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Adult rats were orally administered with a single dose of sanguinarine (10 mg SA per 1 kg body weight) in 1.0 ml water. In the plasma and the liver, dihydrosanguinarine (DHSA) was identified as a SA metabolite by high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC/ESI-MS). Significantly higher levels of DHSA were found in both the plasma and the liver in comparison with those of SA. SA and DHSA were not detected in the urine. The formation of DHSA might be the first step of SA detoxification in the organism and its subsequent elimination in phase II reactions. Benz[c]acridine (BCA), in the literature cited SA metabolite, was found neither in urine nor in plasma and liver.

Capillary zone electrophoresis was applied to the investigation of the interaction of sanguinarine and chelerythrine with mercapto compounds including albumins at pH 7.4. Mercaptoethanol and L-cysteine were chosen for the identification of the type of interaction and for the identification of the interacting chemical form of these alkaloids. It was evidenced that sanguinarine and chelerythrine do not react chemically with these mercapto compounds at pH 7.4 and that non-covalent products form in this interaction. Their interaction is a fast and reversible complexation and is based on non-bonding intermolecular interactions. Conditional binding constants measured at pH 7.4 and 5.0 indicate that only uncharged forms of sanguinarine and chelerythrine (pseudobases) participate in complexation. A negatively charged group, either bound to the mercapto ligand or supplied by the solution, enters in the complexation. The simple 1 : 1 interaction scheme holds, therefore, only for mercapto compounds bearing an anionic group. Constants corrected for the abundance of the uncharged alkaloid form are of the order of magnitude of 104 l mol−1 and depend on the chemical composition of buffer. Interaction of sanguinarine and chelerythrine with human or bovine serum albumins is qualitatively identical with interaction of these alkaloids with simple mercapto compounds. Constants for the binding of uncharged form of sanguinarine with human and bovine serum albumins in sodium phosphate buffer at pH 7.4, corrected for abundance of the interacting uncharged form, are 332 000±38 400 and 141 000±14 400 l mol−1, respectively. The former agrees well with the value K=385 000 (or log K=5.59) reported from static experiments. For the uncharged form of chelerythrine, the constants are 2 970 000±360 000 and 1 380 000±22 600 l mol−1 for human and bovine serum albumins, respectively. Copyright © 2003 John Wiley & Sons, Ltd.

Using the 32P-postlabeling assay, we investigated the ability of quaternary benzo[c]phenanthridine alkaloids, sanguinarine, chelerythrine and fagaronine, to form DNA adducts in vitro. Two enhanced versions of the assay (enrichment by nuclease P1 and 1-butanol extraction) were utilized in the study. Hepatic microsomes of rats pre-treated with beta-naphthoflavone or those of uninduced rats, used as metabolic activators, were incubated in the presence of calf thymus DNA and the alkaloids, with NADPH used as a cofactor. Under these conditions sanguinarine and chelerythrine, but not fagaronine, formed DNA adducts detectable by 32P-postlabeling. DNA adduct formation by both alkaloids was found to be concentration dependent. When analyzing different atomic and bond indices of the C11-C12 bond (ring B) in alkaloid molecules we found that fagaronine behaved differently from sanguinarine and chelerythrine. While sanguinarine and chelerythrine showed a preference for electrophilic attack indicating higher potential to be activated by cytochrome P450, fagaronine exhibited a tendency for nucleophilic attack. Our results demonstrate that sanguinarine and chelerythrine are metabolized by hepatic microsomes to species, which generate DNA adducts.

Capillary electrophoresis was employed to determine the principal quaternary benzo[ c ]phenanthridine alkaloids, sanguinarine and chelerythrine, in two plant extracts and one oral hygiene product. Phosphate–Tris buffer of pH 2.5 was used as a background electrolyte, limits of detection were 3 μmol l −1 (sanguinarine) and 2.4 μmol l −1 (chelerythrine) using UV detection at 270 nm. The method, which correlated well with HPLC, is suitable for serial determination of sanguinarine and chelerythrine in plant products and pharmaceuticals.

