CHROMATOGRAPHIC TECHNIQUES IN PHARMACEUTICAL ANALYSIS IN POIAND: HISTORY AND THE PRESENCE ON THE BASIS OF PAPERS PUBLISHED IN SELECTED POLISH PHARMACEUTICAL JOURNALS IN XX CENTURY.

For a long time, chromatographic techniques and techniques related to them have stimulated the development of new procedures in the field of pharmaceutical analysis. The newly developed methods, characterized by improved metrological parameters, allow for more accurate testing of, among others, the composition of raw materials, intermediates and final products. The chromatographic techniques also enable studies on waste generated in research laboratories and factories producing pharmaceuticals and parapharmaceuticals. Based on the review of reports published in Polish pharmaceutical journals, we assessed the impact of chromatographic techniques on the development of pharmaceutical analysis. The first chromatographic technique used in pharmaceutical analysis was a so-called capillary analysis. It was applied in the 1930s to control the identity of pharmaceutical formulations. In the 1940s and 1950s, the chromatographic techniques were mostly a subject of review publications, while their use in experimental work was rare. Paper chromatography and thin layer chromatography were introduced in the 1960s and 1970s, respectively. These new analytical tools have contributed to the intensive development of research in the field of phytochemistry and the analysis of herbal medicines. The development of colunm chromatography-based techniques, i.e., gas chromatography and high performance liquid chromatography took place in the end of 20th century. Both aforementioned techniques were widely applied in pharmaceutical analysis, for example, to assess the stability of drugs, test for impurities and degradation products as well as in pharmacokinetics studies. The first decade of 21" century was the time of new detection methods in gas and liquid chromatography. The information sources used to write this article were Polish pharmaceutical journals, both professional and scientific, originating from the interwar and post-war period, i.e., "Kronika Farmaceutyczna", "Farmacja Wspólczesna", "Wiadomosci Farmaceutyczne", "Acta Poloniae Pharmaceutica", "Farmacja Polska", "Dissertationes Pharmaceuticae", "Annales UMCS sectio DDD Phamacia". The number of published works using various chromatography techniques was assessed based on the content description of individual issues of the journal "Acta Poloniae Pharmaceutica".

Anthropogenic impact of urban settlements on inorganic anions content in selected watercourses in the Subcarpathian Region of Poland.

BACKGROUND: Legislation for environmentally protecting surface waters in Poland and the EU is considered a priority because of the large human impact on this environmental feature in both highly industrialised countries as well as those that are agriculturally well developed. The biggest threats are regarded as being sewage arising from economic, industrial and agricultural pollution along with rain water run-off from fields treated with fertilizers. One of the most characteristic indicators of pollution exposure in surface waters are inorganic anions which form the principal components of town sew- age and fertilizers. OBJECTIVES: The estimate the effect that six selected sites of human settlement have on variously sized watercourses running through. The human environmental impact was based on determination of chlorides, nitrates and sulphates concentrations in such waters. MATERIALS AND METHODS: Water samples were obtained from the following rivers and towns, respectively; the Nil in Kolbuszowa, the Mleczka in Przeworsk, the San in Jaroslaw, the Wislok in Rzeszow, the Bystrzyca in Olimpow and an unnamed watercourse in Niwiska. Sampling sites were chosen at 4-6 points along each watercourse for a given locality. Analyte levels were measured by ion chromatography using the Dionex ICS 1000 instrument. RESULTS: Mean chlorides concentrations were found to vary from 8.52 (±0.17, n=3) mg/L to 78.41 (±0.19, n=3) mg/L, mean nitrates were 6.76 (±0.00, n=3) mg/L to 23.97 (±1.50, n=3) mg/L and mean sulphates from 29.89 (±1.57, n=3) mg/L to 62.48 (±2.99, n=3) mg/L. The clearest environmental effect of settlements on watercourses were observed for the small to medium sized towns of Kolbuszowa, Przeworsk and Jaroslaw in the form of frequently elevated chlorides levels from sewage. CONCLUSIONS: By designating various sampling locations, along the watercourses for measuring the human environmental impact of nearby settlements, it is possible to identify sources of river pollution and thus take appropriate remedial action, as and when required.

Analysis of selected chemical parameters in Piemontese wines.

