Levbid
Determination of enantiomeric purity of hyoscyamine from scopolia extract using HPLC-CD system without chiral separation.

Kudo K, Iwaya K, Yomota C, Morris S, Saito M.

JASCO Corporation, 2967-5 Ishikawa-cho, Hachioji, Tokyo 192-8537, Japan.

Enantiomeric ratio of hyoscyamine from Scopolia extract was determined by chiral HPLC-CD analysis. It was found that circular dichroism (CD) detection allowed the analysis of the sample without any special pretreatment whereas UV detection required an ammonia-ether extraction. To obtain a shorter analysis time for the determination, reversed-phase HPLC-CD analysis was applied by using a g-factor calibration curve (EE% vs. CD/UV). The analysis time was shortened from 35 to 18 min. EE% values obtained were consistent with those by chiral HPLC analysis.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11126877&dopt=Abstract hyoscyamine Levbid SL



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A rapid densitometric method for the analysis of hyoscyamine and scopolamine in solanaceous plants and their transformed root cultures.

Berkov S, Pavlov A.

Institute of Botany, Bulgarian Academy Sciences, 23 Acad. G. Bonchev Sir., 1113 Sofia, Bulgaria. berkov iph.bio.bas.bg

A rapid and convenient method for sample preparation and simultaneous densitometric quantification of hyoscyamine (1) and scopolamine (2) has been developed. The alkaloid profiles of plant samples, as well as the content of 1 and 2, determined by TLC-densitometry corresponded well with those obtained using a GC method. The proposed method is simple and sensitive and can be used for the routine assay of 1 and 2, and for screening for alkaloid-rich and biochemically-interesting transformed root cultures and intact solanaceous plants.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15202596&dopt=Abstract hyoscyamine Levbid SL



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Datura metel: in vitro production of tropane alkaloids.

Cusido RM, Palazon J, Pinol MT, Bonfill M, Morales C.

Unidad de Fisiologia Vegetal, Facultad de Farmacia, Universidad de Barcelona, Spain.

Hairy root lines of Datura metel were established following infection of aseptic stem segments with Agrobacterium rhizogenes strain A4 and cultured in hormone-free B5 solid medium. The growth and production of hyoscyamine and scopolamine (mg/g dry wt.) of these root cultures was encouraged by using B5 liquid medium with half-strength salts. In these culture conditions, the capacity of the highest productive root line 25 to excrete scopolamine into the culture medium rose from 8.7% to 70% when the permeabilizing agent Tween 20 was added for 24 h to the medium, after 2 and 4 weeks of culture. Using an airlift bioreactor (41) with modifications in order to increase root anchorage, the Tween 20 treatment encouraged both growth and alkaloid productivity of cultured root line 25. After 4 weeks their biomass yield was 2.3 mg/l/day and 0.84 mg/l/day of scopolamine was produced (70% extracellular). The scopolamine released into the culture medium was separated with an Amberlite XAD-2 column located in the media exit.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10193206&dopt=Abstract hyoscyamine Levbid SL



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Rapid in vitro adventitious shoot propagation of Scopolia parviflora through rhizome cultures for enhanced production of tropane alkaloids.

Kang YM, Min JY, Moon HS, Karigar CS, Prasad DT, Lee CH, Choi MS.

Division of Forest Science, Gyeongsang National University, 900 Gajwa-dong, 660-701, Jinju, Gyeongnam, South Korea.

A rapid micropropagation system for Scopolia parviflora Nakai (Solanaceae), a rare medicinal plant native to Korea, was established using rhizome cultures. Shoots that originated from adventitious shoots of the rhizome were multiplied when the rhizomes were cultured on half-strength B5 liquid medium supplemented with various growth regulators. Optimum shoot multiplication was observed in half-strength B5 medium containing 3% (w/v) sucrose and 5.77 microM gibberellic acid (GA(3)). Each rhizome gave rise to an average of 12 shoots. Shoot elongation and root induction from multiple shoots occurred on growth regulator-free half-strength B5 solid medium. Healthy plantlets were transferred to a peat moss:vermiculite mixture for acclimatization, which was successful. The concentrations of tropane alkaloids, hyoscyamine and scopolamine were determined in different tissues of native growing plants, in vitro-propagated plants and acclimatized plants by high-performance liquid chromatography. The analysis revealed that the levels of hyoscyamine and scopolamine were higher in in vitro-propagated plants than in the native growing plants. When the rhizome was cut into segments and transferred to optimal culture conditions for multiple shoot propagation, only 12 weeks were required to produce a mature plant. We conclude that in vitro propagation techniques through rhizome cultures provide an efficient and rapid method for shoot propagation of S. parviflora.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15221279&dopt=Abstract hyoscyamine Levbid SL



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An Atropa belladonna hyoscyamine 6beta-hydroxylase gene is differentially expressed in the root pericycle and anthers.

Suzuki K, Yun DJ, Chen XY, Yamada Y, Hashimoto T.

Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Japan.

The AbH6H gene for hyoscyamine 6beta-hydroxylase (H6H), which converts hyoscyamine to scopolamine, was isolated from Atropa belladonna. This plant also possesses a related sequence, Ab psiH6H, which appears to be a non-functional pseudo-gene. AbH6H RNA was detected in cultured root, native root and anther, but not in stem, leaf, pistil, petal, and sepal tissues. In situ hybridization, immunohistochemistry and promoter::GUS transgene analysis showed that AbH6H is expressed specifically in root pericycle cells, and in tapetum and pollen mother cells. A 671 bp 5'-upstream region from AbH6H was sufficient for pericycle-specific expression in hairy roots of A. belladonna and Hyoscyamus niger, which both produce scopolamine, but cell-specific regulation was severely compromised in tobacco hairy roots, which do not produce scopolamine.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10394953&dopt=Abstract hyoscyamine Levbid SL