Anthropogenic contamination in Sanka stream determination of the caffeine concentration

Cracow University of Technology, Poland From the SelectedWorks of Witold Zukowski 2011 Anthropogenic contamination in Sanka stream determination of the caffeine concentration Agnieszka Jagoda Witold Żukowski Available at: http://works.bepress.com/witold_zukowski/7/

AGNIESZKA JAGODA, WITOLD ŻUKOWSKI Anthropogenic contamination in Sanka stream derermination of the caffeine concentration Key words watercourse caffeine water pollution Abstract Caffeine is a widely consumed pharmaceutical. Despite it's easily metabolized in the organism, caffeine is present in wastewaters generated by households. When untreated wastewaters are discharged into surface waters caffeine levels can increase. Therefore, its detection in waterways can be considered as an indicator of anthropogenic pollution. This paper presents the results of determination of caffeine in Sanka and its tributaries. The Sanka stream has its origin in the areas lying to the north-west of the city of Krakow, and an outlet in the western part of the city (Bielany). Sanka is a left-bank tributary of the Vistula. Sanka is a drinking water supply for the city. Analyses were performed with applied the techniques of solid phase extraction and gas chromatography combined with a mass spectrometry. The concentration of caffeine in samples ranged from 57.9 to 488.0 ng/dm 3. 1. Introduction Caffeine is a substance with psychoactive effects. It has stimulating properties, reduces fatigue and sleepiness. It is widely used in households as an ingredient of drinks, drugs and cosmetics. It is easily metabolized according to various sources to 20% of the caffeine is removed from the body in an unchanged form [5, 11]. Caffeine travels into wastewater by throwing out unused drugs, coffee grounds, leftover tea, washing dirty dishes and coffee machines. When the sewage is discharged without further purification into the environment caffeine pollutes soil and water. There is no dominant method to mark caffeine in water samples. Every research center involved with this issue develops a unique execution technique. However, there are common elements in the various methods. The first stage of preparing a sample is extraction. It is one mgr inż., Cracow University of Technology, Faculty of Chemical Engineering and Technology, Institute of Chemistry and Inorganic Technology, e-mail: ajagoda@chemia.pk.edu.pl dr hab. inż., Cracow University of Technology, Faculty of Chemical Engineering and Technology, Institute of Chemistry and Inorganic Technology, e-mail: pczukows@pk.edu.pl 575

