Analytical method for determination of florfenicol was developed for estimate veterinary drug residue of unestablished MRLs in meat. The method was validated in correspondence with the CODEX guideline for florfenicol residue in meat. The samples mixed with sodium sulfate were extracted with ethyl acetate. After clean-up, the residue was dissolved in mobile phase and analyzed using high-performance liquid chromatography with fluorescence detector. The calibration curve showed good linearity($r^2=0.9997$) within the concentration range of $0.05{\sim}1.0\;mg/kg$. The limit of detection(LOD) and limit of quantification(LOQ) were validated at 0.012 and 0.039 mg/kg, respectively. The recoveries in fortified meat ranged from 85.6 to 95.6%($1.1{\sim}5.3%$ RSD) at the 0.05 to 0.4 spiking levels. We monitored 150 samples of meats that were purchased in Korea(Seoul, Busan, Daegu, Daejeon and Gwangju). Among tested samples, florfenicol was detected in 1 of pig at the level of 0.040 mg/kg, and below LOQ in 1 of cattle, 2 of pig and 2 of chicken. The residues of florfenicol in the tested samples were within the MRLs.
The Journal of the Korean Society for Microbiology
/
v.35
no.2
/
pp.97-108
/
2000
Two-dimensional gel electrophoresis (2-DE) is an essential tool of proteomics to analyse the entire set of proteins of an organism and its variation between organisms. Helicobacter pylori was tried to identify differences between strains. As the first step, whole H. pylori was lysed using high concentration urea contained lysis buffer [9.5 M Urea, 4% CHAPS, 35 mM Tris, 65 mM DTT, 0.01% SDS and 0.5% Ampholite (Bio-Rad, pH 3-10)]. The extract ($10\;{\mu}g$) was rehydrated to commercially available immobilised pH gradient (IPG) strips, then the proteins were separated according to their charges as the first dimensional separation. The IPG strips were placed on Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) to separate according to molecular mass of the proteins as the second dimension. The separated protein spots were visualised by silver staining in order to compare different expression of proteins between strains. Approximately 120 spots were identified in each mini-protein electrophoresised gel, furthermore about 65 to 75 spots were regarded as identical proteins in terms of pI value and molecular weight between strains used. In addition, distinct differences were found between strains, such as 219-1, Y7 and Y14, CH150. Two representative strains were examined using strips which had pH range from 4 to 7. This strips showed a number of isoforms which were considered large spots on pH range 3-10. Furthermore, the rest of spots on pH 4-7 IPG strips appeared very distinctive compared to broad range IPG strips. 2-DE seems to be an excellent tool for analysing and identifying variations between H. pylori strains.
Proceedings of the Korean Radioactive Waste Society Conference
/
2009.06a
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pp.84-85
/
2009
New approaches for detecting, preventing and remedying environmental damage are important for protection of the environment. Procedures must be developed and implemented to reduce the amount of waste produced in chemical processes, to detect the presence and/or concentration of contaminants and decontaminate fouled environments. Contamination can be classified into three general types: airborne, surface and structural. The most dangerous type is airborne contamination, because of the opportunity for inhalation and ingestion. The second most dangerous type is surface contamination. Surface contamination can be transferred to workers by casual contact and if disturbed can easily be made airborne. The decontamination of the surface in the nuclear facilities has been widely studied with particular emphasis on small and large surfaces. The amount of wastes being produced during decommissioning of nuclear facilities is much higher than the total wastes cumulated during operation. And, the process of decommissioning has a strong possibility of personal's exposure and emission to environment of the radioactive contaminants, requiring through monitoring