Hypoglycin A Content in Blood and Urine Discriminates Horses with Atypical Myopathy from Clinically Normal Horses Grazing on the Same Pasture.

Hypoglycin A (HGA) in seeds of Acer spp. is suspected to cause seasonal pasture myopathy in North America and equine atypical myopathy (AM) in Europe, fatal diseases in horses on pasture. In previous studies, this suspicion was substantiated by the correlation of seed HGA content with the concentrations of toxic metabolites in urine and serum (MCPA-conjugates) of affected horses. However, seed sampling was conducted after rather than during an outbreak of the disease. The aim of this study was to further confirm the causality between HGA occurrence and disease outbreak by seed sampling during an outbreak and the determination of i) HGA in seeds and of ii) HGA and MCPA-conjugates in urine and serum of diseased horses. Furthermore, cograzing healthy horses, which were present on AM affected pastures, were also investigated. AM-pastures in Germany were visited to identify seeds of Acer pseudoplatanus and serum (n = 8) as well as urine (n = 6) from a total of 16 diseased horses were analyzed for amino acid composition by LC-ESI-MS/MS, with a special focus on the content of HGA. Additionally, the content of its toxic metabolite was measured in its conjugated form in body fluids (UPLC-MS/MS). The seeds contained 1.7-319.8 µg HGA/g seed. The content of HGA in serum of affected horses ranged from 387.8-8493.8 µg/L (controls < 10 µg/L), and in urine from 143.8-926.4 µg/L (controls < 10 µg/L), respectively. Healthy cograzing horses on AM-pastures showed higher serum (108.8 ± 83.76 µg/L) and urine concentrations (26.9 ± 7.39 µg/L) compared to control horses, but lower concentrations compared to diseased horses. The range of MCPA-carnitine and creatinine concentrations found in diseased horses in serum and urine were 0.17-0.65 mmol/L (controls < 0.01), and 0.34-2.05 µmol/mmoL (controls < 0.001), respectively. MCPA-glycine levels in urine of cograzing horses were higher compared to controls. Thus, the causal link between HGA intoxication and disease outbreak could be further substantiated, and the early detection of HGA in cograzing horses, which are clinically normal, might be a promising step in prophylaxis.

A validated method for quantifying atractyloside and carboxyatractyloside in blood by HPLC-HRMS/MS, a non-fatal case of intoxication with Atractylis gummifera L.

Atractyloside (ATR) and carboxyatractyloside (CATR) are diterpene glycosides that are responsible for the toxicity of several Asteraceae plants around the world. Mediterranean gum thistle (Atractylis gummifera L.) and Zulu impila (Callilepis laureola DC.), in particular, are notoriously poisonous and the cause of many accidental deaths, some suicides and even some murders. There is no current method for measuring the two toxins in biological samples that meet the criteria of specificity required in forensic medicine. We have endeavored to fill this analytical gap. Analysis was carried out using a solid-phase extraction and a high-performance liquid chromatography coupled with high-resolution tandem mass spectrometry detection. The method was validated in the whole blood with quantification limits of 0.17 and 0.15 µg/L for ATR and CATR, respectively. The method was applied to a non-fatal case of intoxication with A. gummifera. To the best of the authors' knowledge, this is the first time that a concentration of ATR and CATR in blood (883.1 and 119.0 µg/L, respectively) and urine (230.4 and 140.3 µg/L, respectively) is reported. ATR and CATR were quantified in A. gummifera roots by the standard method addition (3.7 and 5.4 mg/g, respectively).

Identification of taxine derivatives in biological fluids from a patient after attempted suicide by ingestion of yew (Taxus baccata) leaves.

The yew tree (Taxus baccata) is an evergreen conifer that is widespread over central and southern Europe. The toxic effects of this conifer and its leaves have been known since ancient times. The seeds are generally responsible for accidental intoxications in childhood, whereas the bark and the leaves are mainly used for homicidal or suicidal attempts. We investigated the metabolic pattern of taxines in a healthy 44-year-old male farmer who was admitted to Bergamo Emergency Department after attempting suicide. High-performance liquid chromatography was used to separate and identify taxine metabolites. Data reported in this paper confirmed that the patient attempted suicide by ingesting Taxus baccata leaves, which had been suggested by clinical examination. The most abundant free and conjugated taxine metabolites were characterized. The high concentration of conjugated metabolites found in urine underscores the critical role that conjugation in the liver plays in eliminating taxines and increasing the probability of the patient's survival.