BACKGROUND: Piemontese wines are well known and valued all over the world. The most popular of them are Barolo and Barbaresco wines. However, in Poland, they are still little known and only now are being gradually introduced to a wider range of consumers. OBJECTIVE: The aim of this study was to evaluate the content of inorganic anions, minerals, sugars and glycerol of Piemontese wines from micro-region Langhe, classified as DOCG ("Denominazione di Origine Controllata e Garantita", ie. controlled designation of origin guaranteed) and DOC ("Denominazione di Origine Controllata", ie. controlled designation of origin) products. MATERIAL AND METHODS: Seven types of red wines and one type of white wine were tested. High Performance Ion Chromatography with conductometric detection (HPLC-CD) was used to measure the content of inorganic anions, ie. fluorides, chlorides, sulfates and phosphates. Flame atomic absorption spectrometry (F-AAS) was used to measure the content of minerals, ie. magnesium, calcium, sodium, copper, potassium, zinc and iron, while High Performance Liquid Chromatography with charged aerosol detection (HPLC-CAD) was used to measure the content of glycerol and sugars, ie. fructose, glucose and sucrose. RESULTS: Our studies show that although Piemontese wines are characterized by a relatively low content of minerals in comparison with the wines from other regions, they contain a lot of ingredients that have beneficial effects for human health. Moreover, we observed that the studied wines contain particularly high concentration of inorganic ions--phosphates and fluorides. Furthermore, all tested red wines show far reaching similarities in their chemical properties, which is possibly a direct consequence of using in their production locally cultivated grape varieties. CONCLUSIONS: Analysis of the wines from the Piemont region, classified as DOCG, DOC, confirmed that these are dry wines of a high quality.

[Nitrates and nitrites content in the samples taken from the dug and drilled wells from the area of Podkarpacie region as a methemoglobinemia risk factors]. / Azotany (III) i (V) w wodzie pitnej studni kopanych i wierconych z terenu Podkarpacia jako czynniki ryzyka methemoglobinemii.

The aim of the study was to determine the nitrates and nitrites content in water samples taken from fourteen dug and drilled wells from the area of Podkarpacie, as well as a summary of the previously performed analysis. Private water intakes are not under the supervision of the State Sanitary Inspection. So in the case of exceeding the standards provided by the Regulation of the Minister of Health, regulating the requirements for drinking water, private water intakes can be a serious threat to the health of consumers. Particularly at risk are infants, in whom nitrates and especially nitrites can cause, among others, methemoglobinemia. The analysis was performed by ion chromatography method, making it possible to simultaneously determining the concentrations of nitrates and nitrites. As it turned out there was no presence of nitrites in the water of the tested wells. In five samples taken from the dug wells nitrates concentration exceeding the norm of 50 mg/L have been reported. In two cases, exceeding the nitrate concentrations were significant: 96.53 mg L and 204.65 mg/L.

Sugar and inorganic anions content in mineral and spring water-based beverages.

BACKGROUND: Carbonated and non-carbonated beverages manufactured based on mineral and spring waters have been present at the Polish market shortly, and their production and sales are regularly growing. The products have become commonly known as flavoured waters. OBJECTIVE: The aim of the work was to identify and assess the content of carbohydrates used for sweetening mineral and spring water-based beverages and to estimate a concentration of inorganic anions. MATERIAL AND METHODS: The study was undertaken for 15 mineral and spring water-based beverages subject to an analysis contents of fructose, glucose and sucrose with the high-performance liquid chromatography method with ELSD detection) and chlorides, nitrates and sulphates contents using the ion chromatography method. RESULTS: A chromatographic analysis has confirmed the total contents of sugar declared by the manufacturers. The carbohydrates identified included fructose, glucose and sucrose (added sugar). Chlorides and sulphates were found in the content of all the analysed beverages while nitrates were not determined in only one of the 15 examined beverages. CONCLUSIONS: Mass consumption of mineral and spring water-based beverages should be considered as an important source of sugar and their excessive consumption may be disadvantageous for human health. A consumer should be informed by a manufacturer about a daily dose of sugar in a portion of a drink in per cents, and the easiest way to do it is to provide GDA marks on the label. Mineral and spring water-based beverages do not pose threats to consumer health in terms of their contents of inorganic ions: chlorides, nitrates and sulphates.

The effect of tree age, daily sap volume and date of sap collection on the content of minerals and heavy metals in silver birch (Betula pendula Roth) tree sap.

