Qualitative Evaluation of Bottled Water Stored in Polyethylene Terephtalate Based on Organic Chemical Compounds

Polyethylene terephtalate (PET) is commonly used for bottling drinking water. PET must be harmless in the sense of the migration potentially unsafe materials into its content. The quality determination of migrated organic chemicals in 15 bottled water stored in PET was performed by gas chromatography-mass spectrometry technique. Most of the organic chemical compounds including phthalate, alkyl phenol, higher alkene and organic acid were detected in the samples. However, no carcinogens and hormones were recognized in the analyzed waters. The most migrated compounds identified between 13 to 100% of bottled water. The findings of present study could be alarming for the food safety legislative establishments in Iran due to the existence of some organic compounds with adverse influence on human wellbeing. Further investigation is recommended to evaluate the risk assessment of the public health arising from the presence of these toxic contaminants in the bottled water consumed by the people.


Introduction
The production of water stored in Polyethylene terephtalate (PET) containers is steadily growing up in last decades because of the shortage in safe drinking tap water (Nawrocki et al., 2002;Leivadara et al., 2008).Regarding with the consumption increment of this product, paying attention to the quality of bottled water is necessary for consumer health (Dévier et al., 2013).PET bottles can be responsible for the development of unwanted compounds that migrate into its content (Dąrowska et al., 2003).Some degradation reactions may occur with high temperature between 200 and 300 °C in PET bottle manufacturing and sub-products such as organic chemical compounds appear in this process (Bach et al., 2012).Ingestion of hundreds of migrated packing materials may occur every day based on the consumption of water stored in PET (Grob, 2002).These chemicals cause several serious health sideeffects such as skin and sensory irritation, central nervous system depression, respiratory problems, leukemia and cancer, disturbance in kidney, liver and blood systems (Ceretti et al., 2010;Bina et al., 2012).Some studies have proved the existence of organic compounds in bottled water (Leivadara et al., 2008;Al-Mudhaf et al., 2009;Pinto and Reali 2009;Ikem, 2010;Jin et al., 2010;Amiridou and Voutsa 2011).For this reasons, international and national authorities established regulation to monitor unwanted material migrated (somehow toxic) to water (Castle, 1994;Salehi et al., 2014).In a previous study done in Isfahan region of Iran, chemical quality of 21 brands of bottled water determined, the results compared with label information on package, showed the significant difference between chemical concentration and labeling and water quality guidelines (Moazeni et al., 2013).Furthermore, in a recent study nitrate and nitrite content found in bottled water in Isfahan (Moazeni et al., 2014).To author's knowledge, no study has been conducted to examine the chemical organic compounds in bottled water distributed in this region.Therefore, we aimed a study to determine the qualitative evaluation of bottled water based on organic chemical compounds.

Sampling
The study was conducted in Isfahan, central part of Iran.This city is the most noticeable national cultural and tourist centers.A total of 15 water stored in PET bottles (two samples for each) with different brands of water packaging plants were purchased on random basis from the supermarkets and stores.The samples were opened in the Environment Research Center laboratory in Isfahan University of Medical Sciences and analyzed on the day of collection.

Chemicals and Reagents
Methanol and MTBE with GC-grade were purchased from Merck Company (Germany).A C 18 cartridge (500 mg) (Chromabond, Germany) with silica sorbent was applied for solid phase extraction.

Sample Preparation
Solid phase extraction was applied to prepare the mineral water samples.The cartridge was conditioned by 10 ml Millipore water and 5 ml methanol (99% Merck) to eliminate possible contamination and enhance the binding between organic compound and sorbent.Water sample (100 ml) was passed through cartridge under pressure 20 inches/Hg.Silica sorbent was washed with methanol under pressure 10 inches/Hg.Finally methanol was collected in specific tubes and dried by nitrogen gas in ambient temperature.Extraction was dissolved in 100 µl MTBE solvent and injected to GC/MS for analysis.All experiments were repeated two times.Blank solution was prepared in similar way without bottled water ensuring the compounds in the chromatogram of samples not contaminated with releasing of the cartridge, other materials and equipment used for extraction.Classification of identified compounds was based on the structure similarity including PEs, APs, organic acids and higher alkenes.The group "others" in our study refers to diverse compounds that was not set in definite group.

Apparatus
The Agilent Technology system consisting of a 5975C inert MSD, triple axis, detector equipped 7890A gas chromatograph and a split/ splitless injector was used for the confirmation of organic matter in samples.A fused silica column, HP-5 ms (5% phenyl-95% di methyl polysiloxane; 30 m × 0.25 mm I.D, 0.25 µm film thickness) was employed with helium (purity 99.995%) as the carrier gas at a flow rate of 1 ml/min.The column temperature was The MSD was operated with electron impact ionization in selected full scan mode.The injector port was maintained at 280°C with injection 3 ml of extracted sample in splitless mode within 2 min.The effluent of GC column was passed via a transfer line held at 290°C and fed into a 70eV electron impact ionization source at 280°C (Moghadam et al., 2012).The data were acquired and processed by data analysis software.
The glassware containers were washed and soaked with acid detergent for 24 hours and rinsed several times with distilled water to remove absorbance.

