A new reversed-phase high performance liquid chromatography method was developed to quantitate the activity of xanthine oxidase involved in milk fat globule membrane with xanthine as the substrate and the separation o...A new reversed-phase high performance liquid chromatography method was developed to quantitate the activity of xanthine oxidase involved in milk fat globule membrane with xanthine as the substrate and the separation of product (uric acid). The increment of uric acid in the reaction system was used to calculate the total activity of XO. The optimized assay conditions, linearity of detection, recovery of uric acid and chromatogram were developed in text, indicating this method is simple, rapid and efficient. It is an alternative potential method for the determination of the activity of XO in milk.展开更多
Isolation and comparison of uremic sera and urine and normal sera and urine were performed by gel permeation chromatography, anion exchange chromatography and re-versed-phase high performance liquid chromatography. Tw...Isolation and comparison of uremic sera and urine and normal sera and urine were performed by gel permeation chromatography, anion exchange chromatography and re-versed-phase high performance liquid chromatography. Two uremic middle molecular fractions (A and B) were obtained from uremic sera and urine and normal urine by gel permeation chromatography, but not from normal sera. The anion exchange chromatographic results of fraction A from different origins demonstrate that subfraction A-3 could be excreted in urine by healthy subject, but accumulated in uremic serum for renal failure of patient with uremia. After desalinization subfraction A-3 was analyzed by MALDI-TOF-MS. The results show that subfraction A-3 consists of six compounds with molecular weight 839, 873, 1007.94, 1106, 1680 and 2015 respectively. Finally, by reversed-phase high performance liquid chromatography, subfraction A-3 was further resolved into six independent fractions. Thus, the isolation and purification of six middle molecular展开更多
[Objectives]To establish a method to simultaneously determine the content of paederosidic acid in Paederia scandens dried by four different methods.[Methods]Reversed-phase high performance liquid chromatography(RP-HPL...[Objectives]To establish a method to simultaneously determine the content of paederosidic acid in Paederia scandens dried by four different methods.[Methods]Reversed-phase high performance liquid chromatography(RP-HPLC)was adopted.The chromatographic column:Thermo SCIENTIFIC Hypersil GOLD Dim.(mm);mobile phase:acetonitrile-0.1%phosphoric acid aqueous solution;gradient elution;column temperature:30℃;flow rate:1 mL/min;detection wavelength:236 nm;injection volume:10μL.[Results]The linear range of the detection injection volume of paederosidic acid was 0.64-9.60μg(R=0.9992);the limit of quantity(LOQ)was 5.10 ng,and the limit of detection(LOD)was 1.36 ng;the RSD of the precision,stability and reproducibility test was all less than 3%;the sample recovery rate was 98.87%,RSD<3.00,n=6.The results show that the content of paederosidic acid in the shade-dried P.scandens is the highest,and the order of the content is P.scandens(dry in shade)>P.scandens(dried at 50℃)>P.scandens(dried at 60℃)>P.scandens(dried at 70℃).[Conclusions]This method is sensitive,reliable and reproducible,and can be used to simultaneously determine the content of paederosidic acid in P.scandens dried by four different methods.展开更多
[ Objective] The aim was to establish effective method for endogenous hormone extraction and explore conditions of chromatographic analysis for three endogenous hormones in rhizome of Alhagi sparsifolia. [ Methed ] Ac...[ Objective] The aim was to establish effective method for endogenous hormone extraction and explore conditions of chromatographic analysis for three endogenous hormones in rhizome of Alhagi sparsifolia. [ Methed ] Activol （GA3 ）, zeatin （ZR） and indole-3-acetic acid （IAA） in rhizome were separated and measured as per RP-HPLC. [Result] The average recovery rates of GA3, ZR and IAA were 98.3%, 90.3% and 101.3%, respectively, indicating that the method is suitable for quantitative analysis with little errors. The chromatographic conditions were as follows： methanol/0. 75% of acetic acid at 45：55 （mobile phase） ; flow speed at 0.7 ml/min; wavelength at 254 nm; column temperature at 25 ℃. [Conclusion] The research preliminarily established HPLC conditions for separation of endogenous hormones in rhizome of Alhagi sparsifolia.展开更多
文摘A new reversed-phase high performance liquid chromatography method was developed to quantitate the activity of xanthine oxidase involved in milk fat globule membrane with xanthine as the substrate and the separation of product (uric acid). The increment of uric acid in the reaction system was used to calculate the total activity of XO. The optimized assay conditions, linearity of detection, recovery of uric acid and chromatogram were developed in text, indicating this method is simple, rapid and efficient. It is an alternative potential method for the determination of the activity of XO in milk.
