为研究抗球虫新化合物沙咪珠利在鸡盲肠中的药物代谢和消除情况,建立检测鸡盲肠中沙咪珠利的高效液相色谱法。检测方法以妥曲珠利亚砜为内标,X TERRA MS C18 5μm(Waters,4.6×250 mm)柱为分析柱,乙腈∶0.2%磷酸水溶液(v/v,35∶65)...为研究抗球虫新化合物沙咪珠利在鸡盲肠中的药物代谢和消除情况,建立检测鸡盲肠中沙咪珠利的高效液相色谱法。检测方法以妥曲珠利亚砜为内标,X TERRA MS C18 5μm(Waters,4.6×250 mm)柱为分析柱,乙腈∶0.2%磷酸水溶液(v/v,35∶65)为流动相等度洗脱,C18柱分离后,在251 nm波长紫外检测器检测,内标法定量。鸡盲肠样品经匀浆化后,乙腈提取、己烷除脂。结果表明,该方法在0.2~40 mg/kg添加浓度范围内检测鸡盲肠中沙咪珠利具有很好的线性关系(r2=0.994);平均相对回收率为90%~107%,日内变异系数2.37%~8.72%;日间变异系数4.09%~6.80%,具有特异性好、灵敏度高、准确性和重现性良好、操作简单等优点,能满足鸡盲肠中沙咪珠利的检测需求。展开更多
A rapid, sensitive and specific ultra-high-performance liquid chromatography tandem mass spectrometry （UPLC-MS） method was developed for the analysis of tetracycline antibiotics, including tetracycline （TC）, oxyte...A rapid, sensitive and specific ultra-high-performance liquid chromatography tandem mass spectrometry （UPLC-MS） method was developed for the analysis of tetracycline antibiotics, including tetracycline （TC）, oxytetracycline （OTC）, chlortetracycline （CTC） and their 4-epimers （4-epiTCs） in agricultural soil fertilized with swine manure. Soil samples were extracted and cleaned-up with 10 mL EDTA-McIlvaine buffer solution （pH 4.0）, then cleaned-up and pre-concentrated using the Oasis MAX cartridge and then eluted with 1 mL solution by mixing formic acid, methanol and water at a ratio of 2：15：83 （v/v/v）. The purified samples were separated by an ACQUITY UPLC BEH C18 column using acetonitrile and water containing 0.1% formic acid mobile phase and detected by a single quadrupole MS. The limits of detection for the soil extraction method （LODsoil） ranged from 0.6-2.5 lag kg-~ with recoveries from 23.3-159.2%. Finally, the method was applied to an agricultural field in an area with intensive pig-fattening farming. Tetracyclines were detected in soil from 2.8 to 42.4 μg kg-1 soil. These results demonstrate that soil from swine farms can become severely contaminated with tetracycline antibiotics and their metabolites.展开更多
Quality control and screening of active substances in traditional Chinese medicines have been performed using fingerprint analysis. The spectrum-effect relationship between chromatography fingerprints and efficacy of ...Quality control and screening of active substances in traditional Chinese medicines have been performed using fingerprint analysis. The spectrum-effect relationship between chromatography fingerprints and efficacy of herbal drugs is considered as a potentially useful method for determining active ingredients in complex mixtures. The study was designed to develop a method for determining the bioactive components of a compound Chinese medicine called Tiefeng based on spectrum-effect relationships between high-performance liquid chromatography （HPLC） fingerprints and anticoccidial activities. Four peaks of the established HPLC fingerprint indicate the main bioactive components of this medicine. In addition, pharrnacodynamic atlas was defined and used to assess the anticoccidial activity of Tiefeng from different sources for the first time. We found that the level of anticoccidial activity of Tiefeng was consistent with the degree of similarity between the pharmacodynamic atlas and chromatogram of any sample. Furthermore, effect of this medicine was related with the main active constituents, along with the origin and the harvesting time.展开更多
Lactate dehydrogenase(LDH),a vital enzyme in anaerobic glycolysis,is closely associated with the survival of parasites.Previous studies of some parasites have shown that LDH exhibits unique physicochemical properties ...Lactate dehydrogenase(LDH),a vital enzyme in anaerobic glycolysis,is closely associated with the survival of parasites.Previous studies of some parasites have shown that LDH exhibits unique physicochemical properties and molecular structures and may be an ideal potential target for diagnosis and drug screening.In this study,we aimed to investigate the effects of acetamizuril,a novel anticoccidial compound,on LDH in the second-generation merozoites of Eimeria tenella(mz-LDH).Quantitative real-time PCR,Western blot,immunofluorescence and enzyme activity assays were each applied to detect the changes of mz-LDH.Our results indicated that the mRNA and protein levels of mz-LDH were reduced upon acetamizuril treatment.Immunolocalization of mz-LDH demonstrated that considerable amount of mz-LDH was distributed around or in the nuclei of second-generation merozoites within the untreated group;in contrast,the acetamizuril-treated group had very low level of mz-LDH.Meanwhile,LDH enzyme activity assay suggested that a decreased LDH enzyme activity in both cytoplasm and nucleus of merozoites in the acetamizuril-treated group.Moreover,the induced apoptosis in second-generation merozoites by the acetamizuril was evaluated by detecting caspase 3 activity,and acetamizuril was found to significantly increase caspase 3 activity.The above findings show that LDH may play an important role in the mediating the activity of acetamizuril against coccidiosis,and further investigation into this aspect might contribute to new light on the pathogenesis of E.tenella during its interaction with acetamizuril.展开更多
