The Development of New Methodology for Determination of Vincristine (VCR) in Human Serum Using LC-MS/MS-Based Method for Medical Diagnostics.

In this article, we have presented the development and validation of a rapid and sensitive reversed-phase liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for the determination of vincristine (VCR) in patient serum samples. Chromatographic separation was achieved on a Kinetex® (Singapore) column using a mobile phase consisting of 25 mM acetic acid and 0.3% formic acid (A) and methanol (B) in a gradient elution mode at a flow rate of 0.3 mL/min. The VCR and internal standard (vinblastine) were monitored using the multiple reaction monitoring mode under positive electrospray ionization. The lower limit of quantification (LLOQ) was 0.67 ng/mL, and the upper limit of quantification (ULOQ) was 250 ng/mL for VCR. The calculated values of LOD and LOQ for VCR were 0.075 and 0.228 ng/mL, respectively. The calibration curve was linear over the VCR concentration range of 1.0−250 ng/mL in serum. The intra- and inter-day precision and precision were within the generally accepted criteria for the bioanalytical method (<15%). The method was successfully applied to the analysis of serum samples in clinical practice.

Copper and Zinc as Potential Biomarkers of Mood Disorders and Pandemic Syndrome.

The diagnosis of affective disorders has been the subject of constant research by clinicians from all over the world for many years. Making an appropriate diagnosis among patients suffering from mood disorders is sometimes problematic due to the personality-changing nature of patients and the similarity in the clinical picture of episodes in affective disorders. For this reason, there is a need to develop rapid and effective methods of determining biological markers that differentiate these diseases. The research was carried out with blood taken from 15 patients and 15 volunteers. The analysis of biological material for trace concentrations of zinc and copper was carried out with the use of ultrasensitive triple-quadrupole inductively coupled plasma mass spectrometry (TQ ICP-MS). The obtained results prove that the concentration of copper in the test group was lower than in the control group. For the zinc concentrations, the inverse relationship was observed. The group of patients was characterized by a higher concentration of this element than the group of healthy volunteers. Summarizing the obtained results and comparing them with the results of studies by other authors, it was found that zinc and copper may be potential biomarkers of affective disorders and pandemic syndrome.

A fully validated HPLC-UV method for determination of sulthiame in human serum/plasma samples.

Sulthiame is an old antiepileptic medicine with controversial history, whose effectiveness and safety in use have been stated in some current studies. However, there is still a need for further clinical examinations for confirmation of its usefulness and tolerability in monotherapy and add-on therapy for epilepsy of various etiologies. A fully validated RP HPLC-UV method for determination of sulthiame in serum/plasma samples using desethylatrazine as the internal standard was developed. The biological fluid was prepared for analysis by a simple precipitation method with acetonitrile. The following validation parameters of the method were determined: selectivity/specificity, linearity range (0.2-50.0 µl/ml, R2 > 0.9999), limits of detection (0.19 µl/ml) and quantification (0.58 µl/ml), precision (intra-day CV 1.06% and inter-day CV 1.25%), extraction recovery (~100%), accuracy (bias, -4.61-0.80%), carryover and ruggedness. Moreover, the stability of the medicine in plasma samples under different storage conditions was also tested. The usability of the method for clinical examinations was checked by analysis of serum samples originating from 19 patients treated with sulthiame. The proposed method is appropriate for determination of sulthiame in serum/plasma samples for drug monitoring purposes, as well as for pharmacokinetic studies.

Essential and Toxic Metals in Oral Fluid-a Potential Role in the Diagnosis of Periodontal Diseases.

Recently, many studies have investigated the relationship between the level of metals in the body and various diseases. The objective of this study was to examine any possible influence of periodontal disease upon the concentration of metals in oral fluid and blood and to explore the usability of applying cluster analysis coupled with the analysis of selected elements in oral fluid, calcium (Ca), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn), zinc (Zn), cadmium (Cd) and lead (Pb), for effectively distinguishing people affected by periodontitis from healthy individuals. The quantification of eight metals in oral fluid and blood samples was performed by two inductively coupled plasma techniques-inductively coupled plasma mass spectrometry (ICP-MS) and inductively coupled plasma optical emission spectrometry (ICP-OES). Most of the examined elements were detected at elevated concentration in the oral fluid of periodontal patients. However, the differences were statistically significant in the case of three metals: Cu, Mg and Mn (p < 0.05). Approximately, fivefold increase in the concentration of Cu, threefold-elevated levels of Mn and a twofold increase in the concentration of Mg were found in the oral fluid of the periodontal patients compared to the controls. Cluster analysis confirmed the statistical significance of the differences in the level of metals in the oral fluid between the two groups in most cases, plus enabled the correct classification of the subjects into patients and controls. The relationship between concentrations of metals and periodontal disease may in the future serve to prevent the development of such disease.

