Method development for determination of isoflavones in kudzu was achieved by HPLC/UV/ESI-MSD. Us- ing three kudzu species of Pueraria lobata, P. thomsonii and P. edulis, and analyzing the isoflavones sepa- rately by s...Method development for determination of isoflavones in kudzu was achieved by HPLC/UV/ESI-MSD. Us- ing three kudzu species of Pueraria lobata, P. thomsonii and P. edulis, and analyzing the isoflavones sepa- rately by species and from different plant tissues (roots, stems, leaves, flowers and fruits) in each species, a total of 25 isoflavones were identified by their molecular ions and characteristic fragment ion peaks using LC/MSD under MS and MS/MS mode, and in comparison with standard isoflavones. Two main chemical groups were identified: 1) 8-C-glycosyl isoflavone of puerarin and the analogues of 5-OH puerarin, 3’-OH puerarin, 3’-OMe puerarin, and their glycosides;and 2) daidzein, genistein, glycitein and their glycosyl and malonyl derivatives, which are similar to those known in soy. To accurately quantitate total isoflavones, acidic hydrolysis during extraction of kudzu samples was applied to convert the oxygen glycosides into their respective isoflavone aglycones of daidzein, genistein and glycitein, or non-hydrolyzed carbon glycosides of puerarin, 5-OH puerarin, 3’-OH puerarin and 3’-OMe puerarin. Under the multiple optimized conditions, all seven isoflavones in acidic hydrolyzed kudzu extracts were successfully separated within 30 min and quanti- fied individually with calycosin used as internal standard by both UV and MS detectors. For the quantitative study, several standards e.g. 5-OH puerarin, 3’-OH puerarin and 3’-OMe puerarin are not commercially available. Using polyamide, sephdex-LH20 chromatography and Prep-HPLC, we purified these three stan- dards from kudzu extracts and then elucidated their structures by UV, MS and NMR spectrometric methods. This is the first method to simultaneously quantitate all the isoflavones in kudzu.展开更多
As a part of the ongoing search for new constituents of Stemona species in China, chemical investigation of Stemona sessilifolia, a plant used in the traditional Chinese medicine to treat respiratory disorders, was ca...As a part of the ongoing search for new constituents of Stemona species in China, chemical investigation of Stemona sessilifolia, a plant used in the traditional Chinese medicine to treat respiratory disorders, was carried out. To identify the chemical components rapidly, a selected sample of S. sessilifolia containing bibenzyls was tested using LC-ESIMS and analyzed further using stop-flow LC-UV-NMR, which was sensitive for the detection of the main constituents. LC microfractions were collected using the LC-UV-NMR technique and HR-EIMS off-line analysis was cartied out on the collected fractions. This chemical screening strategy allowed for the on-line identification of the main constituents of S. sessilifolia and provided information that was useful for a further peak-guided isolation procedure. Using these methods, four bibenzyls were isolated： two known compounds, 3,5-dihydroxy-4-methyl bibenzyl（1） and 3, 5-dihydroxy-2＇-methoxy-4-methyl bibenzyl （ 2 ）, and two novel compounds, 3,3＇-dihydroxy-5, 6＇-dimethoxy bibenzyl（3） and 3,5-dihydroxy-2＇, 5＇-dimethoxy bibenzyl （4）.展开更多
Five different quorum sensing peptides(QSP) were iodinated using different iodination techniques. These iodinated peptides were analyzed using a C18 reversed phase HPLC system, applying a linear gradient of water and ...Five different quorum sensing peptides(QSP) were iodinated using different iodination techniques. These iodinated peptides were analyzed using a C18 reversed phase HPLC system, applying a linear gradient of water and acetonitrile containing 0.1%(m/v) formic acid as mobile phase. Electrospray ionization(ESI)ion trap mass spectrometry was used for the identification of the modified peptides, while semi-quantification was performed using total ion current(TIC) spectra. Non-iodinated peptides and mono-and diiodinated peptides(NIP, MIP and DIP respectively) were well separated and eluted in that order. Depending on the used iodination method, iodination yields varied from low(2%) to high(57%).展开更多
