Alkaloids Ergotamine Research Articles

Effects of ergotamine on the central nervous system using untargeted metabolomics analysis in a mouse model

The ergot alkaloid ergotamine is produced by Claviceps purpurea, a parasitic fungus that commonly infects crops and pastures of high agricultural and economic importance. In humans and livestock, symptoms of ergotism include necrosis and gangrene, high blood pressure, heart rate, thermoregulatory dysfunction and hallucinations. However, ergotamine is also used in pharmaceutical applications to treat migraines and stop post-partum hemorrhage. To define its effects, metabolomic profiling of the brain was undertaken to determine pathways perturbed by ergotamine treatment. Metabolomic profiling identified the brainstem and cerebral cortex as regions with greatest variation. In the brainstem, dysregulation of the neurotransmitter epinephrine, and the psychoactive compound 2-arachidonylglycerol was identified. In the cerebral cortex, energy related metabolites isobutyryl-L-carnitine and S-3-oxodecanoyl cysteamine were affected and concentrations of adenylosuccinate, a metabolite associated with mental retardation, were higher. This study demonstrates, for the first time, key metabolomic pathways involved in the behavioural and physiological dysfunction of ergot alkaloid intoxicated animals.

Bovine lateral saphenous veins exposed to ergopeptine alkaloids do not relax12

The ergot alkaloid ergovaline has demonstrated a persistent and sustained contractile response in several different vascular models. It was hypothesized that different alkaloids isolated from tall fescue (Lolium arundinaceum) will contribute to this contractile response differently. The objective was to compare contractile-response patterns of single additions of the ergoline alkaloids lysergic acid, lysergol, and ergonovine and the ergopeptine alkaloids ergotamine, ergocristine, ergocryptine, ergocornine, and ergovaline (provided as tall fescue seed extract). Lateral saphenous veins were collected from 6 Holstein steers (BW = 397 ± 28 kg) immediately after slaughter, sliced into cross-sections, and suspended in myograph chambers containing oxygenated Krebs-Henseleit buffer (95% O2/5% CO2; pH = 7.4; 37°C). Treatments were added at 0 min and buffer was replaced in 15-min intervals for a 120-min incubation. In addition to maximum tension and time to reach maximum tension, percent relaxation and rate of relaxation were determined following maximum tension for each treatment. All compounds tested produced significant contractile responses (P < 0.05). All ergoline alkaloids reached maximum response in less time (P < 0.05) than the remaining compounds and began to relax immediately after first buffer change. Lysergic acid had the greatest (P < 0.05) percent relaxation and ergonovine had the greatest (P < 0.05) rate of relaxation. The ergopeptine alkaloids ergovaline, ergotamine, ergocristine, ergocryptine, and ergocornine had slower developing contractile responses with a longer (P < 0.05) interval until maximum tension was achieved compared to the ergoline alkaloids. Maximal responses to all the ergopeptine alkaloids, however, all persisted for the 120-min duration with negligible relaxation occurring. The different classes of alkaloids differed greatly in the type of contractile response generated in the lateral saphenous vein. Persistence of contractile response is thought to be the primary contributing factor to the vasoconstriction observed in animals demonstrating signs of fescue toxicosis, where different ergot alkaloids can contribute differently.

Herbs and natural products in prevention and treatment of migraine

Migraine headaches are extremely common and it is considered that, on average, one of ten people suffer from migraine. The pathogenesis of migraine is not fully understood. There are two theories, vascular and neurogenic. The treatment of migraine involves the prevention and treatment of acute attacks. Preventive therapy is recommended when migraine attacks are extremely frequent (2 or more attacks per month), intensive, long, and resistant to acute therapy. One of the most important herbs in the treatment of migraine is species Tanacetum parthenium, Asteraceae, feverfew. The most important active compounds in aboveground parts of the plant are sesquiterpene α-methylenebutyrolactones. The dominant compound of this group is parthenolide - sesquiterpene lactone. Parthenolide is considered to be responsible for antimigraine activity. It was shown that parthenolide contains highly electrophilic α-methylene-γ-lactone ring and resin, which directly interact with the nucleophilic groups of biological molecules. Among other, (inhibition of platelet aggregation, the transcription of NF-kappaB and IKC-a), parthenolide inhibits the release of serotonin. Also, ergo-peptide alkaloid ergotamine, product of the metabolism of the fungus principal rye ( Claviceps purpurea ) shows antimigraine activity. It acts as a partial agonist of α-adrenergic receptors, which explains vasoconstriction. On 5-HT receptors it acts as partial agonist and that can explain usage in migraine therapy.

