Autumn Crocus Research Articles

Clinical case of colchicum poisoning

Introduction. Autumn colchicum or crocus is a popular plant among gardeners, characterized by early beautiful flowering, but few people know that it is poisonous. Acute colchicum poisoning is a dangerous urgent clinical situation with high mortality rates. By disrupting cell division, colchicum venom leads to multiple organ failure and death in just a few days. However, colchicum extract in small doses is often used in modern medicine to treat various diseases, due to its high antimitotic and moderate anti-inflammatory activity. The article describes the clinical observation of a patient admitted to the toxicology department after a suicide attempt using a crocus flower, containing the alkaloid colchicine. The results of laboratory and pathological studies are presented. Clinical observation. A clinical case of acute crocus poisoning with the development of hemodynamic disorders, dyspeptic syndrome, respiratory failure, cytolysis syndrome, and oliguria is presented. Treatment was aimed at restoring organ functions and consisted of infusion, detoxification, and hemostatic therapy. Limitations. To carry out this scientific work, a study was conducted within the framework of only one clinical case committed on the territory of the Udmurt Republic. In this case report, there was no comparison group or observation group. The described documents were provided by the Budgetary Health Institution of the Udmurt Republic "City Clinical Hospital no. 6" of the Ministry of Health of the Udmurt Republic and the Budgetary Health Institution of the Udmurt Republic “Bureau of Forensic Medical Examination of the Ministry of Health of the Udmurt Republic” by prior agreement. Conclusion. Colchicum poisoning is a rare and extremely severe pathology, leading to multiple organ failure, agranulocytosis and death. Considering the rapid absorption in the gastrointestinal tract and the initial symptoms in the form of gastrointestinal lesion syndrome, early diagnosis in the absence of anamnesis is difficult, therefore, it is necessary to highlight the issues of diagnosing this poisoning, as well as the study and implementation of modern methods of treating colchicine poisoni

Common autumn crocus in the cultural plantings of South Karelia

In 2024, due to sanctions restrictions, the availability of medical raw materials may be seriously limited, which acutely raises the question of the availability of its own, Russian base of medicinal raw materials. The concept of pharmaceutical safety of the country involves expanding the range of cultivated medicinal plants, primarily in the northern regions, where special soil and climatic conditions (long daylight hours, low temperatures of the growing season, soil conditions) allow plants to accumulate a greater amount of biologically active substances compared to plants cultivated in the southern regions. One of the most valuable, but sparsely distributed species of medicinal plants is the common autumn crocus (Colchicum autumnale), which contains a sparsely distributed alacloid – colchicine. Colchicine is a valuable alkaloid used in medicine and in plant breeding. Previously, it was found that the common autumn crocus containing the alkaloid colchicine tolerates winter in South Karelia and can be used as a local source of colchicine. In this regard, the purpose of our research was to study the features of the introduction of the common autumn crocus on sod-podzolic mineral soils of South Karelia.

Anatomical and phytochemical studies and ethnomedicinal uses of Colchicum autumnale L.

