Determination Of Eugenol Residue In Fish Meat By SPE-UPLC-MS/MS

Eugenol is a plant spices, with strong clove aroma that is volatile, except for the preparation of the fragrance, cosmetics, soap flavor outside, its pharmacological action can used as analgesic agents, preservatives, and narcotics and so on. Because of its low cost, obvious effect, in recent years as an anesthetic widely used in the transportation of aquatic products. The Australia, New Zealand, intelligence and other countries have eugenol column is no residual period of legitimate live fish transportation with the anesthetic, however, the National Toxicology Program(NTP) released the toxicological data show that eugenol on rodents are carcinogenic or potential carcinogens, its safety is questionable. Japan's positive list of eugenol in in aquatic products in maximum residue levels also have limitations, and the provisions of the eugenol residue limit of 0.05 mg / kg. World Health Organization under the food additive Joint Expert Committee(JECFA) released in 1980 risk assessment results, eugenol ADI value(2.5 mg/kg BW), and our country for eugenol residue limit is still in a blank stage. Therefore, it is urgent to build detection of eugenol, and accurate pretreatment method, qualitative and quantitative methods. The determination of residues in fish eugenol tandem mass spectrometry with solid phase extraction high performance liquid chromatography. Methods: the fish can be eaten part of homogeneous treatment, sampling 2.00 g in 15 mL centrifuge tube, adding 2.00 g anhydrous sodium carbonate, 8mL methanol, vortex shock 2min, ultrasonic extraction 10 min, 10000r/min centrifugal 5min. Remove the extract liquid to another 50 mL centrifuge tube; in the residue to add 8mL methanol, repeated extraction, and the extraction of liquid. 10 mL was added to n-hexane in the centrifuge tube, centrifugal concussion. Taking the lower solution 2mL, diluted with water to 6mL, to C18 solid phase extraction purification treatment. Add 5 mL of methanol, 5 mL water washing, activation of the column. Column dilution to be cleaned, the control flow rate of 3 mL/min. With 10 mL water leaching column after adding 5 mL methanol elution velocity control 2mL/min, collecting the eluent. 40 C nitrogen blowing to nearly dry, with methanol volume to 1ml, for LC-MS/MS analysis. UPLC BEH C18 Waters chromatographic column(50mm * 2.1mm, 1.7 mu m); sample volume of 5 m; column temperature 40?; flow rate of 0.2ml/min; mobile phase A for methanol, mobile phase B 0.1% ammonia. Negative ion scan mode(ESI) multiple reaction monitoring detection(MRM), capillary voltage-3.00 kV, cone voltage of-20 V, lens voltage 0.1V, source temperature of 110?, to solvent gas temperature of 350?, to the solvent gas flow 650L/hr, anti blowing gas flow 50L/hr. The qualitative ion pair was 148 m/z and 121m/z, and the quantitative ion was 148 m/z. The results showed that the standard solution of eugenol in the 2.0 ~ 100.0ng/mL concentration range, the linear correlation coefficient r=0.9986, method detection limit of 2 g/kg, RSD was 1.02% ~ 2.85% and the recovery rate was 88.4% to 104.7% between.