基金This work was supported by the State Key Fundamental R & D Project (Grant No. G1999064707)the National Natural Science Foundation of China (Grant No. 59873011) Trans-Century Training Program Foundation for the Talents by the Ministry of Education,
文摘Isolation and comparison of uremic sera and urine and normal sera and urine were performed by gel permeation chromatography, anion exchange chromatography and re-versed-phase high performance liquid chromatography. Two uremic middle molecular fractions (A and B) were obtained from uremic sera and urine and normal urine by gel permeation chromatography, but not from normal sera. The anion exchange chromatographic results of fraction A from different origins demonstrate that subfraction A-3 could be excreted in urine by healthy subject, but accumulated in uremic serum for renal failure of patient with uremia. After desalinization subfraction A-3 was analyzed by MALDI-TOF-MS. The results show that subfraction A-3 consists of six compounds with molecular weight 839, 873, 1007.94, 1106, 1680 and 2015 respectively. Finally, by reversed-phase high performance liquid chromatography, subfraction A-3 was further resolved into six independent fractions. Thus, the isolation and purification of six middle molecular
基金The First Batch High-level Talent Scientific Research Project of The Affiliated Hospital of Youjiang Medical University for Nationalities in 2019(Y20196311)Project for Improving Basic Research Ability of Young and Middle-aged and Young Teachers in Colleges and Universities of Guangxi in 2020(2020KY13034)Program of Youjiang Medical University for Nationalities(yy2018ky018).
文摘[Objectives]To establish a method to simultaneously determine the content of paederosidic acid in Paederia scandens dried by four different methods.[Methods]Reversed-phase high performance liquid chromatography(RP-HPLC)was adopted.The chromatographic column:Thermo SCIENTIFIC Hypersil GOLD Dim.(mm);mobile phase:acetonitrile-0.1%phosphoric acid aqueous solution;gradient elution;column temperature:30℃;flow rate:1 mL/min;detection wavelength:236 nm;injection volume:10μL.[Results]The linear range of the detection injection volume of paederosidic acid was 0.64-9.60μg(R=0.9992);the limit of quantity(LOQ)was 5.10 ng,and the limit of detection(LOD)was 1.36 ng;the RSD of the precision,stability and reproducibility test was all less than 3%;the sample recovery rate was 98.87%,RSD<3.00,n=6.The results show that the content of paederosidic acid in the shade-dried P.scandens is the highest,and the order of the content is P.scandens(dry in shade)>P.scandens(dried at 50℃)>P.scandens(dried at 60℃)>P.scandens(dried at 70℃).[Conclusions]This method is sensitive,reliable and reproducible,and can be used to simultaneously determine the content of paederosidic acid in P.scandens dried by four different methods.
基金Initial Funding of Doctor in Xinjiang University(7020428027)National Innovation Experiment Program for University Students of China (101075512)
文摘[ Objective] The aim was to establish effective method for endogenous hormone extraction and explore conditions of chromatographic analysis for three endogenous hormones in rhizome of Alhagi sparsifolia. [ Methed ] Activol （GA3 ）, zeatin （ZR） and indole-3-acetic acid （IAA） in rhizome were separated and measured as per RP-HPLC. [Result] The average recovery rates of GA3, ZR and IAA were 98.3%, 90.3% and 101.3%, respectively, indicating that the method is suitable for quantitative analysis with little errors. The chromatographic conditions were as follows： methanol/0. 75% of acetic acid at 45：55 （mobile phase） ; flow speed at 0.7 ml/min; wavelength at 254 nm; column temperature at 25 ℃. [Conclusion] The research preliminarily established HPLC conditions for separation of endogenous hormones in rhizome of Alhagi sparsifolia.
