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Supercritical carbon dioxide extraction of colchicine and related alkaloids from seeds of Colchicum autumnale L.

A method for the extraction of the alkaloids colchicine, 3-demethylcolchicine and colchicoside from seeds of Colchicum autumnale by supercritical carbon dioxide has been established. Several parameters such as pressure, temperature, percentage of modifier and extraction time have been examined. Two extraction steps with constant carbon dioxide density (0.90 g/mL) and flux (1.5 mL/min) were required to extract the alkaloids in 110 min using 3% methanol as modifier. The quantitative determination of the alkaloids was performed by HPLC; the percentages of recovery were higher than 98% for the three alkaloids. This extraction procedure was compared with a conventional method involving maceration and sonication, and the same levels of alkaloids were obtained in each case. The supercritical carbon dioxide method is, however, very efficient, more rapid and more environmentally friendly than conventional methods.

Alkaloids↗

Determination of colchicine in Colchicum steveni and C. hierosolymitanum (Colchicaceae): comparison between two analytical methods.

The amounts of colchicine present in two Jordanian species of Colchicum, namely, C. steveni Kunth and C. hierosolymitanum Feibrun (Colchicaceae), have been determined. An HPLC-UV (photodiode array) method employing gradient elution was developed and the results compared with those obtained using a simple TLC-spectrophotometric method. The levels of colchicine as measured by these methods were not significantly different (p < 0.05) indicating that the spectrophotometric method is an acceptable alternative to HPLC. With respect to C. steveni, the leaves contained the largest amount of colchicine (0.204/100 g), whilst in C. hierosolymitanum corms showed the highest colchicine content (0.126/100 g). As a source of colchicine, the two investigated species showed levels comparable with those found in C. autumnale, the traditional source of colchicine.

Chromatography, High Pressure Liquid↗

Acute poisoning with autumn crocus (Colchicum autumnale L.).

INTRODUCTION: Colchicum autumnale, commonly known as the autumn crocus or meadow saffron, contains the antimitotic colchicine, which binds to tubulin and prevents it forming microtubules that are part of the cytoskeleton in all cells. CASE REPORT: A 71-year-old woman ate a plant she thought to be wild garlic (Allium ursinum). Ten hours later she arrived at the emergency department complaining of nausea, vomiting and watery diarrhea. Ingestion of a poisonous plant was suspected and she was treated with gastric lavage, oral activated charcoal and an infusion of normal saline. Toxicology analysis with gas chromatography and mass spectrometry revealed colchicine in the patient's gastric lavage, blood (5 microg/l) and urine (30 microg/l). She developed arrhythmias, liver failure, pancreatitis, ileus, and bone marrow suppression with pancytopenia. Alopecia began in the third week. Treatment was supportive only. Five months later she had no clinical or laboratory signs of poisoning. DISCUSSION: The patient mistakenly ingested autumn crocus instead of wild garlic because of their great similarity. Colchicine primarily blocks mitosis in tissues with rapid cell turnover; this results in gastroenterocolitis in the first phase of colchicine poisoning, bone marrow hypoplasia with pancytopenia in the second and alopecia in the third, all of which were present in our patient. Colchicine toxicity in tissues without rapid cell turnover caused arrhythmias, acute liver failure and pancreatitis. CONCLUSION: Colchicine poisoning can result in gastroenterocolitis followed by multi-organ dysfunction syndrome. In unexplained gastroenterocolitis after ingestion of wild plants as a salad or spice, especially when wild garlic is mentioned, we should always consider autumn crocus. Diagnosis could be confirmed only by toxicology analyses. Management of colchicine poisoning is restricted to supportive therapy.

Aged↗

Characterisation of a new potyvirus species infecting meadow saffron Colchicum autumnale).

The alkaloids contained in Colchicum autumnale seeds are used in numerous medicines. Good quality seeds are difficult to obtain from this undomesticated plant. Therefore, a research program was set up aiming to cultivate C. autumnale in order to improve alkaloid contents and seed yields. In this context, a collection was established in 1999 by transplanting corms from twelve different locations in Eastern France. However, serious symptoms of necrosis and decay have appeared in this collection since 2001. Electron microscopic observations of plants showing symptoms revealed the presence of filamentous particles and pinwheel-like structures characteristic of the Potyviridae family. Leaves and corms from symptomatic plants were assayed with potyvirus-specific Enzyme-Linked Immunosorbent Assay test. Positive reactions were obtained with plants from all the geographic origins, which exhibited flower breaking symptoms on petals. RT-PCR tests with family Potyviridae-specific primers confirmed the ELISA results and showed that the virus can be detected in corms, roots and flowers of symptomatic plants. The 3' region of the genome was cloned, sequenced and compared to other potyvirus species. Phylogenetic analyses suggest the presence of a new viral species tentatively named Meadow saffron breaking virus (MSBV) in C. autumnale.

Colchicum↗

Cornigerine, a potent antimitotic Colchicum alkaloid of unusual structure. Interactions with tubulin.

