis to improve the assay sensitivity to meet the requirement for pharmacokinetic evaluation after topical application. In addition, SEPA is a very volatile compound. Any extraction procedures involving evaporation will result in a poor recovery. The application of solid-phase extraction (SPE) makes it possible to achieve a selective and a 10-fold concentrated extract with an absolute extraction recovery of approximately 90%, which greatly improved the assay sensitivity. This method involved the extraction of SEPA and the internal standard (2-n-heptyl-1,3-dioxolane) from serum (0.1-1 ml) with 100 microliter of hexane-chloroform (1:1, v:v) using a 50 mg 1.0 ml-1 phenyl SPE column (Varian, Harbor City, CA, USA), followed by direct GC-FID analysis on a fused-silica column chemically bonded with cross-linked methyl silicone gum phase (Hewlett Packard Ultra-1, 12 m x 0.2 mm x 0.33 micron, Avondale, PA, USA). The assay demonstrated a lower limit of quantitation of 2.5 ng ml-1 and a linear range of 2.5 to 250 ng ml-1 with intra- and inter-assay precision and accuracy of < or = 10%.
Zhong
W Z
WZ
Drug Metabolism Research, Pharmacia and Upjohn, Kalamazoo, MI 49001-4940, USA. wei-zhu.zhong@am.pnu.com
D000042:Absorption; D000277:Adjuvants, Pharmaceutic / Q000378:metabolism*; D000287:Administration, Topical; D000505:Alopecia / Q000188:drug therapy; D000818:Animals; D002849:Chromatography, Gas / Q000379:methods*; D004148:Dioxolanes / Q000097:blood*; D016503:Drug Delivery Systems; D004355:Drug Stability; D006801:Humans; D008914:Minoxidil / Q000008:administration & dosage / Q000097:blood / Q000627:therapeutic use; D011817:Rabbits; D051381:Rats; D012680:Sensitivity and Specificity
D000277:Adjuvants, Pharmaceutic; D004148:Dioxolanes; C111109:2-n-nonyl-1,3-dioxolane; D008914:Minoxidil