Within-country variation in the ability of ruminants to degrade DHP following the ingestion of Leucaena leucocephala--a Thailand experience.

Goats fed Leucaena leucocephala (leucaena) at an experimental site in Thailand were shown to be excreting DHP in their urine. This was unexpected as earlier results from another site had shown that goats and cattle fed leucaena did not excrete DHP and so possessed DHP-degrading bacteria. Goats sampled near the earlier sample site excreted no DHP in their urine. Rumen fluid taken from these goats was successfully used to transfer DHP--degrading ability to the goats at the Experimental site some 350 km away that did not show the presence of DHP-degrading bacteria. Degradation of mimosine in-vitro and excretion of DHP in the urine ceased 72 hr after addition of rumen fluid and infusion with rumen fluid from protected goats, respectively. The situation in Thailand may not be unique. Countries where leucaena is fed should check that animals are protected. Fortunately, the ferric chloride urine test is simple to use and effective in detecting the problem and also the recovery after transfer of rumen fluid from protected animals.

Analytical aspects of diterpene alkaloid poisoning with monkshood.

A sensitive and specific method for aconitine extraction from biological samples was developed. Aconitine, the main toxic alkaloid from plants belonging to Aconitum species (family Ranunculaceae), was determined in plant material by an external standard method, and by a standard addition calibration method in biological fluids. Described here is one fatal case and five intoxications of accidental aconitine poisoning following the ingestion of aconite mistaken for an edible grass, Aruncus dioicus (Walt.) Fernald, "mountain asparagus", and Cicerbita alpina (L.) Wallroth. The aconitine content in urine was in the range 2.94 microg/mL (dead patient)-0.20 microg/mL (surviving patients), which was almost two to four times higher than that in plasma.

Analysis of toxic alkaloids in body samples.

Many plants contain toxic alkaloids which may be dangerous to humans. Despite the large number of poisonous plants, cases of fatal plant poisonings are relatively rare. The frequencies of poisonings and the plants involved are often regionally specific. Plant poisonings can be aggregated into three categories: unintended ingestions, intended ingestions, and poisoning due to abuse of plant material. Unintended ingestions often occur in children or from a mix-up of plants and mushrooms in adults. Intended ingestions are common in homicides and suicides. Increasingly common is the abuse of plants for hallucinogenic reasons. Toxicological analysis of such alkaloids may help in diagnosis of poisoning or abuse cases. This review describes the toxic alkaloids aconitine, atropine, coniine, colchicine, cytisine, dimethyltryptamine, harmine, harmaline, ibogaine, kawain, mescaline, scopolamine, and taxine, which are often involved in fatal and non-fatal poisonings. The paper summarizes the symptoms of the intoxications and reviews the methods of detection of their toxic constituents in biological fluids.

Simultaneous determination of three aconitum alkaloids in urine by LC-MS-MS.

A sensitive method has been developed for the identification and quantification of the toxic alkaloids yunaconitine, crassicauline A, and foresaconitine in urine specimens. After solid-phase extraction using Oasis MCX cartridges, the extracts were analyzed by LC-MS-MS. The limit of detection is 0.03 ng/mL urine for yunaconitine and 0.05 ng/mL urine for crassicauline A and foresaconitine; the limit of quantitation is 0.15 ng/mL urine for yunaconitine and 0.20 ng/mL urine for crassicauline A and foresaconitine. The method was employed in the analysis of the urine of a 55-year-old male who died after ingestion of herbal medicine powder made from the roots of aconite. Yunaconitine, crassicauline A and foresaconitine were identified in the urine. Crassicauline A and foresaconitine were thus identified in a biological specimen for the first time.

Confirmed Datura poisoning in a horse most probably due to D. ferox in contaminated tef hay.

