[Determination of streptomycin and dihydrostreptomycin residues in tomato ketchup by UPLC-MS/MS].

OBJECTIVE: To develop a method for simultaneous determination of streptomycin and dihydrostreptomycin residues in tomato ketchup by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). METHODS: The sample was dissolved with phosphorus solution (pH 2) and extracted by ultrasonic. The pigment was removed with n-hexane. Then, the sample was cleaned up by HLB SPE. The HILIC chromatographic column (100 mm x 2.1 mm, 1.7 µm) was used to complete the separation under gradient elution. The mixed solution of 0.1% formic acid solution and acetonitrile was used as mobile phase. The detection of streptomycin and dihydrostreptomycin were carried out by MS/MS under multiple reaction monitoring (MRM) mode. The external standard method was used for quantitative analysis. RESULTS: The calibration curves for streptomycin and dihydrostreptomycin were indicated in the range of 0.005 - 0.100 mg/kg, and the detection limits were both 0.005 mg/kg. The recoveries of streptomycin and dihydrostreptomycin were ranged from 79.5% to 93.9% with relative standard deviations no more than 10%. CONCLUSION: The method is simple and accurate to meet the requirements for determination of streptomycin and dihydrostreptomycin residues in tomato ketchup.

Indirect competitive enzyme-linked immuno-sorbent assay (ELISA) for nitroimidazoles in food products.

Ronidazole was used as the starting material to prepare an immunogen and coating antigen. An anti-nitroimidazole monoclonal antibody was produced and an indirect competitive ELISA was established to detect nitroimidazole compounds in food products. The IC(50) values were determined to be 0.20 ng/ml for metronidazole, 4.0 ng/ml for tinidazole, 0.17 ng/ml for dimetridazole and 0.24 ng/ml for ornidazole. Considering that nitroimidazoles were commonly used as veterinary drugs, nitroimidazole residues in food products of animal origin were detected by the method. The coefficient of variation for nitroimidazoles determination in contaminated chicken, chicken liver and shrimp were all <14% and the recovery rate was in the range 74.0-90.6%. The results proved that the developed method was successful in detecting nitroimidazoles in food products.

Generation of anti-trenbolone monoclonal antibody and establishment of an indirect competitive enzyme-linked immunosorbent assay for detection of trenbolone in animal tissues, feed and urine.

Trenbolone (TRE) is a steroid used by veterinarians on livestock to increase appetite and body weight. The use of TRE has been restricted because of its harmful side effect for consumers. To effectively control TRE residue in food and food product, a rapid and convenient immunoassay was developed by preparing an anti-TRE monoclonal antibody. The immunogen and coating antigen were prepared by coupling TRE hapten with carrier proteins via 1-ethyl-3-(dimethylaminopropyl)carbodiimide hydrochloride (EDC) method. The optimized method gave an average IC(50) value of 0.323 ng mL(-1) towards TRE and an average detection limit (LOD) of 0.06 ng mL(-1), which is much lower than the maximum residue levels (2.0 ng g(-1)) accepted by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). The specificity of the antibody was evaluated by measuring cross-reactivity of six structurally related compounds, including 19-nortestosterone (9.7%), testosterone (0.13%), methyltestosterone (<0.01%), methandrostenolone (<0.01%), (+)-dehydroisoandrosterone (<0.001%) and ß-estradiol (<0.001%). The recovery rates of the test in detection of TRE-fortified animal tissue, urine and animal feed samples were in the range of 81.3-89.4%, while the intra- and inter-assay coefficients of variation were less than 12.0%.

Preparation of anti-Sudan red monoclonal antibody and development of an indirect competitive enzyme-linked immunosorbent assay for detection of Sudan red in chilli jam and chilli oil.

Sudan dyes are banned to be used in food additives because of the carcinogenicity of their metabolites. A rapid and sensitive indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed to detect the residues of Sudan dyes. Novel immunogen and coating antigen were synthesized via glutaraldehyde linking. The hapten-bovine serum albumin (BSA) was applied as immunogen and the hapten-ovalbumin (OVA) was served as coating antigen. The monoclonal antibody obtained showed high sensitivity to Sudan I with an IC(50) value of 1.7 µg L(-1) in buffer and was suitable to detect the residues of Sudan red in food products. The specificity of the assay was studied by measuring cross-reactivity of the antibody with the structurally related compounds of Sudan II (<1%), Sudan IV (<1%) and para red (120%). Chilli jam and chilli oil samples spiked with Sudan dyes were analyzed by the method. The detection limit (LOD) of the ELISA method applied in chilli jam and chilli oil was 9.0 µg L(-1) and 19.6 µg L(-1), respectively. The recovery rates of Sudan-I in chilli oil and chilli jam were in the range of 80%-110% with coefficients of variation <25%. The intra-assay variation and inter-assay variation in buffer were both <9%.