Background: In the world, great importance is attached to the intensification of plants. To date, most medicinal plants are only partially used. In case of Colchicum autumnale L. currently mainly the corms are used for the extraction of alkaloids. The purpose of this study was to investigate the intensification of the use of this perspective plant reported from the study area. This article examines the anatomy and phytochemistry of leaf and corm of C. autumnale, which is grown as a medicinal crop in Uzbekistan. Methods: The leaves were fixed in 70% ethanol according to the generally accepted method; transverse and paradermal sections were examined on the fixed material. When studying the anatomical features of the leaf, preparations was formed by manually cutting a transverse section of the leaf were observed by a MOTIC V1 microscope. The slides were stained with glycerin jelly having safranin. Cells and tissues were measured using a micrometer MOB-15. To determine the amount of alkaloids contained in the leaves and in corms, a chemical analysis was carried out in the laboratory of alkaloids of the Institute of Chemistry of Plant Substances. Results: On the transverse section of the stomata of the leaves of Colchicum autumnale anomocytic type, 2-3 rows of parenchyma and palisade cells, the characteristic size of the stomata. The root structure of C. autumnale consists of 4-5 rows of corolla, 4 protoxylems and 1 metaxylem, and the presence of rhomboid calcium oxalate crystals in its parenchyma is a characteristic feature of the species. As a result of studying the chemical composition of the plant in the conditions of the Tashkent region, it was observed that the amount of alkaloids in the aerial part of the colchicum, in the leaves was 0.4% and in the corm 0.14%. Conclusion: The current work sought to resolve taxonomic issues involving species that share the majority of their physical characteristics and to clarify relationships between important taxa by utilizing leaf epidermal characteristics. As a result of chemical analysis, it was revealed that the amount of alkaloids in the leaves of Colchicum autumnale, introduced in the conditions of the Tashkent region (Uzbekistan), is higher than in natural regions. This proves that in the conditions of Uzbekistan it is possible to grow Colchicum and obtain high-quality raw materials. Key words: autumn crocus, meadow saffron, anatomy, phytochemistry, ethnomedicine.

Colchicine may cause elevated serum levels of CA72-4: Case Series

Colchicine is used in the treatment of Behçet's disease, familial Mediterranean fever, gout arthritis, pericarditis, some inflammatory and fibrotic diseases by extracting it as an alkaloid from the plant named Autumn Crocus, which is from the lily family. CA 72-4, which has a glycoprotein structure, is especially useful in the diagnosis and follow-up of gastrointestinal adenocarcinomas as well as it's serum level can be increased in pancreatic, ovarian, endometrial, and breast cancers. No drugs that cause CA 72-4 rise have been reported to date. In this article, four patients who were referred to our clinic because of CA 72-4 serum elevation and who used colchicine for various reasons are presented.

Updated review on gout

First identified by the Egyptians in 2640 BC, podagra (acute gout occurring in the first metatarsophalangeal joint) was later recognized by Hippocrates in the fifth century BC, who referred to it as 'the unwalkable disease'. The term is derived from the Latin word gutta (or 'drop'), and referred to the prevailing medieval belief that an excess of one of the four 'humors' - which in equilibrium were thought to maintain health - would, under certain circumstances, 'drop' or flow into a joint, causing pain and inflammation. Throughout history, gout has been associated with rich foods and excessive alcohol consumption. Because it is clearly associated with a lifestyle that,atleast in the past, could only be afforded by the affluent, gout has been referred to as the 'disease of kings'. Although there is evidence that colchicine, an alkaloid derived from the autumn crocus (Colchicum autumnale), was used as a powerful purgative in ancient Greece more than 2000 years ago, its first use as a selective and specific treatment for gout is attributed to the Byzantine Christian physician Alexander of Tralles in the sixth century AD.

Poisoning by Plants.

Questions on poisoning by plants are a common reason for inquiries to poison information centers (PIC). Over the years 2011-2020, plant poisoning was the subject of 15% of all inquiries to the joint poison information center in Erfurt, Germany (Gemeinsames Giftinformationszentrum Erfurt, GGIZ) that concerned poisoning in children (2.3% in adults). In this patient collective, plant poisoning occupied third place after medical drugs (32%) and chemical substances (24%), and was a more common subject of inquiry than mushroom poisoning (1.5%). This review is based on pertinent publications retrieved by a selective literature search in PubMed/TOXLINE on plant poisoning and on 12 epidemiologically and toxicologically relevant domestic species of poisonous plants in risk categories 2 and 3 (up to 2021). Medical personnel should have basic toxicological knowledge of the following highly poisonous plants: wolfsbane (aconitum), belladonna, angel's trumpet, cowbane (cicuta virosa), autumn crocus, hemlock, jimson weed, henbane, castor bean (ricinus), false hellebore, foxglove (digitalis), and European yew. The intoxication is evaluated on the basis of a structured history (the "w" questions) and the clinical manifestations (e.g., toxidromes). Special analysis is generally not readily available and often expensive and time-consuming. In case of poisoning, a poison information center should be contacted for plant identification, risk assessment, and treatment recommendations. Specimens of plant components and vomit should be obtained, if possible, for further testing. Measures for the elimination of the poisonous substance may be indicated after a risk-benefit analysis. Specific antidotes are available for only a few types of plant poisoning, e.g., physostigmine for tropane alkaloid poisoning or digitalis antibodies for foxglove poisoning. The treatment is usually symptomatic and only rarely evidence-based. Individualized medical surveillance is recommended after the ingestion of large or unknown quantities of poisonous plant components. The clinician should be able to recognize dangerous domestic species of poisonous plants, take appropriate initial measures, and avoid overdiagnosis and overtreatment. To improve patient care, systematic epidemiological and clinical studies are needed.