Two out of a group of 23 mares exposed to tef hay contaminated with Datura ferox (and possibly D. stramonium) developed colic. The 1st animal was unresponsive to conservative treatment, underwent surgery for severe intestinal atony and had to be euthanased. The 2nd was less seriously affected, responded well to analgesics and made an uneventful recovery. This horse exhibited marked mydriasis on the first 2 days of being poisoned and showed protracted, milder mydriasis for a further 7 days. Scopolamine was chemically confirmed in urine from this horse for 3 days following the colic attack, while atropine could just be detected for 2 days. Scopolamine was also the main tropane alkaloid found in the contaminating plant material, confirming that this had most probably been a case of D. ferox poisoning. Although Datura intoxication of horses from contaminated hay was suspected previously, this is the 1st case where the intoxication could be confirmed by urine analysis for tropane alkaloids. Extraction and detection methods for atropine and scopolamine in urine are described employing enzymatic hydrolysis followed by liquid-liquid extraction and liquid chromatography tandem mass spectrometry (LC/MS/MS).

Hidden aconite poisoning: identification of yunaconitine and related aconitum alkaloids in urine by liquid chromatography-tandem mass spectrometry.

Poisoning from aconite occurs worldwide as a result of misuse of the potent plant. Laboratory investigation into suspected intoxication cases is challenging because the content of toxic aconitum alkaloids varies depending on the plant source, market processing, dosing protocol, hydrolytic degradation, and metabolic transformation. Using a triple-quadrupole tandem mass spectrometer, a group screening method was developed based on the mass-fragmentographic scheme of common aconitum alkaloids. The precursor-ion scans of m/z 105 and 135 permitted selective profiling of 14-O-benzoyl-norditerpenoids and the 14-O-anisoyl-norditerpenoids, respectively. Gradient reversed-phase liquid chromatography minimized coelution of isobaric compounds. The screening protocol was applied to a clinical investigation of suspected herbal poisoning. In total, 15 urine samples were thus screened positive for aconitum alkaloid over 5 years. The diagnoses of aconite poisoning in 11 patients were firmly established based on the known prescription history and the positive urine finding. In four patients, without aconitum herbs being listed in the herbal prescriptions, contamination of the herbal remedies by aconite was suspected to be the hidden cause of their acute poisoning. Yunaconitne, a highly toxic aconitum alkaloid, was thus identified in human urine for the first time. The group screening method of aconitum alkaloids in urine is an important diagnostic aid for acute poisoning by aconites of an unclear origin.

Epidemic dropsy: observations on pathophysiology and clinical features during the Delhi epidemic of 1998.

Epidemic dropsy results from the consumption of edible oils adulterated with Argemone mexicana oil by unscrupulous traders. Twenty consecutive 'in-door' patients of dropsy were intensively studied during the recent Delhi epidemic. Samples of edible oil used by them, their urine and their serum samples tested positive for sanguinarine on thin layer chromatography. The illness starts as a gastro-enteric illness followed by oliguria and pedal oedema. The following are often observed: cutaneous erythema with blanching and tenderness on pressure; violacious pigmentation of the skin; shortness of breath with orthopnoea; right-sided heart failure with normal left ventricle (LV) functions; as well as severe anaemia and hypoalbuminaemia. Renal function tests showed: bland urinary sediments; decreased glomerular filtration rate (GFR); mild to moderate azotaemia; acute tubular necrosis; patchy pneumonitis; moderate hypoxia with respiratory alkalosis; and restrictive ventilatory defects on blood gas analysis; and spirometry suggestive of interstitial pulmonary oedema of non-cardiogenic origin. 99mTc colloid sulphur liver scans showed colloid shift. There was marked dilatation and proliferation of dermal capillaries in the absence of significant inflammation in the biopsy specimens. Toxic alkaloids of Argemone mexicana oil induce widespread capillary dilatation and permeability causing leakage of protein rich plasma into the interstitial tissues of various organs. A hypovolaemic state is thus induced producing renal hypoperfusion which may progress to acute tubular necrosis. Interstitial fluid in alveoli causes restrictive ventilatory dysfunction with hypertension and right-sided failure with well-preserved LV function. The hepatic venous congestion induces Kupffer's cell dysfunction, which results in colloid shift on a radionuclide liver scan.