文摘为研究抗球虫新化合物沙咪珠利在鸡盲肠中的药物代谢和消除情况,建立检测鸡盲肠中沙咪珠利的高效液相色谱法。检测方法以妥曲珠利亚砜为内标,X TERRA MS C18 5μm(Waters,4.6×250 mm)柱为分析柱,乙腈∶0.2%磷酸水溶液(v/v,35∶65)为流动相等度洗脱,C18柱分离后,在251 nm波长紫外检测器检测,内标法定量。鸡盲肠样品经匀浆化后,乙腈提取、己烷除脂。结果表明,该方法在0.2~40 mg/kg添加浓度范围内检测鸡盲肠中沙咪珠利具有很好的线性关系(r2=0.994);平均相对回收率为90%~107%,日内变异系数2.37%~8.72%;日间变异系数4.09%~6.80%,具有特异性好、灵敏度高、准确性和重现性良好、操作简单等优点,能满足鸡盲肠中沙咪珠利的检测需求。
文摘A rapid, sensitive and specific ultra-high-performance liquid chromatography tandem mass spectrometry （UPLC-MS） method was developed for the analysis of tetracycline antibiotics, including tetracycline （TC）, oxytetracycline （OTC）, chlortetracycline （CTC） and their 4-epimers （4-epiTCs） in agricultural soil fertilized with swine manure. Soil samples were extracted and cleaned-up with 10 mL EDTA-McIlvaine buffer solution （pH 4.0）, then cleaned-up and pre-concentrated using the Oasis MAX cartridge and then eluted with 1 mL solution by mixing formic acid, methanol and water at a ratio of 2：15：83 （v/v/v）. The purified samples were separated by an ACQUITY UPLC BEH C18 column using acetonitrile and water containing 0.1% formic acid mobile phase and detected by a single quadrupole MS. The limits of detection for the soil extraction method （LODsoil） ranged from 0.6-2.5 lag kg-~ with recoveries from 23.3-159.2%. Finally, the method was applied to an agricultural field in an area with intensive pig-fattening farming. Tetracyclines were detected in soil from 2.8 to 42.4 μg kg-1 soil. These results demonstrate that soil from swine farms can become severely contaminated with tetracycline antibiotics and their metabolites.
基金funded by the National Key Technology R&D Program, China (2008 BADB4B05-02)
文摘Quality control and screening of active substances in traditional Chinese medicines have been performed using fingerprint analysis. The spectrum-effect relationship between chromatography fingerprints and efficacy of herbal drugs is considered as a potentially useful method for determining active ingredients in complex mixtures. The study was designed to develop a method for determining the bioactive components of a compound Chinese medicine called Tiefeng based on spectrum-effect relationships between high-performance liquid chromatography （HPLC） fingerprints and anticoccidial activities. Four peaks of the established HPLC fingerprint indicate the main bioactive components of this medicine. In addition, pharrnacodynamic atlas was defined and used to assess the anticoccidial activity of Tiefeng from different sources for the first time. We found that the level of anticoccidial activity of Tiefeng was consistent with the degree of similarity between the pharmacodynamic atlas and chromatogram of any sample. Furthermore, effect of this medicine was related with the main active constituents, along with the origin and the harvesting time.
基金supported by the National Natural Science Foundation of China(31101857,31272607 and 31472235)。
文摘Lactate dehydrogenase(LDH),a vital enzyme in anaerobic glycolysis,is closely associated with the survival of parasites.Previous studies of some parasites have shown that LDH exhibits unique physicochemical properties and molecular structures and may be an ideal potential target for diagnosis and drug screening.In this study,we aimed to investigate the effects of acetamizuril,a novel anticoccidial compound,on LDH in the second-generation merozoites of Eimeria tenella(mz-LDH).Quantitative real-time PCR,Western blot,immunofluorescence and enzyme activity assays were each applied to detect the changes of mz-LDH.Our results indicated that the mRNA and protein levels of mz-LDH were reduced upon acetamizuril treatment.Immunolocalization of mz-LDH demonstrated that considerable amount of mz-LDH was distributed around or in the nuclei of second-generation merozoites within the untreated group;in contrast,the acetamizuril-treated group had very low level of mz-LDH.Meanwhile,LDH enzyme activity assay suggested that a decreased LDH enzyme activity in both cytoplasm and nucleus of merozoites in the acetamizuril-treated group.Moreover,the induced apoptosis in second-generation merozoites by the acetamizuril was evaluated by detecting caspase 3 activity,and acetamizuril was found to significantly increase caspase 3 activity.The above findings show that LDH may play an important role in the mediating the activity of acetamizuril against coccidiosis,and further investigation into this aspect might contribute to new light on the pathogenesis of E.tenella during its interaction with acetamizuril.
