A pharmacokinetic model of the adjustment of phenprocoumon (Falithrom) dosage

The model presented here is based on a phenprocoumon (PPC) standard dose calculated per kg body weight by adapting it to experimentally obtained mean values of biological half-life (t1/2) and relative volume of distribution (Vrel) of PPC taken from the literature. Moreover, this dose is apt to produce a desired plasma PPC concentration leading to an optimum pharmacodynamic effect. This dose which amounts to 35.2 micrograms PPC per kg body weight will nearly be equal to the clinically used 3-mg dose related to a normal subject of 70 kg, if an absorption loss of 15 per cent is taken into consideration. By means of mathematical equations derived for this purpose, the course of plasma PPC concentration versus time curves is simulated over a time period from the administration of a standard loading dose of 376.8 micrograms PPC per kg body weight until steady state is reached. The simulation of curves is based on the presupposition that the values of t1/2 and Vrel of PPC in an individual subject do not correspond to the values of these parameters underlying the calculation of the standard dose. More than 50 per cent of the combinations of t1/2 and Vrel tested leads to the desired plasma PPC concentration at steady state. The model is suited to estimate the extent of extreme deviations of plasma PPC concentration, i.e., further accumulation of PPC or decrease in plasma PPC level, and to obtain information about the respective combinations of t1/2 and Vrel causing these deviations. Application of the basic principles of the present model to clinical practice would allow to further optimize and individualize the adjustment of multiple dosing regimen of the oral anticoagulant PPC.     

Estrone sulfate and dehydroepiandrosterone sulfate concentrations in normal subjects and men with cirrhosis.

Circulation levels of estrone sulfate (E1S) and dehydroepiandrosterone sulfate (DHAS) have been measured in plasma using a radioimmunoassay for estrone and dehydroepiandrosterone following extraction and hydrolysis of the sulfate. The mean +/- SE concentrations of E1S and DHAS in normal men were 458 +/- 25 pg/ml and 1.45 +/- 0.19 micrograms/ml, respectively. In normal women the values for days 5-7 of the cycle were 880 +/- 117 pg/ml and 1.25 +/- 0.12 micrograms/ml which were not different than the values for days 20-22 of 1195 +/- 176 pg/ml and 1.58 +/- 0.29 micrograms/ml. The mean values in post-menopausal women were 250 +/- 33 pg/ml and 0.47 +/- 0.07 micrograms/ml, both lower than the values in young women. In a group of cirrhotic men the mean values were 325 +/- 55 pg/ml and 0.38 +/- 0.12 micrograms/ml, both significantly lower than the normal values. This suggests a defect in sulfurylation in men with hepatic cirrhosis.     

Pharmacokinetics of sulfadimethoxine in cats

Pharmacokinetic parameters which describe distribution and elimination of sulfadimethoxine were determined in cats. Following intravenous administration of a single dose (55 mg/kg), disposition of the drug was described in terms of the biexponential expression: Cp = Ae-alphat + Be-betat. Based on total (free and bound) sulfonamide levels in the plasma, pseudodistribution equilibrium was slowly attained and the half-time of elimination (half-life) was 10.16 h +/- 2.50 (S.D., n = 6). Body clearance, which is the sum of all clearance processes, was 18.8 +/- 4.6 ml kg-1 h-1. Plasma protein binding, measured by equilibrium dialysis at sulfonamide concentration of 50 microgram/ml, was extensive (87.5% +/- 6.3, n =10). Computer-generated curves for an animal representative of the group, based on individual rate constants associated with the two-compartment open model, showed that 12% and 5% of the dose were present in the central and peripheral compartments, respectively, 24 h after administering the drug. A satisfactory dosage regimen might consist of a priming dose (55 mg/kg) and maintenance dosage (27.5 mg/kg at 24 h dosage intervals). Predicted plasma sulfadimethoxine concentrations would oscillate between 125 and 25 microgram/ml during the steady state. Influence of bacterial disease and febrile states on predicted levels remains to be verified experimentally.     