Metal concentrations in plants and urine from patients treated with traditional remedies.

A number of traditional South African herbal remedies which are associated with morbidity and mortality were analysed for selenium, manganese, copper, lead, zinc and mercury content. Few showed high levels of toxic metals, but in these the concentrations were sufficiently high as to cause concern since there is no quality control on the production of these remedies, which are the accepted form of medication for over 80% of the population. Urine samples, obtained from 65 patients admitted to hospital following treatment with a traditional herbal remedy were also analysed for metals. Only a small number of the patients had abnormally high levels of metal excretion. These data suggest that, in contrast to experience with traditional Chinese and Indian preparations, metal contamination from plants seems not to be a problem in traditional South African remedies.

Macrozamia toxicosis in a dog.

A case of Macrozamia riedlei seed poisoning is described in a young Dachshund. Vomiting and depression commenced within 6 h of ingestion; other signs that developed included severe hepatopathy, jaundice, abdominal pain that was unresponsive to analgesics, severe gastro-intestinal haemorrhage and thrombocytopenia as well as crystalluria and marrow dyserythropoiesis. The dog was euthanased 6 days after ingestion of the seeds.

Plasma and urine concentrations of atropine after the ingestion of cooked deadly nightshade berries.

BACKGROUND: Adult intoxications due to ingestion of deadly nightshade berries is uncommon. CASE REPORTS: Collective intoxication of eight persons occurred after accidental ingestion of ripened Atropa belladonna berries. Three of the four adults displayed delirious states with visual hallucinations; one patient fell into a coma and required mechanical ventilation. Four children and one adult exhibited mild peripheral anticholinergic symptoms. Kinetic data were obtained on the three hospitalized adults. DISCUSSION: The optimal intensive care for such patients is discussed.

Datura stramonium poisoning. Identification of tropane alkaloids in urine by gas chromatography-mass spectrometry.

A case of acute poisoning by ingestion of Datura stramonium infusion is reported. The patient presented with a typical anticholinergic syndrome (dryness of mouth, mydriasis, flushing, tachycardia, agitation, hallucinations) and was treated with symptomatic and supportive measures. The presence of tropane belladona alkaloids in a urine sample was demonstrated by gas chromatography-mass spectrometry.

Jamaican vomiting sickness in Toledo, Ohio.

The endemic illness of Jamaica known as ackee poisoning is reported for the first time in the United States. The toxic exposure resulted from the consumption of canned ackee. The epidemiology, diagnosis, theoretical mechanism, and possible therapy of this disease are discussed.

Determination of metoclopramide in the serum and urine of cattle.

Metoclopramide, a dopamine-2 agent, has been shown to be useful in the antagonism of fescue toxicosis in grazing steers. The determination of this drug is described for the purpose of pharmacokinetic study and consideration of potential delivery devices to combat this economically significant condition.

Metabolic and cardiac effects of Clistanthus collinus poisoning.

A prospective study of 11 cases of Clistanthus collinus leaf poisoning was undertaken to evaluate the mechanism of hypokalaemia that occurs in these cases. Continuous cardiac monitoring was done. Ventricular ectopics, ventricular tachycardia and fibrillation were the important arrhythmias. No atrioventricular blocks or other significant bradyarrhythmias were noted. The renal potassium loss was found to be very high (120 +/- 87.95 mEq/L) even in the presence of hypokalaemia and in spite of continuing drop in serum potassium concentration. Metabolic study established that renal potassium leak is the mechanism by which hypokalaemia was mediated.

Identification of the phenolic substances in bovine urine associated with oak leaf poisoning.

Six phenolic substances in bovine urine associated with oak leaf poisoning were identified by means of the ferric chloride reaction, paper chromatography and gas-liquid chromatography. The results indicate that these are neither oak tannin nor tannic acid but small molecular weight phenolic compounds. The concentration of free volatile phenols in the urine of cattle affected by oak leaves (n = 9, 26.57 +/- 11.20 mg litre-1) was significantly higher than in normal cattle (n = 9, 2.59 +/- 0.03 mg litre-1).