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Determination of colchicine content in Colchicum hierosolymitanum and Colchicum tunicatum under cultivation.

Corms of Colchicum hierosolymitanum and Colchicum tunicatum were collected, identified and planted under field condition. We report here and for the first time the presence of colchicine in an appreciable amount in both species. The effect of different NPK fertilizer levels on colchicine content of the two Colchicum species at different growth stages were evaluated by HPLC. Results indicated that increasing NPK fertilizer levels significantly improve colchicine content in different plant parts and stages. The highest colchicine content observed in corms was at maturity stage 0.766 mg/g and 0.688 mg/g dry weight with C. hierosolimitanum and C. tunicatum, respectively.

Alkaloids↗

Phytochemical studies and cytotoxicity evaluations of Colchicum tunicatum Feinbr and Colchicum hierosolymitanum Feinbr (Colchicaceae): two native Jordanian meadow saffrons.

As a part of our continuing investigation of Jordanian Colchicum species, the biologically active components of Colchicum hierosolymitanum Feinbr and Colchicum tunicatum Feinbr (Colchicaceae) were pursued. The brine shrimp lethality test (BSLT) was used to direct the fractionation and isolation of active components. Five and four known colchicinoids were isolated and characterized from C. tunicatum and C. hierosolymitanum, respectively. The known colchicinoids, reported for the first time from these two species are: (-)-colchicine (I), 3-demethyl-(-)-colchicine (II), (-)-cornigerine (III), beta-lumicolchicine (IV), and (-)-androbiphenyline (V) from C. tunicatum, and (-)-colchicine (I), 2-demethyl-(-)-colchicine (VI), (-)-cornigerine (III), and beta-lumicolchicine (IV) from C. hierosolymitanum. The chemical structures of the isolated compounds have been elucidated using a series of spectroscopic and spectrometric techniques principally; 1D-NMR (1H and 13C) and low resolution EI-MS and APCIMS. All pure compounds were evaluated for cytotoxicity against three human cancer cell lines; MCF-7 human breast carcinoma, NCI-H460 human large cell lung carcinoma, and SF-268 human astrocytoma. (-)-Colchicine (I) and (-)-cornigerine (III) were found to be the most bioactive of the identified compounds with EC50 values in the range of 0.016-0.097 microM.

Animals↗

Seasonal variation of colchicine content in Colchicum brachyphyllum and Colchicum tunicatum (Colchicaceae).

As part of our continuing investigation of Jordanian Colchicum species, (-)-colchicine content in C. brachyphyllum Boiss. & Haussk. ex Boiss and Colchicum tunicatum Feinbr (Colchicaceae), growing wild in Jordan, was determined during different growth stages. Using external reference standard, a reverse-phase gradient photo-diode array high performance liquid chromatography (PDA-HPLC) method was adapted. Underground parts in both species and during different growth stages, always showed higher (-)-colchicine content than the above ground parts. In C. brachyphyllum total (-)-colchicine content of underground parts during flowering stage was found to be about 0.15% (wt/wt), while that of aerial parts was only about 0.04% (wt/wt). In C. tunicatum total (-)-colchicine content of underground parts was found to be 0.12% (wt/wt), and 0.13% (wt/wt) during flowering and vegetating stages, respectively, while that of aerial parts was only about 0.04% (wt/wt) and 0.02% (wt/wt), respectively. In C. tunicatum, stems, roots and unripe seeds are the main storages of (-)-colchicine at flowering, vegetating and seeding stages, respectively, while in C. brachyphyllum, corms are the main storage of (-)-colchicine at flowering and seeding stages.

Chromatography, High Pressure Liquid↗

Colchicine poisoning by accidental ingestion of meadow saffron (Colchicum autumnale): pathological and medicolegal aspects.

