Simultaneous Analysis Of Residues Of Non-steroidal Anti-inflammatory Drugs (Nsaids) By HPLC

The non-steroidal anti-inflammatory drugs(NASIDs)had the curative effect on anti-phlogosis, pain relief and treat rheumatic diseases. In recent years, due to the breeding tends to intension and large scale; the Non-steroidal anti-inflammatory drug had been used widely in the breeding. Because of flood using the NSAIDs, the water and the water organisms surrounding farms were subjected to a certain degree of pollution, the Non-steroidal anti-inflammatory drugs could cause a series of side effects such as the gastric ulcer, liver and kidney damage, blood and the nervous system disorders, there were about thirty million people worldwide used this kind of medicine to alleviate pain, including 0.2% ~ 0.4% percent of patients to death. Therefore, many countries and regions stipulated the Maximum residual, At present, In China detecting the NSAIDs mainly according to the Entry-Exit Inspection and Quarantine standard (SN/T 2190-2008) and the European standard (2002/1352 / EC), But haven't formulated corresponding the National standards, In order to guarantee the animal-derived food export trade and protect consumers' health, ensuring the NSAIDs residues in animal-derived food was safety in to eat range. So establishing a simple, quick, reliable and sensitive detection method is the foundation to control the residual quantity.At present, the methods for detecting the NSAIDs including GC, GC/MS, HPLC, HPLC/MS, CE etc.This paper has chosen four kinds of non-steroidal anti-inflammatory drugs to research the residual detect method by its properties; they were flunixin meglumine, meloxicam, and ketoprofen and diclofenac sodium respectively. The NSAIDs were extracted by the acidified ethanol assisted with ultrasonic-microwave, purified with diatomite SPE and detected by HPLC, Established simultaneously detection methods of the four kinds of NSAIDs residue in the animal-derived foods and aquatic organism, the research results were as follows.1. Optimization of sample extraction conditions: with the mutton as research object, Compared the acidified organic solvents such as acetonitrile, methanol and ethanol on extraction effect, the results showed that acidified ethanol and acetonitrile had higher recovery rate than acidified methanol , given the acidified acetonitrile was toxic, harmful to the environment, so this experiment selected acidified ethanol as extraction agents, On this basis, also studied the assistant extraction process by ultrasonic wave, microwave and vortex concussion, the results showed that the effect of the ultrasonic-microwave extraction is better,2. Optimization of purification conditions: the mutton and carp compositions were more complex, it released huge amounts of polarity (such as amino acids) and non-polarity (such as lipid) impurities by ultrasonic-microwave, the samples were purified through liquid-liquid extraction and solid-phase extraction method. This experiment used hexane to remove the non-polar compositions, compared different SPE fillers on 4 NSAIDs purified effect such as C18, diatomite, alumina and silica gel. The results showed that the diatomite solid phase extraction column on the effect of target objects retain was better.3. Optimization of chromatography conditions: used Hypersil ODS C18 as chromatographic column, it was reported that acetonitrile-0.1% acid (with 0.5mmol/L ammonium acetate) as mobile phase ,but the peaks'shape were more wide and delayed, this experiment optimized mutton tissue samples analysis'chromatography condition, chosen acetonitrile-0.2% triethlamine (40:60, v/v, pH=3.5 adjusted by phosphate acid) as mobile phase at flow rate of 0.8 ml/min at 30℃, the detected wavelength was set at 255nm; for the carp tissue samples, the mobile phase was slightly adjusted as acetonitrile - 0.1% triethlamine (40:60, v/v, pH=3.5 adjusted by phosphate acid),it made the sample analysis time within 10min.4. Establishment of 4 NSAIDs residues simultaneous detection methods: with HPLC(secondary array detector), simultaneously detected flunixin meglumine, meloxicam, ketoprofen and diclofenac sodium in mutton sample, the limits of detection (LOD, S/N = 3) were 5μg/kg~10μg/kg, the recoveries were in the range of 65.3 ~ 99.6%; for carp sample, the limits of detection (LOD, S/N = 3) were 5μg/kg~10μg/kg, the recoveries were in the range of 72.5 ~ 93.8%. The method was easy, rapid, sensitive, specific and safe, could meet the requirement for the qualitative and quantitative analysis.