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Hermanto, Bambang
- High-Performance Liquid Chromatography Ultraviolet-Photodiode Array Detection Method for Aflatoxin B1 in Cattle Feed Supplements
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Materials and Methods: Aflatoxin B1 certified reference grade from Trilogy Analytical Laboratory dissolved acetonitrile (ACN) at 10 μg/mL was used for standard assessment. HPLC instruments such as ultraviolet-PDA detector used a Shimadzu LC-6AD pump with DGU-20A5 degasser, communication module-20A, and PDA detector SPD-M20A with FRC-10A fraction collector. The HPLC was set isocratic method at 354 nm with a reverse-phase ODS C18 column (LiChrospher® 100 RP-18; diameter, 5 μm) under a 20°C controlled column chamber. Rheodyne® sample loops were performed in 20 μL capacities. The mobile phase was performed at fraction 63:26:11 H2O: methanol:ACN at pH 6.8. A total of 1 kg of feed contained 10% bread crumbs and 30% concentrated, 40% forage, and 20% soybean dregs were using commercials samples. Samples were extracted by ACN and separated with solid phase extraction ODS 1 mL than elution with mobile phase to collect at drying samples performed. The samples were ready to use after added 1 mL mobile phase than injected into the system of HPLC.
Results: We found that the retention time of aflatoxin B1 was approximately 10.858 min. Linearity of 0.01-0.08 μg/mL aflatoxin B1 dissolved in mobile phase was obtained at R2=0.9. These results demonstrate that these methods can be used to analyze aflatoxin B1 and gain 89-99% recovery. The limit of detection of this assay was obtained at 3.5×10-6 μg/mL.
Conclusion: This method was easy to apply and suitable to analyzing at small concentrations of aflatoxin B1 in formulated product of feed cattle.
Authors
Affiliations
1 Department of Basic Science, Veterinary Pharmacy Subdivision, Airlangga University, Surabaya, ID
2 Department of Pharmacology, Airlangga University, Prof. Dr. Moestopo 47, Pacar Kembang, Surabaya, ID
1 Department of Basic Science, Veterinary Pharmacy Subdivision, Airlangga University, Surabaya, ID
2 Department of Pharmacology, Airlangga University, Prof. Dr. Moestopo 47, Pacar Kembang, Surabaya, ID
Source
Veterinary World, Vol 10, No 8 (2017), Pagination: 932-938Abstract
Aim: The objective of the current study is to determine the concentration of aflatoxin B1 using high-performance liquid chromatography (HPLC) with a photodiode array (PDA) detector.Materials and Methods: Aflatoxin B1 certified reference grade from Trilogy Analytical Laboratory dissolved acetonitrile (ACN) at 10 μg/mL was used for standard assessment. HPLC instruments such as ultraviolet-PDA detector used a Shimadzu LC-6AD pump with DGU-20A5 degasser, communication module-20A, and PDA detector SPD-M20A with FRC-10A fraction collector. The HPLC was set isocratic method at 354 nm with a reverse-phase ODS C18 column (LiChrospher® 100 RP-18; diameter, 5 μm) under a 20°C controlled column chamber. Rheodyne® sample loops were performed in 20 μL capacities. The mobile phase was performed at fraction 63:26:11 H2O: methanol:ACN at pH 6.8. A total of 1 kg of feed contained 10% bread crumbs and 30% concentrated, 40% forage, and 20% soybean dregs were using commercials samples. Samples were extracted by ACN and separated with solid phase extraction ODS 1 mL than elution with mobile phase to collect at drying samples performed. The samples were ready to use after added 1 mL mobile phase than injected into the system of HPLC.
Results: We found that the retention time of aflatoxin B1 was approximately 10.858 min. Linearity of 0.01-0.08 μg/mL aflatoxin B1 dissolved in mobile phase was obtained at R2=0.9. These results demonstrate that these methods can be used to analyze aflatoxin B1 and gain 89-99% recovery. The limit of detection of this assay was obtained at 3.5×10-6 μg/mL.