Although intoxications with colchicine, the alkaloid of Colchicum autumnale (meadow saffron), are well known, in most cases the intoxications are evoked by oral or parenteral preparations traditionally used as medication against gout. The accidental ingestion of Colchicum autumnale, on the other hand, is a rare event and has to our knowledge only twice been described in detail. We report a further case in which two persons confused this highly poisonous plant with wild garlic (Allium ursinum), a popular spice in the Central European cuisine. While one person merely complained about a 3-day episode of nausea, vomiting and watery diarrhea, the second person died of multi-organ system derangements 48 h after the ingestion of the colchicum leaves. At autopsy hemorrhagic lung oedema, hypocellular bonemarrow, centrilobular fatty necrosis of the liver and necrosis of the proximal convoluted tubuli of the kidneys were observed. A colchicine concentration of 7.5 micrograms/ml was found in the bile whereas no substance was detected in the postmortem blood.

Accidents↗

Colchicum autumnale agglutinin activates all murine T-lymphocytes but does not induce the proliferation of all activated cells.

Plant lectins with mitogenic properties for T-lymphocytes have been particularly useful for the study of T-cell activation and effector functions. In the search for mitogenic lectins possessing activation features different from the ones associated with the already known mitogens, we found that an agglutinin isolated from Colchicum autumnale tubers, Colchicum autumnale agglutinin (CAA), possesses interesting properties. First, contrasting with the classical mitogens, CAA induces the proliferation of a fraction of the CD4+ and CD8+ mouse T-lymphocytes. Second, the CAA-induced proliferation requires MHC class II and CD4 molecules. Third, although only a fraction of T-cells enters into the cell cycle, all T-lymphocytes are activated and express high levels of the activation markers CD69 and CD44. Finally, CAA-stimulation is characterized by a particular pattern of the cytokine gene expression, reflected by the transcription of the IL2, IL5, and IFN-gamma genes, while the IL4 and IL10 genes remained silent. Taken together these data demonstrate that CAA activation does not conform to the pathway of T-cell triggering observed with classical mitogenes and represents a new tool for the analysis of T-cell activation.

Animals↗

Development and optimization of a stability indicating method on a monolithic reversed-phase column for Colchicum dry extract.

This article presents a method developed on a monolithic reversed-phase column for separation of related alkaloids and degradation products from colchicine in dry extract preparations from Colchicum seeds. Optimization was performed using an optimization software with variation of the pH of the mobile phase and the percentage of organic modifier. This method permits for the first time a fast separation of all alkaloids including colchiceine without prior derivatization or complexation with excellent linearity, precision and low limits of quantitation which makes it a suitable method for stability testing of extracts or drug formulations containing extracts of Colchicum seeds.

Chromatography, High Pressure Liquid↗

New colchicinoids from a native Jordanian meadow saffron, colchicum brachyphyllum: isolation of the first naturally occurring dextrorotatory colchicinoid.

As part of our continuing investigation of Jordanian Colchicum species, the biologically active components of Colchicum brachyphyllum were pursued. Using bioactivity-directed fractionation, nine colchicinoids were isolated and characterized. One of these has a novel ring system, to which we have ascribed the trivial name (+)-demecolcinone (9), and it represents the first naturally occurring dextrorotatory colchicinoid. Another isolated compound was a new colchicinoid analogue, (-)-2,3-didemethyldemecolcine (8), while the remaining seven known colchicinoids were new to the species: (-)-colchicine (1), (-)-3-demethylcolchicine (2), (-)-cornigerine (3), beta-lumicolchicine (4), (-)-androbiphenyline (5), (-)-demecolcine (6), and (-)-3-demethyldemecolcine (7). The brine shrimp lethality test was used to direct the isolation of these colchicinoids. Moreover, all pure compounds were evaluated for cytotoxicity against a human cancer cell panel, for antimicrobial activity in an array of bacteria and fungi (including yeast), and for their potential to be allosteric modulators of the gamma-aminobutyric acid type A receptor.

Alkaloids↗

Phytochemical study and cytotoxicity evaluation of Colchicum stevenii Kunth (Colchicaceae): a Jordanian meadow saffron.