Age-related characteristics of ceftazidime pharmacokinetics in children]

To reveal possible age-dependent variations in the ceftazidime pharmacokinetics, the drug plasma concentrations were determined by HPLC in 10 children aged 2 to 13 years with peritonitis. The blood specimens were collected 0.25, 0.5, 1, 3, 6 and 8 hours after intravenous bolus administration of ceftazidime (Kefadym, Eli Lilly) in a single dose of 20 mg/kg. The mean values of the model-independent parameters were: total clearance (Cl), 3.3 +/- 0.8 ml/min.kg; steady-state distribution volume, 0.32 +/- 0.06 ml/kg; mean residence time, 1.7 +/- 0.4 hours. The C-coordinate of the gravity center was equal to 26 +/- 7 mg/l. A noticeable age-dependent decrease in Cl was detected by comparing the Cl estimates in our study for the children aged 7.0 +/- 3.0 years with earlier findings in children aged 12 years as well as in adults (18 and 26 years) and elderly patients (77 years): 2.5, 2.2, 2.0 and 1.1 ml/min.kg, respectively. A similar trend was observed for the ceftazidime volume of distribution (Varea). Due to the described reduction in Cl and Varea the age-induced changes in the half-life of ceftazidime were negligible. The age-dependent differences in ceftazidime pharmacokinetics should be taken into account in designing rational dosage regimens for the drug administration.     

Determination of procarbazine in human plasma by liquid chromatography with electrospray ionization mass spectrometry

Procarbazine is a cytotoxic chemotherapeutic agent used in the treatment of lymphomas and brain tumors. Its pharmacokinetic behavior remains poorly understood even though more than 30 years have elapsed since the drug was approved for clinical use. To characterize the pharmacokinetics of procarbazine in brain cancer patients during a phase I trial, a method for determining the drug in human plasma by reversed-phase high-performance liquid chromatography (HPLC) with electrospray ionization mass spectrometry (ESI-MS) was developed and thoroughly validated. Plasma samples were prepared for analysis by precipitating proteins with trichloroacetic acid and washing the protein-free supernatant with methyl tert-butyl ether to remove excess acid. The solution was separated on a Luna C-18 analytical column using methanol-25 mM ammonium acetate buffer, pH 5.1 (22:78, v/v) as the mobile phase at 1.0 ml/min. A single-quadrupole mass spectrometer with an electrospray interface was operated in the selected-ion monitoring mode to detect the [M+H](+) ions at m/z 222.2 for procarbazine and at m/z 192.1 for the internal standard (3-dimethylamino-2-methylpropiophenone). Procarbazine and the internal standard eluted as sharp, symmetrical peaks with retention times (mean+/-S.D.) of 6.3+/-0.1 and 9.9+/-0.3 min, respectively. Calibration curves of procarbazine hydrochloride in human plasma at concentrations ranging from 0.5 to 50 ng/ml exhibited excellent linearity. The mean absolute recovery of the drug from plasma was 102.9+/-1.0%. Using a sample volume of 150 microl, procarbazine was determined at the 0.5 ng/ml (1.9 nM) lower limit of quantitation with a mean accuracy of 105.2% and an interday precision of 3.60% R.S.D. on 11 different days over 5 weeks. During this same time interval, the between-day accuracy for determining quality control solutions of the drug in plasma at concentrations of 2.0, 15 and 40 ng/ml ranged from 97.5 to 98.2% (mean+/-S.D., 97.9+/-0.4%) and the precision was 3.8-6.2% (mean+/-S.D., 5.1+/-1.2%). Stability characteristics of the drug were thoroughly evaluated to establish appropriate conditions to process, store and prepare clinical specimens for chromatographic analysis without inducing significant chemical degradation. The sensitivity achieved with this assay permitted the plasma concentration-time profile of the parent drug to be accurately defined following oral administration of standard doses to brain cancer patients.     

