Analysis of trace elements in blood sera of breast cancer patients by particle induced X-ray emission

Trace elemental imbalance in human beings is postulated to exert action, directly or indirectly, on the carcinogenic process. The objective of this study was to evaluate the levels of trace elements in blood sera of breast cancer patients and analyze their alteration with respect to healthy controls. This work was also intended to establish the role played by the trace elements in carcinogenic process. Particle induced X-ray emission (PIXE) technique was used for trace elemental analysis of blood sera of breast cancer patients and healthy controls. The PIXE measurements were carried out using a 2.5?MeV collimated proton beam from the 3 MV Tandem Pelletron accelerator at Institute of Physics, Bhubaneswar, India. On comparing the trace elemental content in the sera of breast cancer patients with those of control subjects, significant variations were observed in the levels of most of the trace elements. The serum levels of almost all the elements except Fe and Cu were observed to be depressed in cancer patients with respect to normal subjects. However, this variation was significant only for Ti (P?<?0.00005), Cr (P?<?0.005), Mn (P?<?0.0005), Ni (P?<?0.01), Zn (P?<?0.000001), and Se (P?<?0.05). On the other hand, significant elevations were observed in serum Fe (P?<?0.05) and Cu (P?<?0.005) levels in cancer patients. The findings presented in this paper give guidelines for future study into the possible roles and interactions of essential trace elements in the breast carcinogenic process.     

Analysis of blood serum of lung cancer patients using particle induced X-ray emission

Trace elemental imbalance in human beings is postulated to exert action, directly or indirectly, on the carcinogenic process. The objective of this study was to evaluate the levels of trace elements in blood serum samples of lung cancer patients and analyze their alteration with respect to healthy controls. Particle induced X-ray emission (PIXE), a well established method for elemental analysis, was used to identify and quantify trace elements in the blood serum samples of the studied groups. The PIXE measurements were carried out using 2.5 MeV collimated proton beam from the 3 MV Tandem Pelletron Accelerator at Ion Beam Laboratory, Institute of Physics, Bhubaneswar, India. The serum of the cancerous group displayed increased concentrations of Ti, Ni, and Cu but lowered concentrations of V, Cr, Mn, Fe, Co, Zn, and Se. Statistically significant differences were found for serum Cr (p < 0.01), Fe (p < 0.0005), Ni (p < 0.05), Cu (p < 0.00005), and Zn (p < 0.0005) between the two studied groups. The copper to zinc ratio for the lung cancer group was 2.24 ± 0.39, which was almost three times the value for normal subjects (0.77 ± 0.14). The observed alterations are discussed with respect to the possible mechanisms by which these elements might influence the carcinogenic process. Significant reduction in mortality from lung cancer can be achieved by advances in early diagnosis and implementation of multidisciplinary treatment programmes leading to improvement of survivorship and better quality of life. It is expected that similar studies from all corners of the world would ultimately lead to the development of novel therapeutic agents to treat lung cancer.     

Levels of mercury,cadmium, lead and other selected elements in canned tuna fish commercialised in Jordan

During 2010, the concentrations of mercury and eight other trace elements in 90 canned tuna samples commercialised in Jordan were determined using mercury analyser (Hydra C®) and the inductively coupled plasma-optical emission spectrometer (ICP-OES). The mean concentrations and ranges for elements analysed in mg?kg ^{? 1} (wet base) were as follows: total Hg (0.21; 0.06–0.57), Cd (0.06;?<?0.01–0.63), Pb (0.09; <0.04–0.24), total As (0.74; 0.11–1.56), Ni (0.51; 0.03–2.85), V (0.04; <0.03–0.1), Al (0.26; 0.08–1.63), Ba (0.13; 0.05–0.42) and Ag was not detected in any of the analysed samples (<0.02?mg?kg^?1). The data obtained in the present study compared well with data obtained from similar studies carried out in different parts of the world. Few samples had the mercury and cadmium levels slightly exceeding the Codex Committee on Food Additives and Contaminants draft guidelines. However, the estimated weekly intakes of these metals showed that there was no health risk associated with the consumption of the analysed canned tuna samples.     