The cardiovascular effects and safety of colchicine

Colchicine is an alkaloid obtained from the bulb and seed of the Crocus (Colchicum autumnale L. (Liliaceae), Autumn Crocus) plant with a history of 2000 years and is currently prescribed for the treatment of rheumatic diseases. In 2015, the European Society of Cardiology supported and recommended colchicine as a first-choice treatment agent that can be added to traditional anti-inflammatory treatments to improve response to treatment, increase remission rates and reduce relapses in acute and recurrent pericarditis in the diagnosis and treatment guidelines of pericardial diseases. In addition, the use of colchicine in the Secondary Prevention of Atherosclerosis is recommended with evidence level A in the European Society of Cardiology Guidelines published in 2021. It was concluded that colchicine is a safe and effective treatment modality to prevent AF recurrence after catheter ablation. It has been noted that colchicine, used secondary to the currently used preventive drugs against CHD, has lower rates of cardiovascular death, MI, stroke, or emergency hospitalization due to angina requiring coronary revascularization, and the risk of acute cardiovascular events compared to placebo. In addition, it has been reported that the use of colchicine in patients with MI reduces the risk of ischemic cardiovascular events. Colchicine is an effective and safe option as a first-choice treatment agent that can be added to conventional anti-inflammatory treatments in patients with recurrent cardiovascular events despite optimal medical therapy and in the secondary prevention of atherosclerotic cardiovascular diseases where other risk factors cannot be adequately controlled.

Cultivation of Saffron (Crocus sativusL.) in cold climates

Saffron, an autumn crocus that produces a highly valuable spice, is grown mainly in Mediterranean climates. Nevertheless, saffron farms have been established recently in the province of Quebec. This led us to test cultivation practices that could influence plant phenology, saffron yield, and corm growth, including planting depth, planting period, and the application of fertilizers, mycorrhizal fungi, and biostimulants at planting. Soil temperature was monitored at the different planting depths throughout the year. Floral initiation was also monitored during spring and summer. Shoot emergence was delayed and final emergence reduced as planting depth increased; however, more shoots were produced by shallow-planted corms, which could lead to the production of corms too small to flower. The best time for planting saffron corm is between the end of July and the third week of August. Mineral fertilization hastened leaf emergence and improved corm production and their nutrient content. Neither the addition of mycorrhizal fungi or of biostimulants had any significant impact on saffron growth or flowering. Floral induction likely took place in July as flower bud appeared in early August. In most years, flower and saffron production was low in this location. It appears that soil temperature did not remain high for long enough during the summer to promote floral induction and autumn temperatures decreased too fast, limiting shoot and flower emergence most years. However, these climatic conditions did not affect corm production; corms could thus be sold to secure revenues for producers.

Colchicine, terug van nooit weggeweest

Colchicine: ancient and brand new Colchicine is an alkaloid initially extracted from an autumn crocus. Its use dates back to the 6th century BC, with the first indication being gout. In the last 50 years, further insights into its mechanism of action emerged along with an expansion of the number of indications. Colchicine inhibits the chemotaxis of leukocytes. Recently, it was discovered that in addition to its action on the microtubules, there is also an effect on the NLRP3 inflammasome. Colchicine has a narrow therapeutic index and caution is advised in patients with renal and hepatic failure. It’s a CYP3A4 and P-gp substrate. Therefore, concomitant use with CYP3A4 and P-gp inhibitors is contraindicated given the greatly increased risk of toxicity. Its most frequent side effect is diarrhea and severe intoxications can be lethal. The main rheumatologic indications are the treatment of gout in the acute phase, attack prevention and amyloidosis in familial Mediterranean fever, and ulcerations in Behçet’s disease. The cardiovascular use of colchicine in acute and relapsing/chronic pericarditis has been well established. Recent studies also show a beneficial effect in case of acute and chronic coronary artery disease. Additional studies for the implementation of colchicine for these indications are ongoing.

