Development and validation of a sensitive LC-MS/MS method for the determination of fenoterol in human plasma and urine samples.

Due to the lack of sensitivity in current methods for the determination of fenoterol (Fen), a rapid LC-MS/MS method was developed for the determination of (R,R')-Fen and (R,R';S,S')-Fen in plasma and urine. The method was fully validated and was linear from 50pg/ml to 2000pg/ml for plasma and from 2.500ng/ml to 160ng/ml for urine with a lower limit of quantitation of 52.8pg/ml in plasma. The coefficient of variation was <15% for the high QC standards and <10% for the low QC standards in plasma and was <15% for the high and low QC standards in urine. The relative concentrations of (R,R')-Fen and (S,S')-Fen were determined using a chirobiotic T chiral stationary phase. The method was used to determine the concentration of (R,R')-Fen in plasma and urine samples obtained in an oral cross-over study of (R,R')-Fen and (R,R';S,S')-Fen formulations. The results demonstrated a potential pre-systemic enantioselective interaction in which the (S,S')-Fen reduces the sulfation of the active (R,R')-Fen. The data suggest that a non-racemic mixture of the Fen enantiomers may provide better bioavailability of the active (R,R')-Fen for use in the treatment of cardiovascular disease.

Metabolites of ephedrines in human urine after administration of a single therapeutic dose.

Ephedrine (EPH), pseudoephedrine (PEPH), phenylpropanolamine (PPA), methylephedrine (MEPH) and cathine are sympathomimetic amines. These drugs are commonly found in over-the-counter (OTC) cold medicines and some dietary supplements. In Taiwan, the misuse of these drugs has resulted in an increase in athletic violations. Excretion studies of the ephedrine-related drugs have been performed to better understand the metabolic yields of ephedrines in urine. After consuming a single clinical dose of each of these drugs, urine samples from volunteers (n=3 for each drug) were subjected to tert-butyl-methyl-ether (TBME) extraction and trifluoroacetic acid (TFAA) derivatization before gas chromatography-mass spectrometry (GC-MS) analysis. Most ephedrines were excreted unchanged in urine, including EPH (40.9%), PEPH (72.2%), and PPA (59.3%). However, only a relatively small amount of MEPH (15.5%) was excreted unchanged in urine. In addition, a trace amount of PPA (1.6%) and cathine (0.7%) was found to be the metabolites of EPH and PEPH, respectively. Urinary EPH, PEPH, and PPA reached peaks at 2-6h and disappeared in urine at approximately 24-48 h post-administration. For MEPH, the peaks of excretion extended from 4 to 12h post-administration and were undetectable at approximately 48 h. A single clinical dose of EPH (25 mg) may exceed threshold level (10 microg/mL) in sport drug testing if the urine samples are tested within approximately 8h post-administration. However, a single dose of MEPH (20 mg) never reached the threshold value (10 microg/mL).

Urinary catecholamine levels and bruxism in children.

This study was performed to test the hypothesis that emotionally stressful states measured by the urinary catecholamines may affect the development of bruxism. Three hundred and fourteen children, boys and girls, aged 6-8 years were included in this study. Bruxism was recorded by a clinical examination and an interview. Positive evidence of this parafunction was defined as the presence of both historical and clinical indicators. Information concerning systemic and socio-economic factors was collected by a questionnaire. A 24-h urine sample was collected for each subject and analysed by the high performance liquid chromatography technique to assay the catecholamine content. Of the total of 273 children who had a complete 24-h urine sample, 167 were identified to be with and without positive evidence of bruxism. The logistic multiple-regression analysis was carried out to test whether the presence of bruxism was affected by the variables studied; 95% probability was used. The results showed that epinephrine and dopamine had a significant and strong association with bruxism. The data therefore provide support for the concept that emotional stress is a prominent factor in the development of bruxing behaviour.

Cardiovascular changes during subgingival debridement.