Influence of diet sequence on intake of foods containing ergotamine d tartrate, tannins and saponins by sheep

Plant secondary compounds (SCs) play crucial roles in plant survival and defense against herbivores and in appropriate amounts SCs provide health and nutritional benefits to herbivores. The presence of SCs in plants is important, and means need to be found to enhance their benefits while minimizing their negative impacts on herbivores. One approach to this challenge is to offer herbivores mixtures of forages with diverse and complementary SCs such that a combination of SCs becomes less toxic than each of its components in isolation. We determined if a diversity of SCs (tannins, saponins, and alkaloids) offered in different temporal sequences influenced intake of food by sheep. In Experiment 1, three groups of sheep (n=8/group) received a food containing the alkaloid ergotamine d tartrate (EDT) and the same food containing condensed tannins (TAN) in three different sequences. Group 1 received TAN for 30min followed by EDT for 1h (TAN→EDT), Group 2 received EDT for 1h followed by TAN for 30min (EDT→TAN), and Group 3 (Control Group) received EDT for 30min followed by EDT for 1h (EDT→EDT). Experiment 2 was similar to Experiment 1 except a new group of sheep was used and saponins (SAP) replaced tannins. Sheep had greater intakes of EDT when supplemented with TAN (17 vs. 19g EDT/kg BW/d for TAN→EDT and EDT→TAN, respectively; P=0.0002) or SAP (SAP→EDT vs. EDT→SAP – 10 vs. 12g/kg BW/d; P=0.0979) than when fed only EDT (9 and 7g/kg BW/d, respectively), regardless of the sequence in which the foods were presented. Intake of TAN was greater for sheep in TAN→EDT than for sheep in EDT→TAN on 4 or the 7 days of the trial (P=0.0336). Likewise, intake of SAP was greater for the sequence SAP→EDT than for EDT→SAP (6 vs. 2g SAP/kg BW/d; P=0.0162). Regardless of sequence, sheep ate consistently more food with EDT than with either TAN or SAP. Sheep had higher total food intake, and they also ate more EDT, when they were fed TAN as opposed to SAP. Under the conditions of our experiment, sheep ate more food with alkaloids (EDT), and perhaps benefitted more nutritionally, when they had tannins (TAN) rather than saponins (SAP) in their diet.

Interaction of ergotamine with liver Cytochrome P450 3A in rats

This study was conducted to investigate the effect of the ergot alkaloid, ergotamine (ET), on the induction of CYP3A and the interaction in vivo and in vitro with ET. Sprague-Dawley rats were treated intraperitoneally for 4 days as follows: control (injecting with 0.5 ml of only corn oil); dexamethasone treatment (injecting with 100mg/kg of dexamethasone in corn oil); and ergotamine treatment (injecting with 100mg/kg of ergotamine in corn oil). Liver tissues were collected from each group (n = 5, total of 30 rats) and liver microsomes were prepared. Cytochrome CYP3A activity was evaluated using ET and its isomer as substrates in medium containing liver microsomes and NADPH at 37℃ for 30 min. HPLC was used to measure the disappearance of the substrate and the appearance of the metabolites. Liver microsomes from rats pretreated with dexamethasone were five times more (P 0.05) in activity of CYP3A when compared to the control group (5.2 vs. 7.0 nM ET/min/mg protein; SE = 4.83) or ET isomer (1.5 vs. 4.7 nM ET isomer/ min/mg protein; SE = 1.70). When ketoconazole was used as specific inhibitor of CYP3A, ergotamine metabolisms were inhibited in a dose dependent fashion reaching a maximum at an inhibitor to substrate ratio of greater than one and LD50 at 0.5 nM of ketoconazole/mg protein. The data presented in this study suggest that although the ergot alkaloids ergotamine and its isomer are ideal substrates for the isozyme CYP3A, these compounds have no effect on the induction of CYP3A after 4 days of treatment.

