Project: WETLAB-USE (B1.33.03) Network of reference Labs assessing environmental quality for water uses

European Territorial Cooperation Operational Programme Greece Bulgaria 2007-2013 Project: WETLAB-USE (B1.33.03) Network of reference Labs assessing environmental quality for water uses WP3: Joint Methodological framework for inter-laboratory Calibration ACTIONS 3.1 and 3.3 Inventory of current water monitoring capacity Inter-laboratory calibration & debugging methodology Deliverables 3.1.2 and 3.3.2 Thessaloniki March 2012 1

Preface This deliverable was prepared within the framework of the WETLABUSE project and more specifically refers to the contribution of Region of Central Macedonia in the implementation of Actions 3.1 and 3.3. The deliverable consists of two parts. The first part includes the Inventory of current water monitoring capacity of the Laboratory of Monitor & Certification of Hygiene & Safety of Shellfish in the Region of Central Macedonia (deliverable 3.1.2 of the project WETLABUSE) while the second part refers to the Inter-laboratory calibration & debugging methodology (deliverable 3.3.2 of the project WETLABUSE). The WETLABUSE project is co-financed by the European Union (ERDF) and National Funds of Greece and Bulgaria. 2

TABLE OF CONTENTS PART I... 4 Action 3.1: Inventory of current water monitoring capacity 5 3.1.1 General information and chart... 6 3.1.2 Sampling, equipment and methods... 12 3.1.3 Reports and communication... 32 PART II... 35 Action 3.3: Inter- laboratory calibration & debugging methodology... 36 Work Instruction 1.1: Heavy Metal Analysis... 37 Work Instruction 1.2: Physicochemical Parameter Estimation by UV-VIS spectrophotometer... 48 Work Instruction 1.3: Ion Analysis by Ion Chromatography... 55 Work Instruction 1.4: Water ph / EC Measurement... 62 Work Instruction 1.5: Phytoplankton Quality and Quantity Estimation... 66 Work Instruction 2: Critical Material Control... 72 Work Instruction 3: Laboratories Internal Quality Control... 73 Work Instruction 4: Testing Uncertainty Estimation... 77 Work Instruction 5: Method Validation... 80 Work Instruction 6: Standard Weights Handling and Control Balance Control... 84 Work Instruction 7: Use and Clearness of Glass Volumetric Equipment... 86 Work Instruction 8: Sampling from Rivers / Streams... 88 ΕΚΤΕΤΑΜΕΝΗ ΕΛΛΗΝΙΚΗ ΠΕΡΙΛΗΨΗ... 97 3

PART I 4

Action 3.1: Inventory of current water monitoring capacity In the past years, a consistent and systematic research work in the chemical analysis field took place, something that is keeping since today. Many innovative technologies have been produced while methods of analysis have been developed. All the above lead to continuous increasing accuracy and reliability in parallel with less errors and lower detecting limits. Thus, any chemical laboratory has to improve operation and decrease problems and deficiency of human resources, equipment, methods and any other parameter related to the laboratory operation. For that reason, a detailed inventory should be carried out so as every partial detail related to laboratory operation and performance to be recorded. The daily certification of the good operation of the laboratory via the recording of the methods used and the comparison with the international certified methods, the training of the personnel, the accuracy and the reliability of the results etc. can prove a laboratory s quality. The above procedure can make easily detectable the better and the weaker operations of the laboratory and consequently the chances for improving. So a detailed inventory of the Laboratory of Monitor & Certification of Hygiene & Safety of Shellfish in the Region of Central Macedonia was performed, so as the desired continuous improvement be achieved. 5

3.1.1 General information and chart The Laboratory of Monitor & Certification of Hygiene & Safety of Shellfish in the Region of Central Macedonia (R.C.M.), was financed under the Operational Programme for Fisheries 2000-2006 (Measure 4.4.: Measures implemented by professionals) under the Ministry No. 155802/5-5-2006 decision to bring the project the Directorate of Fisheries Application of General / Directorate of Fisheries Ministry of Rural Development and Food. The Laboratory of Monitor & Certification of Hygiene & Safety of Shellfish in the Region of Central Macedonia was established so as to ensure the hygiene conditions that rule the production and sell of shellfish in Greece and al over Europe. The aim of the Region of Central Macedonia was to structure an organization so as to support the productivity of shellfish, to secure the public health and to protect consumers via a self controlling routine system used by producers and dealers. Moreover, the Region of Central Macedonia could support the productive activity of shellfish, which in recent years faced several problems with biotoxins, protect public health and protect consumers, through the elimination of pathological risks posed by shellfish in some cases and improve the management capacity of producers in tackling crucial situations, such as the presence of toxic phytoplankton. The specific laboratory was considered to be the tool for this self controlling routine system so as to support the producers and dealers services and any other complementary work needed by the Region of Central Macedonia or other authorities. So, the aim is to create a structure which will contribute significantly in: Supporting the productive activity of shellfish, which in recent years faced several problems with biotoxins as well as to Protect public health and protect consumers, through the elimination of pathological risks posed by shellfish in some cases The reasons for creating the Laboratory are: Improving the health and safety of products produced of shellfish To improve the production and the competitiveness of products To improve the management capacity of producers in tackling crucial situations, such as the presence of toxic phytoplankton To generate added value The complementarities and synergy with other existing projects and projects that are designed to develop within the sector 6

Laboratory was installed in an existed building of the Ministry of Rural Development and Food, into the Port of Thessaloniki zone, at 26 th Oktovriou Street (picture 1). Picture 1: Map view The building was used as to insecticide imported rural products. Today, a part of the building is used by the quality control department of the Region of Central Macedonia. Region of Central Macedonia is represented according to the Greek low by a General Manager, responsible for the laboratories operation. The administrative department of the organization is located at Vasilisis Olgas 198 Street. An external view and the label of the laboratory can be seen in pictures 2,3. 7