Isolation, characterization, and biological evaluation of active components of Colchicum stevenii Kunth (Colchicaceae) are described. Colchicum stevenii is an unexplored Jordanian specie with toxic reputation. Directed by brine shrimp lethality test (BST), methanolic extraction, liquid-liquid partition, preparative TLC, and semi-preparative HPLC, it resulted in the isolation of six cytotoxic compounds. The compounds, reported for the first time from this specie, are: (-)-colchicine (1), 2-demethyl-(-)-colchicine (2), (-)-cornigerine (3), beta-lumicolchicine (4), (-)-isoandrocymbine (5) and (-)-O-methylandrocymbine (6). A new, in-house developed, acidic-based reverse-phase gradient semi-preparative HPLC method for the separation of colchisides is presented here. Structural elucidation was based on spectroscopic techniques principally; 1H-NMR and low resolution EIMS. Based on BST results, reported as LC50 values in microg mL(-1) (ppm) with 95% confidence intervals, (-)-colchicine (2.5 ppm) and (-)-cornigerine (2.7 ppm) were the most potent.

Animals↗

Self-poisoning with Colchicum autumnale L. flowers.

UNLABELLED: A 44-year-old man ingested about 40 flowers of Colchicum autumnale L. The patient presented with nausea, vomiting, and abdominal pain 2 hours after ingestion and had diarrhea 14 hours after ingestion. Hematological values remained within normal range. Treatment was mainly supportive. The outcome was favorable. The intoxication was confirmed by high-performance liquid chromatography-mass spectrometry. Maximal colchicine levels were 4.34 ng/mL at 13 hours in plasma and 5.43 ng/mL at 16 hours in erythrocytes. CONCLUSION: We report one of the few symptomatic cases of Colchicum autumnale L. poisoning confirmed by toxicological analysis.

Adult↗

Case report: fatal poisoning with Colchicum autumnale.

INTRODUCTION: Colchicum autumnale, commonly known as the autumn crocus, contains alkaloid colchicine with antimitotic properties. CASE REPORT: A 76-year-old man with a history of alcoholic liver disease and renal insufficiency, who mistakenly ingested Colchicum autumnale instead of wild garlic (Aliium ursinum), presented with nausea, vomiting and diarrhea 12 hours after ingestion. On admission the patient had laboratory signs of dehydration. On the second day the patient became somnolent and developed respiratory insufficiency. The echocardiogram showed heart dilatation with diffuse hypokinesia with positive troponin I. The respiratory insufficiency was further deteriorated by pneumonia, confirmed by chest X-ray and later on by autopsy. Laboratory tests also revealed rhabdomyolysis, coagulopathy and deterioration of renal function and hepatic function. The toxicological analysis disclosed colchicine in the patient's urine (6 microgram/l) and serum (9 microgram/l) on the second day. Therapy was supportive with hydration, vasopressors, mechanical ventilation and antibiotics. On the third day the patient died due to asystolic cardiac arrest. DISCUSSION AND CONCLUSION: Colchicine poisoning should be considered in patients with gastroenterocolitis after a meal of wild plants. Management includes only intensive support therapy. A more severe clinical presentation should be expected in patients with pre-existing liver and renal diseases. The main reasons for death are cardiovascular collapse, respiratory failure and leukopenia with infection.

Aged↗

A New Lectin from Meadow Saffron (Colchicum automnale).

A lectin has been isolated from tubers of the meadow saffron (Colchicum autumnale). It is an octameric protein (M(r) 100,000) composed of 4A- and 4B-subunits of M(r) 15,000 and 10,000, respectively. It is a glycoprotein with 4.4% carbohydrate, the main sugars are (N-acetyl-) glucosamine, mannose, fucose, and xylose. Although the Colchicum autumnale agglutinin (CAA) agglutinates human red blood cells, it has a much higher activity with rabbit erythrocytes. With respect to its carbohydrate-binding specificity CAA behaves rather unusually as it is inhibited by lactose, galactose, N-acetylgalactosamine and related sugars when assayed with human red blood cells but not in assays with rabbit erythrocytes.

Journal Article↗

Changes in colchicine and demecolcine content during vegetation period of colchicum autumnale L.

Colchicine and demecolcine were determined in raw and dried leaves, stems, mother and daughter corms of Colchicum autumnale L. in four stages of its ontogenesis. The colchicine content in raw material varies during plant growth. The content of both alkaloids decreases with drying. The HPLC method used is suitable both for the phytochemical analysis of Colchicum and for the toxicological evaluation in cases of intoxication by these plants.

Chromatography, High Pressure Liquid↗