Cre-lox66/lox71-Based Elimination of Phosphotransacetylase or Acetaldehyde Dehydrogenase Shifted Carbon Flux in Acetogen Rendering Selective Overproduction of Ethanol or Acetate

Acetogen strain Clostridium sp. MT1121 produced 300?mM acetate (p?<?0.005) and 321?mM ethanol (p?<?0.005) from synthesis gas (syngas) blend 60?% CO and 40?%?H₂. Clostridium sp. MT1121 was metabolically engineered to eliminate production of either acetate or acetaldehyde during syngas fermentation. We used Cre-lox66/lox71-based gene removal system to eliminate either phosphotransacetylase (pta), or acetaldehyde dehydrogenase (aldh). The resulted biocatalyst with eliminated pta increased ethanol yield to 610?mM (p?<?0.005). Inactivation of pta rendered only 502?mM of ethanol (p?<?0.005). The acetogen biocatalyst with eliminated aldh produced 450?mM acetate (p?<?0.005). The role of cell energy pool preservation for re-directed carbon flux is discussed. This is the first report on time- and cost-efficient gene elimination in acetogens using lox66/lox71 gene elimination system.     

Application of an ETV-ICP system for the determination of elements in human hair

When determining element contents in hair samples without sample digestion it is necessary to analyze large sample volumes in order to minimize problems of inhomogeneity of biological sample materials. Therefore an electrothermal vaporization system (ETV) is used for solid sample introduction into an inductively coupled plasma (ICP) for the determination of matrix and trace elements in hair. This paper concentrates on the instrumental aspects without time consuming sample preparation. The results obtained for optimization tests, ETV operating parameters and ICP operating parameters, are shown and discussed. Standard additions are used for calibration for the determination of Zn, Mg, and Mn in human hair. Studies including reproducibility and detection limits for chosen elements have been carried out on certified reference materials (CRMs). The determination of reproducibility (relative standard deviation (RSD) of n = 10) and detection limits (DLs) of Zn (RSD < 8.5%, DL < 0.8 μ g⁻¹), Mn (RSD < 14.1%, DL < 0.3 μ g⁻¹), and Mg (RSD < 7.4%, DL < 6.6 μ g⁻¹) are satisfactory. The concentration values found show good agreement with the corresponding certified values. Further sample preparation steps, including hair sampling, washing procedure and homogenization for hair, relating to measurements of real hair samples are described.     

Toxic mineral elements in Mytilus galloprovincialis from Sicilian coasts (Southern Italy)

Abstract

We assessed the relationship between V, Cr, Mn, Hg, As, Cd, Sn, Sb and Pb concentrations in Mytilus galloprovincialis samples from the coasts of Sicily and the expression of metallothioneins. Toxic mineral elements assessment was carried out by A.A. Spectrometry and ICP-MS. The metallothioneins expression was performed by q-PCR method. Low metals’ levels were found in the mussel samples examined, in comparison with what was reported in literature. The highest mean values of toxic mineral elements were found in Gela (Cr 0.178?±?0.03?mg/Kg, Mn 4.325?±?0.012?mg/Kg, As 3.706?±?0.009?mg/Kg, Sn 0.148?±?0.014?mg/Kg, Sb 0.009?±?0.004?mg/Kg e Pb 0.364?±?0.01?mg/Kg). Significant levels of Hg were found in samples from Catania (0.014?±?0.005?mg/Kg). Only vanadium and lead concentrations showed significant differences between sampling areas (p?<?0.05). Molecular analysis verified a basal expression of Mt1 and the absence of over-expression of Mt2, confirming the low mineral’s concentrations found in the samples examined.     

Optimisation of Batch Culture Conditions for Cell-Envelope-Associated Proteinase Production from Lactobacillus delbrueckii subsp. lactis ATCC? 7830?

Using a combination of conventional sequential techniques, the batch growth conditions for the production of cell-envelope-associated proteinases have for the first time been studied and optimised in Lactobacillus delbrueckii subsp. lactis 313 (ATCC 7830; LDL 313). Concentrations of inoculum (0.1?<?X?<?10?% vol/vol), agitation speed (0?<?S?<?200?rpm), varying incubation temperature (30?<?T?<?50?°C), starting pH (4.5?<?pH?<?7) and carbon/nitrogen ratio of production medium (0.2?<?r?<?5) had an individual effect on proteinase yield (p?<?0.01). Optimal conditions for proteinase production included an initial pH of 6.0, 45?°C incubation temperature, 2?% (v/v) inoculum size of OD₅₆₀?=?1, 150?rpm agitation speed, and growth medium carbon/nitrogen ratio of 1.0. Maximum proteinase activity obtained for whole cells was 0.99 U/ml after 8?h of incubation. The variables studied are very relevant due to their significance in improving the productivity of proteinase synthesis from LDL 313, under process and, likely, economic optimum conditions.     