Effective and simple approach for colchicine determination in saffron parts

Crocus sativus (Iridaceae) or saffron is important for its flavoring properties in the food industry and its medicinal properties in the pharmaceutical industry. Real saffron must meet the quality requirements set to FDA or ISO specifications, and in addition, must be safe and non-toxic. An “autumn crocus” or scientific name Colchicum spp. (Colchicaceae) is a plant that contains a highly toxic alkaloid colchicine. The problem is that Colchicum autumnale is often mistaken for C. sativus. This study aim is to develop the high-performance liquid chromatography method for determining the presence of colchicine in C. sativus stigma, flowers and corms, and then for solving the issue of saffron possible falsification and toxicity. The chromatographic separation was performed on ACE C18 column (5.0 μm, 250 mm × 4.6 mm), by using 0.1% acetic acid in water and acetonitrile as the mobile phase and UV detector (350 nm). Current analysis showed that all Crocus raw materials didn’t contain colchicine. This confirms the safety of saffron and its parts for using in the development of substances based on them. The proposed method was successfully applied on C. autumnale corms and showed good separation and colchicine identification. The presence of tricyclic proto-alkaloid colchicine in plant products is not a new phenomenon, however, the need for new sensitive analytical methods that can detect it in food and medicine still exists. The identification and confirmation of colchicine absence in saffron raw materials are important for further herbal drug development and for the food industry.

Colchicine: old medication with new benefits : Use in rheumatology and beyond

Colchicin, das Gift der Herbstzeitlosen, hat verschiedene antiinflammatorische Effekte. Aus diesem Grund kommt es zur Behandlung von rheumatologischen Erkrankungen aus dem autoinflammatorischen Formenkreis, wie z. B. der Arthritis urica oder dem familiären Mittelmeerfieber (FMF), zum Einsatz. Darüber hinaus gibt es erste Daten, die einen positiven Nutzen von Colchicin bei kardiovaskulären Erkrankungen nahelegen. Des Weiteren werden aktuell verschiedene antiinflammatorische Therapieansätze in der COVID-19-Behandlung in Studien erprobt. Hier gibt es ebenfalls erste Publikationen, die einen potenziellen Nutzen von Colchicin in bestimmten Krankheitsphasen der Virusinfektion nahe legen. Dieser Beitrag will einen Überblick über die Wirkweise, den Nutzen und Nebenwirkungen sowie die verschiedenen Einsatzmöglichkeiten von Colchicin in der Rheumatologie geben. Weiterhin soll ein kurzer Ausblick in neue Einsatzgebiete dieses Medikamentes gegeben werden.

Gout and ‘Podagra’ in medieval Cambridge, England

ObjectiveTo estimate the prevalence rate of gout and to explore the social factors that contributed to its development in the various sub-populations in medieval Cambridge. Materials177 adult individuals from four medieval cemeteries located in and around Cambridge, UK. MethodsLesions were assessed macroscopically and radiographically. Elements with lytic lesions were described and imaged using micro-computed tomography (μCT) to determine their morphology. ResultsGout was identified in 3 % of the population. Individuals buried in the friary had highest prevalence (14 %), with low prevalence rates in the Hospital (3 %) and town parish cemetery (2 %), with no cases in the rural parish cemetery. Gout was more prevalent during the 14th–15th centuries than the 10th–13th centuries. ConclusionThe high prevalence rate of gout in the friary is at least partly explained by the consumption of alcohol and purine-rich diets by the friars and the wealthy townsfolk. Medieval medical texts from Cambridge show that gout (known as podagra) was sometimes treated with medications made from the root of the autumn crocus. This root contains colchicine, which is a medicine that is still used to treat gout today. SignificanceThis is one of the first studies to assess the epidemiology of gout in medieval England and suggests that gout varied with social status. LimitationsOur sample size precludes statistical analysis. Suggestions for further researchAdditional studies that assess the epidemiology of gout in medieval Europe is needed in order to be able to fully contextualize these findings.