and estimation of radiation and radioactivity. So, it is important to monitor the radioactive contamination level of the nuclear facilities for the determination of the decontamination method, the establishment of the decommissioning planning, and the worker's safety. But it is very difficult to measure the surface contamination of the floor and wall in the highly contaminated facilities. In this study, the poly(styrene-ethyl acrylate) [poly(St-EA)] core-shell composite polymer for measurement of the radioactive contamination was synthesized by the method of emulsion polymerization. The morphology of the poly(St-EA) composite emulsion particle was core-shell structure, with polystyrene (PS)as the core and poly(ethyl acrylate) (PEA) as the shell. Core-shell polymers of styrene (St)/ethyl acrylate (EA) pair were prepared by sequential emulsion polymerization in the presence of sodium dodecyl sulfate (SOS) as an emulsifier using ammonium persulfate (APS) as an initiator. The polymer was made by impregnating organic scintillators, 2,5-diphenyloxazole (PPO) and 1,4-bis[5-phenyl-2-oxazol]benzene (POPOP). Related tests and analysis confirmed the success in synthesis of composite polymer. The products are characterized by IT-IR spectroscopy, TGA that were used, respectively, to show the structure, the thermal stability of the prepared polymer. Two-phase particles with a core-shell structure were obtained in experiments where the estimated glass transition temperature and the morphologies of emulsion particles. Radiation pollution level the detection about under using examined the beta rays. The morphology of the poly(St-EA) composite polymer synthesized by the method of emulsion polymerization was a core-shell structure, as shown in Fig. 1. Core-shell materials consist of a core structural domain covered by a shell domain. Clearly, the entire surface of PS core was covered by PEA. The inner region was a PS core and the outer region was a PEA shell. The particle size distribution showed similar in the range 350-360 nm.
Lee, Han Sol;Park, Ji-Su;Lee, Su Jung;Shin, Hye-Sun;Kim, Ji-Young;Yun, Sang Soon;Jung, Yong-hyun;Oh, Jae-Ho
Korean Journal of Environmental Agriculture
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v.39
no.3
/
pp.246-252
/
2020
BACKGROUND: Pesticides are broadly used to control weeds and pests, and the residues remaining in crops are managed in accordance with the MRLs (maximum residue limits). Therefore, an analytical method is required to quantify the residues, and we conducted a series of analyses to select and validate the quick and simple analytical method for tolpyralate in five agricultural products using QuEChERS (quick, easy, cheap, effective, rugged and safe) method and LC-MS/MS (liquid chromatography-tandem mass spectrometry). METHODS AND RESULTS: The agricultural samples were extracted with acetonitrile followed by addition of anhydrous magnesium sulfate, sodium chloride, disodium hydrogencitrate sesquihydrate and trisodium citrate dihydrate. After shaking and centrifugation, purification was performed with d-SPE (dispersive-solid phase extraction) sorbents. To validate the optimized method, its selectivity, linearity, LOD (limit of detection), LOQ (limit of quantitation), accuracy, repeatability, and reproducibility from the inter-laboratory analyses were considered. LOQ of the analytical method was 0.01 mg/kg at five agricultural products and the linearity of matrix-matched calibration were good at seven concentration levels, from 0.0025 to 0.25 mg/L (R2≥0.9980). Mean recoveries at three spiking levels (n=5) were in the range of 85.2~112.4% with associated relative standard deviation values less than 6.2%, and the coefficient of variation between the two laboratories was also below 13%. All optimized results were validated according to the criteria ranges requested in the Codex Alimentarius Commission (CAC) and Ministry of Food and Drug Safety (MFDS) guidelines. CONCLUSION: In conclusion, we suggest that the selected and validated method could serve as a basic data for detecting tolpyralate residue in imported and domestic agricultural products.