During dental hygiene sessions, systolic and diastolic blood pressure and heart frequency were registered continuously with a finger manometer to investigate the possible cardiovascular effects of root planing/scaling. Ultrasonic subgingival debridement was performed for an average period of 10.6 +/- 2.1 min. Analysis of variance showed significant changes of systolic and diastolic blood pressure, heart rate and the rate pressure product during debridement. The increases in systolic and diastolic blood pressure both correlated significantly with the length of the debridement. After completion of the dental hygiene session, the urinary excretion of adrenaline was increased. The data from this study suggest that painful stimuli during ultrasonic subgingival debridement have extensive cardiovascular effects.

Metabolic and ergogenic effects of carbohydrate and caffeine beverages in tennis.

OBJECTIVE: Metabolic and ergogenic effects of carbohydrate and caffeine concentrations, common in commercial available beverages, were investigated in 16 tournament players (8 males and 8 females) during a 4 hrs interrupted tennis match (30 min rest after 150 min). METHODS: On three double-blind occasions players ingested a placebo (PLA), carbohydrate (CHO) or caffeine drink (CAF) at court changeover and during the resting period. In men (women) total intake consisted of 2.8 l (2.0 l) fluid, supplemented with 243 g (182 g) carbohydrates (CHO) or with 364 mg (260 mg) caffeine (CAF), respectively. Postexercise all players performed a ball-machine test (BMT) and a tennis-sprint test (TST). RESULTS: During match play blood glucose (GLU) was higher in CHO and did not differ between CAF and PLA. Immediately after the resting period GLU temporary declined in CHO and PLA, while no significant changes occurred in CAF. Increases of serum FFA and glycerol as well as the decrease of insulin were similar during the PLA and CAF trials and less pronounced in CHO. Postexercise urine concentrations of epinephrine and caffeine were significantly higher in CAF. Perception ratings and hitting accuracy (BMT) were not affected by treatment. CHO resulted in higher blood lactate levels during match play and a better post-exercise sprint performance (TST). Under CAF women won significantly more games than during both other treatments. CONCLUSIONS: CHO enhances tennis-specific running-speed but has no ergogenic effect on tennis performance under the conditions of our study. CAF improves glucose homeostasis at the beginning of work load after rest and may increase tennis success in women.

MB fraction of cumulative creatine kinase correlates with insulin secretion in patients with acute myocardial infarction: insulin as a possible determinant of myocardial MB creatine kinase.

To test whether insulin is a regulatory factor of myocardial MB creatine kinase content, we investigated the correlation between the ability of insulin secretion and the MB fraction of cumulative CK released in patients with acute myocardial infarction. We analyzed 18 patients who underwent successful direct angioplasty within 10 hours of the onset of their first myocardial infarction. Exclusion criteria were age more than 75 years, heart failure, severe obesity, multivessel disease, and history of diabetes mellitus. Cumulative activity of serum MB CK divided by that of total CK was defined as MB%, which was considered to represent myocardial MB CK content. Two weeks or more after the onset of myocardial infarction, 75 gm oral glucose tolerance test with serial determination of plasma glucose and serum insulin (0, 0.5, 1, 2, 3 hours) was done. Urinary and plasma catecholamines and echocardiographic left ventricular (LV) mass were measured. MB% significantly correlated with insulinogenic index (r = 0.564, p = 0.019), insulin area (r = 0.594, p = 0.012), insulin area/glucose area (r = 0.630, p = 0.007), and urinary adrenaline (r = -0.542, p = 0.025) and tended to correlate with plasma adrenaline (r = -0.431, p = 0.084). Age, body mass index, infarct size, glucose metabolism, and LV mass were not significant univariate predictors of MB%. Multivariate analysis showed that the ability of insulin secretion contributed to MB% more than catecholamines did and that insulin area/glucose area was the strongest independent predictor of MB% (t = 3.01, p = 0.015). Thus MB fraction of cumulative CK released, indicative of Myocardial MB CK distribution, strongly related to the ability of insulin secretion in subjects without overt insulin resistance. Regulation by insulin of myocardial MB CK is suggested.