Results and Discussion
Results obtained in the present study revealed the existence of unwanted compounds from PET to water.In Table 1 the name of detected chemical compounds in water tested samples are listed by their chemical classification.Chemical compounds belonging to the classes of PEs material, APs, higher alkanes and organic acids were permanently identified in the examined water samples; however, a diverse group of chemicals were detected individually.The most common organic compounds detected between 13 to 100% of the samples.Some other compounds were rarely presented in few samples.PEs are esters of phthalic acid and mostly used as plasticizers in PET.Since PEs molecules are weakly bounded to polymers, they are susceptible to be liberated into water and beverages contacted with the packaging polymer (Amiridou & Voutsa, 2011).The probable consequence of phthalates within endocrinemediated unpleasant effects in wildlife and humans is reported over the past few decades.There is a variation among the results of PET migration studies (Amiridou & Voutsa, 2011).It was found BBP, BPA, DBP, DEHP, DEP, DMP and DNOP in bottled waters stored in polycarbonate container in Greece.Iranian data are limited with the report migration of BPA in baby bottles in Isfahan urban society (Moghadam et al., 2012;Moghadam et al., 2013).Traces of pharmaceutical substances, alkylphenols and some phthalates including DEHP were detected in two French brands of bottled natural mineral waters.Devier et al., indicated the complexity of reliable measure to qualify the contamination of mineral water samples (Dévier et al., 2013).Some derivatives of APs have already been identified in the samples analyzed of our study (Table 1).These compounds generate by oxidation of an additive called TNPP or degradation of APEOs as washing agents in bottle manufacturing.The other sources of APs could be the antioxidants used in laboratory equipment and materials such as vessels, tubes, detergents, the water itself or contamination through washing stages of bottles (Bach et al., 2012).For example, NP is usually spread in the environment because of its usage in industry as raw material or additive in epoxy resins to improve polymerization, drying and plasticity properties.Hebert et al., reported the existence of disinfection by products (N-nitrosodimethylamine, halofuranones, chlorate, formaldehyde and acetaldehyde, 2,4,6-trichlorophenol and pentachlorophenol, hydrazine and two unregulated halomethanes, dichloromethane and tetrachloromethane) with greatest potential risk like critical mutagenic, genotoxic, carincogenic and cytotoxic properties in drinking water of France.The aforementioned authors emphasized the importance of improving drinking water treatment and distribution (Hebert et al., 2010).Here we found higher alkenes compounds in samples.These compounds are defined with nine or further carbon atoms.Mixes of the normal alkenes could be measured in boiling point by gas chromatography (Hillen et al., 1982).We identified different kinds of carboxylic acids.We haven't found information on the contamination of bottled water with those compounds in literature review.Extant of haloacetic acids in drinking water as carboxylic acids which a hydrogen atom replaced by halogen reported (Hebert et al., 2010).Furthermore it is mentioned disinfection by-products occurring during the disinfection process by the reaction of disinfectants (haloacetic acids, cyanogen halides, halogenated acetonitriles, chlorinated ketones and furanones) and organic matter in water (Niri et al., 2008).VOCs in group "others" are one of the main subjects in the environment (Figure 1).The wide spread of aforementioned compounds due to highly volatile nature has raised major concerns, in particular with environmental sciences.US-EPA defines VOCs as a contributor in the creation of photochemical ozone (Chary & Fernandez-Alba, 2012).VOCs can lead to disorder in liver, kidney,

Figure 1
Figure 1 Chromatogram for one of the samples showing the presence of group "others" in current study

Qualitative Evaluation of Bottled Water Stored in Polyethylene Terephtalate Based on Organic Chemical Compounds
Afshin Ebrahimi; MalliheMoazeni; Zahra Esfandiari; Fateme Estaki; Amir Masoud Samani Majd;  Maryam Mirlohi; Zohreh Abdi Moghadam; Majid Falahati & HamidrezaPourzamani scheduled with 100 • C for 1 minute, increasing rate of 10 • C/ min to 210 • C as well as 5 • C/ min to 250 • C with ultimate temperature of 280 • C at 30 • C/ min.Overall run time of extraction was 25 min.

Qualitative Evaluation of Bottled Water Stored in Polyethylene Terephtalate Based on Organic Chemical Compounds Afshin
Ebrahimi; Mallihe Moazeni; Zahra Esfandiari; Fateme Estaki; Amir Masoud Samani Majd;  Maryam Mirlohi; Zohreh Abdi Moghadam; Majid Falahati & Hamidreza Pourzamaniin bottle waters of Kuwait in the concentration of much lower than established as safe by WHO guidelines and US-EPA maximum contaminants levels (Al-Mudhaf

Qualitative Evaluation of Bottled Water Stored in Polyethylene Terephtalate Based on Organic Chemical Compounds
Afshin Ebrahimi; MalliheMoazeni; Zahra Esfandiari; Fateme Estaki; Amir Masoud Samani Majd;  Maryam Mirlohi; Zohreh Abdi Moghadam; Majid Falahati & Hamidreza Pourzamanithe importance of bottled water stored in PET as a source of transmission of migrated organic chemical compounds to human.More studies are needed to define the risk assessment of the public health in Isfahan society through consumption of contaminated bottled water.