Microwave-assisted extraction was optimized with response surface methodology for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii.The optimized extraction procedure was achieved b...Microwave-assisted extraction was optimized with response surface methodology for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii.The optimized extraction procedure was achieved by soaking the sample with 70% methanol（1∶15,v/v）for 30 min,and then microwave irradiation for 11 min at a power of 600 W.Coupling the extraction process with HPLC-fluorescence presented good recovery,satisfactory precision,and good linear relation.Compared with a method from the Chinese Pharmacopoeia,the proposed method enables higher extraction efficiency and more accurate analytical results.It can be of potential value in quality assessment of Radix Puerariae thomsonii medicinal materials.展开更多
A highly sensitive HPLC method for the detection of amino acids and oligopeptides with 9-acridine formyl chloride by pre-column fluorescence derivatization has been developed. Glycine, glycylglycine, histidine, trigly...A highly sensitive HPLC method for the detection of amino acids and oligopeptides with 9-acridine formyl chloride by pre-column fluorescence derivatization has been developed. Glycine, glycylglycine, histidine, triglycine and glutathione were separated on a reversed-phase C-18 column with methanol-water-triethylamine eluent, derivatization and chromatographic conditions were optimized. The five derivatives were eluted in 28 min with a good reproducibility. Linear range of the calibration graph was 0.08-260 nmol/ml(-1). The relative standard deviations(n=6) are < 5%. Detection limits (signal-to-noise ratio=3) for the five derivatives are 20-40 fmol.展开更多
Aim To develop a sensitive and accurate HPLC method for the determination of fleroxacin in human plasma, and study its pharmacokinetics in healthy subjects. Methods The analytes were isolated fi＇om plasma by simple p...Aim To develop a sensitive and accurate HPLC method for the determination of fleroxacin in human plasma, and study its pharmacokinetics in healthy subjects. Methods The analytes were isolated fi＇om plasma by simple protein precipitation with methanol, separated on a Diamonsil C18 column by isocratic elution with the mobile phase consisted of 1% triethylamine at pH 4.8 （adjusted with phosphoric acid） and acetonitrile （80/20, V/V） at a flow rate of 1.0 mL.min^-1 and analyzed by fluorescence detector with an excitation at 290 nm and emission 458 nm. The pharmacokinetic study of fleroxacin was performed according to a double period crossover design. Results The weighted （1/x） calibration curve was linear over the plasma concentration range of 0.025 - 8.00 μg.mL^-1 The inter- and intra-day precisions （RSD/%） were no more than 5.16%, and the method accuracies and extraction recoveries at three concentrations ranged firom 99.1% to 100.9%, and 86.7% to 92.0%, respectively. Following oral administration at a dose of 400 mg fleroxacin, the main pharmacokinetic parameters for test and reference capsules were Cmax5.08 ± 0.78 and 5.38 ± 1.40 μg.mL^-1, tmax 1.72 ±0.79 and 1.82 ± 0.78 h, t1/2 11.68 ± 1.27 and 11.38 ± 1.51 h^-1, AUC0-∞ 78.44 ± 11.44 and 76.53 ± 13.24 μg.mL^-1.h, respectively. Conclusion The method is sensitive and accurate, and suitable for human pharmacokinetic study of fleroxacin.展开更多
Aim To develop an HPLC method with fluorescence detection for the assay ofDL111-IT in rabbit plasma. Methods DL111-IT and internal standard glybenzcyclamide in rabbit plasmawere extracted with chloroform. The determin...Aim To develop an HPLC method with fluorescence detection for the assay ofDL111-IT in rabbit plasma. Methods DL111-IT and internal standard glybenzcyclamide in rabbit plasmawere extracted with chloroform. The determination was performed on a Diamonsil ODS-C_(18) column(150 mm x 4.6 mm, 5 μm) with a mobile phase of acetonitrile and 0.025 mol·L^(-1) diammoniumhydrogen phosphate buffer (pH 5.0, adjusted by phosphoric acid) (60:40, V/V) at a flow-rate of 1.0mL·min^(-1) . Fluorescence detector was operated at excitation wavelength of 250 nm and emissionwavelength of 332 nm. Results The calibration curve in plasma was linear from 1.00 - 20.00ng·mL^(-1) ( r = 0.999 6, n = 5). The method afforded average extracting recoveries of 85.3% ±1.3%, 84.9% ± 2.7% and 85.8% ± 1.8%, and the average method recoveries were 99.5% ±0.4%, 102.1%± 1.8% and 101.3% ± 2.4% for the high (20.00 ng·mL^(-1)) , middle (10.00 ng·mL^(-1)) and low (1.00 ng·mL^(-1)) check samples, respectively. The intra-day (n = 5) and inter-day (n = 5) precisions(RSD) were less than 3.0% and 7.0%, respectively. The limit of detection and quantitation for themethod were 0.3 ng·mL^(-1) (S/N = 3) and 1 ng·mL^(-1) (S/N = 10, RSD<7.0%) plasma sample,respectively. Conclusion The developed method was accurate, sensitive, simple and could be used forpharmacokinetic study of DL111- IT.展开更多