Determination of the rare earth elements La, Eu, and Yb using solidified floating organic drop microextraction and electrothermal vaporization ICP-MS

We have developed a method for the determination of trace levels of the rare earth elements La, Eu, and Yb in biological and environmental samples. It is based on solidified floating organic drop microextraction using 1-(2-pyridylazo)-2-naphthol (PAN) as a chelator, followed by electrothermal vaporization (ETV) and quantification by inductively coupled plasma mass spectrometry. PAN also acts as a modifier in ETV. The effects of pH, amount of PAN, extraction time, stirring rate, volume of sample solution, and temperature program were examined. Under optimized conditions, the detection limits are 2.1, 0.65 and 0.91 pg mL^?1 for the elements La, Eu and Yb, respectively. The relative standard deviations are <6.0 % (c?=?0.1 ng mL^?1, n?=?9). When applied to the analysis of (spiked) natural water samples, the recoveries range from 92 to 105 %. The accuracy was validated with certified reference materials (combined sample of branch and leaf of shrub: GBW 07603 and human hair: GBW 07601), and the results were in good agreement with the certified values.

Determination of trace elements in blood samples of patients affected by multiple sclerosis from Iran by neutron activation analysis

Multiple sclerosis (MS) is a neurological autoimmune disease in which the immune system attacks the central nervous system for unknown reasons and causes several damages to human body by demyelinating the nerve cells. One of the possible cause of this disease is the abnormality levels of some trace elements such as Br, Fe, Rb, Sb, As, and Zn in human body. This study attempts to measure the levels of four trace elements of Br, Fe, Rb, and Zn in the patients?? blood samples and compare them with control samples from healthy individuals. It should be noted that the objectives set out partly met. According to the obtained results, the differences between the levels of Br, Fe, and Rb in patients?? blood samples and control was not significant (P?>?0.05). However, the average level of Zn between samples and controls showed a significant difference (P?<?0.05). Therefore the lower level of Zn in blood is likely to be a major cause of MS emergence. Furthermore, by using the concentration level of Zn as indicator, it was revealed that the risk of MS infection rises as the number of pregnancies increase.     

A new approach for determination of crustal and trace elements in airborne particulate matter

An ICP-OES procedure was developed for fast and accurate determination of various crustal (Al, Ca, Fe, Mg, Si) and trace elements (Ba, Cu, Mn, Na, K, Sr, Ti, Zn) in airborne particulate matter. The method is based on a preliminary treatment of the aerosol samples with a mixture of nitric acid and hydrogen peroxide at elevated temperature leading to a mineralization of the organic sampling substrate, dissolution of soluble material and homogeneous suspension of the remaining non-soluble fraction. After dilution the derived slurry solutions were measured using ICP-OES. The reproducibility of analysis given as the relative standard deviation (% RSD) varied between 3.2 and 6.8% for bulk constituents such as Al, Ca, Fe, Mg and Si whereas values ranging from 3.5 to 9.1% were obtained for trace metals present with distinctly lower abundance in PM10 (e.g. Ba, Cu, Mn, Sr, Zn). The limits of detection (LOD) calculated as three times the standard deviation (3σ) of the signal derived from filter blank samples ranged from approximately 1?ng?m^?3 (Sr) to 71?ng?m^?3(Ca). The developed procedure was evaluated by comparing the obtained results with the findings derived for the same set of aerosol samples analyzed using a microwave procedure for sample dissolution with subsequent ICP-OES analysis. Finally the developed procedure was applied for the analysis of crustal and trace elements in PM10 samples collected at an urban site (Getreidemarkt, Vienna) and a rural site (Hartberg, Styria), in Austria. The concentrations of the investigated crustal elements varied between some hundred ng?m^?3 and few µg?m^?3 with highest concentrations for Fe and Si, distinctly reduced concentrations ranging from some ng?m^?3 (Sr) to more than hundred ng?m^?3 (K) were found for trace elements. Observed PM10 concentrations were found to be in accordance to literature findings from urban sites in central Europe.     