The aim of this study was to determine the effect of the age of trees, daily sap volume as well as the term of tapping birch sap collected in the forest environment on the content of selected minerals (zinc, copper and manganese) and heavy metals (lead, nickel, chromium and cadmium). The study was performed on material taken from two stands (aged 34 and 84 years) in a moist broadleaved forest habitat with a dominant share of silver birch (Betula pendula Roth). The research results confirmed the presence of both nutritional essential minerals and hazardous heavy metals in the birch sap. At the same time, the content of minerals and heavy metals was found to be very variable and the differences between their concentrations, recorded on the same day of collecting in several trees of the same age group, can be even several dozen times higher. Depending on the examined elements, the factors influencing their content vary. The age of the trees determines only the manganese content; daily sap volume significantly affects the content of manganese and copper, and date of collection differentiates the content of zinc, lead, nickel and cadmium. The results may be interesting in the context of developing procedures for collecting birch sap for the purpose of obtaining raw material with beneficial nutritional values and a high level of health safety. For this reason, our recommendation for guaranteeing the health safety and high nutritional value of birch sap is to combine batches of raw material taken from as many trees as possible, and at the same time to publicize the fact that collecting birch sap from just one single tree may result in a raw material that is both dangerous and has no nutritional benefits.

Fusion of Taq DNA polymerase with single-stranded DNA binding-like protein of Nanoarchaeum equitans-Expression and characterization.

DNA polymerases are present in all organisms and are important enzymes that synthesise DNA molecules. They are used in various fields of science, predominantly as essential components for in vitro DNA syntheses, known as PCR. Modern diagnostics, molecular biology and genetic engineering need DNA polymerases which demonstrate improved performance. This study was aimed at obtaining a new NeqSSB-TaqS fusion DNA polymerase from the Taq DNA Stoffel domain and a single-stranded DNA binding-like protein of Nanoarchaeum equitans in order to significantly improve the properties of DNA polymerase. The DNA coding sequence of Taq Stoffel DNA polymerase and the nonspecific DNA-binding protein of Nanoarchaeum equitans (NeqSSB-like protein) were fused. A novel recombinant gene was obtained which was cloned into the pET-30 Ek/LIC vector and introduced into E. coli for expression. The recombinant enzyme was purified and its enzymatic properties including DNA polymerase activity, PCR amplification rate, thermostability, processivity and resistance to inhibitors, were tested. The yield of the target protein reached approximately 18 mg/l after 24 h of the IPTG induction. The specific activity of the polymerase was 2200 U/mg. The recombinant NeqSSB-TaqS exhibited a much higher extension rate (1000 bp template in 20 s), processivity (19 nt), thermostability (half-life 35 min at 95°C) and higher tolerance to PCR inhibitors (0.3-1.25% of whole blood, 0.84-13.5 µg of lactoferrin and 4.7-150 ng of heparin) than Taq Stoffel DNA polymerase. Furthermore, our studies show that NeqSSB-TaqS DNA polymerase has a high level of flexibility in relation to Mg2+ ions (from 1 to 5 mM) and KCl or (NH4)2SO4 salts (more than 60 mM and 40 mM, respectively). Using NeqSSB-TaqS DNA polymerase instead of the Taq DNA polymerase could be a better choice in many PCR applications.

[Sugar content in non-alcoholic beverages and dietary recemmendations for children and adolescents]. / Zawartosc cukrów prostych i sacharozy w napojach bezalkoholowych a zalecenia dietetyczne dla dzieci i mlodziezy.

INTRODUCTION: Increase the intake of sugars among the inhabitants of developed countries is related to, among others, increasing consumption of non-alcoholic beverages, for which the relationship with the epidemic of obesity, particularly among children and adolescents, has been proven. The most frequently cited are non-alcoholic beverages, sweetened glucose-fructose syrup, ie. colas, tonics, ice teas, lemonades. Fruit drinks, fruit juices and nectars are commonly cited as a healthy alternative to non-alcoholic beverages and, however, we do not pay attention to the high content of sugars in these products. OBJECTIVE: Determine the content of sugars in non-alcohollic beverages popular among children and adolescents. MATERIAL AND METHODS: 80 non-alcoholic beverages such as cola, tonic, lemonade, ice tea, flavored waters, fruit juices, fruit nectars and fruit drinks. Evaluation of the content of monosaccharides and sucrose was performed by high performance liquid chromatography method (HPLC). RESULTS: In the tested non-alcohollic beverages, monosaccharides ie. glucose and fructose and the disaccharide sucrose were detected in different proportions. The product with the lowest content of the total sugars content was flavored water with lemon flavor based on the mineral water (2.72 g/100 ml). In the group of fruit juices, fruit nectars and fruit drinks highest sugars content have been reported (12.94 g/100 ml for aronia nectar and 12.76 g/100ml for the juice of pomegranate and grapes). CONCLUSIONS: Significant monosaccharides and sucrose content in the tested non-alcohollic beverages tends to claim that their manufacturers should be obliged to place warnings on the labels addressed to patients suffering from disorders of carbohydrate metabolism. Educational programs for children and adolescents with diabetes should include information about the content of a large amount of sugars in fruit products: fruit juices, fruit drinks and fruit nectar.