Multiphoton microscopy(MPM)is a powerful imaging technology for brain research.The imaging depth in MPM is partly determined by emission wavelength of fluorescent labels.It has been demonstrated that a longer emission...Multiphoton microscopy(MPM)is a powerful imaging technology for brain research.The imaging depth in MPM is partly determined by emission wavelength of fluorescent labels.It has been demonstrated that a longer emission wavelength is favorable for signal detection as imaging depth increases.However,there has been no comparison with near-infrared(NIR)emission.In order to quantitatively analyze the effect of emission wavelength on 3-photon imaging of mouse brains in vivo,we utilize the same excitation wavelength to excite a single fluorescent dye and simultaneously collect NIR and orange-red emission fluorescence at 828 nm and 620 nm,respectively.Both experimental and simulation results show that as the imaging depth increases,NIR emission decays less than orange-red fluorescent emission.These results show that it is preferable to shift the emission wavelength to NIR to enable more e±cient signal collection deep in the brain.展开更多
目的建立同时测定马齿苋中10个指标成分含量的高效液相色谱法(high performance liquid chromatography,HPLC),用化学计量学和熵权-逼近理想解排序法(entropy weight-technique for order preference by similarity to an ideal solutio...目的建立同时测定马齿苋中10个指标成分含量的高效液相色谱法(high performance liquid chromatography,HPLC),用化学计量学和熵权-逼近理想解排序法(entropy weight-technique for order preference by similarity to an ideal solution,EW-TOPSIS)对马齿苋的质量进行综合分析和评价,为该产品质量控制和标准提升提供参考依据。方法用高效液相色谱法同时测定5省18批马齿苋样品中柠檬酸、琥珀酸、咖啡酸、染料木苷、木犀草素、槲皮素、东莨菪亭、6,7-二羟基香豆素、异茴香内酯和佛手柑内酯的含量,用化学计量学方法对含量测定结果进行综合分析,挖掘影响马齿苋质量的主要潜在标志物,联合熵权-逼近理想解排序法对不同产地来源的马齿苋进行质量评价。结果10种成分分别在各自线性范围内线性关系和准确度良好;化学计量学分析结果显示,18批马齿苋聚为3类,呈现一定的产区差异;木犀草素、槲皮素、琥珀酸和柠檬酸是影响马齿苋产品质量的主要潜在标志物;熵权-逼近理想解排序法分析结果显示四川和贵州地区所产马齿苋质量最优,其次为河南、辽宁和吉林。结论所建立的高效液相色谱法能同时测定马齿苋中10个指标成分的含量,化学计量学和熵权-逼近理想解排序法可用于马齿苋的综合质量评价。展开更多
Chromosome aberrations are distinctive features of human malignant tumors. Analysis of chromosomal changes can illuminate the molecular mechanisms underlying the development and progression of cancer. To establish the...Chromosome aberrations are distinctive features of human malignant tumors. Analysis of chromosomal changes can illuminate the molecular mechanisms underlying the development and progression of cancer. To establish the technique of multicolor fluorescence in situ hybridization （M-FISH） for identifying chromosome aberrations in esophageal carcinoma cell line KYSE 410-4, four pools of 6-color whole-chromosome painting probes have been designed and hybridized on the same metaphase spread by four rounds of repetitive FISH. Repetitive 6-color M-FISH was successfully established and the cytogenetic abnormalities in KYSE 410-4 cells were characterized. Chromosome gains occurred at 2q, 3, 8, 17p, and X. An isochromosome 3q was visualized in the cell line, which might be one intermediate mechanism leading to 3p losses and/or 3q gains. Furthermore, 16 structural arrangements were detected, including four derivative chromosomes. The rearrangement of the centromeric regions accounted for approximately 44% of all rearrangements. The results added a more complete and accurate information of the genetic alterations to the classical cytogenetic description of KYSE 410-4 and provided a detailed cytogenetic background data for appropriate use of the cell line. The established 6-color M-FISH was useful for analyzing chromosomes in the whole genome of human tumors.展开更多
基金the National Natural Science foundationof China(No.20875060) for financial support
文摘Microwave-assisted extraction was optimized with response surface methodology for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii.The optimized extraction procedure was achieved by soaking the sample with 70% methanol（1∶15,v/v）for 30 min,and then microwave irradiation for 11 min at a power of 600 W.Coupling the extraction process with HPLC-fluorescence presented good recovery,satisfactory precision,and good linear relation.Compared with a method from the Chinese Pharmacopoeia,the proposed method enables higher extraction efficiency and more accurate analytical results.It can be of potential value in quality assessment of Radix Puerariae thomsonii medicinal materials.
