Acute intraperitoneal and oral toxicity of the leaf glycosidic extract of Trigonella foenum-graecum in mice.

The present study was carried out to determine the acute toxicity of the leaf glycosidic extract of Trigonealla foenum-graecum by estimation of its medium lethal dose (LD(50)) after oral and intraperitoneal administration to mice and also to identify the target organs for its possible toxic effects. The main target organ affected among the four organs studied (liver, kidney, stomach, small and large intestine) was the liver, where early degeneration with infiltration of mononuclear and mild hepatitis was found in some animals treated with toxic doses of glycosidic extract. It is concluded that the glycosidic extract of T. foenum-graecum leaves is considered to be safe and have minimal adverse effect.

Hypoglycaemic effect of aqueous extract of the leaves of Trigonella foenum-graecum in healthy volunteers.

The safety and efficacy of Trigonella foenum-graecum extract was investigated using 20 male volunteers aged 20-30 years. They were randomly treated with either 40 mg/kg aqueous extract powder in 10 mL distilled water or 10 mL distilled water in which coffee simulated the extract. The extract significantly lowered blood glucose level by 13.4% 4 hours after ingestion. A significant change of 14.1% was observed in potassium levels. No significant alteration in serum cholesterol, total serum protein and blood urea occurred. Approximately one-third experienced feelings of hunger, frequency of micturition or dizziness during the 24 hours after ingestion. The aqueous extract effectively reduced blood glucose in normal subjects safely. Its hypokalaemic effect merits further investigation.

Hypoglycaemic and antihyperglycaemic effects of Trigonella foenum-graecum leaf in normal and alloxan induced diabetic rats.

The aqueous and alcoholic extracts of Trigonella foenum-graecum leaf were tested for hypoglycaemic activity in normal and alloxan-diabetic rats. Graded amounts (0.06, 0.2, 0.5, 1 g/kg, i.p. and 1, 2, 8 g/kg, p.o.) of the aqueous extract of Trigonella foenum-graecum leaf when given to both normal and alloxan-diabetic rats, a significant reduction of blood glucose concentration was noticed. On the other hand ethanolic extract of Trigonella foenum-graecum leaf produced no reduction in blood glucose concentration in normal rats but intra-peritoneal administration of 0.8 g/kg of the ethanolic leaf extract to diabetic rats produced a significant reduction of blood glucose concentration (p < 0.02) at 2 and 24 h only. Intraperitoneal and oral acute toxicity (LD50) and target organ effects were studied for the aqueous extract of Trigonella leaf in mice. LD50 of i.p. and oral administration were 1.9 and 10 g/kg respectively. The main organ affected after i.p. administration of the aqueous extract was the liver while oral administration of the aqueous extract of Trigonella did not produce any sign of organ damage. These results suggest that the aqueous extract of Trigonella foenum-graecum leaves given both orally and intraperitoneally possesses a hypoglycaemic effect in normoglycaemic and alloxan induced hyperglycaemic rats.

Pharmacokinetics of small doses of methotrexate in patients with psoriasis.

The pharmacokinetics of methotrexate was studied in 23 patients with chronic plaque psoriasis. Ten patients received a single oral does and 13 patients a single intramuscular injection of 25 mg methotrexate. Serum methotrexate was measured for 24 hours following administration of the drug using a magnetizable solid-phase radio-immunoassay. The disappearance of methotrexate from the serum fitted a two-compartment model with a distribution phase half-life of 1.18 +/- 0.12 h and an elimination phase half-life of 5.35 +/- 0.62 h following the oral dose and 1.45 +/- 0.22 h and 4.71 +/- 0.32 h, respectively following the intramuscular dose. Peak serum methotrexate concentrations varied greatly between patients but the mean area below the curve for the two routes of administration did not differ significantly. In a further 18 psoriatic patients receiving long-term maintenance methotrexate therapy there was no consistent relationship between salivary and serum methotrexate levels.

Fluoroimmunoassay of digitoxin in serum.