Ciprofloxacin concentrations in serum, bone and bone marrow of rabbits

Ciprofloxacin concentrations were determined in serum, bone and bone marrow of rabbits. Four experimental groups of animals were examined: group A (n = 6) received a dosage of 60 mg/kg/day intramuscularly for 4 weeks, groups B (n = 6), C (n = 15) and D (n = 15) received dosages of 120 mg/kg/day subcutaneously for 2 days, 2 weeks, and 4 weeks, respectively. In the kinetic portion of the study, peak serum concentrations of ciprofloxacin measured at the 15 min sampling time were: 2.61 +/- 0.27 micrograms/ml in the 60 mg/kg/day group (group A) and 3.24 +/- 0.78 micrograms/ml in the 120 mg/kg/day group (group B). At necropsy, rabbits in group A had mean ciprofloxacin concentrations of 3.60 +/- 2.27 micrograms/ml in serum, 2.24 +/- 1.19 micrograms/g in marrow and 1.19 +/- 0.44 micrograms/g in bone. Rabbits in group B achieved mean levels of 4.02 +/- 1.23 micrograms/ml in serum, 2.48 +/- 0.79 micrograms/g in marrow, and 1.35 +/- 0.40 micrograms/g in bone. Rabbits in group C achieved mean levels of 5.65 +/- 2.16 micrograms/ml in serum, 3.74 +/- 1.33 micrograms/g in marrow and 1.92 +/- 0.94 micrograms/g in bone. Rabbits in group D achieved mean levels of 7.24 +/- 2.50 micrograms/ml in serum, 4.48 +/- 1.68 micrograms/g in marrow, and 1.93 +/- 0.54 micrograms/g in bone. Differences between mean values for the four experimental groups were not statistically significant.     

The inter- and intra-subject variabilities of plasma GH response to human growth hormone-releasing hormone (1-44) NH2 in men

The effects of intravenously given human growth hormone-releasing hormone (1-44) NH2 (hGRH-44) on growth hormone (GH) secretion were studied in normal men. A wide variability of intersubject GH response to hGRH-44 was observed. The peak plasma GH levels in response to 50, 100 and 200 micrograms hGRH-44 in 7 normal men were 9.1 +/- 3.2 ng/ml (Mean + SEM), 19.3 +/- 3.3 ng/ml and 22.4 +/- 4.0 ng/ml, respectively. Both the mean peak values for plasma GH response to 100 and 200 micrograms were significantly greater than that for 50 micrograms hGRH-44 injection (p less than 0.01), although there was no significant difference of the mean peak plasma GH values and mean concentrations at each time point, except for those at 120 min, when 100 or 200 micrograms hGRH-44 was administered. A significant difference in the mean amount of plasma GH secreted in response to hGRH-44 was observed only between 50 and 200 micrograms hGRH-44 injection (p less than 0.01). Furthermore, a dose-related plasma GH increase in response to hGRH-44 was not always observed in each subject. In contrast to the wide intersubject variability, the difference among responses of plasma GH to 100 micrograms or 200 micrograms of hGRH-44 given at multiple times separated by intervals of at least 1 week in each individual was relatively small.(ABSTRACT TRUNCATED AT 400 WORDS)     

Direct radioimmunoassay for human atrial natriuretic peptide (hANP) and its clinical evaluation

A direct radioimmunoassay for the rapid and accurate detection of human ANP from unextracted plasma is described. The sensitivity was approximately 50 pg/ml, respectively 2.5 pg/tube, the intra-assay variation 4%, and the inter-assay variation less than 12%. Rat ANP (1-28, 5-25, 5-27 and 5-28), oxydized and reduced hANP as well as plasma samples from various patients run in parallel to the 1-28 hANP standard curve. These findings imply, that the antibody primarily recognizes the mid-region (amino acids 6-25) of the intact ANP, that the C-terminal portion further increases the immunoreactivity, and that circulating plasma hANP is reliably measured. Plasma hANP ranged from 50-166 pg/ml (mean +/- SD: 98.3 +/- 44.6) in healthy individuals, there was no significant difference between samples were drawn in upright or lying position, the apparent half-life of injected hANP was 5.65 minutes. Patients with liver cirrhosis revealed significantly higher hANP levels of 244.5 +/- 173.5 pg/ml. Patients with various forms of cardiac disease had hANP concentrations ranging from 50 to 1744 pg/ml, depending at least partially on the right atrial pressure. No difference was observed if the samples were drawn from either right or left intracardial locations. Our findings with this system demonstrate that hANP is reliably measured even without prior extraction.     