Preparation of monoclonal antibody for melamine and development of an indirect competitive ELISA for melamine detection in raw milk, milk powder, and animal feeds.

Melamine (MEL) has been involved in several food recalls after the discovery of severe kidney damages in children and pets poisoned by melamine-adulterated food. To detect MEL residue in foods and animal feeds, an indirect competitive ELISA (cELISA) method was developed in this study based on preparation of monoclonal antibodies (MAbs) to MEL. The immunogen was prepared by linking MEL hapten with carrier protein via carbodiimide method. The method is applicable in the range of 5.0-135.0 microg L(-1) MEL in buffer solution, with an IC(50) value of 22.6 +/- 1.9 microg L(-1). The MAbs showed high specificity with low cross-reactivity (< or =1%) toward cyanurate, ammelide, and ammeline. The method was utilized in the detection of MEL in raw milk, milk powder, and animal feeds, with detection limits of 0.1 mg L(-1) for milk, 0.2 mg kg(-1) for milk powder, and 0.5 mg L(-1) for feeds. The recovery ratio was 79-110% for all matrices. The intra-assay and interassay coefficients of variation were <12.0 and <13.0%, respectively. Finally, the application of the cELISA in quantity evaluation of MEL in various feeds from local markets was evaluated and discussed.

Preparation of a monoclonal antibody and development of an indirect competitive ELISA for the detection of chlorpromazine residue in chicken and swine liver.

BACKGROUND: Chlorpromazine is a typical antipsychotic drug used to make food-producing animals calm and promote growth as feed additives. Accumulation of chlorpromazine in animal bodies would cause side effects in the circulatory and nervous systems, and have adverse effects on blood cells, the skin and the eye. To detect the chlorpromazine residue in food producing animals, an indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed based on preparation of an anti-chlorpromazine monoclonal antibody. RESULTS: The antibody generated from immunogen of cationic bovine serum albumin (cBSA) coupled with chlorpromazine showed high sensitivity toward chlorpromazine with an IC(50) value of 0.73 ppb. The ELISA method was applied to detect swine liver and chicken samples spiked by chlorpromazine and satisfactory results were obtained. The recovery rates in chicken and swine liver were in the range of 88-95% and 86-95%, respectively; the intra-assay coefficients of variation were both < 15.3% and < 13.5%, respectively. CONCLUSION: An indirect competitive ELISA method based on a monoclonal antibody towards chlorpromazine with excellent sensitivity and specificity has been successfully developed. The immunoassay provided in this study was a hopeful alternative to chromatography spectrometry for regulatory analysis of chlorpromazine residue in food-producing animals.

[One stage repair of scrotal and perineal hypospadias with prepuce island flap].

OBJECTIVE: To explore an effective method to repair penile-scrotal or perineal hypospadias in one stage with prepuce island flap. METHODS: Different prepuce island flaps were designed according to the different pathological anatomy of the penile-scrotal or perineal hypospadias. The prepuce island flaps were thus translocated and sutured to form the urethra. Thirty-one cases of hypospadias (21 cases of peinil-scrotal type, 10 cases of perineal type) were repaired with prepuce island flap. The biggest length and the width of the prepuce island flap were 7.5 cm and 1.5-1.8 cm respectively. RESULTS: All the cases resulted in a good contour of the penis and a normal anatomic position of urethral meatus without any redundancy or tortuosity. The urination was perfect and acceptable. CONCLUSION: One stage repair of penile-scrotal or perineal hypospadias with prepuce island flap can be considered as an acceptable effective surgical technique.

[Repair of foot defect with a reverse fascial pedicled posterolateral lower leg flap].

OBJECTIVE: To explore the applications of ipsilateral reverse fascial pedicled posterolateral lower leg flap in foot defect repair. METHODS: A fasciocutaneous flap was designed over the gastrocnemius muscle to repair the defects in the foot or the lower third of the leg. The flap was based on a perforator branch between the lateral malleolus and the Achilles tendon, coming from the vessel network of the ankle as well as the deep fascia. RESULTS: This method was used in 6 cases, including 2 children of 5-6 years old. All the flaps survived without necrosis and venous congestion. CONCLUSIONS: The operation is simple and a large flap over the whole gastrocnemius muscle can be raised without sacrifice of a main blood vessel. It is an excellent method for repair of the defects in the heel, medial and lateral malleolus, dorsum pedis and the lower third of the leg.