Determination of minor, trace and toxic elements in chewing tobacco products by instrumental neutron activation analysis and identification of glutamic acid

Tobacco smoking/chewing has been a cause of concern because of it being related with oral cancer. It causes stimulation and ill physiological effects. Ten different brands of spit tobacco, eight gutkaas and five paan masalas have been analyzed for seven minor (Al, Na, K, Ca, Cl, Mg, and P) and 17 trace (As, Ba, Br, Co, Cr, Cs, Cu, Eu, Fe, Hg, La, Mn, Rb, Sb, Sc, Th, and Zn) elements by instrumental neutron activation analysis. Also Ni and Pb were determined by atomic absorption spectrophotometry. Concentration of Cd was below detection limit (<10?mg?kg^?1) in the tobacco samples. Mg, generally added as MgCO₃ to prevent caking, is present as minor constituent in spit tobacco and gutkaas but is below detection limit (<1?g?kg^?1) in paan masalas. Most elemental concentrations vary in a wide range depending on the nature of chewing tobacco. Spit tobacco has been found to be more enriched in essential elements (Ca, K, Na, P, Mn, and Rb), whereas gutkaas contain higher concentrations of Fe, Cr, Cu, and Zn. Paan masalas contain lower contents of other elements but higher content of Hg. Gutkaas also contain higher amounts of As and Pb. Further glutamic acid has been separated from tobacco leaves and characterized as it might bind with some elements.     

Occurrence of synthetic hormones in sewage effluents and Langat River and its tributaries,Malaysia

This study investigated for the first time the occurrence of selected synthetic hormones including 17α-ethinylestradiol, levonorgestrel, norethindrone and cyproterone acetate in Malaysian tropical waters. Samples were collected from the effluents of five sewage treatment plants (STPs) and at seven stations along the Langat River in Selangor, Malaysia, and its main tributaries. Samples were extracted by solid phase extraction (SPE) and analyzed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). In Langat river samples, only levonorgestrel and cyproterone acetate were found at 50% and 3.3% frequency. The concentrations detected for levonorgestrel ranged from less than the method detection limit (<MDL) to 213?ng?L^?1 and from?<?MDL to 76?ng?L^?1 for cyproterone acetate. For the STP effluent samples, levonorgestrel was <MDL in all samples and the concentrations detected were in the range?<?MDL – 262?ng?L^?1 for cyproterone acetate,?<?MDL ?11336?ng?L^?1 for norethindrone and?<?MDL – 1898?ng?L^?1 for 17α-ethinylestradiol.     

Trace element concentration in hair of Bangladesh children under normal and malnutrition conditions

The trace element concentrations of eight elements (Ca, Cr, Mn, Fe, Ni, Cu, Zn and Pb) in the scalp hair of 49 normal and 55 malnourished children of the Bangladesh population, in the age group of 1 to 5 years were analyzed by the external beam PIXE method to establish their baseline values in normal children for pre-diagnostic screening of deficiency and excess in malnutrition. The results of this investigation show that the ranges of trace elements in hair are somewhat wide. For example, the normal Zn level has the range of 63–377 g·g^–1 for 49 subjects. The reasons for these large individual deviations are unknown till now. The average of the elemental concentrations between normal and malnourished groups are compared (t-test,p<0.05). A correlation study has been done on each pair of original data by a non-parametric Speaman's rank test. The frequency distributions of the elements are presented and the results are compared with the data reported in the literature. A substantive correspondence between the present data in hair of both the study groups and the literature values from different countries have been observed but they did not show any regular dependence on the nutritional status of the subjects. Only the low Cu content in the hair of malnourished group can possibly be linked with nutritional disorders as evidenced from this study.Part of a Ph.D. Thesis.     

Oral fluid and plasma 3,4-methylenedioxymethamphetamine (MDMA) and metabolite correlation after controlled oral MDMA administration