Toxicokinetics and tissue distribution of titanium in ionic form after intravenous and oral administration.

Titanium is widely used both in food and cosmetics, as well as in surgery and industry. Contrary to most studies, the present work focused on the determination of the toxicokinetic parameters of titanium in ionic form, as well as on its tissue biodistribution in rats. The animals were administered either a single intravenous dose of 6 mg Ti/kg b.w., or received the same dose orally every day for 30 days. The concentration of titanium in the serum and organs was measured by a graphite furnace atomic absorption spectrometry. Metal rapidly distributed from the circulation to the investigated organs after both routes of administration, and kidney was identified as the main target tissue, followed by liver and spleen. One month of oral exposure to Ti led to the increase of its concentration in liver, kidneys, spleen, and heart. In the intravenous study, both the highest area under concentration-time curves and the longest elimination half-life time were recorded in the kidney followed by serum, spleen and liver. The present study contributes to the knowledge of the toxicokinetics of titanium in ionic form, which may be especially useful when assessing the health risks of long-term exposure to titanium alloy implants in patients.

Metal concentrations in hair of patients with various head and neck cancers as a diagnostic aid.

Head and neck cancers are one of the most frequent cancers worldwide. This paper attempts to evaluate disturbances of homeostasis of the necessary elements (calcium, magnesium, zinc, copper, iron, manganese) and changes in the levels of toxic metals (lead, cadmium, cobalt, chromium VI) in hair of patients with head and neck cancers, as well as people without a diagnosed neoplastic disease. In order to quantify the necessary elements and toxic metals, a method using ICP-MS and ICP-OES techniques had been developed and validated. The studies have shown that patients with head and neck cancer used to drink alcohol and smoked much more frequently than healthy individuals, both in the past and presently. Statistically significant differences in concentrations of average metal content in the group of patients with head and neck cancers compared to the control group were confirmed. Significant differences in metal content between the group of patients with head and neck cancers and healthy individuals were found which enabled distinguishing between the study groups. To this end, a more advanced statistical tool, i.e. chemometrics, was used. The conducted research analyses and the use of advanced statistical techniques confirm the benefits of using alternative material to distinguish the patients with head and neck cancers from the healthy individuals.

Essential metals profile of the hair and nails of patients with laryngeal cancer.

Trace elements have an impact on numerous physiological processes. The monitoring of their levels in the organism allows you to detect not only their deficiencies, but also several illnesses. The aim of this study was to compare the levels of essential elements (calcium, magnesium, zinc, copper, iron, manganese) in hair, nails and serum of both patients with laryngeal cancer and healthy people. The determination of six metals was performed by an inductively coupled plasma mass spectrometry (ICP-MS) and an inductively coupled plasma optical emission spectrometry (ICP-OES). The concentration of essential elements in hair and nails of the control group was statistically significantly higher than in the group of patients with laryngeal cancer. In the case of serum, differences were found between the patients and controls in respect of the level of three metals. The results of principal component analysis (PCA) revealed the strong and similar clustering behavior of essential elements in hair and nails. The metals did not correlate between two alternative materials. The present study indicated that, using the level of essential elements in hair and nails as a basis, it is possible to distinguish cancer patients from healthy people. The alternative materials are independent of homeostasis and therefore seem to be more useful in the detection of diseases and mineral deficiencies in human than the classical biological materials, such as blood.

Classification models based on the level of metals in hair and nails of laryngeal cancer patients: diagnosis support or rather speculation?

The etiology of cancer is complex, and the disturbances in toxic and essential metals homeostasis are among many of the factors that lead to the development of malignancy. The aim of this study is to investigate the relationship between cancer risk and element status as well as cancer risk and external factors, such as diet, smoking and drinking habits, in order to support diagnosis of cancer. The samples of hair and nails obtained from patients with larynx cancer and healthy subjects were analyzed. Essential elements (Ca, Cr, Mg, Zn, Cu, Mn, and Fe), besides toxic metals (Cd, Co, and Pb), were determined using inductively coupled plasma atomic emission spectrometry (ICP-OES) and mass spectrometry (ICP-MS) techniques. The concentration of essential elements was from 1.5- (Zn) to 4.7-fold (Fe) higher in hair and from 2.4- to 3.3-fold higher in the nails of the control group compared to the patients, while the opposite trend was observed for the heavy metals. The differences between two groups in the level of metals (except for Zn) were statistically significant (p < 0.05). The association of cancer with metals and other factors was evaluated using various statistical methods, for which the best predictions were obtained using logistic regression, artificial neural networks and canonical discriminant analysis. The classifiers constructed using the data from a survey of diet and lifestyle, and analysis of elements in hair and nails, can be useful tools for estimating cancer risk and early screening of the disease.

