Investigation of tropane alkaloids in genetically transformed Atropa belladonna L. cultures

H. Hank, E. Szőke, K. Tóth, I. László, L. Kursinszki

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The purpose of the present study was to determine the tropane alkaloid content of genetically transformed hairy root cultures of Atropa belladonna L. Determination of alkaloids was performed by HPLC method. Samples were extracted with chloroform - methanol - cc. ammonia 15:5:1 (v/v/v). Crude extracts were purified on Extrelut columns. HPLC separation was performed on Luna C8 reversed phase column. An isocratic mixture of acetonitrile - 30 mM phosphate buffer - methanol 12.2:79.7:8.1(v/v/v) was used as eluent. Peaks were identified by addition of standards and diode-array detection. Hyoscyamine, scopolamine and apoatropine were determined by external standard method at 210 nm. We measured the alkaloid content of genetically transformed in vitro cultures (hairy roots and reorganised plants) cultivated on Gamborg B5 basic media. The highest hyoscyamine and scopolamine content was found in hairy root clone #K5 (0,223 m/m%) and in hairy root clone #K4 (0,018 m/m%) respectively. Alkaloid contents were higher in the hairy roots than in the reorganised plants.

Original languageEnglish
JournalChromatographia
Volume60
Issue numberSUPPL.
Publication statusPublished - 2004

Fingerprint

Atropa belladonna
Tropanes
Alkaloids
Hyoscyamine
Scopolamine Hydrobromide
Methanol
Clone Cells
High Pressure Liquid Chromatography
Plant Roots
Chloroform
Complex Mixtures
Ammonia
Buffers
Diodes
Phosphates

Keywords

  • Atropa belladonna L.
  • Column liquid chromatography
  • Hairy root culture
  • Hyoscyamine
  • Scopolamine

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry

Cite this

Investigation of tropane alkaloids in genetically transformed Atropa belladonna L. cultures. / Hank, H.; Szőke, E.; Tóth, K.; László, I.; Kursinszki, L.

In: Chromatographia, Vol. 60, No. SUPPL., 2004.

Research output: Contribution to journalArticle

Hank, H, Szőke, E, Tóth, K, László, I & Kursinszki, L 2004, 'Investigation of tropane alkaloids in genetically transformed Atropa belladonna L. cultures', Chromatographia, vol. 60, no. SUPPL..
Hank, H. ; Szőke, E. ; Tóth, K. ; László, I. ; Kursinszki, L. / Investigation of tropane alkaloids in genetically transformed Atropa belladonna L. cultures. In: Chromatographia. 2004 ; Vol. 60, No. SUPPL.
@article{757bebeeb4b5451a911f0b9197d91a8b,
title = "Investigation of tropane alkaloids in genetically transformed Atropa belladonna L. cultures",
abstract = "The purpose of the present study was to determine the tropane alkaloid content of genetically transformed hairy root cultures of Atropa belladonna L. Determination of alkaloids was performed by HPLC method. Samples were extracted with chloroform - methanol - cc. ammonia 15:5:1 (v/v/v). Crude extracts were purified on Extrelut columns. HPLC separation was performed on Luna C8 reversed phase column. An isocratic mixture of acetonitrile - 30 mM phosphate buffer - methanol 12.2:79.7:8.1(v/v/v) was used as eluent. Peaks were identified by addition of standards and diode-array detection. Hyoscyamine, scopolamine and apoatropine were determined by external standard method at 210 nm. We measured the alkaloid content of genetically transformed in vitro cultures (hairy roots and reorganised plants) cultivated on Gamborg B5 basic media. The highest hyoscyamine and scopolamine content was found in hairy root clone #K5 (0,223 m/m{\%}) and in hairy root clone #K4 (0,018 m/m{\%}) respectively. Alkaloid contents were higher in the hairy roots than in the reorganised plants.",
keywords = "Atropa belladonna L., Column liquid chromatography, Hairy root culture, Hyoscyamine, Scopolamine",
author = "H. Hank and E. Szőke and K. T{\'o}th and I. L{\'a}szl{\'o} and L. Kursinszki",
year = "2004",
language = "English",
volume = "60",
journal = "Chromatographia",
issn = "0009-5893",
publisher = "Springer Vieweg",
number = "SUPPL.",

}

TY - JOUR

T1 - Investigation of tropane alkaloids in genetically transformed Atropa belladonna L. cultures

AU - Hank, H.

AU - Szőke, E.

AU - Tóth, K.

AU - László, I.

AU - Kursinszki, L.

PY - 2004

Y1 - 2004

N2 - The purpose of the present study was to determine the tropane alkaloid content of genetically transformed hairy root cultures of Atropa belladonna L. Determination of alkaloids was performed by HPLC method. Samples were extracted with chloroform - methanol - cc. ammonia 15:5:1 (v/v/v). Crude extracts were purified on Extrelut columns. HPLC separation was performed on Luna C8 reversed phase column. An isocratic mixture of acetonitrile - 30 mM phosphate buffer - methanol 12.2:79.7:8.1(v/v/v) was used as eluent. Peaks were identified by addition of standards and diode-array detection. Hyoscyamine, scopolamine and apoatropine were determined by external standard method at 210 nm. We measured the alkaloid content of genetically transformed in vitro cultures (hairy roots and reorganised plants) cultivated on Gamborg B5 basic media. The highest hyoscyamine and scopolamine content was found in hairy root clone #K5 (0,223 m/m%) and in hairy root clone #K4 (0,018 m/m%) respectively. Alkaloid contents were higher in the hairy roots than in the reorganised plants.

AB - The purpose of the present study was to determine the tropane alkaloid content of genetically transformed hairy root cultures of Atropa belladonna L. Determination of alkaloids was performed by HPLC method. Samples were extracted with chloroform - methanol - cc. ammonia 15:5:1 (v/v/v). Crude extracts were purified on Extrelut columns. HPLC separation was performed on Luna C8 reversed phase column. An isocratic mixture of acetonitrile - 30 mM phosphate buffer - methanol 12.2:79.7:8.1(v/v/v) was used as eluent. Peaks were identified by addition of standards and diode-array detection. Hyoscyamine, scopolamine and apoatropine were determined by external standard method at 210 nm. We measured the alkaloid content of genetically transformed in vitro cultures (hairy roots and reorganised plants) cultivated on Gamborg B5 basic media. The highest hyoscyamine and scopolamine content was found in hairy root clone #K5 (0,223 m/m%) and in hairy root clone #K4 (0,018 m/m%) respectively. Alkaloid contents were higher in the hairy roots than in the reorganised plants.

KW - Atropa belladonna L.

KW - Column liquid chromatography

KW - Hairy root culture

KW - Hyoscyamine

KW - Scopolamine

UR - http://www.scopus.com/inward/record.url?scp=3943074404&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3943074404&partnerID=8YFLogxK

M3 - Article

VL - 60

JO - Chromatographia

JF - Chromatographia

SN - 0009-5893

IS - SUPPL.

ER -