文摘A highly sensitive HPLC method for the detection of amino acids and oligopeptides with 9-acridine formyl chloride by pre-column fluorescence derivatization has been developed. Glycine, glycylglycine, histidine, triglycine and glutathione were separated on a reversed-phase C-18 column with methanol-water-triethylamine eluent, derivatization and chromatographic conditions were optimized. The five derivatives were eluted in 28 min with a good reproducibility. Linear range of the calibration graph was 0.08-260 nmol/ml(-1). The relative standard deviations(n=6) are < 5%. Detection limits (signal-to-noise ratio=3) for the five derivatives are 20-40 fmol.
文摘Aim To develop a sensitive and accurate HPLC method for the determination of fleroxacin in human plasma, and study its pharmacokinetics in healthy subjects. Methods The analytes were isolated fi＇om plasma by simple protein precipitation with methanol, separated on a Diamonsil C18 column by isocratic elution with the mobile phase consisted of 1% triethylamine at pH 4.8 （adjusted with phosphoric acid） and acetonitrile （80/20, V/V） at a flow rate of 1.0 mL.min^-1 and analyzed by fluorescence detector with an excitation at 290 nm and emission 458 nm. The pharmacokinetic study of fleroxacin was performed according to a double period crossover design. Results The weighted （1/x） calibration curve was linear over the plasma concentration range of 0.025 - 8.00 μg.mL^-1 The inter- and intra-day precisions （RSD/%） were no more than 5.16%, and the method accuracies and extraction recoveries at three concentrations ranged firom 99.1% to 100.9%, and 86.7% to 92.0%, respectively. Following oral administration at a dose of 400 mg fleroxacin, the main pharmacokinetic parameters for test and reference capsules were Cmax5.08 ± 0.78 and 5.38 ± 1.40 μg.mL^-1, tmax 1.72 ±0.79 and 1.82 ± 0.78 h, t1/2 11.68 ± 1.27 and 11.38 ± 1.51 h^-1, AUC0-∞ 78.44 ± 11.44 and 76.53 ± 13.24 μg.mL^-1.h, respectively. Conclusion The method is sensitive and accurate, and suitable for human pharmacokinetic study of fleroxacin.
文摘Aim To develop an HPLC method with fluorescence detection for the assay ofDL111-IT in rabbit plasma. Methods DL111-IT and internal standard glybenzcyclamide in rabbit plasmawere extracted with chloroform. The determination was performed on a Diamonsil ODS-C_(18) column(150 mm x 4.6 mm, 5 μm) with a mobile phase of acetonitrile and 0.025 mol·L^(-1) diammoniumhydrogen phosphate buffer (pH 5.0, adjusted by phosphoric acid) (60:40, V/V) at a flow-rate of 1.0mL·min^(-1) . Fluorescence detector was operated at excitation wavelength of 250 nm and emissionwavelength of 332 nm. Results The calibration curve in plasma was linear from 1.00 - 20.00ng·mL^(-1) ( r = 0.999 6, n = 5). The method afforded average extracting recoveries of 85.3% ±1.3%, 84.9% ± 2.7% and 85.8% ± 1.8%, and the average method recoveries were 99.5% ±0.4%, 102.1%± 1.8% and 101.3% ± 2.4% for the high (20.00 ng·mL^(-1)) , middle (10.00 ng·mL^(-1)) and low (1.00 ng·mL^(-1)) check samples, respectively. The intra-day (n = 5) and inter-day (n = 5) precisions(RSD) were less than 3.0% and 7.0%, respectively. The limit of detection and quantitation for themethod were 0.3 ng·mL^(-1) (S/N = 3) and 1 ng·mL^(-1) (S/N = 10, RSD<7.0%) plasma sample,respectively. Conclusion The developed method was accurate, sensitive, simple and could be used forpharmacokinetic study of DL111- IT.