Chronic treatment with zinc hydroaspartate induces anti-inflammatory and anti-ulcerogenic activity in rats.

BACKGROUND: The previous study indicated the enhancement of the anti-inflammatory effect of ketoprofen by acute and sub chronic administration of zinc hydroaspartate. METHODS: The present study examined anti-inflammatory, anti-ulcerogenic and analgesic activity induced by chronic (14 days) administration of ZHA (30 mg/kg, po), with a combination of a single administration of ketoprofen, in rats. Moreover, the zinc concentration in serum and stomach mucosa was also determined. RESULTS: Chronic ZHA po administration exhibits anti-inflammatory activity and enhanced the effect induced by ketoprofen. Likewise, ZHA administration demonstrated anti-ulcerogenic activity. While ZHA alone did not exhibit analgesic action, it enhanced the effect of ketoprofen. CONCLUSIONS: The present study demonstrated for the first time that chronic treatment with zinc salt exhibits anti-inflammatory activity. Besides, anti-ulcerogenic activity and the enhancing properties of zinc to ketoprofen induced anti-inflammatory and analgesic activity were also shown.

Urine as a material for evaluation of exposure to manganese in methcathinone users.

Chronic exposure even to low doses of manganese may lead to development of neurological syndrome similar to parkinsonism. The aim of this research is to assess the possibility of manganese poisoning based on the level of metal in the urine of long-term methcathinone users from Poland. Graphite furnace atomic absorption spectroscopy (GFAAS) was used to determine manganese in urine, while the detection of the psychoactive drugs was performed by high-performance liquid chromatography (HPLC). Results of survey on longitudinal patterns of drug use showed that users of traditional illicit drugs now turn to cheaper alternatives, such as methcathinone. Parkinsonian features were observed in almost half of methcathinone users. The subjects had a higher mean level of Mn in their urine (8.68±9.27 µg L(-1)) than the controls (4.27±1.91 µg L(-1)). The presence of numerous psychoactive substances (in unchanged forms and their metabolites) was confirmed in all of the samples, with only one exception. The elevated level of manganese in urine (in 29.2% of patients) can be used as a primary marker of recent methcathinone administration, especially in the case of long time intravenous drug users where blood sampling is complicated.

[Physiological metals in the serum, hair and nails of patients with head and neck cancer]. / Metale fizjologiczne w surowicy, we wlosach i paznokciach pacjentów z nowotworami w obrebie glowy i szyi.