Jagoda A., Żukowski W.: Anthropogenic contamination in Sanka stream derermination... of the most important stages. It aims to isolate, enrich and purify the sample. The most common method is solid phase extraction. Sometimes liquid-liquid extraction is used. Solid phase extraction is done by using cartridges filled with: divinylbenzene/n-vinylpyrrolidone copolymer (Oasis HLB), octadecyl bonded silica gel (C18) or polystyrene-divinylbenzene copolymer (HR-P). The mass of the cartridges' load varies from 60 mg to 1000 mg. Acetone, dichloromethane, methanol and ethyl acetate or mixtures thereof are most commonly used for elution. The liquid-liquid extraction is done with dichloromethane and n-pentane. For separation and qualitative and quantitative determinations gas chromatography and liquid chromatography is used. Mass spectrometry can confirm the identification of caffeine in the trials [2]. In Europe caffeine was detected in the Danube River (average concentration 137 ng/dm 3, maximum concentration 1467 ng/dm 3 ) and its tributaries (average concentration 406 ng/dm 3, maximum concentration 6798 ng/dm 3 ), the Seine (from 3.2 ng/dm 3 to 186.9 ng/dm 3 ), Lipper River Germany (10 ng/dm 3 to 420 ng/dm 3 ), Hoje River Sweden (up to 430 ng/dm 3 ) [1, 4, 6]. Analysis of the waters from the Krakow rivers: Vistula, Rudawa and Wilga conducted in November and December 2010 also revealed the presence of caffeine. Concentration in the Vistula River was within the range from 522 to 974 ng/dm 3 whereas Wilga and Rudawa amounted to 367 and 326 ng/dm 3 [3]. The aim of this study is to present the results of the analysis of caffeine content in the waters streams that are sources of drinking water for residents of the city of Krakow. Sanka and its tributaries (Brzoskwinka and Potok Kaszowski) were analyzed. 2. Study area Sanka is an 18.3 km long stream with a catchment area of approx. 90 km 2 [10]. It flows through the area in the Krakow district. It is formed by combining a couple of smaller streams located over a dozen kilometers north-west of Krakow's city limits near the village Frywałd. Over its entire length it s fed by numerous small watercourses. The biggest ones are: Potok Czułowski, Potok Kaszowski and Brzoskwinka. Sanka's mouth to Vistula is located in the city of Krakow in Bielany, 1 km below the Kościuszko Barrage (Figure 1). Although the river basin lies in five communes (Czernichów, Liszki, Krzeszowice, Zabierzów and Krakow), the vast majority of the land belongs to the Liszki commune (Figure 2, Table 1). Eleven villages of the community are within the catchment area. Only three (Jeziorzany, Rączna and Ściejowice) are not included. According to The Operational Programme Infrastructure and Environment 2007-2013 on the area of Sanka s catchment is realized a project called "Providing the correct wastewater management in the basin in the Liszki community ". A sewer system has already been constructed in Liszki village. A similar system is being built and expanded in Kaszów, Mników, Morawica and Chrosna. The sewage system in Piekary is scheduled for expansion and modernization. Completion of construction works is planned for 2014 [8]. Figure 2 shows the development in Liszki s sewer system. Despite the continuous increase in the number of residents using the sewer, only 31.2% of residents have used it in 2009. Facts describing the villages located in Sanka s catchment area are collected in Table 1. Data on population and the sewer system s development was collected from the Municipal Offices. Villages located in the lower reaches of Sanka have a full sewer system available. In the middle and upper reach of the stream sewage system is only partial or not present at all. This poses a significant risk of pollution of Sanka s catchment with substances of anthropogenic origin. 576

VI Krakowska Konferencja Młodych Uczonych, Kraków 2011 Sanka has been used as a source of Krakow s drinking water since 1957. Water intake for Bielany waterworks is located a few hundred meters from the mouth of Sanka into Vistula. Initially Sanka was a complementary source of water but due to the growing contamination of Vistula in 1988 Sanka has become the only drinking water source in Krakow. This was possible when water demand in Bielany was reduced due to supply of drinking water from the Raba River [7]. Thanks to the efforts of the Municipal Water and Sewage Company in Cracow, Sanka s basin area is under environmental protection. It is prohibited dump untreated sewage into the soil or into the water within a protective belt 200 m on both sides of the river [12]. The Regional Inspectorate for Environmental Protection in Krakow monitors the water quality in Sanka. The classification is based on the Regulation of the Minister of Environment dated 27 November 2002 on requirements to be met by surface water used in water supply for human consumption (Dz.U. No. 204, pos. 1728). Table 2 includes information about Sanka water quality in 2005-2010. Unfortunately, it was classified as A3 or did not meet the requirements for categories [9]. According to the Regulation A3 rating means that the water requires a high level of physical and chemical treatment, in particular oxidation, coagulation, flocculation, decantation, filtration, adsorption on activated carbon, disinfection (ozone, final chlorination). Common indicators of poor water quality were: the number of coliforms and the number of bacteria of faecal coliform. These pollutants are anthropogenic origin, which indicate Sanka s contamination with sewage generated by households. Figure 1. Map with system of surface water and samples locations points. Rysunek 1. Mapa przedstawiająca sieć wód powierzchniowych i miejsca pobierania próbek. 577

