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Zeng Y.,Guangxi Key Laboratory of Subtropical Bioresource Conservation and Utilization
Se pu = Chinese journal of chromatography / Zhongguo hua xue hui | Year: 2010

A method for the simultaneous determination of berberine, palmatine, matrine, catechin and baicalin in Funing Shuan was established using micellar electrokinetic capillary chromatography-electrospray ionization mass spectrometry (MEKC-ESI MS). The compounds were separated on an uncoated capillary (80 cm x 50 microm) with the operating voltage of 25 kV and the running buffer of 40 mmol/L lauric acid-100 mmol/L ammonia mixture containing 25% acetonitrile (pH 9.5). The baseline separation of the five compounds was achieved within 16 min with a satisfactory repeatability and sensitivity. The solution of 50% 2-propanol/water solution (containing 3 mmol/L acetic acid) was used as the sheath liquid for the ESI MS analysis. The results showed that the linear ranges for berberine, palmatine, matrine, catechin and baicalin were 0.03 - 15, 0.05 - 15, 0.2 - 250, 1.5 - 300 and 2.0 - 500 mg/L, respectively, and the detection limits were 0.01, 0.02, 0.05, 0.5 and 0.6 mg/L, respectively. The average recoveries of the five components were between 94.0% - 104.0% with the relative standard deviations (RSDs) of 0.3% - 3.2%. The developed method is simple, rapid, and accurate, and it is suitable for the routine analysis of the five effective components in Funing Shuan. Source


Zeng Y.,Guangxi Key Laboratory of Subtropical Bioresource Conservation and Utilization | Zeng Y.,China University of Technology | Wei X.,China University of Technology | Xu Y.,Guangxi Key Laboratory of Subtropical Bioresource Conservation and Utilization | Xu Y.,China University of Technology
Chemistry Bulletin / Huaxue Tongbao | Year: 2010

An online micellar electrokinetic chromatography-electrospray ionization mass spectrometry method for simultaneous determination of forsythin, chrysophanol and emodin in Lianqiao Baidu Pills was established. The samples were separated by an uncoated capillary (50μm × 78cm) on the operating voltage of 25kV using 40 lauric acid - 100 mmol/L ammonia mixture containing 20% acetonitrile (pH =9. 0) as the running buffer. The effect of experimental conditions, such as selection of bases, concentration of lauric acid, addition of organic solvent, applied voltage and the composition of sheath liquid on the separation and ionization of herbicides was investigated. The baseline separation of three components was achieved within 14 minutes with satisfactory repeatability and sensitivity. The linear ranges of forsythin, chrysophanol and emodin were 0. 100 - 100, 2. 50 - 250, 0. 0500 - 50. 0mg/L with detection limits of 0.0200, 0.800, 0. 00500mg/L, respectively. The average recoveries ranged from 95% - 104% with all relative standard deviations less than 4.0%. The developed method is rapid, simple, and accurate and is suitable for the quality control of the three effective components in Lianqiao Baidu Pills. Source


Zeng Y.,Guangxi Key Laboratory of Subtropical Bioresource Conservation and Utilization | Zeng Y.,Guangxi University | Huo P.,Guangxi Key Laboratory of Subtropical Bioresource Conservation and Utilization | Xu Y.,Guangxi Key Laboratory of Subtropical Bioresource Conservation and Utilization | Xu Y.,Guangxi University
Chinese Journal of Chromatography (Se Pu) | Year: 2010

A method for the simultaneous determination of b erbe rine, palmatine, matrine, catechin and baicalin in Funing Shuan was established using micellar electrokinetic capillary chromatography- electrospray ionization mass spectrometry (MEKC-ESI MS). The compounds were separated on an uncoated capillary (80 cm x 50|xm) w ith the operating voltage of 25 kV and the running b uffer of 40 m m o l/L lauric acid-100 m m o l/L ammonia mixture containing 25 % acetonitrile (pH 9.5). The baseline separation of the five compounds was achieved with in 16 min with a satisfactory repeatability and sensitivity. The solution of 50% 2-prop an ol/water solution (containing 3 mm ol/L acetic acid) was used as the sheath liquid for the ESI MS analysis. The results showed that the linear ranges for berberine, palmatine, matrine, catechin and baicalin were 0.03 - 15, 0.05 -15, 0.2- 250, 1. 5 - 300 and 2.0 - 500 m g /L, respectively, and the detection lim its were 0.01, 0.02, 0.05, 0.5 and 0.6 m g /L, respectively. The average recoveries of the five components were between 94. 0% - 104. 0% with the relative standard deviations (RSDs) of 0. 3% - 3. 2%. The developed method is simple, rapid, and accurate, and it is suitable for the routine analysis of the five effective components in Funing Shuan. Source


Tang M.,Guangxi University of Technology | Shen P.,Guangxi University of Technology | Shen P.,Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering | Shen P.,Guangxi Key Laboratory of Subtropical Bioresource Conservation and Utilization | And 12 more authors.
Chinese Journal of Applied and Environmental Biology | Year: 2010

A put A gene was isolated from the genomic library of Sinorhizobium fredii HN01 by molecular and biotechnological methods, and it had 82% and 86% identities with the sequence of the proline dehydrogenase from S.meliloti 1021 at nucleotide and amico acid level, respectivly. With the suicide plasmid pK18mob, the polar mutant GXHNPB and non-polar mutant GXHNPA inactivated in putA gene were constructed through homologous recombination. The putA gene was cloned into pLAFRJ vetor resulting in a complemention plasmid pGXHN37. The complement strains GXHNPHA and GXHNPHB were constructed by transforming pGXH37 into host strains GXHNPA and GXHNPB, respectively. The results of growth test showed that there was no difference among HN01, mutant and complement strains when were cultivated on completemedium YMB, but the mutants showed no growth in the MM medium containing proline as the sole carbon and nitrogen source. Plant tests revealed that mutants had lower nodulation efficiency ability and nitrogen-fixation, and nodulated 1 day later on soybean compared with the wild-strain HN01. Fig 6, Tab 1, Ref 19. Source


Zhang W.,Guangxi University | Zhang W.,Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering | Zhang W.,Guangxi Key Laboratory of Subtropical Bioresource Conservation and Utilization | Tian R.,Guangxi University | And 17 more authors.
Chinese Journal of Applied and Environmental Biology | Year: 2011

In order to know function of cysDN and its effect on plant nodulation, a random insertion mutant library of Sinorhizobium fredii 15142 was constructed by three parental hybridization with the plasmid pTnMod-RKm' as transposon vector. A mutant strain which could not metabolize sulfate while could use cysteine as sulfur source, was selected by MM medium with different sulfur sources from the library. Molecular cloning and sequence analysis showed that this operon had 92% and 96% similarity with the cysDN gene sequence of Sinorhizobium sp. BR816 at nucleotide and amino acid level, respectively. With the suicide plasmid pK18mob, the polar mutant cysDF/15142, cysNF/15142 and non-polar mutant cysDR/15142, cysNR/15142 inactivated in cysD and cysN were constructed through homologous recombination. A recombinant plasmid pLAFRJ+cysDN was constructed by using plasmid pLAFRJ ligated with cysDN, and transferred into above mutants to obtain complement strains. The results of growth tests found that the mutants had no growth on the MM medium with sulfate as sulfur source but the complement strains could grow normally, indicating that cysDN operon was related to sulfate assimilation. The plant tests indicated that the mutant strains delayed 1-2 d to nodulate than the wild strain, and nitrogenase activity and nodulation efficiency in mutants were slightly lower than those of the original strain, but there were no differences in the number and weight of nodules and the dry weight of plant. Source

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