Oral fluid (OF) offers a noninvasive sample collection for drug testing. However, 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) in OF has not been adequately characterized in comparison to plasma. We administered oral low-dose (1.0 mg/kg) and high-dose (1.6 mg/kg) MDMA to 26 participants and collected simultaneous OF and plasma specimens for up to 143 h after dosing. We compared OF/plasma (OF/P) ratios, time of initial detection (t _first), maximal concentrations (C _max), time of peak concentrations (t _max), time of last detection (t _last), clearance, and 3,4-methylenedioxyamphetamine (MDA)-to-MDMA ratios over time. For OF MDMA and MDA, C _max was higher, t _last was later, and clearance was slower compared to plasma. For OF MDA only, t _first was later compared to plasma. Median (range) OF/P ratios were 5.6 (0.1–52.3) for MDMA and 3.7 (0.7–24.3) for MDA. OF and plasma concentrations were weakly but significantly correlated (MDMA: R ²?=?0.438, MDA: R ²?=?0.197, p?<?0.0001). Median OF/P ratios were significantly higher following high dose administration: MDMA low?=?5.2 (0.1–40.4), high?=?6.0 (0.4–52.3, p?<?0.05); MDA low?=?3.3 (0.7–17.1), high?=?4.1 (0.9–24.3, p?<?0.001). There was a large inter-subject variation in OF/P ratios. The MDA/MDMA ratios in plasma were higher than those in OF (p?<?0.001), and the MDA/MDMA ratios significantly increased over time in OF and plasma. The MDMA and MDA concentrations were higher in OF than in plasma. OF and plasma concentrations were correlated, but large inter-subject variability precludes the estimation of plasma concentrations from OF.

Neutron activation analysis for determination of metal ions in biological fluids of patients after CoCrMo arthroplasty

Determination of metals and trace elements in patients with total knee or hip arthroplasty with CoCrMo alloy was performed. Blood, urine and cerebrospinal fluid (CSF) samples were analyzed and compared with samples from healthy people. Levels of Co, Cr as well as Na, Ca, Fe, Zn, Se, Rb, Sb and Br were determined by means of neutron activation analysis. The values of Cr and Co of the blood and urine measurements were elevated in patients with replacement, and according to the statistical analysis, significant differences of the elements Zn, Br, Co and Sb were found in the CSF (p?<?0.05).

     

Heart Failure and PAHs,OHPAHs, and Trace Elements Levels in Human Serum: Results from a Preliminary Pilot Study in Greek Population and the Possible Impact of Air Pollution

Cardiovascular diseases (CVDs) have been associated with environmental pollutants. The scope of this study is to assess any potential relation of polycyclic aromatic hydrocarbons (PAHs), their hydroxylated derivatives, and trace elements with heart failure via their direct determination in human serum of Greek citizens residing in different areas. Therefore, we analyzed 131 samples including cases (heart failure patients) and controls (healthy donors), and the respective demographic data were collected. Significantly higher concentrations (p < 0.05) were observed in cases’ serum regarding most of the examined PAHs and their derivatives with phenanthrene, fluorene, and fluoranthene being the most abundant (median of >50 μg L⁻¹). Among the examined trace elements, As, Cd, Cu, Hg, Ni, and Pb were measured at statistically higher concentrations (p < 0.05) in cases’ samples, with only Cr being significantly higher in controls. The potential impact of environmental factors such as smoking and area of residence has been evaluated. Specific PAHs and trace elements could be possibly related with heart failure development. Atmospheric degradation and smoking habit appeared to have a significant impact on the analytes’ serum concentrations. PCA–logistic regression analysis could possibly reveal common mechanisms among the analytes enhancing the hypothesis that they may pose a significant risk for CVD development.     

Determination of macro and trace elements in canned marine products by inductively coupled plasma—optical emission spectrometry (ICP-OES) and ICP—mass spectrometry (ICP-MS)

This study determined the concentration of eight macroelements (Na, K, Mg, P, S, Ca, Fe, Zn) and nineteen trace elements (Li, Be, Cr, Mn, Co, Ni, V, Cu, Ga, Se, Rb, Sr, In, Sn, Cs, Ba, Tl, Bi, U) in commonly consumed canned marine products from South Korea. The samples were wet-digested using nitric acid and hydrogen peroxide by a microwave system and analyzed for macroelements using inductively coupled plasma—optical emission spectrometry (ICP-OES) and for trace elements by inductively coupled plasma—mass spectrometry (ICP-MS). The analytical methods were validated by the correlation coefficients, limits of detection and quantification, correlation variance, spiking recovery tests, and analyzing a NIST 1566?b oyster tissue certified reference material. The concentrations of macro and trace elements varied among the canned marine products. The macroelements were present in the order of Na?>?K?>?P?>?S> Mg?>?Ca?>?Fe?>?Zn. In general, the concentrations of macro and trace elements were within the specified limits of Food and Nutrition Board, Joint Food and Agriculture Organization/World Health Organization Expert Committee on Food Additives, and Ministry of Food and Drug Safety, Korea. The results suggest that the analyzed canned marine products are safe in terms of the analyzed elements and their consumption therefore does not cause any threat to human health.     