Indole Prenyltransferases from Fungi: A New Enzyme Group with High Potential for the Production of Prenylated Indole Derivatives

Prenylated indole derivatives are hybrid natural products containing both aromatic and isoprenoid moieties and are widely spread in plants, fungi and bacteria. Some of these complex natural products, e.g. the ergot alkaloids ergotamine and fumigaclavine C as well as the diketopiperazine derivative fumitremorgin C and its biosynthetic precursors tryprostatin A and B, show a wide range of biological and pharmacological activities. Prenyl transfer reactions catalysed by prenyltransferases represent key steps in the biosynthesis of these compounds and often result in formation of products which possess biological activities distinct from their non-prenylated precursors. Recently, a series of putative indole prenyltransferase genes could be identified in the genome sequences of different fungal strains including Aspergillus fumigatus. The gene products show significant sequence similarities to dimethylallyltryptophan synthases from fungi. We have cloned and overexpressed six of these genes, fgaPT1, fgaPT2, ftmPT1, ftmPT2, 7-dmats and cdpNPT from A. fumigatus in E. coli and S. cerevisiae. The overproduced enzymes were characterised biochemically. Three additional prenyltransferases, DmaW-Cs, TdiB and MaPT were identified and characterised in a Clavicipitalean fungus, Aspergillus nidulans and Malbranchea aurantiaca, respectively. Sequence analysis and alignments with known aromatic prenyltransferases as well as phylogenetic analysis revealed that these enzymes belong to a new group of "aromatic prenyltransferases". They differ clearly from membrane-bound aromatic prenyltransferases from different sources and soluble prenyltransferases from bacteria. The characterised enzymes are soluble proteins, catalyse different prenyl transfer reactions on indole moieties of various substrates and do not require divalent metal ions for their enzymatic reactions. All of the enzymes accepted only dimethylallyl diphosphate as prenyl donor. On the other hand, they showed broad substrate specificity towards their aromatic substrates. Diverse tryptophan derivatives and tryptophan-containing cyclic dipeptides were accepted by these enzymes, providing a new strategy for convenient production of biologically active substances, e.g. by chemoenzymatic synthesis.

Sheep foraging behavior in response to interactions among alkaloids, tannins and saponins

AbstractBACKGROUND: A mixture of plant species adds biochemical diversity to pastures that may enhance productivity while decreasing reliance on herbicides and insecticides. All plants contain secondary metabolites (PSMs) that interact in plant communities in a variety of ways. Our objective was to determine if PSM diversity enhanced food intake when sheep were allowed to select from foods that varied in concentrations of tannins, saponins, and alkaloids. We hypothesized that intake of foods containing alkaloids would increase when sheep were offered supplemental foods with tannins and saponins. The alkaloid gramine is proteinaceous in nature, tannins bind to proteins in the gut, which enhances excretion of proteins, so we hypothesized tannins would bind to gramine and enhance its excretion from the body. The alkaloid ergotamine is steroidal, saponins have a high affinity for binding to lipid and steroidal compounds in the gut and then being excreted, so we hypothesized saponins would bind to ergotamine and enhance its excretion from the body.RESULTS: We found that sheep fed either gramine or ergotamine in combination with tannin‐ or saponin‐containing foods had higher intakes of food than sheep offered only foods with gramine or ergotamine.CONCLUSION: These findings are consistent with the hypothesis that animals can neutralize the negative effects of alkaloids by ingesting tannins and saponins. Our results show that PSMs can complement one another to increase a herbivore's intake of unpalatable plant species, with the potential to maintain healthy animals and bio‐diverse landscapes. Copyright © 2008 Society of Chemical Industry

Growth and hepatic in vitro metabolism of ergotamine in mice divergently selected for response to endophyte toxicity

This study investigated if genetic differences exhibited in endophyte-resistant and -susceptible mouse lines had persisted after 13 generations in which the integrity of lines was maintained yet selection ceased. Experimental groups were mouse lines fed an endophyte-free (E−) or -infected (E+) diet. The in vitro metabolism of the ergot alkaloid ergotamine in mouse liver microsomes was characterized by LC-MS/MS and compared between both lines before and after exposure to E+ feed. No difference in the average daily weight gain of pups between resistant and susceptible mice was observed on the E+ diet. Thus, for the weight gain selection criterion under study, the difference established between the two lines appears not to have persisted over the extended period of relaxed selection. Microsomal incubations produced nine predominate peaks in the HPLC assay. The peaks were confirmed by LC-MS/MS to be ergotamine, ergotamine epimer, monohydroxylated metabolites (M1, M2, M1e, M2e) and dihydroxylated metabolites (M3–5). A gender difference for metabolite formation was observed on the E− diet, in that females produced a greater amount of M1, M1e and M3–5 than males. When challenged with the E+ diet, mice showed differences in concentration of M3 for line (resistant > susceptible) and gender (female > male) and of M4 and M5 for gender (female > male). Gender differences in the metabolism of ergotamine have not been shown before in these lines of mice or other species used to study ergot alkaloid metabolism. This adds a potential source of variation in the susceptibility to fescue toxicity not explored previously and would be of value to investigate further.

