In order to establish an HPLC-MS/MS method for detecting progesterone in raw milk,raw milk was extracted with methanol,and then determined by HPLC-MS/MS after the purification with a C18 solid phase extraction column....In order to establish an HPLC-MS/MS method for detecting progesterone in raw milk,raw milk was extracted with methanol,and then determined by HPLC-MS/MS after the purification with a C18 solid phase extraction column. Qualitative determination was performed according to retention time and selective ion abundance ratio,and quantification was performed by external standard method. The results showed that progesterone had a good linear relation in the range of0. 1-5. 0 ng/ml,with a correlation coefficient of 0. 999 9. The detection limit of the established method was 0. 02 μg/kg,and the quantification limit was0. 04 μg/kg. In the standard addition range of 0. 1-5. 0 ng/ml,the recovery ranged from 79. 6% to 105. 1%,with relative standard deviation≤8. 9%. Compared with the industry standard,the detection limit and the quantification limit of the established method are significantly reduced,and the sensitivity of the method is enhanced,so this method is suitable for the detection of progesterone residue in fresh milk in pastures.展开更多
Reversed-phase high-performance liquid chromatography （RP-HPLC） method with C4 column and C18 column for analyzing β-lactoglobulin and α-lactalbumin in bovine milk was developed and the performance and characteris...Reversed-phase high-performance liquid chromatography （RP-HPLC） method with C4 column and C18 column for analyzing β-lactoglobulin and α-lactalbumin in bovine milk was developed and the performance and characteristic of two columns were compared. Shiseido Proteonavi C4 column （250 mm×4.6 mm×5μm） and Shiseido CAPCELL PAK SG 300 C18 column （250 mm× 4.6 mm×5 μm） were used in the experiment. Phase A was composed of 0.1% （V/V） trifluoroacetic acid （TFA） in ultrapure water and Phase B （organic phase） was composed of 0.1% TFA in acetonitrile. Gradient elution was taken. Flow rate was 1 mL min-1. The detection wavelength was 215 nm. The injection volume was 20 μL and the column temperature was 30℃. The results showed that linear relationship was good and recovery of α-lactalbumin and β-lactoglobulin was 86.12%-104.38%, C18 column had stronger ability to resist acid and more stable, and the method with C4 column had excellent sensitivities and good separation.展开更多
A high performance liquid chromatographic method for the simultaneous determination of four fat soluble vitamins: retinol (vitamin A), cholecalciferol(vitamin D), tocopherol(vitamin E) and phylloquinone(vitamin K_1) i...A high performance liquid chromatographic method for the simultaneous determination of four fat soluble vitamins: retinol (vitamin A), cholecalciferol(vitamin D), tocopherol(vitamin E) and phylloquinone(vitamin K_1) in fortified milk powders and egg yolk has been developed. The method requires enzymatic hydrolysis of lipid component of the sample as a pretreatment. Several factors which influence the enzymatic hydrolysis were studied-Separation was achieved using μ-Bondapak C-18 column(3.9×300mm), 98% methanol as mobile phase, a double channel detection was selected; vitamins D_3 E_1 K_1 were detected by UV spectrophotometry (265 nm) first, then vitamin A by fluorometry (EX 325nm, EM 480mm). The retention times of vitamin A_1 D_3, E and K_1 4.87, 9.00, 10.58 and 15.45 min respectively. Detection limit were 0.64, 0.25, 0.50 and 0.07 ng; and the recoveries were 90.5%～103.6% 90.0%～95.6%, 91.7%～98.8%, 91.5%～98.6%, respectively. The vitamins A, D_0, E, K_1 contents in foods were determined satisfactorily.展开更多
Here we propose a method for determination of Avermectin residual content in milk and beef content. After extracting and processing the sample, use high performance liquid chromatography （HPLC） for determination. Mo...Here we propose a method for determination of Avermectin residual content in milk and beef content. After extracting and processing the sample, use high performance liquid chromatography （HPLC） for determination. Mobile phase is： V（Methanol）： V（Water）=90： 10; determination wavelength is 245 nm; flow rate is 0.8 mL/min. Avermectin is completely separated in 17min, linear range is 5 - 0.3 g/L, correlation coefficient is 0.9997, average recovery rates of Avermectin in milk and beef are respectively 91.20 %- 100.32 % and 89.93 %-99.15 %, the relative standard deviations are respectively less than 7% and 4%. This method is simple, sensitive and accurate, so is suitable for determination of Avermectin residues in milk and beef, and it can meet domestic and foreign testing requirements.展开更多
Tetracycline and analogues are among the most used antibiotics in the dairy industry. Besides the therapeutic uses, tetracyclines are often incorporated into livestock feed as growth promoters. A considerable amount o...Tetracycline and analogues are among the most used antibiotics in the dairy industry. Besides the therapeutic uses, tetracyclines are often incorporated into livestock feed as growth promoters. A considerable amount of antibiotics is released unaltered through milk from dairy animals. The presence of antibiotic residues in milk and their subsequent consumption can lead to potential health impacts, including cancer, hypersensitivity reactions, and the development of antibiotic resistance. Thus, it is important to monitor residual levels of tetracyclines in milk. The purpose of this study is to develop a quick and simple method for simultaneously extracting five tetracycline analogues from bovine milk. Specifically, five tetracycline analogues: Chlortetracycline (CTC), demeclocycline (DEM), doxycycline (DC), minocycline (MC), and tetracycline (TC) were simultaneously extracted from milk using trifluoroacetic acid. Subsequently, the extracted analogues were separated by reverse-phase high-performance liquid chromatography (RP-HPLC) and detected at 355 nm using UV/Vis. Calibration curves for all five tetracycline analogues show excellent linearity (r2 value > 0.99). Percent recovery for MC, TC, DEM, CTC, and DC were: 31.88%, 96.91%, 151.29, 99.20%, and 85.58% respectively. The developed extraction method has good precision (RSD < 9.9% for 4 of the 5 analogues). The developed method with minimal sample preparation and pretreatment has the potential to serve as an initial screening test.展开更多
基金Supported by Science and Technology Open Cooperation Project of Henan Province(162106000017)Science and Technology People-benefiting Plan Project of Henan Province(152207110004)+1 种基金Major Science and Technology Program of Henan Province(16110051020)Puyang Science and Technology Plan Project(160215)
文摘In order to establish an HPLC-MS/MS method for detecting progesterone in raw milk,raw milk was extracted with methanol,and then determined by HPLC-MS/MS after the purification with a C18 solid phase extraction column. Qualitative determination was performed according to retention time and selective ion abundance ratio,and quantification was performed by external standard method. The results showed that progesterone had a good linear relation in the range of0. 1-5. 0 ng/ml,with a correlation coefficient of 0. 999 9. The detection limit of the established method was 0. 02 μg/kg,and the quantification limit was0. 04 μg/kg. In the standard addition range of 0. 1-5. 0 ng/ml,the recovery ranged from 79. 6% to 105. 1%,with relative standard deviation≤8. 9%. Compared with the industry standard,the detection limit and the quantification limit of the established method are significantly reduced,and the sensitivity of the method is enhanced,so this method is suitable for the detection of progesterone residue in fresh milk in pastures.