The optimum conditions for measuring cadmium content of less than 0.2ppm by flame atomic absorption spectrophotometry were investigated. The cadmium in urine was extracted by APDC-MIBK for the analysis by atomic absorption spectrophotometry after ashing them by a wet method. 1. Optimum conditions by APDC-MIBK and DDTC-MIBK extractions. The acidic aqueous solution was prepared with appropriate amount of 0.IN nitric acid, 5ml of 25% (W/V) sodium potasstum tartarate, 10ml of saturated ammonium sulfate, and 2ml of 2% APDC(or 1 ml of 5% DDTC) chelating agent. The total volume of solution was adjusted to 55 ml and pH to $2{\sim}10$ (or$7{\sim}10$). The aqueous solution was extracted with 10ml MIBK. Concentration of Triton X-100 did not effect the absorbance for APDC-MIBK extraction of cadmium, but absorbance decreased as the concentration increased for DDTC-MIBK extraction. The sensitivity and detection limits for the cadmium determination from APDC-MIBK extraction were 0.0038ppm and 0.0102, 0.0022ppm and 0.0116 for DDTC-MIBK, and 0.0132ppm and 0.0034 for 0.1N nitric acid. APDC-MIBK and DDTC-MIBK extractions were 3 times higher than 0.1N nitric acid for the sensitivity. 2. Excretion of cadmium in 24-hour urine by APDC-MIBK extraction. Determination of cadmium in urine by atomic absorption spectrophotometry of A.A. (Cd=2 mA) mode and B.C. (Cd=4 mA) mode and B.C. (Cd=4mA, $D_2=20mA$) mode showed some difference (p<0.05). The difference of cadmium determination and recovery according to method of standard additions and standard calibration curve method in urine was not significant (p>0.05, $93.48{\pm}11.78%,\;94.83{\pm}22.00%$). Excretion of cadmium in 24-hour urine collection from normal person and variance analysis within measurement variation was not significant (p>0.05), but between interindividual was significant (0.05). Determination of cadmium content by two different methods of flame atomic absorption spectrophotometry and dithizone colorimetry showed that the results from the two methods can be described by a regression line with a good correlation (y=1.0153x-0.2927, x=Cd by D.C., y=Cd by A.A.S., $r=0.8651^*$, p<0.01).
This study has been carried out to examine the feasibility of washing technique for reducing the heavy metal contamination level of tailing wastes and agricultural soil surrounding abandoned metal mines. Some organic acids with low molecular weight were used as washing solution. Initial contamination levels of copper and lead for some soil samples were found to exceed the standard levels of countermeasure and concern, and those of cadmium to approach the standard level of countermeasure. Experimental results using sequential extraction method revealed that more than half of copper and lead existing in tailing wastes are adsorbed forms available for plants. There are some proportional relationships between metal concentrations determined by using 0.1N HCI solution and those determined by sequential extractions. Citric acid was turned out to be superior to oxalic acid and acetic acid with low molecular weight in washing above three metals. When citric acid is used for washing heavy metals from soil, it is desirable to operate at pH less than 5.5 for better washing effect. Metal removal effect by citric acid solution has been proved to depend upon solution concentration and the mass ratio of solution to soil. Addition of SDS(Sodium Dodecyl Sulfate) to citric acid improved the washing effect of cadmium among three metal most significantly. while copper removal did not change. Washing technique using citric acid for removal of heavy metals from agricultural soil or tailing wastes is recognized to be an effective remediation method.
Serodiagnosis of parasitic infections is widely used, since parasites or their eggs are not always detected by ordinary methods. The sensitive tests such as ELISA are highly dependent on the purity of antigens used. To solve this problem. many workers have tried to find species-specific components of antigens, The present study was performed to determine the antigenic profile of crude saline extracts of 3, 5, 8 and 12-week old p. westermani worms, which were collected from experimentally infected cats, based on SDS-PAGE and immunoblot technique. The results were as follows: 1. The SDS-PAGE showed at least 30 Protein bands ranging from 229 kDa to 10 kDa molecular weight. The protein components of p. wsstermani changed chronologically during its developmental period. The 229 kDa band was recognized only in 12-week old worms ($$SEP_{l2}$). 2. Analysis by ELISA showed a significant increase in antibody levels at 3 weeks in infected cats using crude saline extract antigens ($SEP_3,{\;}SEP_5,{\;}SEP_8,{\;}SEP_{l2}$). 3. By EITB using $SEP_3$ and $SEP_5$ infected cats recognisea major protein bands with molecular weight of 60, 35, 28, 25 or 21 kDa at 3~12 weeks of infection, and 3 additional antigens, 19, 13 and 10 kDa, were detected at 8~12 weeks of infections. 4. Using $SEP_8$ 5 antigens, 91, 85, 31, 25 and 21 kDa, were consistently detected by all infected sera tested. In addition, 3 antigens of lg. 13 and 10 kDa were detected at 8~12 weeks of infection. Using $SEP_12$, similar results were obtained with that by using $SEP_8$ and 1 additional antigen of 229 kDa, specifically reacting with the sera from 12 weeks of in(traction, was recognized.
