Preparation and determination of arprinocid solution
Author(s): YAN Ming, ZHANG Lifang, XUE Feiqun, WANG Xiaoyang, WU Hao, ZHANG Chong, JIANG Shanxiang, College of Veterinary Medicine, Nanjing Agricultural University, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences
Pages: 107-111
Year: 2014 Issue: 3
Journal: Journal of Nanjing Agricultural University
Keyword: aprinocid; solution; orthogonal design; HPLC; determination;
Abstract: This paper developed a new formulation of arprinocid,and established a mothod for the determination of arprinocid solution by using high performance liquid chromatography(HPLC). These results contributed to provide a reference for further application of arprinocid solution. Orthogonal designing method was used to filter out solutions optimal formula,which was based on univariate tests. The best combination of the solution was 40% solvent and 50% co-solvent and 0. 2% antioxidant,and pH value was approximately 3. 0. The content of the solution was determined by HPLC. An C18column(4. 6 mm×250 mm,5. 0 μm) was used,and the proportion of acetonitrice and water which was added with 0. 1% phosphoric and 0. 1% triethanolamine was 18 ∶82 by the mobile phase. Flow rate was 1. 0 mL·min-1,and the column temperature was 30 ℃,and the detection wavelength was 261 nm,with injection volume of 10 μL. Acetic acid was heated to about 40 ℃,then added the dissolved tartaric acid and polyethylene glycol 400,and then filtratted and packed in 100 mL brown glass bottles after being added arprinocid until dissolved. Blank accessories was taken as a blank control,and pipetted for the appropriate amount arprinocid solution,diluted with methanol,with filterring by 0. 45 μm membrane,then injected to detection. Accessories interference test results showed that arpronocid was undisturbed in the peak position. And specificity test revealed that solutions could be separated from degradation products. The linear range of the calibration curve for arprinocid was 50-100 μg·mL-1(R2= 0. 999 3,n = 5),taking concentration(x) as the abscissa and peak area(y) for the vertical axis linear regression,and regression equation showed y = 34 772x +14 371. Pipette exactly 1 mL of arprinocid solution to 100 mL volumetric flask,dissolve it in methanol to the mark,and then place the solution 1 mL to 10 mL volumetric flask with methanol,finally injecte six times continuously. The relative standard deviation(RSD) of precision was 0. 4%(n = 6). Prepare nine blank accessories,and then add for the equivalent concentration of the test solution arprinocid reference 80,100 and 120 μg·mL-1 three copies of each. At the spiked levels of 80,100,120 μg·mL-1,the average recovery rate was 98. 56%(RSD=0. 53%). The solution was gradually diluted with methanol,according to the signal to noise ratio(SNR) 3 ∶ 1 counted as the limit of detection(LOD),and according to SNR 10 ∶1 calculated as the limit of quantification(LOQ). The LOQ and LOD were 64 and 24 ng·mL-1. Three batches of solution were diluted with methanol to 80 μg·mL-1,filterred by the organic membrane,and then injected todetection. Using the external standard method,the content of arprinocid was changed to the indicated value content of the sample. The average concentration of solution was 40. 08 mg·mL-1and its signal content was 100. 19%. The prescription was reasonable and the preparation technology was reliable. The HPLC method has high accuracy and sensitivity and can be used for arprinocid solution test.
Pages: 107-111
Year: 2014 Issue: 3
Journal: Journal of Nanjing Agricultural University
Keyword: aprinocid; solution; orthogonal design; HPLC; determination;
Abstract: This paper developed a new formulation of arprinocid,and established a mothod for the determination of arprinocid solution by using high performance liquid chromatography(HPLC). These results contributed to provide a reference for further application of arprinocid solution. Orthogonal designing method was used to filter out solutions optimal formula,which was based on univariate tests. The best combination of the solution was 40% solvent and 50% co-solvent and 0. 2% antioxidant,and pH value was approximately 3. 0. The content of the solution was determined by HPLC. An C18column(4. 6 mm×250 mm,5. 0 μm) was used,and the proportion of acetonitrice and water which was added with 0. 1% phosphoric and 0. 1% triethanolamine was 18 ∶82 by the mobile phase. Flow rate was 1. 0 mL·min-1,and the column temperature was 30 ℃,and the detection wavelength was 261 nm,with injection volume of 10 μL. Acetic acid was heated to about 40 ℃,then added the dissolved tartaric acid and polyethylene glycol 400,and then filtratted and packed in 100 mL brown glass bottles after being added arprinocid until dissolved. Blank accessories was taken as a blank control,and pipetted for the appropriate amount arprinocid solution,diluted with methanol,with filterring by 0. 45 μm membrane,then injected to detection. Accessories interference test results showed that arpronocid was undisturbed in the peak position. And specificity test revealed that solutions could be separated from degradation products. The linear range of the calibration curve for arprinocid was 50-100 μg·mL-1(R2= 0. 999 3,n = 5),taking concentration(x) as the abscissa and peak area(y) for the vertical axis linear regression,and regression equation showed y = 34 772x +14 371. Pipette exactly 1 mL of arprinocid solution to 100 mL volumetric flask,dissolve it in methanol to the mark,and then place the solution 1 mL to 10 mL volumetric flask with methanol,finally injecte six times continuously. The relative standard deviation(RSD) of precision was 0. 4%(n = 6). Prepare nine blank accessories,and then add for the equivalent concentration of the test solution arprinocid reference 80,100 and 120 μg·mL-1 three copies of each. At the spiked levels of 80,100,120 μg·mL-1,the average recovery rate was 98. 56%(RSD=0. 53%). The solution was gradually diluted with methanol,according to the signal to noise ratio(SNR) 3 ∶ 1 counted as the limit of detection(LOD),and according to SNR 10 ∶1 calculated as the limit of quantification(LOQ). The LOQ and LOD were 64 and 24 ng·mL-1. Three batches of solution were diluted with methanol to 80 μg·mL-1,filterred by the organic membrane,and then injected todetection. Using the external standard method,the content of arprinocid was changed to the indicated value content of the sample. The average concentration of solution was 40. 08 mg·mL-1and its signal content was 100. 19%. The prescription was reasonable and the preparation technology was reliable. The HPLC method has high accuracy and sensitivity and can be used for arprinocid solution test.
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