INCREASED PROINFLAMMATORY CYTOKINES PRODUCTION BY ERGOTAMINE IN MALE BALB/c MICE

The ergopeptine alkaloid ergotamine (ET) mimics the effects of ergopeptine alkaloids found in endophyte-infected (E+) fescue forage considered causative for fescue toxicosis. Altered immune capacity, compromised intake and thermoregulation, and inflammatory changes are observed in fescue toxicosis. Taken together, these suggest the cytokine pattern may be altered by ergot alkaloids. Thus, the objective of this study was to determine whether major splenocyte-derived cytokines-interleukin 2 (IL-2), interleukin 4 (IL-4), interferon gamma (IFN-gamma)-and macrophage-derived cytokines-interleukin 1, (IL-1), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-alpha)-were affected by ergotamine. Two sets of male BALB/c mice (n = 5/treatment) were treated with ergotamine tartrate (sc) for 10 d at doses of 0 (control), 0.4, 2, 10, or 50 mg/kg body weight. Twentyfour hours after the last treatment, splenocytes (S) were isolated from one set of animals and macrophages (MO) from the other set for determination of IL-2, IL-4, INF-gamma, and IL 1beta, IL-6, TNF-alpha, respectively. Following activation with 5 mug/ml concanavalin A (Con A) (S) and 10 mug/ml lipopolysaccharide (LPS) (MO), cells were incubated for 48 and 24 h, respectively, and supernatants were collected and assayed for respective cytokines by enzyme-linked immunosorbent assay (ELISA). Additionally, differential white blood cell (WBC) counts were performed and the neutrophil (N) :lymphocyte (L) ratio calculated. Ergotamine treatment significantly increased IL-6 levels at the 2.0 mg/kg dose and greater and TNF-alpha at the highest dose. There was no treatment effect on IL-1 , IL-2, IL-4, and IFN-gamma. Also, no effect was observed upon total and differential WBC counts as well as N:L ratio. Ergotamine affected the proinflammatory cytokine IL-6, and this increase may contribute to fescue tosicosis.

REGION-SPECIFIC DECREASE OF DOPAMINE AND ITS METABOLITES IN BRAINS OF MICE GIVEN ERGOTAMINE

Ergot alkaloids (EA) such as the ergopeptine alkaloid ergotamine (ET) are adrenergic, dopaminergic, and serotonergic agents. The objective of this experiment was to investigate regional brain neurotransmitter alterations caused by EA. Male BALB/ c mice were treated sc daily with ergotamine tartrate for 10 d at 0 (saline), 0.4, 2, 10, or 50 mg/ kg body weight. Twenty-four hours after the last treatment, animals were sacrificed and brains dissected. Regional concentrations of norepinephrine (NE), dopamine ( DA), serotonin (5-HT), and metabolites 3-methoxy-4-hydroxyphenylethylene glycol (MHPG), dihydroxyphenylacetic acid (DOPAC) , homovanillic acid (HVA), and 5-hydroxyindole3-acetic acid (5-HIAA) were measured by high-performance liquid chromatography (HPLC) . Moreover, selected organ weights and plasma prolactin (PRL) were determined. Dopamine concentration was significantly reduced by ET at all doses in the striatal and hypothalamic regions. A reduction of the DA metabolite HVA occurred in striatum at only the highest dose, whereas in the hypothalamus both HVA and DOPAC were markedly reduced. Concentrations of NE, MHPG, 5-HT, and 5-HIAA were not affected by treatment in these regions. In the cerebellum, MHPG was significantly elevated at the 50 mg/ kg dose. No effect of treatment was observed in the cerebrum, medulla, and midbrain. Further, no treatment-related differences in plasma PRL and organ weights other than a significant liver weight decrease at intermediate doses were found. Therefore, the effects of ET were predominantly upon DA metabolism in the corpus striatum and hypothalamus. The reductions in DA, HVA, and DOPAC indicate decreased DA synthesis.