基金Supported by the Project of Science & Technology Plans in Heilongjiang Province in the 11th Five-year Period (GB07B407)
文摘Reversed-phase high-performance liquid chromatography （RP-HPLC） method with C4 column and C18 column for analyzing β-lactoglobulin and α-lactalbumin in bovine milk was developed and the performance and characteristic of two columns were compared. Shiseido Proteonavi C4 column （250 mm×4.6 mm×5μm） and Shiseido CAPCELL PAK SG 300 C18 column （250 mm× 4.6 mm×5 μm） were used in the experiment. Phase A was composed of 0.1% （V/V） trifluoroacetic acid （TFA） in ultrapure water and Phase B （organic phase） was composed of 0.1% TFA in acetonitrile. Gradient elution was taken. Flow rate was 1 mL min-1. The detection wavelength was 215 nm. The injection volume was 20 μL and the column temperature was 30℃. The results showed that linear relationship was good and recovery of α-lactalbumin and β-lactoglobulin was 86.12%-104.38%, C18 column had stronger ability to resist acid and more stable, and the method with C4 column had excellent sensitivities and good separation.
文摘A high performance liquid chromatographic method for the simultaneous determination of four fat soluble vitamins: retinol (vitamin A), cholecalciferol(vitamin D), tocopherol(vitamin E) and phylloquinone(vitamin K_1) in fortified milk powders and egg yolk has been developed. The method requires enzymatic hydrolysis of lipid component of the sample as a pretreatment. Several factors which influence the enzymatic hydrolysis were studied-Separation was achieved using μ-Bondapak C-18 column(3.9×300mm), 98% methanol as mobile phase, a double channel detection was selected; vitamins D_3 E_1 K_1 were detected by UV spectrophotometry (265 nm) first, then vitamin A by fluorometry (EX 325nm, EM 480mm). The retention times of vitamin A_1 D_3, E and K_1 4.87, 9.00, 10.58 and 15.45 min respectively. Detection limit were 0.64, 0.25, 0.50 and 0.07 ng; and the recoveries were 90.5%～103.6% 90.0%～95.6%, 91.7%～98.8%, 91.5%～98.6%, respectively. The vitamins A, D_0, E, K_1 contents in foods were determined satisfactorily.
文摘Here we propose a method for determination of Avermectin residual content in milk and beef content. After extracting and processing the sample, use high performance liquid chromatography （HPLC） for determination. Mobile phase is： V（Methanol）： V（Water）=90： 10; determination wavelength is 245 nm; flow rate is 0.8 mL/min. Avermectin is completely separated in 17min, linear range is 5 - 0.3 g/L, correlation coefficient is 0.9997, average recovery rates of Avermectin in milk and beef are respectively 91.20 %- 100.32 % and 89.93 %-99.15 %, the relative standard deviations are respectively less than 7% and 4%. This method is simple, sensitive and accurate, so is suitable for determination of Avermectin residues in milk and beef, and it can meet domestic and foreign testing requirements.
文摘Tetracycline and analogues are among the most used antibiotics in the dairy industry. Besides the therapeutic uses, tetracyclines are often incorporated into livestock feed as growth promoters. A considerable amount of antibiotics is released unaltered through milk from dairy animals. The presence of antibiotic residues in milk and their subsequent consumption can lead to potential health impacts, including cancer, hypersensitivity reactions, and the development of antibiotic resistance. Thus, it is important to monitor residual levels of tetracyclines in milk. The purpose of this study is to develop a quick and simple method for simultaneously extracting five tetracycline analogues from bovine milk. Specifically, five tetracycline analogues: Chlortetracycline (CTC), demeclocycline (DEM), doxycycline (DC), minocycline (MC), and tetracycline (TC) were simultaneously extracted from milk using trifluoroacetic acid. Subsequently, the extracted analogues were separated by reverse-phase high-performance liquid chromatography (RP-HPLC) and detected at 355 nm using UV/Vis. Calibration curves for all five tetracycline analogues show excellent linearity (r2 value > 0.99). Percent recovery for MC, TC, DEM, CTC, and DC were: 31.88%, 96.91%, 151.29, 99.20%, and 85.58% respectively. The developed extraction method has good precision (RSD < 9.9% for 4 of the 5 analogues). The developed method with minimal sample preparation and pretreatment has the potential to serve as an initial screening test.