Toxicity and Sublethal Effects of Autumn Crocus (Colchicum autumnale) Bulb Powder on Red Imported Fire Ants (Solenopsis invicta).

Autumn crocus (Colchicum autumnale L.) is a medicinal plant as it contains high concentrations of colchicine. In this study, we reported that the ground powder of autumn crocus bulb is highly toxic to invasive Solenopsis invicta Buren, commonly referred to as red imported fire ants (RIFAs). Ants fed with sugar water containing 5000 mg/L of bulb powder showed 54.67% mortality in three days compared to 45.33% mortality when fed with sugar water containing 50 mg/L of colchicine. Additionally, the effects of short-term feeding with sugar water containing 1 mg/L of colchicine and 100 mg/L of autumn crocus bulb powder were evaluated for RIFAs’ colony weight, food consumption, and aggressiveness, i.e., aggregation, grasping ability, and walking speed. After 15 days of feeding, the cumulative colony weight loss reached 44.63% and 58.73% due to the sublethal concentrations of colchicine and autumn crocus bulb powder, respectively. The consumption of sugar water and mealworm (Tenebrio molitor L.) was substantially reduced. The aggregation rates decreased 48.67% and 34.67%, grasping rates were reduced to 38.67% and 16.67%, and walking speed decreased 1.13 cm/s and 0.67 cm/s as a result of the feeding of the two sublethal concentrations of colchicine and autumn crocus bulb powder, respectively. Our results for the first time show that powder derived from autumn crocus bulbs could potentially be a botanical pesticide for controlling RIFAs, and application of such a product could be ecologically benign due to its rapid biodegradation in the environment.

Accidental poisoning with autumn crocus (Colchicum autumnale): a case series

Introduction Colchicum autumnale (autumn crocus) is a plant that contains highly toxic alkaloid colchicine. The aim was to evaluate accidental C autumnale poisoning and assess serum troponin as a prognostic parameter. Methods In this study, we retrospectively included all adult patients with a history of accidental C autumnale ingestion and serum colchicine confirmation during the study period from 2000 to 2019. The medical files of enrolled patients were reviewed. Literature search of accidental ingestions of C autumnale was done. Results Over the study period of 20 years, 16 adult patients were admitted to the University Medical Centre Ljubljana due to acute colchicine poisoning after ingestion of C autumnale. They all mistakenly ingested C autumnale’s leaves instead of Allium ursinum in the spring and had confirmed colchicine in serum by GC–MS or LC–MS/MS (15.5 µg/L (0.5–80 µg/L)). They developed vomiting and diarrhoea within 1–9 h after the meal. Vomiting within 2 h was associated with lethality (p=.04). Bone marrow suppression developed in 15 patients (94%). Acute myocardial injury with positive troponin I (>0.10 µg/L) developed in five patients; lethal cardiogenic shock with decreased cardiac output and hypotension occurred in four of these patients despite supportive therapy. Positive troponin I ultra (>0.10 µg/L) was associated with need for intensive support therapy (p=.01), decreased cardiac output (p=.01) and death (p=.01). The mortality was 4/16 (25%). On review, we found 58 cases; 95% cases accidently ingested leaves of C autumnale instead of A ursinum. Troponin I was reported in 3% cases. The lethality of this and reviewed cases was 35% (26/74). Conclusions In unexplained gastroenterocolitis after ingestion of wild plants as a salad or spice in the spring, especially when wild garlic is mentioned, we should always consider C autumnale poisoning. Cardiogenic shock can be predicted by a positive serum troponin I measurement.

What kind of poison was used in "The Name of the Rose"?