Enzyme-linked immunoelectrotransfer blot (EITB) using crude worm antigen of adult Paragonimus westermani was performed for human patients sera to identify the species-specific components. Crude antigen was obtained by homogenizing and centrifuging 24-week old adult worms at 10,000 rpm for 60 minutes in phosphate buffered saline (PBS, PH 7.2) containing: Phenyl methyl sulfonyl auoride (PMSF). Gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) was performed and blotted electrophoretically onto a sheet of nitrocellulose paper. The sheet was cut into strips and exposed to sera diluted 1 : 200 with PBS. SDS-PAGE showed 26 protein bands ranging 229 to 10 kDa. Of them 229, 91, 60, 50, 35∼31, 27, 25, 21, 17, 11 and 10 kDa components showed positive reaction with serum antibody of patients with p. westermani. Sera of patients infected with Clcnorchis sinensis reacted with 35∼31, 19, and 11 kDa bands. Human sera from cysticercosis and diphyllobothriasis cases showed non-specific cross reactions with 229, 35∼31, 27, 25 and 17kDa bands. Protein bands of 91, 60, 21 and 10kDa showed strong positive reaction without cross reactions with sera from other helminthic infections.
To observe antibody changes after praziquantel treatment in paragonimiasis, a total of 46 serum samples from 13 serologically diagnosed patients was collected for 4~28 months. The specific antibody (IgG) levels were measured by enzyme- linked immunosorbent assay (ELISA). All but one patient who needed retreatment became symptom-free within a week. Antibody levels were dropped near to or below a cut-off absorbance (abs.) of 0.25 in varying intervals from 4 to 18 months. Of 9 patients who were retested within 3 months, 5 revealed temporary elevation of antibody level. After the elevation, the levels be낙an to decline slowly to negative ranges. If treated earlier after symptoms developed, the temporary elevation did not occur and intervals to negative conversion were shorter. By sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) /immunoblot, antigen-antibody reactions in individual patient faded gradually without significant changes in reacting antigen bands.
Kim, Sung-Hwan;Kong, Yoon;Kim, Suk-Il;Kang, Shin-Yong;Cho, Seung-Yull
Parasites, Hosts and Diseases
/
v.26
no.4
/
pp.239-244
/
1988
In order to observe the antigenic fractions in saline extract of adult Paragonimus westermani, proteins in the crude extract were separated by sodium dodecyl sulfate-polyacylamide gel electrophoresis(SDS.PAGE) in reducing conditions. The separated protein fractions were transferred to nitrocellulose paper on which 20 sera from human paragonimiasis were reacted and immunoblottrd. Out of 15 stained protein bands in SDS-PAGE, 7 reacted with infected sera while 8 did not. Additionally, 7 unstained protein bands in SDS-PAGE reacted with the sera. Of 14 reacted bands, 30 kilodalton(kDa) band was the most frequently reacted (95%) and was a strong antigen. Protein bands of 23 and 46 kDa were also strong antigens. Bands of over 150 kDa, 120 kDa, 92 kDa, 86 kDa, 74 kDa, 62 kDa, 51 kDa, 32 kDa, 28 kDa, 16.5 kDa and 15.5 kDa were also reactive but their frequencies of the reaction were variable. Key words: Paragonimus westermani, human paragonimiasis, antigenic proteins, SDS-PAGE/ immunoblot
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