基金work is funded by the National Natural Sci-ence Foundation of China(Grant/Award Numbers 62075135 and 61975126)Shenzhen Science and Technology Planning Project(ZDSYS2021-0623092006020)+2 种基金Key R&D Program of Shandong Province(Grant Number 2021CXGC010202)the Science and Technology Innovation Commission of Shenzhen(Grant/Award Numbers JCYJ201908-08174819083 and JCYJ20190808175201640)and Natural Science Foundation of Shandong Province(Grant Number ZR2022MA046)Major Innovation Projects for Integrating Science,Education&Industry of Qilu University of Technology(Shan-dong Academy of Sciences,Grant Number 2022JBZ01-04).
文摘Multiphoton microscopy(MPM)is a powerful imaging technology for brain research.The imaging depth in MPM is partly determined by emission wavelength of fluorescent labels.It has been demonstrated that a longer emission wavelength is favorable for signal detection as imaging depth increases.However,there has been no comparison with near-infrared(NIR)emission.In order to quantitatively analyze the effect of emission wavelength on 3-photon imaging of mouse brains in vivo,we utilize the same excitation wavelength to excite a single fluorescent dye and simultaneously collect NIR and orange-red emission fluorescence at 828 nm and 620 nm,respectively.Both experimental and simulation results show that as the imaging depth increases,NIR emission decays less than orange-red fluorescent emission.These results show that it is preferable to shift the emission wavelength to NIR to enable more e±cient signal collection deep in the brain.
文摘目的建立同时测定马齿苋中10个指标成分含量的高效液相色谱法(high performance liquid chromatography,HPLC),用化学计量学和熵权-逼近理想解排序法(entropy weight-technique for order preference by similarity to an ideal solution,EW-TOPSIS)对马齿苋的质量进行综合分析和评价,为该产品质量控制和标准提升提供参考依据。方法用高效液相色谱法同时测定5省18批马齿苋样品中柠檬酸、琥珀酸、咖啡酸、染料木苷、木犀草素、槲皮素、东莨菪亭、6,7-二羟基香豆素、异茴香内酯和佛手柑内酯的含量,用化学计量学方法对含量测定结果进行综合分析,挖掘影响马齿苋质量的主要潜在标志物,联合熵权-逼近理想解排序法对不同产地来源的马齿苋进行质量评价。结果10种成分分别在各自线性范围内线性关系和准确度良好;化学计量学分析结果显示,18批马齿苋聚为3类,呈现一定的产区差异;木犀草素、槲皮素、琥珀酸和柠檬酸是影响马齿苋产品质量的主要潜在标志物;熵权-逼近理想解排序法分析结果显示四川和贵州地区所产马齿苋质量最优,其次为河南、辽宁和吉林。结论所建立的高效液相色谱法能同时测定马齿苋中10个指标成分的含量,化学计量学和熵权-逼近理想解排序法可用于马齿苋的综合质量评价。
基金Acknowledgements This study was supported by the National Science Foundation （No. 30630067）；the State Key Basic Research Grant of China （No. 2004CB518705）； the Program for Changjiang Scholars and Innovative Research Team in University （No. IRT0416）.
文摘Chromosome aberrations are distinctive features of human malignant tumors. Analysis of chromosomal changes can illuminate the molecular mechanisms underlying the development and progression of cancer. To establish the technique of multicolor fluorescence in situ hybridization （M-FISH） for identifying chromosome aberrations in esophageal carcinoma cell line KYSE 410-4, four pools of 6-color whole-chromosome painting probes have been designed and hybridized on the same metaphase spread by four rounds of repetitive FISH. Repetitive 6-color M-FISH was successfully established and the cytogenetic abnormalities in KYSE 410-4 cells were characterized. Chromosome gains occurred at 2q, 3, 8, 17p, and X. An isochromosome 3q was visualized in the cell line, which might be one intermediate mechanism leading to 3p losses and/or 3q gains. Furthermore, 16 structural arrangements were detected, including four derivative chromosomes. The rearrangement of the centromeric regions accounted for approximately 44% of all rearrangements. The results added a more complete and accurate information of the genetic alterations to the classical cytogenetic description of KYSE 410-4 and provided a detailed cytogenetic background data for appropriate use of the cell line. The established 6-color M-FISH was useful for analyzing chromosomes in the whole genome of human tumors.