In the novel "The Name of the Rose" by Umberto Eco, afanatical monk laced the pages of abook with apoison, which led to the death of several monks in amedieval monastery. Based on modern toxicological knowledge, an attempt can be made to determine whether there exists asubstance meeting the criteria of the poison described in the novel. To this end, toxicological literature on the lethal doses of plant-derived poisons and their duration of action was reviewed. Cowbane (Cicuta virosa), aplant used for preparing poisonous potions, contains cicutoxin which kills at adose of 500 mg. Similar toxicity is displayed by water dropwort (Oenanthe crocata) and Cerbera odollam growing in India and Madagascar. Hemlock (Conium maculatum) contains coniine which is able to kill avictim after exposure to adose as low as 100 to 200 mg. Morphine found in opium poppy (Papaver somniferum) and colchicine contained in autumn crocus (Colchicum autumnale) produce lethal effects at similar doses. So do more exotic plants such as Calabar bean (Physostigma venenosum) which contains physostygmine. Two other exotic plants that need to be mentioned in this context are far more poisonous. They include Saint Ignatius's bean (Strychnos ignatii) and strychnine tree (Strychnos nux-vomica). Both contain strychnine, the lethal dose of which is 30 mg. Death cap (Amanita phalloides) might also be considered in this context, but despite being lethal at adose of 20 mg, amanitin contained in this fungus takes afew days to kill avictim, so the effect is considerably slower than that experienced by the monks in Eco's novel. Adose of 10 mg is sufficient to kill ahuman with any of four other plant-derived poisons: antiarin, atropine, digoxin and strophantin. Aconitum (Aconitum napellus), also known as monkshood, contains aconitine and is an even more deadly plant, with the lethal dose for humans being only 2 mg. Ricin and abrin are ranked even higher in the list of plant-derived toxins, as they are able to kill in doses lower than 1 mg. However, they could not have been used by the murderer in "The Name of the Rose", as they cause death only several days after ingestion. To conclude, the plant that best matches the criteria mentioned in the novel is Aconitum.

Effect of Colchicine administration on the structure of the Spleen of adult male albino rat: Histological and Immunohistochemical study

Background: Colchicine is a natural alkaloid derived from autumn crocus; it is used in the treatment of gout and in preventing familial Mediterranean fever attacks. It accumulates in many organs such as spleen and may lead to their damage.Aim: This work aimed to study short and long duration effects of colchicine administration on the structure of the spleen of the adult male albino rat.Material and Methods: Forty-five adult male albino rats were used in this study, aged from 4-6 months, weighting 150 -180 gm. Rats were equally divided into three groups: Group I: It was further equally subdivided into three subgroups: subgroup IA: Kept as a negative control, subgroup IB: received 1ml of distilled water/day orally for seven days and subgroup IC: received 1ml of distilled water/day orally for 30 days. Group II: received 3mg/Kg body weight/ day of colchicine orally for seven days. Group III: received colchicine as those of group II but for 30 days.Results: The present work revealed that administration of colchicine for short duration induced mild structural alteration of rat’s spleen in the form of reduction in lymphoid follicles size and subcapsular congestion of blood vessels, while, marked structural alteration of rat’s spleen was noticed in group III that received colchicine for long duration in the form of loss of distinction between the white and red pulps with nearly unidentifiable lymphoid follicles and marked parenchymal congested blood vessels.Conclusion: Colchicine administration induced structural alteration of rat’s spleen which was mild with short term use and extensive with long term use.

Autumn poisoning hazards

The main poisoning hazard of autumn is the abundance of fruits and seeds following summer growth, but if the summer has seen extreme weather conditions, then pasture may be poor and horses may eat plants they would normally avoid. Usually the most significant hazard for horses in the autumn is atypical myopathy from repeated ingestion of sycamore seeds. Dried or wilted leaves of a related species, red maple, can cause haemolysis and methaemoglobinaemia in equines. Other potential risks in the autumn include yew; autumn crocus; oak (acorns); horse chestnut (conkers); plants containing cyanogenic glycosides, such as Prunus species; and ragwort. The fruit bodies of fungi (that is, mushrooms) are also abundant in warm, wet weather and although rarely reported (and possibly underreported), mushroom poisoning is a potential risk in horses. Depending on the type of mushroom there is risk of gastrointestinal, neurological, renal and liver effects. Treatment of plant and mushroom poisoning in horses will depend on the clinical signs and the plant or mushroom involved, but should include identification and removal of the plant or mushroom, and symptomatic care.