5000 4000 3000 2000 1000 0 0 60 50 40 30 20 10 Jagoda A., Żukowski W.: Anthropogenic contamination in Sanka stream derermination... 48.9 89 Waste water discharged to sewerage system [dam3] Residential buildings connected to sewerage system [unit] Population using sewerage system [person] Persons using sewerage systems to total population [%] Length of working sewerage system [km] 5.7 10.4 16.7 172 17.4 41.1 204 24.0 7.5 40.7 283 1127 104.9 25.2 13.3 630 2031 106.3 34.9 15.1 720 2344 116.7 44.3 21.3 1043 3328 234.8 49.9 25.1 1258 3944 174.8 53.0 27.0 1368 4266 269 56.6 28.6 1471 4576 317.7 64.0 31.2 1637 5020 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 Figure 2. Chart showing the development of the sewage systems in Liszki commune in 1999-2009 [13]. Rysunek 2. Wykres przedstawiający rozwój sieci kanalizacji gminy Liszki w latach 1999-2009 [13.] Table 1. Characteristic of villages within Sanka s catchment. Tabela 1. Charakterystyka wsi położonych na terenie zlewni Sanki. Village Community Population Sewage (31.12.2010r.) system Aleksandrowic e Zabierzów 738 complete Baczyn Liszki 167 none Budzyń Liszki 332 complete Cholerzyn Liszki 978 complete Chrosna Liszki 492 none Czułów Liszki 1172 none Czółówek Czernichów 469 complete Frywałd Krzeszowic e 261 none Kaszów Liszki 2051 partial Kryspinów Liszki 1434 complete Liszki Liszki 1902 complete Mników Liszki 1161 partial Morawica Liszki 991 partial Piekary Liszki 1483 complete Source: Communal Offices in: Czernichów, Krzeszowice, Liszki, Zabierzów 578

VI Krakowska Konferencja Młodych Uczonych, Kraków 2011 Year Table 2. Water quality class in Sanka in 2005-2010 [9]. Tabela 2. Kategorie jakości wody w Sance w latach 2005-2010 [9]. Class General class according to Indicators of water quality of water quality indicators 2005 A3 2006 2007 2008 A3 2009 2010 A3 A3 A3 A3 3. Experimental 3.1. Sampling Anionic surfactants Suspension Suspension Manganese Suspension ChZT-Cr Total number of fecal coliforms Anionic surfactants Manganese Total number of fecal coliforms Suspension Total number of fecal coliforms Fecal streptococci Total number of fecal coliforms Water from Sanka stream and two of its tributaries (Brzoskwinka and Potok Kaszowski) was analyzed. Sampling points were located within the Liszki community and the Krakow city. The coordinates and description of those points can be found in Table 3. Samples were gathered 29 May 2011 in the afternoon. There was a significant weather breakdown the day before temperature dropped to c.a. 13ºC and there was intense rainfall. However, on the day the samples were collected there was spring weather: temperature at around 20ºC, no rain, and cloudless sky. 2.7 dm 3 amber glass bottles were used to contain the samples 579