This fluoroimmunoassay for digitoxin in serum involves use of a sheep antiserum to digitoxin coupled to magnetizable solid-phase particles and fluorescein-labeled 3-O-succinyl digitoxigenin as tracer. Sodium salicylate blocks binding of the drug by binding proteins, and endogenous fluorophores and other interfering components in serum samples are reliably and completely removed at the separation and wash steps, which are facilitated by magnetic sedimentation. The method is sufficiently sensitive, precise, and specific for application to routine monitoring of digitoxin therapy, and results correlate closely (r = 0.992) with those of an established radioimmunoassay.

Fluoroimmunoassay of digoxin in serum.

A heterogeneous (solid-phase separation) fluoroimmunoassay for digoxin in serum was developed employing antibodies coupled to magnetisable cellulose/iron oxide particles and a fluorescein-labelled digoxin derivative as tracer. Intrinsic fluorophores and other potentially interfering components of serum samples were reliably and completely removed at the separation and wash steps of the assay procedure which were facilitated by magnetic sedimentation. In order to attain adequate sensitivity (detection limit 0.2 micrograms/l (0.26 nmol/l) serum digoxin), a sample volume of 500 microliters was necessary. Assay results for patients' specimens correlated well with those obtained using established charcoal--separation (r = 0.96) and magnetisable solid-phase (r = 0.95) radioimmunoassays. The feasibility of a "stat" adaptation of the fluoroimmunoassay that involved only two standards (0.5 and 4 micrograms/l digoxin) was demonstrated. The stat method would be suitable for the assay of urgent or single specimens.

Development of Fluoroimmunoassays for the determination of individual or combined levels of procainamide and N-acetylprocainamide in serum.

A fluoroimmunoassay has been developed for the simultaneous determination of serum levels of procainamide and its active metabolite N-acetylprocainamide. It employs procainamide linked through its aromatic amino group to fluorescein isothiocyanate as tracer and an antiserum raised against procainamide conjugated to human thyroglobulin through the same position. Separation is rapidly and simply achieved by covalently linking the antiserum to magnetisable microparticles and use of a magnet. Specific magnetisable particle fluorimmunoassays were also developed for procainamide and for N-acetylprocainamide by the use of suitable immunogens and fluorescein-labelled tracers. That for procainamide uses an antiserum raised to a procainamide-enzyme conjugate and fluorescein-labelled p-aminobenzoic acid while the fluoroimmunoassay for N-acetylprocainamide employs an antiserum against a N-acetylprocainamide-enzyme conjugate and fluorescein-labelled p-acetamidobenzoic acid.

Sequential fluoroimmunoassay for measurement of pregnancy specific beta 1 glycoprotein using antibody coupled to magnetisable particles.

A direct, sequential fluoroimmunoassay has been developed for the determination of serum levels of pregnancy specific beta 1 glycoprotein (SP1). The method employs rabbit anti-SP1 serum coupled to magnetisable cellulose-iron oxide particles and fluorescein-labelled SP1. Serum samples or standards are incubated with magnetisable solid phase anti-SP1 for 30 min. After magnetic sedimentation of the particles, the supernate, which includes endogenous fluorophores and other interfering factors, is discarded. Fluorescein-labelled SP1 is then added and incubated for a further 45 min; the particles are again sedimented and the fluorescence of the labelled SP1 remaining in the supernate is estimated. This reading related directly to the SP1 content of the original sample. The entire procedure, including fluorometry, is performed within a single disposable polystyrene test tube and is sufficiently simple and reliable for routine application. The sensitivity, specificity and precision is very similar to that of radioimmunoassay, and the results correlate with those of the radioimmunoassay (r = 0.9887).

Direct determination of propranolol in serum or plasma by fluoroimmunoassay.

A fluoroimmunoassay for the determination of serum of plasma levels of propranolol was developed using antibodies to propranolol coupled to magnetizable solid-phase particles and fluorescein-labeled propranolol as tracer. The method was sufficiently sensitive, precise, and specific for application to routine monitoring of propranolol therapy, and gave good correlation (r = 0.99) with a widely used ultraviolet fluorometric method in the assay of patients' specimens. The fluoroimmunoassay involves the same instrumentation as the fluorometric assay and has practical advantages, including greater sensitivity (only 100 microliters of sample required), avoidance of an extraction step, and visible-wavelength fluorometry, which permits the use of disposable plastic apparatus throughout the entire procedure.