Brain and cerebrospinal fluid uptake of zidovudine (AZT) in rats after intravenous injection

Uptake kinetics of zidovudine into cerebrospinal fluid (CSF) and brain tissue were determined in adult Sprague Dawley male rats after single intravenous injection of 6.7 mg/kg (25 mumol/kg). The drug kinetics in plasma followed biexponential disposition with an initial distribution half-life of approximately 11 minutes and an elimination half-life of 40 minutes. Over the plasma concentration range of 0.2 to 10 micrograms/ml, the cerebrospinal fluid to plasma ratio averaged 14.8 +/- 1.9% whereas the mean brain tissue to plasma ratio was 8.2 +/- 1.2% (uncorrected) or 2.3 +/- 1.8% (corrected) for the brain vascular space contribution. Simultaneous nonlinear regression analysis of brain, CSF and plasma concentration data indicate that the overall rate constant for efflux of drug from brain is approximately 75-fold higher and from CSF is 8-fold higher than the respective rate constants for influx. Thus, the ratio of the efflux to influx appears to be the predominant factor in determining the net accumulation of drug into CSF and brain parenchymal tissue.     

Determination of the antispasmodic agent ethaverine in human plasma by high-performance liquid chromatography

Ethaverine can be measured in the plasma of human subjects by reversed-phase high-performance liquid chromatography employing UV detection. The limit of detection was 2 ng/ml, and the precision was ± 14, ± 6 and ± 2% at concentrations of 5, 25 and 50 ng/ml respectively. A peak mean plasma drug concentration of 20 ng/ml occurred at 1.5 h after single oral doses of a capsule formulation to human subjects, and declined with a half-life of 2.9 h.     

Novel effects of PGF2 alpha on airway function in asthmatic subjects

The effects of inhaled prostaglandin F2 alpha (PGF2 alpha) have been examined in eight subjects with asthma. Incremental PGF2 alpha aerosol concentrations, ranging from 1 to 5,000 micrograms/ml, were administered at 15-min intervals. Plethysmographic specific airway conductance (sGaw), forced expiratory volume at 1 s (FEV1), and maximum expiratory flow at 50% vital capacity breathing air (Vmax50% air) and 80% He-20% O2 (Vmax50% He-O2) were measured after each dose and compared with saline control values. We observed unexpected triphasic dose-response characteristics, i.e., an initial decline in physiological variables at low concentrations (1-100 micrograms/ml), followed by improvement at intermediate concentrations (100-1,000 micrograms/ml) and a subsequent steep decline at high concentrations (1,000-5,000 micrograms/ml). Improvement in FEV1 and Vmax50% air between 100 and 1,000 micrograms/ml was associated with sGaw increases above control levels in six subjects and a significant fall in density-dependent index (Vmax50% He-O2/Vmax50% air) when compared with values before challenge and at low concentrations. Inhaled atropine (5 mg) improved prechallenge lung function but had no effect on PGF2 alpha dose-response characteristics. Intermediate PGF2 alpha concentrations given as a single dose consistently induced greater FEV1 reductions than the same concentration during graded dose challenges. Our findings are consistent with the demonstration of in vivo airway tachyphylaxis and indicate that airway effects of PGF2 alpha are far more complex than previously reported. Moreover, these novel effects suggest that, in addition to its well-known bronchoconstrictor effects, PGF2 alpha directly or indirectly causes airway relaxation, predominantly in large airways.     

Pharmacokinetics of florfenicol in cod Gadus morhua and in vitro antibacterial activity against Vibrio anguillarum

The pharmacokinetic profile of the antibacterial agent florfenicol was studied in plasma after intravenous (i.v.) injection and in plasma, muscle and liver following oral (p.o.) administration to cod Gadus morhua, held in seawater at 8 degrees C and weighing 100 to 200 g. Following i.v. injection, the plasma drug concentration-time profile showed 2 distinct phases. The plasma distribution half-life (t1/2alpha) was estimated to be 1.6 h, the elimination half-life (t1/2beta) to be 43 h, the total body clearance (ClT) to be 0.015 1 kg(-1) h(-1) and mean residence time (MRT) to be 74 h. The volume of distribution at steady state, Vd(ss), was calculated to be 1.1 l kg(-1). Following p.o. administration, the bioavailability was estimated to be 91%, the peak plasma concentrations (Cmax) to be 10.8 microg ml(-1) and the time to peak plasma concentrations (Tmax) to be 7 h. Corresponding Cmax and Tmax values were 13.0 microg g(-1) and 9 h, respectively, in muscle and 12.1 microg g(-1) and 9 h, respectively, in liver. The in vitro minimum inhibitory concentration (MIC) values of florfenicol against 3 Vibrio anguillarum strains isolated from diseased cod (A-21, HI-610, HI-618) were 0.5 microg ml(-1) for all 3 strains.     

