[LC/ESI-MS/MS method for the simultaneous determination of macrocyclic lactone parasiticides in livestock products and fish].

An LC/ESI (positive-mode)-MS/MS method was developed for simultaneous quantification of 8 macrocyclic lactones (abamectin B1a, abamectin 8,9-Z isomer B1a, emamectin benzoate B1a, emamectin benzoate 8,9-Z isomer B1a, ivermectin, eprinomectin B1a, doramectin and moxidectin) in animal tissues, egg, milk and honey. The separation was achieved on a TSK-GEL ODS 100 V column (2.0 x 50 mm, 3 microm) with a mobile phase consisting of 0.1% formic acid in acetonitrile, and 0.1 mM ammonium formate-0.1% formic acid in water, at a flow rate of 0.2 mL/min with gradient elution. Linear calibration plots were obtained with high correlation coefficients (r=0.998-0.999). The LOQ and LOD ranged from 0.02-1.5 ng/mL and 0.1-5 ng/mL, respectively. Average recoveries were in the range of 70.8-117.1% with associated RSD values<15% (n=10) for repeatability and reproducibility. The spiking levels for recoveries and RSDs met the validation criteria for Japanese maximum residue limits (MRLs). Based on these results, the proposed analytical method has been proven to be highly efficient and suitable for routine determinations of macrocyclic lactones in animal food matrices.

Simultaneous determination of avermectins in bovine tissues by LC-MS/MS.

Analytical method for the simultaneous quantification of avermectins (AVMs), abamectin B1a, abamectin 8,9-Z isomer B1a, emamectine benzoate B1a, emamectine benzoate 8,9-Z isomer B1a, ivermectin, eprinomectin B1a, doramectin and moxidectin in bovine tissues (muscle, liver and fat) was developed by LC-MS/MS in electrospray positive ion mode. The separation was achieved on a short TSK-GEL ODS 100V column with the mobile phase consisting of acetonitrile and aquatic 0.1 mM ammonium formate containing 0.1% formic acid v/v at a flow rate of 0.2 mL/min with gradient elution. Liquid-liquid extraction with isooctane was used for the sample extraction/preparation of analytes in bovine samples. The linearity of the calibration curves was excellent in matrix-matched standards, and yielded the coefficients (r(2)=0.997-0.999, range from LOQ to 500, 1000 or 5000 ng/g) of determination of the target analytes. Recoveries were in the range of 87.9-99.8% with associated precision values (within-day: 1.5-7.4%, n=6, and between-day: 1.5-8.4% for 3 days) for repeatability and reproducibility. LC-MS/MS method has been proven to be highly efficient and suitable for the simultaneous determinations of eight AVMs in bovine tissue samples.

Determination of avoparcin in animal tissues and milk using LC-ESI-MS/MS and tandem-SPE.

A highly sensitive and selective method using LC-ESI-MS/MS and tandem-SPE was developed to detect trace amounts of avoparcin (AV) antibiotics in animal tissues and milk. Data acquisition using MS/MS was achieved by applying multiple reaction monitoring of the product ions of [M + 3H](3+) and the major product ions of AV-alpha and -beta at m/z 637 --> 86/113/130 and m/z 649 --> 86/113/130 in ESI(+) mode. The calculated instrumental LODs were 3 ng/mL. The sample preparation was described that the extraction using 5% TFA and the tandem-SPE with an ion-exchange (SAX) and InertSep C18-A cartridge clean-up enable us to determine AV in samples. Ion suppression was decreased by concentration rates of each sample solution. These SPE concentration levels could be used to detect quantities of 5 ppb (milk), 10 ppb (beef), and 25 ppb (chicken muscle and liver). The matrix matching calibration graphs obtained for both AV-alpha (r >0.996) and -beta (r >0.998) from animal tissues and milk were linear over the calibration ranges. AV recovery from samples was higher than 73.3% and the RSD was less than 12.0% (n = 5).