ABSTRACT
A competitive ELISA technique using digoxin-specific antibody has been developed to determine digoxin and its related compounds in hairy root cultures of Digitalis lanata. The ELISA could detect 0.2-2 nM digoxin and closely related cardenolides. Hairy roots cultured in the dark accumulated very small amounts of cardenolides (0.02-0.07 micrograms/g dry wt), while the content of cardenolides in green hairy roots cultured in the light was increased maximally 600-fold (16.5 micrograms/g dry wt) compared to those in the dark.
Subject(s)
Cardenolides/analysis , Digitalis/analysis , Enzyme-Linked Immunosorbent Assay , Plants, Medicinal , Plants, Toxic , Antibodies, Monoclonal/immunology , Cells, Cultured , Chromatography, High Pressure Liquid , Cross Reactions , Digoxin/analysis , Digoxin/immunologyABSTRACT
An analytical method for the determination of secondary cardiac glycosides in Digitalis purpurea leaves by high-performance liquid chromatography (HPLC) is described. The procedure consisted of extraction of dry leaf powder with ethanol-chloroform (2:1) and clean-up by Sep-Pak cartridges prior to HPLC analysis. HPLC was performed on an octylsilyl bonded silica column, using acetonitrile-methanol-water (4:4:5) for trisdigitoxosides and acetonitrile-methanol-water (8:30:43) for strospeside; the effluent was monitored by ultraviolet detection (at 220 nm). Quantitation of these cardiac glycosides was carried out by the internal standard method. The amounts of digitoxin, gitoxin, gitaloxin and strospeside per 100 mg of dry leaf powder were estimated to be 22.6, 14.0, 54.7 and 1.9 micrograms, respectively. The method is sufficiently sensitive and reproducible to assay secondary glycosides in Digitalis purpurea leaves.
Subject(s)
Digitalis Glycosides/analysis , Digitalis/analysis , Plants, Medicinal , Plants, Toxic , Chemical Phenomena , Chemistry , Chromatography, High Pressure Liquid , Spectrophotometry, UltravioletABSTRACT
A rapid extraction method followed by high-performance liquid chromatographic assay was developed for the quantitative determination of the cardioactive glycosides of Digitalis lanata. The leaf samples were extracted with water or aqueous alcohols. The simple extraction method gives a better yield than the methods described previously. Lanatoside C and its metabolites have been separated on a reversed-phase column with various mixtures of acetonitrile, methanol, and water as mobile phases for isocratic elution. Extraction and quantitative determination of lanatoside C and digoxin from a leaf sample require not more than 30 min.
Subject(s)
Digitalis Glycosides/isolation & purification , Digitalis/analysis , Plants, Medicinal , Plants, Toxic , Chromatography, High Pressure Liquid , Crystallization , Lanatosides/isolation & purification , SolventsSubject(s)
Digoxin/blood , Hypertension/etiology , Natriuresis , Animals , Calcium/physiology , Cattle , Digitalis/analysis , Digoxin/isolation & purification , Humans , Hypertension/blood , Muscle Contraction , Plants, Medicinal , Plants, Toxic , Rats , Receptors, Drug/analysis , Sodium-Potassium-Exchanging ATPase/bloodSubject(s)
Cardenolides/analysis , Digitalis/analysis , Plants, Medicinal , Plants, Toxic , Cells, Cultured , Digitalis/cytology , Methods , Species SpecificityABSTRACT
In Digitalis grandiflora, flavonoids are mainly constituted by flavone glycuronides which are found in all aerial organs, except the seeds; in the flowers are supplementary flavonoids, particularly, in the stamens, a kaempferol derivative.