Management of Nonobstetric Pain During Pregnancy and Lactation

Management of Nonobstetric Pain During Pregnancy and Lactation

Management of nonobstetric pain during pregnancy and lactation.

Management of nonobstetric pain during pregnancy and lactation.

Ergopeptine Alkaloids and Neotyphodium lolii-Mediated Resistance in Perennial Ryegrass Against Adult Heteronychus arator (Coleoptera: Scarabaeidae)

Heteronychus arator F. adults are deterred from feeding on perennial ryegrass, Lolium perenne L., infected with the endophytic fungus Neotyphodium lolii, which produces many alkaloids and other mycotoxins in the host grass. This article examines the effects of a number of these compounds on adult H. arator feeding. Pure mycotoxins at several concentrations were incorporated into a carrot-based artificial diet developed specifically for this study. The mycotoxins tested were either compounds commonly found in N. lolii-infected perennial ryegrass (peramine, lolitrem B, paxilline, ergovaline, and ergovalinine), or compounds structurally similar to those occurring in N. lolii/perennial ryegrass associations (ergotamine, alpha-ergosine, alpha-ergocryptine, festuclavine, lysergol, lysergic acid amide, ergonovine, dihydroergotamine, alpha-bromoergocryptine, and dihydroergocrystine). Peramine, a Listronotus bonariensis (Kuschel) feeding deterrent, did not affect adult H. arator feeding. Lolitrem B, a mycotoxin involved in livestock health problems, and its structurally related precursor, paxilline, had no effect. The ergot alkaloids festuclavine, lysergol, and lysergic acid amide also did not deter adult H. arator feeding. Ergonovine was the only nonergopeptine ergot alkaloid to deter adult H. arator, though only moderately. The greatest antifeedant effects were observed with the ergopeptine alkaloids ergotamine, ergovaline, alpha-ergosine, and alpha-ergocryptine, all of which significantly reduced feeding at 5.0 micrograms/g. The ergopeptine epimer ergovalinine, and the ergopeptine alkaloid derivatives dihydroergotamine, dihydroergocrystine, and alpha-bromoergocryptine were also active, although less so than the parent ergopeptines. Thus ergopeptine alkaloids are likely to be a major factor in the N. lolii-mediated resistance to H. arator. It was further observed that perennial ryegrass seed containing an endophyte strain that did not produce ergot alkaloids, but did produce other mycotoxins normally associated with N. lolii infection, did not deter H. arator feeding, whereas an ergot alkaloid producing N. lolii/perennial ryegrass association did. The implications of these findings in relation to the toxicity of ergot alkaloids to mammals, and in relation to the definition of a desirable endophyte, are discussed.

Release of ergotamine from poly (2-hydroxyethyl Methacrylate)

The release of the alkaloid ergotamine from thin discs of poly(2-hydroxyethyl methacrylate) [PHEMA] into an aqueous buffered medium has been effected under sink conditions and monitored by fluorescence spectroscopy. The process was found to be diffusion controlled, the fraction of available drug released being linear in (time)1/2 during the initial stages. The influences of disc thickness, drug load and temperature were investigated and yielded 2.0 (± 1.0) × 10−13 m2 s−1 for the diffusion coefficient of ergotamine through the swollen gel at 310K and a value of 69 ± 5 kJ mol−1 for the activation energy of diffusion. For the uptake of aqueous buffer alone into PHEMA the corresponding values were ca 100 times greater and three times smaller respectively.

ChemInform Abstract: CONFORMATIONAL ANALYSIS OF THE ERGOT ALKALOIDS ERGOTAMINE AND ERGOTAMININE

Conformational analyses by 1H NMR and potential-energy calculations are reported for the ergot alkaloids ergotamine and ergotaminine, both as free bases and as the protonated species. In the neutral forms in CDCl3. two strong intramolecular hydrogen bonds fix the molecules in folded conformations, but the protonated species adopt a more extended conformation, with a single intramolecular hydrogen bond. Of the 24 alternative conformations available to ergotamine, the most likely biologically active species in environments with low dielectric constants, e.g., the presumed ergotamine binding site, is the folded, hydrogen-bonded conformation observed for the neutral molecule in CDCl3 solution.

Conformational analysis of the ergot alkaloids ergotamine and ergotaminine.