Autumn plant poisoning hazards

Potentially poisonous plants are a common part of the environment of many farm animals. Risk of poisoning varies season by season and year by year with variation in the environmental conditions. In the autumn there is risk of poisoning from seeds and fruits and autumn flowering plants. This includes plants containing cyanogenic glycosides (such as cherry laurel) and yew which contains taxine alkaloids. These plants commonly cause sudden death in livestock. Oak, although part of the natural diet of many animals, can cause episodic outbreaks of poisoning, particularly following hot, dry summers. Signs mainly involve the gastrointestinal and renal systems. Poisoning with horse chestnut is not common but can cause neurological and gastrointestinal signs. Ingestion of autumn crocus can also cause gastrointestinal signs in livestock. Prevention of poisoning by recognising the risks and limiting potential circumstances of exposure and ensuring access to forage is essential, as treatment options for management of poisoning in livestock are limited and, in some cases, the only sign of exposure is sudden death.

An engineered lipocalin that tightly complexes the plant poison colchicine for use as antidote and in bioanalytical applications.

Colchicine is a toxic alkaloid prevalent in autumn crocus (Colchicum autumnale) that binds to tubulin and inhibits polymerization of microtubules. Using combinatorial and rational protein design, we have developed an artificial binding protein based on the human lipocalin 2 that binds colchicine with a dissociation constant of 120 pm, i.e. 10000-fold stronger than tubulin. Crystallographic analysis of the engineered lipocalin, dubbed Colchicalin, revealed major structural changes in the flexible loop region that forms the ligand pocket at the open end of the eight-stranded β-barrel, resulting in a lid-like structure over the deeply buried colchicine. A cis-peptide bond between residues Phe71 and Pro72 in loop #2 constitutes a peculiar feature and allows intimate contact with the tricyclic ligand. Using directed evolution, we achieved an extraordinary dissociation half-life of more than 9 h for the Colchicalin-colchicine complex. Together with the chemical robustness of colchicine and availability of activated derivatives, this also opens applications as a general-purpose affinity reagent, including facile quantification of colchicine in biological samples. Given that engineered lipocalins, also known as Anticalin® proteins, represent a class of clinically validated biopharmaceuticals, Colchicalin may offer a therapeutic antidote to scavenge colchicine and reverse its poisoning effect in situations of acute intoxication.

Extraction, development and validation of HPLC-UV method for rapid and sensitive determination of colchicine from Colchicum autumnale L. Bulbs.

Colchicum autumnale L. also known as the autumn crocus, contains colchicine alkaloid having antifungal properties. The tuber of this plant is rich in terms of colchicine. In this research an ultrasound-assisted extraction (UAE) method was optimized for the extraction of colchicine from Colchicum autumnale L. bulbs before high-performance liquid chromatography with UV detection (HPLC-UV). Optimization of various extraction parameters was performed using response surface methodology (RSM) to evaluate the maximum colchicine yield from Colchicum autumnale L. bulbs. The Box-Behnken design (BBD) and RSM were used to investigate the effect of three key parameters (extraction time (20–60 min), extraction temperature (40–80 °C) and ultrasound power (500–700 W) on extraction efficiency. The variance analysis suggested that the dependent response variable of yield of colchicine may be expressed by a quadratic polynomial model. The optimal theoretical extraction conditions were found to be an ultrasonication power of 602.4 W, an extraction time of 42 min and a temperature of 64 °C. Under these conditions, the optimum foreseen yield was 0.237%. The experimental colchicine yield obtained by following the optimized conditions was found to be 0.238%. These values are very well compatible with each other.