Study On The Detection Methods About The Opium Poppy Husk Added In Foods

Among the many food safety problems, the wrongful act about abusing the non-food processing additives bring to a serious food safety risks. The opium poppy husk contains morphine, codeine, papaverine and other alkaloids. Because of the organism effect to the opium poppy husk like the morphine, the harm about eating for a long time is like smoking the heroin. Therefore, the state is included in the control of narcotic drugs, and prohibite illicit supplying, transporting and using. Some lawless elements added them to the food in order to economic interests, and it endanger people's health seriously. Therefore, it is urgent and significant to establish a simple, rapid and selective method to determine the opium poppy husk added in foods.In this work, the methods of HPLC and gene detection have been developed for determination of the opium poppy husk added in foods.Method 1, A method was developed for determination of three alkaloids in the opium poppy husk added in foods by High Performance Liquid Chromatography. The analytes were extracted by ultrasonic-microwave synergistic extraction technique, and partitioned with organic solvents. Separation were carried out on the Agilent XDB-C18(4.6×150 mm) with a binary gradient elution programme of acetonitrile-phosphate buffer salt at a flow-rate of 1.0 mL/min. Detection was performed by UV absorbance at 210 nm by morphine, 213 nm by codeine, 253 nm by papaverine.The method of external standard was used for quantitative analysis. The method of imidazolinone herbicides showed good linearity with correlation coefficients being up to 0.9999 with three alkaloids. The average recovery are 87.87% , 90.80% , 89.47% and the RSD of the precision are 1.276%, 1.523%, 1.926% and the RSD of the reproducibility are 1.268%,1.687%,1.738%, respectively.Method 2, A method was developed for determination of the opium poppy husk added in foods by gene detection. It is used to extract the DNA in samples with the CTAB method that is improved. PVP-40 and VC will be shared to prevent oxidation. In present study, The reaction mixture consisted of 1μL of Taq(5 U/μL of stock solution), 2.5μL of 10×PCR amplification buffer, 3.5μL of dNTPs(1mM each in solution), 1.5μL of each primer(10μM stock solution) , DNA templates 5μL and 10μL of double-distilled water. The reaction was run under the following conditions: Cool start; DNA pre-denaturation at 94℃for 5 min; DNA denaturation at 94℃for 30 s, primer annealing at 60℃for 30 s, and DNA extension at 72℃for 60 s for 35 cycles; the last extension was performed at 72℃for 6 min. The PCR products were examined by electrophoresis with 2% agarosegel.The two methods developed have been used for analyzing the opium poppy husk added in foods. Method 1 had high recovery, sensitivity and precision, that is suitable for the determination of the alkaloid in opium poppy husk in food. Method 2 is only the genetic testing of this method. There are lots of things that need to improve, and will have broad application prospect.