Cornigerine is a natural product analog of colchicine produced by Colchicum cornigerum in which the vicinal 2- and 3-methoxy groups are condensed into a methylenedioxy bridge. This produces a fourth ring and a structure which resembles a hybrid of colchicine, podophyllotoxin, and steganacin. Cornigerine was somewhat more toxic than colchicine with L1210 murine leukemia cells and caused them to accumulate in metaphase arrest. Cornigerine resembled colchicine in its interactions with tubulin in vitro, and it was also somewhat more potent than colchicine in these drug-tubulin interactions. Cornigerine inhibited tubulin polymerization both with and without microtubule-associated proteins, inhibited the binding of radiolabeled colchicine to tubulin, and stimulated tubulin-dependent GTP hydrolysis. Indirect evidence suggested that the binding of cornigerine to tubulin is relatively slow and temperature-dependent, like the binding of colchicine to the protein.

Animals↗

Partial synthesis and antitubulin activity of minor colchicum alkaloids: N-acetoacetyl-deacetylcolchicine and 2-demethylspeciosine (speciocolchine).

Two minor Colchicum alkaloids, N-acetoacetyl-deacetylcolchicine [1] and 2-demethylspeciosine [7], were synthesized. The diacetate 8 of 2-demethylspeciosine was also prepared. The antitubulin activity of these compounds, in comparison to colchicine, was measured. N-Acetoacetyl-deacetylcolchicine [1] has in vitro activity similar to that of colchicine. Both 2-demethylspeciosine [7] and the diacetate 8 were considerably less potent inhibitors of tubulin polymerization.

Alkaloids↗

Inhibition activities of Colchicum luteum baker on lipoxygenase and other enzymes.

In vitro enzymes inhibition activities of the crude methanolic extract and various fractions of Colchicum luteum Baker (Liliaceae) including chloroform, ethyl acetate, n-butanol and aqueous were carried out against actylcholinesterase, butyrylcholinesterase, lipoxygenase and urease enzymes. A significant enzyme inhibition activity (89%) is shown by the crude methanolic extract and its fractions against lipoxygenase, while low to significant activity (32-75%) was evident against butyrylcholinesterase. The crude methanolic extract and its various fractions demonstrated low activity (29-61%) against acetylcholinesterase and no activity against urease.

Acetylcholinesterase↗

Histopathological study of experimental acute poisoning of cattle by autumn crocus (Colchicum autumnale L.).

Crude or dehydrated bulbs of autumn crocus (Colchicum autumnale L.) were fed to eleven calves. All the calves developed severe diarrhea and died or euthanized within 63 hr. At necropsy, the gastro-intestinal mucosa was edematous and hemorrhagic. Histologically, necrosis and degeneration with karyopyknosis and karyorrhexis were shown in the basal cell layer of the tongue, esophagus, forestomach, renal pelvis, urinary bladder, neck cell layer of the abomasal gastric glands, and intestinal cryps. These findings were also seen in Kupffer cells, renal tubular epithelial cells, and lymphocytes in the lymphoid and hemopoietic systems. The lesion of the present acute crocus poisoning of cattle closely resembled those reported in humans with colchicine intoxication. Refined acetone extract of organs of poisoned cattle proved to contain colchicine and demecolcine by high performance liquid chromatography.

Animals↗

Supplementary report on experimental autumn crocus (Colchicum autumnale L.) poisoning in cattle: morphological evidence of apoptosis.

Previously we reported that tissue destruction characterized by the presence of karyopyknotic, karyorrhectic and mitotically arrested cells was seen in alimentary epithelial cells and lymphocytes in the lymphoid and hemopoietic systems of cattle experimentally administered with autumn crocus (Colchicum autumnale L.). This report deals with the mechanism of acute cellular injury following experimental autumn crocus poisoning in cattle as demonstrated by the in situ DNA strand break analysis and electron microscopy. The analyses revealed that cellular injury caused by autumn crocus was closely associated with apoptosis.

Animals↗

Accidental plant poisoning with Colchicum autumnale: report of two cases.

Colchicine poisoning is a rare but serious and potentially fatal event, which results from food poisoning or overdose with drugs containing colchicine, with no currently available antidote. We report two cases of plant poisoning with Colchicum autumnale, in which the patients had identical initial symptoms but developed extremely different clinical courses. One patient recovered after only moderate gastroenteritis and liver injury, whereas the other died of rapid progressive multiple organ failure 52 h after the plant ingestion. We recommend that all patients suspected of colchicine intoxication due to its unpredictable outcome should be managed according to the principles of intensive care, irrespective of the actual degree of poisoning.

Colchicum↗

[Meadow saffron (Colchicum autumnale) in hay and colic in horses].

Following feeding of a new delivery of hay (bought in Southern Germany) 3 horses out of a unit of 17 developed colic within a few days one horse died. Post-mortem there was an intensive accumulation of serous or serous-haemorrhagic fluid in the thorax and abdomen. The hay was heavily contaminated by autumn crocus (colchicum autumnale)--about 1.48% of total mass. Other feedstuffs were of good quality and following removal of the affected hay no further cases of colic occurred. The pathological findings also indicated a colchicine intoxication.

Animal Feed↗

[Transformation of the names of certain herbs by linguistic transfer - colchicum, wormseed, soybean, nandina and wijnruit].

Five examples of transformation of the names of certain herbal medicines by transfer from one language to another are presented: (1) to meadow saffron (colchicum) from Medea's saffron, (2) from semenzina to semen cinae or semen China, (3) from the Japanese condiment sho-yu to soy and soybeans, (4) from the Japanese plant nanten to the genus Nandina, and (5) from the Dutch term wijnruit (wine-ruta) to hen-ruda in Japanese. In most cases, the transformation underwent phonetically and resulted in a departure from the original meaning.

China↗