Application of SPE for the HPLC analysis of taxanes fromTaxus cell cultures

Theodoridis, Georgios; Jong, C.F. de; Laskaris, Gregory; Verpoorte, Robert

doi:10.1007/bf02466782

Cited by 20 publications

(11 citation statements)

References 25 publications

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“…The cleaning of samples taken from root tissue and the aqueous phase of the medium was performed using the method described by Theodoridis et al (1998a). The residues arising from SPE (Solid Phase Extraction) extraction (root tissue and aqueous phase of medium) and the PFD phases were re-dissolved in 100 % MeOH (400 ll) and 20 ll underwent High Performance Liquid Chromatography with Diode Array Detection (HPLC-DAD) analysis using the DIONEX (USA) system with a UVD 340S diode array detector and an automated sample injector (ASI-100).…”

Section: Chemical Analysismentioning

confidence: 99%

Perfluorodecalin-supported system enhances taxane production in hairy root cultures of Taxus x media var. Hicksii carrying a taxadiene synthase transgene

Sykłowska-Baranek

Pilarek

Bonfill

et al. 2014

Plant Cell Tiss Organ Cult

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Enhanced taxane production was observed in a hybrid, two liquid phases containing, cultures of Taxus x media var. Hicksii hairy root carrying a taxadiene synthase transgene, supported with liquid perfluorodecalin (PFD) in degassed or aerated form. The hairy root cultures were elicited with methyl jasmonate (MJ, 100 lM) or coronatine (COR, 1 lM), and fed with sucrose and L-phenylalanine. The root growth was not stimulated by PFD addition, irrespective of the day of its application (day 0 and 14). However, in the cultures elicited with MJ and performed in the presence of PFD the final root biomass accumulation was higher than in cultures performed without PFD while the opposite effect was observed in cultures supplemented with COR. The highest paclitaxel content in root biomass was determined at the end of the cultures elicited with MJ and supplemented with PFD-degassed at day 0 or 14, 1,440.8 and 1,432.5 lg g -1 DW, respectively. The highest total (i.e. intracellular ? extracellular: both in aqueous and PFD phases) paclitaxel yield in flasks (149.15 lg flask -1 ) was noted after the application of PFD-degassed at day 14. The other taxane detected was baccatin III, only in the root biomass, with the highest content (76.9 lg g -1 DW) observed under COR treatment. Although COR stimulated paclitaxel production with less efficiency than MJ, it resulted in higher paclitaxel excretion to the liquid phases of culture medium and PFD.

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Section: Chemical Analysismentioning

confidence: 99%

Perfluorodecalin-supported system enhances taxane production in hairy root cultures of Taxus x media var. Hicksii carrying a taxadiene synthase transgene

Sykłowska-Baranek

Pilarek

Bonfill

et al. 2014

Plant Cell Tiss Organ Cult

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“…The samples were cleaned using the method described by Theodoridis et al (1998a). Prior to HPLC analysis, samples were re-dissolved in 100 % MeOH (400 ll) and 20 ll underwent HPLC-DAD analysis using the DIONEX (USA) system with a UVD 340S diode array detector and an automated sample injector (ASI-100).…”

Section: Taxane Determinationmentioning

confidence: 99%

Paclitaxel production and PAL activity in hairy root cultures of Taxus x media var. Hicksii carrying a taxadiene synthase transgene elicited with nitric oxide and methyl jasmonate

et al. 2015

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The role of sodium nitroprusside (SNP, 10 lM), a nitric oxide (NO) donor, and/or methyl jasmonate (MJ, 100 lM) spiked with L-phenylalanine (PHEN, 100 lM) and additional sucrose (S; 30 g l -1 ), in taxane production and phenyl ammonia lyase (PAL) activity in cultures of two Taxus media x var. Hicksii transgenic root lines (ATMA and ATM) carrying the taxadiene synthase transgene was investigated. SNP addition, when applied together with MJ and/or PHEN, resulted in paclitaxel production only in ATMA cultures. The application of the NO donor gave the highest paclitaxel content (7.56 mg l -1) in the combination of SNP?S?MJ?PHEN, after 2 weeks of treatment in the ATMA root line. In ATM cultures, taxane production was not affected by SNP. In both ATMA and ATM lines the highest total (intra?extracellular) paclitaxel yield was determined when elicited with MJ?PHEN, and amounted to 10.78 mg l -1 at 1 week and 1.63 mg l -1 at 2 weeks of treatment, in cultures of ATMA and ATM lines, respectively. The excretion of paclitaxel was observed only in ATMA cultures, with the highest level (2.34 mg l -1 ) obtained after elicitation with S?MJ?PHEN. The comparison of PAL activity in the two root lines revealed that this enzyme was almost 3-times more active in ATM than ATMA roots. An increase in both PAL activity and paclitaxel production was only observed in ATMA cultures growing in medium supplemented with S?MJ?PHEN.

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“…Taxus ×media suspension culture did not contain any of the lignans in question. After extended duration of methanolic extraction according to Theodoridis et al [35] method used for paclitaxel determination, the pinoresinol content was about 20 percent higher comparing Schmitt and Petersen [29] method employed (Tab. 1).…”

Section: Discussionmentioning

confidence: 99%

“…Samples of all H 2 O and CH 2 Cl 2 phases were analyzed by HPLC [29]. We also extended the duration of plant material MeOH-extraction up to 24 h according to Theodoridis et al [35] method used for paclitaxel determination. A portion of 0.1 g freeze-dried, powdered material was suspended in 1 ml MeOH, mixed, sonicated for 15 min, and next centrifuged on the Gyrotory Shaker at 110 rpm for 24 h. The plant sample was centrifuged at 5°C, 15 500 rpm for 15 min, then the methanolic extract was collected and the residue was again extracted with 1 ml MeOH, mixed, sonicated for 15 min and centrifuged as above.…”

Section: Lignan Extraction and Determinationmentioning

confidence: 99%

Identification and quantitative determination of pinoresinol in Taxus ×media Rehder needles, cell suspension and shoot cultures

et al. 2015

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The aim of our study was to investigate the presence and quantitative contents of lignans in the tissues of Taxus ×media. The presence of the lignans: pinoresinol, matairesinol and secoisolariciresinol was assessed in needles, shoots cultures and suspension culture. Pinoresinol was the only lignan found in the tissue of T. ×media. The total pinoresinol content in the needles and in the shoots was 1.24 mg/g dry weight (dw) and 0.69 mg/g dw, respectively. Most of the pinoresinol identified was appeared glycosidically bound. In needles, the amount of glycosidically bound pinoresinol (0.81 mg/g dw) was about twice as high as that of free pinoresinol (0.43 mg/g dw). The content of free and glycosidically bound pinoresinol showed the level of 0.18 mg/g dw and 0.51 mg/g dw, respectively in the in vitro shoot cultures. In the cell culture, no pinoresinol was found.

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Application of SPE for the HPLC analysis of taxanes fromTaxus cell cultures

Cited by 20 publications

References 25 publications

Perfluorodecalin-supported system enhances taxane production in hairy root cultures of Taxus x media var. Hicksii carrying a taxadiene synthase transgene

Perfluorodecalin-supported system enhances taxane production in hairy root cultures of Taxus x media var. Hicksii carrying a taxadiene synthase transgene

Paclitaxel production and PAL activity in hairy root cultures of Taxus x media var. Hicksii carrying a taxadiene synthase transgene elicited with nitric oxide and methyl jasmonate

Identification and quantitative determination of pinoresinol in Taxus ×media Rehder needles, cell suspension and shoot cultures

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