文摘Method development for determination of isoflavones in kudzu was achieved by HPLC/UV/ESI-MSD. Us- ing three kudzu species of Pueraria lobata, P. thomsonii and P. edulis, and analyzing the isoflavones sepa- rately by species and from different plant tissues (roots, stems, leaves, flowers and fruits) in each species, a total of 25 isoflavones were identified by their molecular ions and characteristic fragment ion peaks using LC/MSD under MS and MS/MS mode, and in comparison with standard isoflavones. Two main chemical groups were identified: 1) 8-C-glycosyl isoflavone of puerarin and the analogues of 5-OH puerarin, 3’-OH puerarin, 3’-OMe puerarin, and their glycosides;and 2) daidzein, genistein, glycitein and their glycosyl and malonyl derivatives, which are similar to those known in soy. To accurately quantitate total isoflavones, acidic hydrolysis during extraction of kudzu samples was applied to convert the oxygen glycosides into their respective isoflavone aglycones of daidzein, genistein and glycitein, or non-hydrolyzed carbon glycosides of puerarin, 5-OH puerarin, 3’-OH puerarin and 3’-OMe puerarin. Under the multiple optimized conditions, all seven isoflavones in acidic hydrolyzed kudzu extracts were successfully separated within 30 min and quanti- fied individually with calycosin used as internal standard by both UV and MS detectors. For the quantitative study, several standards e.g. 5-OH puerarin, 3’-OH puerarin and 3’-OMe puerarin are not commercially available. Using polyamide, sephdex-LH20 chromatography and Prep-HPLC, we purified these three stan- dards from kudzu extracts and then elucidated their structures by UV, MS and NMR spectrometric methods. This is the first method to simultaneously quantitate all the isoflavones in kudzu.
基金Supported by Science and Technology Committee of Shanghai, P R China(No 036505003)
文摘As a part of the ongoing search for new constituents of Stemona species in China, chemical investigation of Stemona sessilifolia, a plant used in the traditional Chinese medicine to treat respiratory disorders, was carried out. To identify the chemical components rapidly, a selected sample of S. sessilifolia containing bibenzyls was tested using LC-ESIMS and analyzed further using stop-flow LC-UV-NMR, which was sensitive for the detection of the main constituents. LC microfractions were collected using the LC-UV-NMR technique and HR-EIMS off-line analysis was cartied out on the collected fractions. This chemical screening strategy allowed for the on-line identification of the main constituents of S. sessilifolia and provided information that was useful for a further peak-guided isolation procedure. Using these methods, four bibenzyls were isolated： two known compounds, 3,5-dihydroxy-4-methyl bibenzyl（1） and 3, 5-dihydroxy-2＇-methoxy-4-methyl bibenzyl （ 2 ）, and two novel compounds, 3,3＇-dihydroxy-5, 6＇-dimethoxy bibenzyl（3） and 3,5-dihydroxy-2＇, 5＇-dimethoxy bibenzyl （4）.
基金the‘Research Foundation–Flanders(FWO)’(Grant no.1S21017N to Nathan Debunne)the‘Institute for the Promotion of Innovation through Science and Technology in Flanders(IWT-Vlaanderen)’(Grant no.131356 to Frederick Verbeke)
文摘Five different quorum sensing peptides(QSP) were iodinated using different iodination techniques. These iodinated peptides were analyzed using a C18 reversed phase HPLC system, applying a linear gradient of water and acetonitrile containing 0.1%(m/v) formic acid as mobile phase. Electrospray ionization(ESI)ion trap mass spectrometry was used for the identification of the modified peptides, while semi-quantification was performed using total ion current(TIC) spectra. Non-iodinated peptides and mono-and diiodinated peptides(NIP, MIP and DIP respectively) were well separated and eluted in that order. Depending on the used iodination method, iodination yields varied from low(2%) to high(57%).
