Research On Rapid Detection Of Clenbuterol

Clenbuterol is a kind of epinephrine, which is being abused more and more toredistribute livestock fat recent years. Clenbuterol in food endangers consumers’health seriously and hit their consuming confidence deeply. It has been found thatthere are lots of methods to detect clenbuterol home and abroad, including HPLC、GC-MS、ELISA、DIGFA、BS and so on. Owning to various disadvantages, methodsmentioned above could not meet requirements of quick、low-priced and batchdetection. Aiming to solve these problems, this thesis focuses on developing a newmethod, using alkaline potassium permanganate to detect clenbuterol in serum.Firstly, the proper colour-display reagent and best parameters are as follows:The best reaction parameters for guaiacol are:0.36mol/L KHSO4,0.72mol/LNaNO₂,0.11mol/L H₂NSO₃NH₄, volume ratio is0.6:0.2:1, detection limit is5.4μg/mL; the best reaction parameters for1-alpha-naphthol are:0.10mol/L NaNO₂,1.50mol/L HCL,0.15mol/L H₂NSO₃NH₄, volume ratio is1:1:1, detection limit is4.0μg/mL; the best reaction parameters for alizarin are:5.0×10^-4mol/L alizarin,1.0×10^-5mol/L acetic acid, volume ratio is1:1, detection limit is4.0μg/mL; the bestreaction parameters for N-（1-naphthalene base）-ethylenediamine are:0.2mol/LHCL,0.1mol/L NaNO₂,0.3mol/L H₂NSO₃NH₄, volume ratio is1:1:1, detection limitis1.2μg/mL; the best reaction parameters for alkaline potassium permanganate are:1.5mol/L NaOH,5.0-×10⁴g/mL KMnO₄, VNaOH：VKMnO₄=10:1, detection limit is1.0ng/mL.We applied alkaline potassium permanganate to detect clenbuterol in serum,optimizing parameters by orthogonal experimental design and general rotatoryanalysis,the best parameters are:0.468mol/L NaOH,0.3273×10-g/mL KMnO₄,VNaOH：VKMnO₄=0.159.With the clenbuterol’s concentration of X-axis and absorbencyof Y-axis, standard curve equation was computed as follow: y=0.6614-0.1057x（R²=0.9973）,detection limit is1ppb.This method could distinguish clenbuterol fromadrenalin hydrochloride, ractopamine and salbutamol sensitively. RSDs in and out ofbatch are respectively2.10%-16.67%and0.71%-7.44%.We made contrast trail of alkaline potassium permanganate and ELISA, both the standard point fitting error andsample recovery measuring error of the former are lower than the later, therefore thestability of alkaline potassium permanganate is better than ELISA.Basing on the reacting principle of Clenbuterol and potassium permanganate, weresearched a type of quick test paper with alkaline-resisting PVC, the detection limitis1ppb; experiments showed that test paper could be stable for3months under4℃,for2months under25℃; when the concentration of adrenaline hydrochloride,ractopamine and salbutamol reached100ng/mL, there was no interference.This thesis researched the best parameters of testing Clenbuterol with potassiumpermanganate, did experiments to test specificity and reproducibility, validatedexperiment principle and researched a new type of quick test paper. This is a novelmethod, which is more quick, sensitive, convenient and inexpensive, it providestheory basis for quick detection of Clenbuterol.