TY - JOUR
T1 - Reversed-phase liquid chromatographic isolation of lubimin and solavetivone from Hyoscyamus muticus "hairy" root cultures
AU - Reddy, G. Ramakrishna
AU - Signs, Mark W.
AU - Flores, H. E.
AU - Curtis, W. R.
N1 - Funding Information:
We are grateful to Dr. Mani Venkatachalam (Supelco, Bellefonte, PA) for carrying out GC-MS and to Dr. Adil Dhalla (The Pennsylvania State University) for performing the NMR spectroscopy. We would like to acknowledgeD r. S. Yella Reddy (The PennsylvaniaS tate University) for his technical assistancea nd helpful comments during preparation of this manuscript. Finally we would like to acknowledge NSF (Grant no. BCS-9110288)f or financial support.
PY - 1993/12/31
Y1 - 1993/12/31
N2 - A high-performance liquid chromatographic reversed-phase procedure was developed to estimate the concentration of sesquiterpene phytoalexins secreted into the media by Agrobacterium transformed root cultures of Hyoscyamus muticus. An isolation procedure based on solid-phase extraction (C18 Sep-pak) results in very rapid analysis and gives full recovery, compared to sequential chloroform extraction. A simple isocratic procedure permits quantification of sesquiterpene production by these root cultures in response to exposure to fungal elicitors. Calibration curves for lubimin and solavetivone content were obtained by fractionating sufficient material to provide a measurable weight. The identity of these compounds was confirmed by TLC, UV, GC-MS, and NMR. This experimental system is being used to examine the regulation of sesquiterpene biosynthesis, and to develop large-scale processing techniques for the production of secondary metabolises from plant roots grown in bioreactor systems.
AB - A high-performance liquid chromatographic reversed-phase procedure was developed to estimate the concentration of sesquiterpene phytoalexins secreted into the media by Agrobacterium transformed root cultures of Hyoscyamus muticus. An isolation procedure based on solid-phase extraction (C18 Sep-pak) results in very rapid analysis and gives full recovery, compared to sequential chloroform extraction. A simple isocratic procedure permits quantification of sesquiterpene production by these root cultures in response to exposure to fungal elicitors. Calibration curves for lubimin and solavetivone content were obtained by fractionating sufficient material to provide a measurable weight. The identity of these compounds was confirmed by TLC, UV, GC-MS, and NMR. This experimental system is being used to examine the regulation of sesquiterpene biosynthesis, and to develop large-scale processing techniques for the production of secondary metabolises from plant roots grown in bioreactor systems.
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U2 - 10.1016/0021-9673(93)80300-W
DO - 10.1016/0021-9673(93)80300-W
M3 - Article
AN - SCOPUS:0027738628
VL - 657
SP - 440
EP - 444
JO - Journal of Chromatography A
JF - Journal of Chromatography A
SN - 0021-9673
IS - 2
ER -