Reversed-phase ion-pair liquid chromatographic determination of chlorophacinone and diphacinone in steam-rolled oat baits and steam-rolled oat/wax baits

A reversed-phase ion-pair liquid chromatographic (LC) method was developed for analysis of steam-rolled oat (SRO) baits fortified with either chlorophacinone or diphacinone. Baits were prepared with and without paraffin wax. Chlorophacinone or diphacinone was extracted from wax-free SRO baits with 5 mM tetrabutylammonium phosphate methanolic ion-pairing solution. Wax baits were initially extracted with petroleum ether and then cleaned up by liquid extraction into methanolic ion-pairing solution containing 20% water. SRO extracts were analyzed with reversed-phase ion-pair LC. Chlorophacinone and diphacinone were quantified by UV absorption at 325 nm. Recoveries from SRO fortified with chlorophacinone at 25 and 150 microgram/g were 90.7 and 90.8%, respectively, whereas for diphacinone at the same levels, recoveries were 93.5 and 92.3%, respectively. Recoveries from wax baits fortified at 25 and 75 microgram/g chlorophacinone were 98.5 and 100%, respectively, whereas for diphacinone at the same levels, recoveries were 93.6 and 98.0%, respectively. Method limits of detection for chlorophacinone and diphacinone in SRO baits were estimated to be 1.0 and 0.76 microgram/g, respectively. Method limits of detection for chlorophacinone and diphacinone in wax baits were estimated to be 42 and 2.8 microgram/g respectively