Sanguinarine, chelerythrine and fagaronine were examined for hepatotoxic effect. Administration of sanguinarine and chelerythrine (10 mg/kg/day; i.p.) produced marked parenchymal injury not found in the livers of fagaronine treated rats. In chronic administration of sanguinarine (0.2 mg/kg/56 days; i.p.) degeneration and necrosis were not observed in the liver. The effect of the alkaloids was tested by enzyme leakage (LDH and ASAT) and changes in glutathione levels in isolated rat hepatocytes. Sanguinarine produced the greatest toxicity followed by lesser toxicity in the case of chelerythrine while no toxicity was displayed with fagaronine. These effects correlate with pK R values for pseudobase formation. Finally, no increased lipid peroxidation or mitochondrial membrane damage were found.

A high-performance liquid chromatographic method for the determination of colchicinoid alkaloids in plant material is described. The determination is performed separately in neutral and basic alkaloid fractions using a C18-bonded silica column. Nine alkaloids, i.e., 3-demethylcolchicine, 2-demethylcolchicine, colchifoline, N-deacetyl-N-formylcolchicine, colchicine, cornigerine, 2-demethyldemecolcine, 3-demethyldemecolcine and demecolcine, in seven Colchicum plants were assayed. For identification of phenolic compounds, a method using gas chromatography-mass spectrometry was elaborated and twenty phenolic compounds were identified in extracts from five Colchicum species. As the presence of luteolin appeared to be of value for chemotaxonomic purposes, its simple densitometric determination was developed.

A rapid prechromatographic sample preparation and a reversed phase high performance liquid chromatographic (RP-HPLC) ion pair method have been developed for the qualitative and quantitative analysis of the main alkaloids of Eschscholtzia californica, namely allocryptopine, californidine, chelerythrine, eschscholtzine, N-methyllaurotetanine, protopine and sanguinarine. The identity and purity of the component HPLC-peaks were established by photodiode array detection and by comparison with standards. The distribution of alkaloids in the organs of the plant was examined: allocryptopine is the main constituent in roots, whilst californidine is the major alkaloid in all aerial organs except flowers where the main compound is eschscholtzine. HPLC fingerprint chromatograms of extracts of E. californica from different commercial sources were found to be similar.

Sanguinarine, a quaternary benzo[c]phenanthridine alkaloid, exhibits antimicrobial and anti-inflammatory activities and for this reason it is used in dental hygiene products and feed additives. Its metabolism and disposition is the subject of constant scientific discourse. In this paper we summarize current knowledge on sanguinarine metabolism. We show in particular that: (i) Sanguinarine is not transformed to 3,4-benzacridine and that the literature reporting this compound as a metabolite of sanguinarine is based on artifacts and misinterpretations that in course of time have created a dogma; (ii) Sanguinarine is converted to dihydrosanguinarine in vivo, the conversion being tentatively a detoxication pathway; (iii) Aryl hydrocarbon receptor metabolic signaling pathways modulate sanguinarine biological activity.

Using mercaptoethanol and ( l )-cysteine as representatives of mercapto compounds and capillary zone electrophoresis as experimental technique, it was evidenced that sanguinarine and chelerythrine do not react with the mercapto group of organic compounds at pH 7.4. Their interaction is fast and reversible complexation based on non-bonding intermolecular interaction in which enter uncharged forms of sanguinarine or chelerythrine. A negatively charged group, either bound to the mercapto ligand or supplied from solution, participates in the complexation. Simple 1:1 interaction scheme reported in literature holds therefore only for mercapto compounds bearing anionic group. Stoichiometric binding constants corrected for the abundance of the uncharged alkaloid form at pH 7.4 are of the order of magnitude of 10 4 l/mol and depend on both cations and anions of the background electrolyte. Interaction of sanguinarine and chelerythrine with human or bovine serum albumins does not qualitatively differ from their interaction with simple mercapto compounds. Stoichiometric binding constants for the binding of sanguinarine with human and bovine serum albumins in sodium phosphate buffer pH 7.4, corrected for the abundance of the interacting uncharged form, are 332 000±38 400 and 141 000±14 400 l/mol, respectively. The former agrees well with the value K =385 000 (or log K =5.59) from static photometric experiments. Constants for the complexation of uncharged chelerythrine with human and bovine serum albumins are 2 970 000±360 000 and 1 380 000±22 600 l/mol, respectively.

The alkaloid content (3-demethylcolchicine, colchifoline, N -deacetyl- N -formylcolchicine, colchicine, cornigerine, 2-demethyldemecolcine, 3-demethyldemecolcine, demecolcine) of corms, leaves and seeds of C. turcicum was determined by HPLC. In addition to these compounds, corms contained β-lumicolchicine, luteolin, and vanillic acid. Complete spectral data of colchifoline are presented.