Conclusion: This method was easy to apply and suitable to analyzing at small concentrations of aflatoxin B1 in formulated product of feed cattle.
Keywords
Feeds Supplements, High-Performance Liquid Chromatography Photodiode Array Detector, Isocratic Methods, Mycotoxin.Full Text
- Calculate of Withdrawal Times of Clenbuterol in Goats to Obtain Safe Times of Slaughter
Abstract Views :360 |
PDF Views:5
Materials and Methods: Five healthy male goats with a mean body weight of 20.64 kg were treated with a single dose of 5.10−3 mg/kg in the BW onto jugular vein. Whole blood samples of approximately 5 mL were taken in a time series at 5, 30, 60, 90, 150, 210, 270, 390, 510, 630, and 750 min. At 24 h posttreatment, all subjects were sacrificed, and 300 g samples of the liver were obtained. The plasma concentration and liver residue of the drug were observed by reverse-phase high-performance liquid chromatography.
Results: The drug reached a maximum concentration of 19.233±0.331 μg/mL at 5 min, and the elimination half-life was at 173.25 min. The limit detection was obtained at 0.053 μg/mL. A one-way analysis of variance between all goats showed that elimination of the clenbuterol in their bodies was similar (p=1.00), with a withdrawal time of 1,479.326 min and no residues in the liver (p<0.05).
Conclusion: Safe times for slaughter were determined to be at 2 days, 13 h, and 12 min as the 2nd safety factor (SF) time and 3 days, 1 h, and 58 min as the 3rd SF time with the liver organ free from residue.
Authors
Affiliations
1 Department of Basic Science, Veterinary Pharmacy Subdivision, Universitas Airlangga, Surabaya, ID
2 Department of Pharmacology, Universitas Airlangga, Surabaya, ID
3 Department of Reproduction, Airlangga University, Surabaya, ID
1 Department of Basic Science, Veterinary Pharmacy Subdivision, Universitas Airlangga, Surabaya, ID
2 Department of Pharmacology, Universitas Airlangga, Surabaya, ID
3 Department of Reproduction, Airlangga University, Surabaya, ID
Source
Veterinary World, Vol 11, No 6 (2018), Pagination: 731-738Abstract
Background and Aim: Clenbuterol as a β2-agonist drug was investigated according to the concentration of the drug available in the bodies of goats and according to the level of sensitivity of the instruments used for detection. The objective of the current study was to determine withdrawal times after giving a therapeutic dose that resulted in safe slaughters.Materials and Methods: Five healthy male goats with a mean body weight of 20.64 kg were treated with a single dose of 5.10−3 mg/kg in the BW onto jugular vein. Whole blood samples of approximately 5 mL were taken in a time series at 5, 30, 60, 90, 150, 210, 270, 390, 510, 630, and 750 min. At 24 h posttreatment, all subjects were sacrificed, and 300 g samples of the liver were obtained. The plasma concentration and liver residue of the drug were observed by reverse-phase high-performance liquid chromatography.
Results: The drug reached a maximum concentration of 19.233±0.331 μg/mL at 5 min, and the elimination half-life was at 173.25 min. The limit detection was obtained at 0.053 μg/mL. A one-way analysis of variance between all goats showed that elimination of the clenbuterol in their bodies was similar (p=1.00), with a withdrawal time of 1,479.326 min and no residues in the liver (p<0.05).
Conclusion: Safe times for slaughter were determined to be at 2 days, 13 h, and 12 min as the 2nd safety factor (SF) time and 3 days, 1 h, and 58 min as the 3rd SF time with the liver organ free from residue.
Keywords
Elimination Half-Life, New Method for Calculating Withdrawal Time, Prescriptions for Obtained β2-Agonist, Residues in Liver.Full Text
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