Jagoda A., Żukowski W.: Anthropogenic contamination in Sanka stream derermination... Table 3. Water sampling spots. Tabela 3. Miejsca pobierania próbek wody. Sampling Geographic coordinates Creek (sampling spots) spot Latitude Longitude 19 42'17.90" Sanka (Mnikowska Valley) P1 50 4'0.23"N E 50 2'46.44" Sanka (road to a sandpit) P2 19 45'3.35"E N 50 2'44.09" 19 44'55.24" Potok Kaszowski (field) P3 N E Sanka (route no. 780, Liszki- 50 2'30.09" P4 19 47'7.51"E Kryspinów) N 50 3'32.39" Brzoskwinka (Cholerzyn) P5 19 46'5.43"E N 19 49'25.83" Sanka (Mirowska St., Kraków) P6 50 2'8.94"N E 3.2. Sample preparation As soon as the samples were transported to the laboratory the ph level (ph-meter CP- 411, ELMETRON) and the water s conductivity (microcomputer conductivity meter CC-317, ELMETRON) was measured. The results are shown in Table 4. An internal standard (Caffeine isotope 13 C 3 in methanol, Sigma Aldrich) was added to the sample. The solutions ph was elevated to 9 ph by adding a concentrated solution of sodium hydroxide (POCH). The solution was divided into three parts proportionally. Solid phase extraction was done by a VISIPRED 24TM DL (Supelco) set with 6 cm 3 cartridges containing 500 mg of C18 sorbent (J.T. Baker). The mixture of solvents ethyl acetate: acetone 1:1 (v/v) (POCH) was used as an eluent. The samples were concentrated until the solvent was completely evaporated using a vacuum concentrator (Concentrator 5301, Eppendorf). Afterwards methanol (POCH) was added and received solutions were analyzed on a gas chromatograph and mass spectrometer. Table 4. The results of measurements of ph and conductivity of water samples. Tabela 4. Wyniki pomiarów ph i przewodności badanych próbek wody. Sampling spot ph Conductivity [ms/cm] P1 8.00 0.541 P2 7.98 0.607 P3 7.79 0.831 P4 7.97 0.594 P5 7.84 0.584 P6 7.96 0.576 3.3. Gas chromatography mass spectrometry The GC/MS analyses were carried out with a Clarus 500 gas chromatograph (PerkinElmer) linked to a Clarus 500 mass spectrometer (PerkinElmer). A 30 m x 0.25 mm ID x 0.25 µm film Rtx -200MS fused silica capillary column (Restek) was used. The GC 580

VI Krakowska Konferencja Młodych Uczonych, Kraków 2011 oven temperature was programmed as follows: initial temperature of 70ºC was held for 120 s, and then raised at 0.2ºC/s to 250ºC with a hold time for 120 s. The 1 mm 3 injection volume was carried out on a splitless mode at temperature of 250ºC. The carrier gas was helium at a flow of 0.025 cm 3 /s. The mass spectrometer was working in: scan mode from 70 to 230 amu with scan time of 40 ms, sir mode m/z=194 amu of 100 ms and sir mode m/z=197 amu of 100 ms. 3.4. Results and discussion Table 5 contains the results of caffeine concentration in the water collected from different sampling spots. Caffeine was detected in all studied samples. The concentration level ranged from 55.0 to 495.2 ng of caffeine per cubic decimeter of water. The lowest concentration was found in sampling spot P1, and the highest one in sampling spot P2. Both spots are located in the Sanka stream. This significant increase of caffeine concentration between two consecutive spots is the result of lack or partial sewage system of the villages located along the stream. Other sampling spots on the Sanka revealed a lower caffeine concentration. This shows that the villages in the lower reaches of Sanka have an efficient sewer management infrastructure. Water in Potok Kaszowski and Brzoskwinka is highly polluted with caffeine. This is also caused by a lack or just partial sewage systems in the villages located near the two waterways. Recovery of the internal standard for a single trial ranged between 61 and 93% and average recovery for all trials was 79%. This proves a good selection of analytical methods used in the study in terms of their efficiency and reliability. Less than 9% relative standard deviation shows the precision of developed method. Samplin g spot Number of samples Table 5. Caffeine concentration in water samples. Tabela 5. Zawartość kofeiny w próbkach wody. Average recovery of standard [%] Caffeine concentration in water samples [ng/dm 3 ] Relative standard deviation [%] P1 3 70 57.9 7.7 P2 2 78 488.0 2.1 P3 3 87 286.3 8.6 P4 3 69 181.7 6.3 P5 3 88 334.1 6.7 P6 3 78 128.6 3.6 4. Conclusions The study of caffeine concentration in Sanka and its two tributaries: Brzoskwinka and Potok Kaszowski were done. Caffeine is an anthropogenic waste and its presence in the streams confirms that untreated sewage is discharged into watercourses. This is unsafe because water from Sanka is used as source of drinking water. The decrease of caffeine concentration in the lower reaches of Sanka is positive information as it confirms the efficiency of the sewage systems located there. 581