Episodic luteinizing-hormone release in the ovariectomized female guinea pig: rapid inhibition by estrogen

Negative feedback of estrogen was investigated in ovariectomized female guinea pigs. Two weeks after ovariectomy, indwelling catheters were inserted into the jugular vein, and 3 days later, blood samples were taken every 10 min to determine the pattern of luteinizing hormone (LH) secretion. LH secretion in these guinea pigs was episodic, with a mean pulse period of 32 min. The mean pulse amplitude was 2.1 ng/ml, with mean plasma LH levels of 1.8 ng/ml. Twenty-five micrograms 17 beta-estradiol (E2), given i.v., caused a pronounced inhibition of pulsatile LH release. Twenty-five microliters of 100% ethanol (vehicle) had no effect on plasma LH values. In a second set of experiments, ovariectomized female guinea pigs were given two injections of luteinizing hormone-releasing hormone (LHRH) (1 microgram/kg BW, i.v.) separated by 30 min. Sharp rises in serum LH values were detected after each injection. A third injection of LHRH was administered after an injection of either 25 micrograms E2 or 25 microliters vehicle. In the presence of E2, the LH response was significantly (p less than 0.005) diminished, whereas the vehicle did not change the LH response to LHRH. These rapid effects of E2 on LH secretion and the pituitary responsiveness to LHRH infusion indicate that in the ovariectomized guinea pig E2 can directly block gonadotropin secretion. These findings are consistent with the hypothesis that negative feedback actions of E2 are directly on the membrane of the gonadotrope.     

Local production of IgG4 in human colostrum

Total IgG4 levels were determined in 27 colostrum and 27 plasma paired samples by using RIA techniques, and total IgG was determined on the same pairs by using radial immunodiffusion. In colostrum, the mean IgG4 level was 4.6 micrograms/ml (0.6 to 19.0), and in the plasma the mean IgG4 level was 170.5 micrograms/ml (30 to 920). IgG averaged 42.3 micrograms/ml (12 to 240) in colostrum and 7980.9 micrograms/ml (3250 to 16,000) in plasma. Of colostral IgG, 15.3% was IgG4, whereas only 3.5% of plasma IgG was IgG4. Specific IgG4 antibodies to beta-lactoglobulin, bovine serum albumin, bermuda grass, and alpha-gliadin were also assayed. In six patients, strong evidence was found for local mammary production of IgG4-specific antibodies.     

Adenosine 5'-triphosphate formation in Thiobacillus ferrooxidans vesicles by H+ ion gradients comparable to those of environmental conditions.

Vesicles prepared from iron-grown Thiobacillus ferrooxidans, and subsequently loaded with adenosine 5'-diphosphate and inorganic phosphate, produced adenosine 5'-triphosphate when subjected to H+ gradients comparable to those in the cells' normal environment (i.e., an internal pH in the range of 6.0 to 8.0 with an optimum of 7.0 to 7.8 and an external pH in the range of 2.1 to 4.1 with an optimum of 2.8). Nigericin, dicyclohexylcarbodiimide, and pentachlorophenol decreased adenosine 5'-triphosphate synthesis. Valinomycin at concentrations of 2.5 and 5.0 micrograms/ml increased adenosine 5'-triphosphate formation by 25 and 30%, respectively.     

In vitro development of rat embryos undergoing organogenesis in heparin-plasma.

This study examines the use of heparin-plasma as a culture medium for mammalian postimplantation whole-embryo culture. The growth and differentiation of head-fold rat embryo explants over 48 hours in a standard serum medium was compared with development of same stage explants over 48 hours in a plasma medium prepared using sodium heparin. Heparin disrupted the morphological differentiation of embryos, in a concentration-dependent manner, from 25 micrograms sodium heparin/ml media (i.e., 5 IU/ml media), with overall embryo growth being adversely affected from a concentration of 200 micrograms sodium heparin/ml media (i.e., 40 IU/ml media). Defects of cranial neural tube development were the first apparent structural anomalies resulting from culture in heparin media. Forebrain development was grossly abnormal and associated with failure of eye development. As the heparin concentration in media increased, the cephalic neural folds remained widely open and the edges became increasingly everted, although differentiation of the heart, otic primordia, and pharyngeal arch persisted. Similar concentration-dependent dysmorphogenic effects were seen when embryos were cultured in the standard serum media with added heparin. A minimum heparin concentration of 100 micrograms sodium heparin/ml media (i.e., 20 IU/ml media) was required to effectively inhibit coagulation of the plasma medium over the 48 hour culture period. Although embryonic growth was not adversely affected at this heparin concentration, morphological differentiation was severely disrupted. Therefore, heparin is not a suitable anticoagulant for the preparation of plasma for use in postimplantation whole-embryo culture.     