Proton microprobe analysis of interplanetary dust particles

Interplanetary dust particles (IDPs) are typically <10 μm in diameter and <10^?9 g in mass on collection. The Heidelberg proton microprobe (beam spot ?2 × 3 μm²) allows non-destructive detectio of trace elements in such samples. Two IDPs (Zodiac and Bounce) were examined. To evaluate the proton-induced x-ray spectra quantitatively, the x-ray production rates for thin samples were calculated with a computer routine; the elemental ratios of the samples normalized to silicon or iron were evaluated. The ratios found for the Zodiac sample agree within a factor of two with those of cosmic abundance. The other IDP showed depletion in Ca, K, Zn and S.     

Effects of low-level gamma irradiation on the characteristics of fermented pork sausage during storage

The effect of gamma irradiation (0.5, 1, 2, and 4 kGy) on the quality of vacuum-packaged dry fermented sausages during refrigerated storage was evaluated. At Day 0 of irradiation, the pH, redness (CIE a^?), yellowness (CIE b^?), 2-thiobarbituric acid-reactive substances (TBARS) and volatile basic nitrogen (VBN) values of samples irradiated at 2 and 4 kGy were higher (p<0.05), but the CIE L^? values (lightness) were lower than those of the non-irradiated control (p<0.05). At<1 kGy irradiation, however, the pH, CIE L^?, CIE a^? and CIE b^?-value of samples were not significantly influenced by irradiation. The CIE a^?, and CIE b^?-values of samples irradiated at 2 and 4 kGy decreased with the increase of storage time. The VBN, TBARS, and CIE L^?-values of samples irradiated at 4 kGy were not changed significantly during refrigerated storage for 90 days (p>0.05). The total plate counts (TPC) and lactic acid bacteria (LAB) in the samples irradiated at 4 kGy were significantly lower (p<0.01) than those with lower irradiation doses. At the end of storage, the TPC, coliform, and LAB in the samples were not increased after irradiation at 1, 0.5 and 1 kGy, respectively. TPC and LAB were not detected in samples irradiated at 4 kGy at Day 90. In addition, no coliform bacteria were found in samples irradiated at 1 kGy during refrigerated storage. Sensory evaluation indicated that the rancid flavor of samples irradiated at 4 kGy was significantly higher, but aroma and taste scores were lower than those of the control at Day 3 of storage. Irradiation of dry fermented sausages at 2 kGy was the best conditions to prolong the shelf-life and decrease the rancid flavor without significant quality deterioration.     

Methadone concentrations in blood, plasma, and oral fluid determined by isotope-dilution gas chromatography–mass spectrometry

Methadone (MTD) is widely used for detoxification of heroin addicts and also in pain management programs. Information about the distribution of methadone between blood, plasma, and alternative specimens, such as oral fluid (OF), is needed in clinical, forensic, and traffic medicine when analytical results are interpreted. We determined MTD and its metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) in blood, plasma, blood cells, and OF by gas chromatography–mass spectrometry (GC-MS) after adding deuterium-labeled internal standards. The analytical limits of quantitation for MTD and EDDP by this method were 20 and 3 ng/mL, respectively. The amounts of MTD and EDDP were higher in plasma (80.4 % and 76.5 %) compared with blood cells (19.6 % and 23.5 %) and we found that repeated washing of blood cells with phosphate–buffered saline increased the amounts in plasma (93.6 % and 88.6 %). Mean plasma/blood concentration ratios of MTD and EDDP in spiked samples (N?=?5) were 1.27 and 1.21, respectively. In clinical samples from patients (N?=?46), the concentrations of MTD in plasma and whole blood were highly correlated (r?=?0.92, p?<?0.001) and mean (median) plasma/blood distribution ratios were 1.43 (1.41). The correlations between MTD in OF and plasma (r?=?0.46) and OF and blood (r?=?0.52) were also statistically significant (p?<?0.001) and the mean OF/plasma and OF/blood distribution ratios were 0.55 and 0.77, respectively. The MTD concentration in OF decreased as salivary pH increased (more basic). These results will prove useful in clinical and forensic medicine when MTD concentrations in alternative specimens are compared and contrasted.