A new bisbenzylisoquinoline alkaloid (−)-(1S,1'R)-O,O'-dimethylgrisabine was isolated from the leaves of Phaeanthus vietnamensis Ban. Its structure was determined on the basis of the extensive 2-D and 1-D nmr long-range heteronuclear correlations. Its antibacterial activity is also described

Fluorescence data for the interaction of acetylcholinesterase (EC 3.1.1.7, ACHE) from Electrophorus electricus with berberine, 13-ethylberberine, coralyne, and chelerythrine have been presented. By a comparison with known inhibitors of acetylcholinesterase, tetramethylammonium and tacrine, the electrostatic interaction of quaternary alkaloids with the γ-anionic site of enzyme could be confirmed. During the interaction, two molecules of the ligand are bound to the molecule of the enzyme.

The isotachophoretic separation of some isoquinoline alkaloids in acid-base partially purified extracts from Bulgarian FUMARIA PARVIFLORA was studied. The electrolyte system for model mixtures of alkaloids (-)-stylopine, (-)-canadine, coptisine, berberine, protopine, cryptopine, chelidonine, bulbocapnine, papaverine, parfumine and for the plant extracts contained the leading ion K + (0.01 M), counter ion CH 3 COO - , pH L 5.5, and the terminating ion H + (acetic acid 0.01 M). In the plant extracts, protopine and parfumine were quantitatively determined during F. PARVIFLORA growth and development in the term May-June 1984.

The structure of N-deacetyl-N-3-oxobutyrylcolchicine (V), based on spectroscopic measurements, was proposed for a minor alkaloid isolated from seeds of Colchicum autumnale L.

The tropolone alkaloids colchicine, colchiceine, demecolcine, demecolceine, isocolchicine, isocolchiceine, N-deacetylcolchicine, N-deacetylcolchiceine, N-deacetyl-N-formylcolchicine, N-deacetyl-N-formylcolchiceine, were separated by thin-layer chromatography on silica gel in a new solvent system. Conditions for quantitative fluorodensitometric determination of tropolone alkaloids were investigated.

FUMARIA DENSIFLORA DC., collected near Varna (Bulgaria) was investigated for its alkaloidal content. Protopine, cryptopine, coptisine, palmatine, adlumidiceine, (+/-)-sinactine, and a new spirobenzylisoquinoline alkaloid densiflorine were isolated. Benzophenanthridine alkaloids were not detected.

A rapid isotachophoretic separation of benzo[ C]phenanthridine alkaloids from CHELIDONIUM MAJUS L. is described. The method allows the determination of sanguinarine and chelerythrine in the presence of other isoquinoline alkaloids in crude alkaloids extracts.

The distributions of the alkaloids colchicine, 2-demethylcolchicine, 3-demethylcolchicine, demecolcine, 2-demethyldemecolcine, and 3-demethyldemecolcine in the corms, stems, leaves, flowers, and seeds of COLCHICUM CILICICUM were analysed by HPLC. In addition, the alkaloids cornigerine, N-formyl- N-deacetylcolchicine, N-methyl-demecolcine, beta-lumicolchicine, gamma-lumicolchicine, beta-lumidemecolcine, gamma-lumidemecolcine, and N-ethoxycarbonyldemecolcine were identified, the last named being reported for the first time from a COLCHICUM species. The flavones luteolin and apigenin were isolated and benzoic, 2-hydroxy-6-methoxybenzoic, and vanillic acids were also detected.

The aerial parts of ANDROCYMBIUM MELANTHIOIDES yielded known alkaloids cornigerine, colchicine, N-formyl- N-deacetylcolchicine, 2-demethylcolchicine, 3-demethylcolchicine, demecolcine, 2-demethyldemecolcine, 3-demethyldemecolcine, beta-lumicornigerine, beta-lumicolchicine, gamma-lumicolchicine, androcymbine, O-methylandrocymbine, and melanthioidine. Of these, O-methylandrocymbine and beta-lumicornigerine are reported for the first, time. The alkaloid, formerly designated as compound AM-3, has been identified as N-deacetyl- N-dimethyl-gamma-lumicornigerine 1. Its spectral data are given.