Page 634 - Veterinary Toxicology, Basic and Clinical Principles, 3rd Edition

Anticoagulant Chapter | 46 599

VetBooks.ir samples, liquid, and solid biological samples are briefly fluorescence detection (Hunter, 1985; Hunter and Sharp,
1988), as well as other fluorescent methods (Mundy and
summarized.
Machin, 1982; AOAC, 1976a,b) for diphacinone (Bullard
Bait formulation and technical material: HPLC meth-
ods to detect chlorophacinone (Vigh et al., 1981; Grant et al., 1975, 1976), bromadiolone (Deepa and Mishra,
and Pike, 1979), difenacoum (Hadler and Shadbolt, 1975), 2005), and brodifacoum (Fu et al., 2006). Diode array
and Rozol (Kawano and Chang, 1980) in bait formulation (Yang et al., 2001) and rapid fluorimetric methods for
have been reported. Recently, NMR has been used to detection of chlorophacinone or diphacinone (Cai et al.,
detect source-to-source and batch-to-batch variations in 2011) in human serum based on formation of histidine
distereomer ratios of brodifacoum (Cort et al., 2012). complexes have also been reported.
Air, soil, or water environmental samples: A recent Increasingly, methods are developed to look for many,
method quantifies brodifacoum in workplace air if not all, anticoagulant rodenticides in a single method.
0.2B10.0 μg/mL, with a detection limit of 0.08 μg/mL An early example extracted and detected eight anticoagu-
and minimum detectable concentration of 0.00067 mg/m 3 lant rodenticides in serum or liver using fluorescence and
(Fu et al., 2013). An LC-ESI-MS analytical method is UV detection with detection of 4-hydroxycoumains to
reported to determine chlorophacinone, bromadiolone, about 1 ng/mL of serum and 1 ng/g of liver; and indane-
pindone, diphacinone, warfarin, coumatetralyl, brodifa- diones down to 10 ng/mL of serum and 10 ng/g of liver
coum, fluocoumafen, and difenacoum residues with limits (Chalermchaikit et al., 1993; Felice et al., 1991; Felice
of detection and quantitation of 0.09 and 2.2 μg/kg in soil and Murphy, 1989). A number of methods for simul-
and 0.08 and 1.7 μg/L in water (Herna ´ndez et al., 2013a). taneous detection of multiple rodenticides in serum
Food: An interesting new method uses HPLC to detect (Kuijpers et al., 1995), with detection limits of 3 12 ng/mL
anticoagulant rodenticides in soft drinks (Dimuccio et al., for fluorescence and 20 75 ng/mL for UV detection
1991), an ion pair LC method to detect chlorophacionone (Kuijpers et al., 1995; McCarthy et al., 1997; Mura
and diphacinone in oats (Primus et al., 1998), and a cell et al., 1992; Feng et al., 1999; Jin et al., 2007)or less
culture/ELISA assay to detect anticoagulant rodenticides than 10 851 ng/L for brodifacoum, difethialone, and
in treated grain (Lawley et al., 2006) have all been difenacoum (Robben et al., 1998), were reported.
reported.

Exposed Animals: Liquid (Serum, Blood, Urine) Solid: (Hair, Feces, Tissues and Food)
Methods to detect anticoagulant rodenticides in exposed A high performance TLC method with an estimated
animals were initially focused on individual compounds, detection limit of 200 ppb and 87% recovery from liver
then expanded to detection of groups of compounds in the has been reported (Berny et al., 1995). HPLC methods for
same method. Matrices from exposed animals are com- chlorophacinone (Hunter, 1984), difethiolone (Goldade
monly either liquids, such as urine, blood or serum, or et al., 1998), difenacoum (Mundy and Machin, 1977),
solids, such as tissues. Methods are presented similarly. brodifacoum (Hagenboom and Rammell, 1983), and
Warfarin: Early fluorimetric and GC methods were bromadiolone (Nahas, 1986) in tissue have been reported.
for serum (Corn and Berberich, 1967; Fasco et al., 1977; Methods for detection of multiple molecules from tis-
Hanna et al., 1978; Lee et al., 1981; Lewis et al., 1970; sue samples, such as a solid phase cartridge extraction
O’Reilly et al., 1962; Vesell and Shivley, 1974; Welling from liver, with recoveries ranging from 52% for difena-
et al., 1970; Mildha et al., 1974). coum to 78% for warfarin, with limits of detection of
Brodifacoum: Numerous HPLC methods for detection about 10 ppb for warfarin and difenacoum and 110 ppb
of brodifacoum (Murphy and Gerken, 1989; Koubek for chlorophacinone (Fauconnet et al., 1997; Jones, 1996;
et al., 1979; Hoogenboom and Rammell, 1983; Keiboom Addison, 1982), were reported. HPLC: method reports
and Rammel, 1981; Ray et al., 1989; Hong et al., 2010; simultaneous determination of bromadiolone, brodifacoum,
Kim et al., 2010; Fu et al., 2012) or bromadiolone coumachlor, coumafuryl, coumatetralyl, difenacoum, flo-
(Subbiah et al., 2005; Hunter and Sharp, 1988) in serum coumafen, and warfarin in liver (Armentano et al., 2012).
or plasma have been reported. One method reports the use Many recent methods use mass spectroscopy. For
of difenacoum as the internal standard (O’Bryan and example, for indandiones diphacinone and chlorophaci-
Constable, 1991). none in whole blood or urine, and the 4-hydroxy coumar-
Difenacoum: An HPLC method distinguishes cis and ins, bromadiolone, brodifacoum, coumachlor,
trans isomers with detection limits of 5 ng/mL (Kelly coumatetralyl, difenacoum, and warfarin, in whole blood
et al., 1993). specimens (Schaff and Montgomery, 2013), or LC-IT/MS
Various detection methods have also been used histor- method with limits of detection of 5 25 ng/mL in
ically, including a postcolumn pH shift to enhance plasma.