Cigarette smoking and excessive alcohol drinking result in the rise of numbers of patients suffering from the head and neck cancer. Addiction to any of these stimulants carry a risk of developing a cancerogenesis process. Using them simultaniously lead not to a summary of each of those risks but multiplies them. Scientific research also indicates the important difference in the incidence of cancer in people who have never smoked cigarettes or drunk alcohol in comparison to those, whose exposure to these stimulatns was longterm - in such case, the former group had a lower percentage of developing the disease. Human body burdened with the ongoing cancer shows disturbances on various levels of the system. One of such disturbances is change of the concetration levels of physiological metals, such as calcium, magnesium, iron, copper, zinc or mangenese. They play key roles in maintaing the hormonal and ionic stability, they act as cofactors in many enzymes in metabolic processes. Diagnostic research of any deviations in levels of those essential elements enables a full estimation of a patient condition. The aim of this study was physiological metal levels evaluation in different kinds of biological material in patients with tumors of larynx, salivary glands and oral cavity and tongue. Hair and nail samples were used as examples of alternative material, beside the serum samples, which is a standard material and often used. Subjects were patients of Otolaryngology and Laryngological Oncology Clinic of Poznan University of Medical Sciences (Samodzielny Publiczny Szpital Kliniczny nr 2 im. Heliodora Swiecickiego Uniwersytetu Medycznego im. Karola Marcinkowskiego w Poznaniu) and The Head and Neck Surgery Ward of The Greater Poland Cancer Centre in Poznan. Subjects were 41 men and 18 women with tumors of larynx, salivary glands and oral cavity and tongue. The control group consisted of patients from the Otolaryngology and Laryngological Oncology Clinic of Poznan University of Medical Sciences (Samodzielny Publiczny Szpital Kliniczny nr 2 im. Heliodora Swiecickiego Uniwersytetu Medycznego im. Karola Marcinkowskiego w Poznaniu), The Head and Neck Surgery Ward of The Greater Poland Cancer Centre in Poznan and patients of Department of Endocrinology, Metabolism and Internal Medicine of Poznan University of Medical Sciences (Samodzielny Publiczny Szpital Kliniczny nr 2 im. Heliodora Swiecickiego Uniwersytetu Medycznego im. Karola Marcinkowskiego w Poznaniu) and Department of Conservative Dentistry and Periodontology Poznan University of Medical Sciences. They gave answers to the questionnaire concerning smoking habits, alcohol consumption and dietary habits, Then the samples of their serum, hair and nails were collected. After careful preparations the biological material has underwent the process of digestion, and then calcium, magnesium, iron, copper, zinc, mangenese were determined quantitatively using the method of ICP-MS. Profile of the patients who took part in the research displayed a strong correlation between tobacco smoking with alcohol drinking and appearance of larynx, salivary gland and oral cavity and tongue cancer as well as between exclusively tobacco smoking and appearance of these types of cancer. There is a higher incidence of larynx, salivary gland and oral cavity and tongue cancer when there is a deficiency of grain products or fibre in everyday diet. A higher level of calcium, magnesium, iron and manganese was found in patients' hair and nails who suffered from salivary gland cancer. According to applied Chemometric Analysis of Principal Component 1 - concentrations of iron, copper and manganese with magnesium and zinc in patients' nail samples showed strong correlation between measured variables. In patiens' hair samples measured correlation between variables was decreased - concentrations of calcium and magnesium as well as of iron and manganese were highlighted as two groups of variables which showed some correlation in this type of biological material. Further research is required to indicate which of alternative biological materials - hair or nail samples - in relation to serum, would provide a better evaluation of physiological metal levels.

[The influence of periodontal diseases on the concentration of selected metals in saliva]. / Wplyw chorób przyzebia na stezenie wybranych metali w slinie.

Analysis of elements (mainly metals) in biological materials provides a challenge for analytics. It results from complex matrix of this kind of samples and strict requirements for purity at all stages of the analytical process. Over the years many effective methods for determination of metals in body fluids have been developed, which link with searching for the association between elemental composition of human body and various diseases. The aim of the investigation was to study the usefulness of available methodology to determination of selected metals in saliva and blood of patients with periodontitis and healthy controls by two techniques" ICP-MS and ICP-OES. Next statistical analysis of the data statistical was carried out. The influence of periodontal disease upon the concentrations of selected metals (Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Pb and Zn) in saliva was examined, as well the attempt to classify samples of patients with periodontitis and healthy individuals correctly was made. Additionally mutual relations between analytes in examined materials were determined by computing the Pearson's correlation coefficient and principal component analysis (PCA).

Versatile flow injection manifold for analytical calibration.

A flow injection calibration system, originally designed and tested in our laboratory, is presented here as a versatile analytical tool serving for calibration purposes. It is characterized by a simple construction, easy operation and the possibility of offering rich measurement information with the use of a single standard solution. It is shown that depending on instrumental conditions and the composition of the calibration solutions the manifold is able to realize various calibration procedures according to interpolative, extrapolative and integration modes. As an experimental example, the determination of calcium in the cabbage sample by flame atomic absorption spectrometry has been calibrated by means of dilution method (DM) in the integration mode.

A new approach to the integrated calibration in flow injection analysis.

A new proposal how to perform the analytical procedure according to the integrated calibration method is presented. An original flow injection system has been designed for this purpose. When using only a single standard solution, the measurement information gathered during a single analytical course permits construction of four calibration graphs and calculation of as many as four independent estimations of the analyte concentration in the sample examined. As the calibration method applied integrates the set of standards method and the standard addition method, the analytical estimations may be obtained in both the interpolative and the extrapolative manner and the final result can be effectively verified in terms of accuracy. The system developed was experimentally tested on the example of spectrophotometric determination of chromium. It has been proved to be capable of saving time and reagents as well as providing reliable analytical results. Owing to the instrumental simplicity and analytical efficiency the system is expected to be useful for routine analysis.