Fibronectin in fulminant hepatic failure.

Thirteen out of 17 patients with fulminant hepatic failure had plasma fibronectin concentrations below the normal range (194--472 micrograms/ml), the mean concentration in all 17 patients being 117.9 +/- SE 19.4 micrograms/ml. There was a significant negative correlation between plasma fibronectin concentration and aspartate aminotransferase activity, suggesting that fibronectin is consumed during clearance of hepatocyte debris. The reduced availability of fibronectin may be an important factor in the impaired function of Kuppfer cells in patients with fulminant hepatic failure.     

Determination of milnacipran, a serotonin and noradrenaline reuptake inhibitor, in human plasma using liquid chromatography with spectrofluorimetric detection

Milnacipran is an antidepressant drug belonging to the class of serotonin and noradrenaline reuptake inhibitors. A sensitive high performance liquid chromatographic during the development method coupled with a fluorimetric detection was set up, validated and then used routinely of the drug. After liquid-liquid extraction, milnacipran and its internal standard were analyzed by reversed-phase liquid chromatography (LC). The drug was derivatized with fluorescamine for fluorescence detection. The identity of the liquid chromatography peaks was controlled using mass spectrometry. The assay linearity was validated up to 1000 ng/ml. The limit of quantification was set at 5 ng/ml. Precision values (relative standard deviations) were lower than 5.4%, whereas the mean accuracy was higher than 95%. The extraction recoveries were higher than 70% for both milnacipran and the internal standard. In clinics, the LC-fluorescence method was routinely used to investigate the pharmacokinetics of milnacipran in patients and proved to be robust and capable of quantifying milnacipran in plasma for at least 36 h (four- to five-fold the elimination half-life).     

Removal of cadmium from aqueous system by shelled Moringa oleifera Lam. seed powder

The present study explores the unexploited sorption properties of the plant Moringa oleifera Lam. for decontamination of Cd at laboratory scale. Sorption studies using standard practices were carried out in batch experiments as functions of biomass dosage, contact time, metal concentrations, particle size and pH. Percentage sorption in each case was computed on the basis of Cd estimation using a planar NaI (TI) detector coupled to a 4K MCA (Canberra Accuspec Card with PC-AT 386). The adsorption data accurately in a Freundlich isotherm. Sorption studies resulted in the standardization of optimum conditions for removal of Cd (85.10%) as follows: biomass dosage (4.0 g), metal concentration (25 microg/ml), contact time (40 min) and volume of the test solution (200 ml) at pH 6.5. Fourier transform infrared (FTIR) spectrometry highlighted amino acid-Cd interactions responsible for sorption phenomenon. The findings open up new avenues in the removal of toxic metals by shelled Moringa oleifera seeds (SMOS) from water bodies as low cost, domestic and environmentally friendly safe technology.     

Blood and placental concentrations of cadmium, lead, and mercury in mothers and their newborns

Placental transfer of cadmium, lead and mercury was studied under the conditions of environmental exposures of pregnant women to these heavy metals. Fifty pregnant women from industrial area and a similar control group from a semirural area were examined. Cadmium, lead and total mercury levels were determined in maternal erythrocytes and plasma, in placenta, and in erythrocytes and plasma of umbilical cord blood using atomic absorption spectrophotometry. Except for the cadmium plasma concentration in the control area, levels of the three metals were higher in maternal than in cord blood. The mean plasma values (arithmetic mean) of cadmium, lead, and mercury in industrial area were 0.53, 6.37, and 0.37 micrograms.100 ml-1 in maternal blood and 0.30, 4.82, and 0.31 micrograms.100 ml-1 in cord blood. Highest values of the correlation coefficients were found between the cadmium and mercury concentrations in maternal and cord blood erythrocytes. No striking effect of the place of residence of pregnant women on the heavy metal concentrations in biological materials could be found.