Jagoda A., Żukowski W.: Anthropogenic contamination in Sanka stream derermination... The values of the caffeine content in analyzed streams are comparable with results for other European rivers. The Danube River is less polluted than their tributaries, in the case of the Vistula River is the opposite. Vistula is more polluted by caffeine than Sanka. The results have been obtained during the analysis of water samples collected only once at the measuring points. It is necessary to repeat the analysis in the future. This will eliminate the influence of weather conditions. Increasing the number of sampling spots will help in the identification of pollution sources. There is also a need to take measurements after the completion of sewer construction project in the commune of Liszki. This will allow checking the efficiency of the system for collection and sewage disposal. References [1] Dsikowitzky L., Schwarzbauer J., Littke R.: The anthropogenic contribution to the organic load of the Lippe River (Germany), Part II: quantification of specific organic contaminants, Chemosphere 57, p. 1289 1300, 2004. [2] Jagoda A., Dąbrowska B., Żukowski W.: Kofeina jako wskaźnik antropogenicznego zanieczyszczenia środowiska - metody oznaczania, V Krakowska Konferencja Młodych Uczonych, Kraków, p. 255 263, 2010. [3] Jagoda A., Żukowski W., Dąbrowska B.: Kofeina w rzekach Krakowa, Czasopismo Techniczne Politechniki Krakowskiej (w druku), 2011. [4] Loos R., Locoro G., Contini S.: Occurrence of polar organic contaminants in the dissolved water phase of the Danube River and its major tributaries using SPE-LC- MS2 analysis, Water Research 44, p. 2325 2335, 2010. [5] Mandel H.G.: Update on caffeine consumption, disposition and action, Food and Chemical Toxicology 40, p. 1231 1234, 2002. [6] Togola A., Budzinski H.: Analytical development for analysis of pharmaceuticals in water samples by SPE and GC MS, Anal Bioanal. Chem. 388, p. 627 635, 2007. [7] Wierzbicki R.: Wodociągi Krakowa 1940-2000, MPWiK S.A. w Krakowie, 2001 [8] www.jrp.liszki.pl/, access: 11 June 2011 [9] www.krakow.pios.gov.pl/access/dostep10/wocena.htm, access: 11 June 2011 [10] www.krakow.pios.gov.pl/raport98/rozdz_06.html, access: 10 June 2011 [11] www.leki-informacje.pl/jamnik/img/galeriaplikow/3/etopiryna_spc.pdf, access: 10 June 2011 [12] www.liszki.pl/informator/studium/studium_zalaczniki_1_3.pdf, access: 11 June 2011 [13] www.stat.gov.pl/bdlen/app/strona.html?p_name=indeks, access: 11 June 2011 AGNIESZKA JAGODA, WITOLD ŻUKOWSKI Antropogeniczne zanieczyszczenia w strumieniu Sanka oznaczenie zawartości kofeiny Słowa kluczowe ciek wodny kofeina zanieczyszczenie wody 582

VI Krakowska Konferencja Młodych Uczonych, Kraków 2011 Streszczenie Kofeina jest powszechnie spożywanym farmaceutykiem. Pomimo dobrego metabolizowania jest składnikiem ścieków powstałych w gospodarstwach domowych. Gdy nieoczyszczone ścieki są odprowadzane do wód powierzchniowych, trafia tam wraz z nimi. Dlatego też, jej wykrycie w ciekach wodnych może być uznane za wskaźnik ich antropogenicznego zanieczyszczenia. Praca poniższa ma na celu przedstawienie wyników oznaczania zawartości kofeiny w Sance. Potok ma początek na terenach leżących na północny - zachód od miasta Krakowa, a ujście w zachodniej części miasta, na Bielanach. Jest lewobrzeżnym dopływem Wisły. Na terenie miasta jest na nim zlokalizowane jedno z ujęć wody pitnej. Analizy przeprowadzono z wykorzystaniem techniki ekstrakcji do fazy stałej i chromatografii gazowej sprzężonej ze spektrometrią mas. Wyniki oznaczeń zawartości kofeiny wyniosły od 57.9 do 488.0 ng/dm 3. 583