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Quantitation of a Dinitro Flavonoid (DNF) in Rat Plasma Using the Varian 1200L LC/MS/MS System

D. Garteiz, R. Morrow, and B. Alexander, TexMS Analytical Services, Houston, TX C.
Schmidt, Varian, Inc., Walnut Creek, CA

Introduction

Flavonoids are compounds that make up the pigments in fruits, vegetables, and herbs. There have been reports of flavonoids having multiple beneficial therapeutic effects for a large number of serious illnesses, such as rheumatoid arthritis, cancer, cataracts, anxiety, and depression. The objective of this work was to carry out a pharmacokinetic evaluation to determine a profile with the formulation being tested at increasing doses where there is pharmacological effect.

A Varian 1200L LC/MS/MS system was used for the quantitation for plasma levels of a novel dinitro-substituted flavone (DNF) in samples from a pharmacokinetic study following oral administration of the drug. Flavone was used as the internal standard. Structures are shown in Figure 1.

Instrumentation

Varian 1200L LC/MS/MS equipped with ESI source

Materials and Reagents

DNF provided by Faber-Kramer Pharmaceuticals, Houston, Texas.

Flavone (Catalog Number F602) from Sigma-Aldrich.

All other chemicals are reagent grade or HPLC grade.

Sample Preparation

A 200 μL aliquot of rat plasma containing DNF is spiked with 10 μL of flavone (0.2 ng) as the internal standard (ISTD). The rat plasma is made basic and extracted with hexane: chloroform (2:1, v/v).

After separation of the phases by centrifugation, the organic layer is transferred by pouring into a silanized conical tube and evaporated to dryness. The dry residue is reconstituted in 50 μL of methanol:water (60:40, v/v).

A 10 μL aliquot is injected into an ESI-LC/MS/MS for analysis using Selected Reaction Monito
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