Conformational analyses by 1H NMR and potential-energy calculations are reported for the ergot alkaloids ergotamine and ergotaminine, both as free bases and as the protonated species. In the neutral forms in CDCl3. two strong intramolecular hydrogen bonds fix the molecules in folded conformations, but the protonated species adopt a more extended conformation, with a single intramolecular hydrogen bond. Of the 24 alternative conformations available to ergotamine, the most likely biologically active species in environments with low dielectric constants, e.g., the presumed ergotamine binding site, is the folded, hydrogen-bonded conformation observed for the neutral molecule in CDCl3 solution.

Radioimmuntests für die Bestimmung der Peptidealkaloide Ergotamin und Ergocristin in Claviceps purpurea

Radioimmunoassays for the separate and quantitative determination of the peptide ergot alkaloids ergotamine and ergocristine were developed. The alkaloids were coupled covalently to bovine serum albumin and antibodies were raised in rabbits. 3H-labelled tracers were used. The antibodies exhibited high specificity and affinity towards these alkaloids [Ka = 0.15 × 109l/mol (ergocristine); Ka = 1.32 × 109 l/mol (ergotamine)]. Simple lysergic acid derivatives and clavines did not cross react. The measuring range was between 3.5-86 pmol (2-50 ng) of ergotamine and 0.8-33 pmol (0.5-20 ng) of ergocristine. With this assay the alkaloid distribution in sclerotia of Claviceps purpurea (Fries) Tulasne was determined, and the time course of alkaloid formation in parasitic Claviceps cultures was followed. Alkaloid rich sclerotia were selected from natural sources and successfully used in the establishment of high alkaloid yielding parasitic strains.

Ergotamine tolerance in patients with migraine.

The use of ergot for the treatment of headache was first recommended in 1898 by Thomson 1 in a medical textbook. In 1922 alkaloid ergotamine tartrate was developed, and several years after this its use in the treatment of migraine was suggested. 2 Since then it has become generally recognized as the treatment of choice in migraine. In view of its widespread and sometimes indiscriminate use, particularly in the treatment of headaches, it seems appropriate to review the value and use of this medicament. In particular we would like to discuss the phenomenon of tolerance in relation to drugs of the ergot family. PHARMACOLOGY AND PHYSIOLOGICAL EFFECTS OF ERGOT Ergot, a fungus (dried sclerotium of Claviceps purpurea) growing on the grain of cereals, is obtained commercially from infected rye. All ergot alkaloids are derived from lysergic acid and are related chemically and with respect to their actions. They occur in

BILATERAL GANGRENE OF FEET DUE TO ERGOTAMINE TARTRATE USED FOR PRURITUS OF JAUNDICE

Ergot is one of the most efficacious and commonly used drugs. Ergot itself (Claviceps purpurea), a fungus which grows on rye and certain grasses, is a variable mixture of various potent and relatively inert substances. The main active constitutents are the alkaloids ergotoxine and ergotamine, and the amines histamine and tyramine. Before the isolation of crystalline ergotamine, the official fluidextract of ergot was the preparation mainly employed. According to Rothlin,¹Nelson and Pattee²and others the alkaloid ergotamine is the most important constituent of ergot and the one the presence of which in ergot preparations should be ensured. Since the isolation of ergotamine (C₃₃H₃₅O₅N₅) by Stoll and Spiro in 1921 this alkaloid has come to be extensively used, mainly as the tartrate.³Its most important uses have been the prevention and control of postpartum hemorrhage from atonic uteri, and the

The present status of the ergot question, with particular reference to the preparations used in obstetrics and gynecology

1.1. The active substances in ergot or its preparations are histamine, tyramine, ergotoxine, and ergotamine.2.2. All of these substances will stimulate the isolated uterus, but there is no satisfactory clinical evidence for the value of tyramine or ergotoxine in obstetrics or gynecology; the evidence for histamine is somewhat questionable; adequate clinical as well as experimental evidence exists to justify the conclusion that the alkaloid ergotamine is the most important constituent of ergot, and the one whose presence in ergot preparations should be insured.3.3. An examination of a number of the ergot preparations available in the market reveals that only the U. S. P. fluid extracts contain important amounts of the active alkaloids.4.4. Only the official fluid extract or preparations definitely shown by proper methods of assay to contain the alkaloids of ergot should be used in medicine.