Picture 2: External view Picture 3: Label pictures 4,5,6. An ichnography of the site and an office view can be seen in the following Picture 4: Ground floor Picture 5: First floor 8

Picture 6: Office view The Laboratory Monitor & Certification of Hygiene & Safety of Shellfish belongs to the Region of Central Macedonia and specifically to the Department of Fisheries Directorate of Agriculture and of Thessaloniki. However there has been no placement of personnel in the respective positions. The administrative structure of the laboratory is listed below: General Manager: G. Oikonomidis, Dr, biologist Administrative Manager: Associate Administrative Manager: Laboratory Manager: Associate Laboratory Manager: Secretariat and Online Support: Financial Management the Laboratory: Sample Preparation and Processing: Physicochemical Analyses and Analyses of Metals / analyst #1: Physicochemical Analyses and Analyses of Metals / analyst #2: Microbiological Analysis / analyst #1: Microbiological Analysis / analyst #2: Phytoplankton analysis /analyst #1: Phytoplankton analysis / analyst #2: 9

Administrative Manager The Administrative Managers is responsible for the logistic operation of the laboratory and cares to ensure the proper operation of it but do not interfere with the technical function. More specifically, he/she is responsible for the publication, revision and distribution of documents, the appropriate storage of all records, the staff training, the evaluation of suppliers, the monitoring of effectiveness, the implementation of the corrective actions and every other factor that affects laboratories good performance and quality. The minimum requirement concerning the qualifications of the administrative manager is an agronomist or biologist or chemist or chemical engineer degree. A good knowledge and use of a computer (word, excel, etc.) and of the English language (level of at least first certificate) is necessary. Also, experience of at least one (1) year in a position of laboratory administrative or five (5) years at a similar laboratory is needed. A good knowledge of ΕΝ ISO/IEC 17025 quality system is desired. Laboratory Manager The laboratory manager is responsible for selecting and supplying the appropriate equipment, materials, methods and services for carrying out the testing in the laboratory in accordance with existing requirements. Also, he/she has to prepare and continuous update the written instructions for the use, control and calibration of equipment, to maintain the necessary calibration/verification of equipment wherever needed and to protect it from damage or misuse. Finally, the overseeing the handling of objects (samples) during tests and the supervision of the work performed by analysts and technical support is necessary. The minimum requirement concerning the qualifications of the laboratory manager is a chemist or chemical engineer degree. A good knowledge and use of a computer (word, excel, etc.) and of the English language is necessary. Also, experience of at least one (1) year in a similar laboratory is needed. A good knowledge of the methods and procedures performed in the laboratory and of handling laboratory equipment is desired. 10

Analysts The analysts are responsible for performing the testing and handling the equipment exactly as described in the Guidelines for Laboratories while completing and signing the work forms for the testing that they are authorized to perform. Also they should perform the preventive maintenance and controlling of the equipment, which they have been authorized, in parallel with the implementation of any other work, which is indirectly related to testing and for which they are authorized. Some of this work could be the management of the items to be tested, the record of the environmental conditions, the transfer and control data in the test reports, the records controlling, etc. The minimum requirement concerning the qualifications of the physicochemical parameters analysts is a chemist or chemical engineer or agronomist or biologist or other technological education degree. For microbiological parameters analysts, a chemist or agronomist or biologist or other technological education degree is necessary. For biological parameters analysts, an agronomist or biologist or an ichthyologist or other technological education degree is required. All analysts should have experience of at least one (1) year in a similar laboratory is needed. A good knowledge of the methods and procedures performed in the laboratory and of handling laboratory equipment is desired. 11

3.1.2 Sampling, equipment and methods The wide range of work carried out during the process of taking samples requires considerable coordination among team members and good planning to avoid errors and omissions. For this reason, the sampling team members should be properly trained and have received this preparation and carrying out guide as well as a description of the sampling area, a site design and information on any special features of the region (e.g., key holders, security and protection.). The selections of the sampling point as well as the time at which the samples are collected are very important parameters. Samples are usually collected directly into sample collecting bottles, as this method poses less risk of contamination of the sample. For the purposes of sampling a series of further auxiliary materials and security materials are used such as plastic sample collecting container, plastic funnel, towing straps, plastic boots, full body rubber suits, rubber boots, respiratory masks, medical gloves and first aid kit. By the end of sampling, the samples should be checked if sealed properly and are placed in the maintenance and transportation refrigerator. Laboratory equipment does not contain any sampling equipment at present, so the type of water bodies, the sampling methods and parameters to be measured cannot be defined. The housing, energy and radiation sources, light, heating and ventilation favor the performance of accurate testing. Rooms are not used for activities that may affect testing accuracy. The laboratory monitors, checks and records the environmental conditions (temperature and/or humidity) that are contained at the standard methods used for testing and affect the quality of the results. Testing is stopped when the environmental conditions downgrade testing results. When two different activities take place, then a defective room separation is necessary. The access in testing rooms is defined and checked while only licensed personnel can access all laboratory rooms. This license can be seen on laboratory notice board or another visible place. An appropriate interior plan is necessary for an easy access to testing equipment used directly or indirectly. The laboratory is designed to measure parameters via samples from coastal waters. Moreover, samples from rivers, lakes, transitional waters and artificial surface water bodies or heavily modified surface water bodies can be also be tested. All tests are carried out in a laboratory base as field equipment or telemetric stations are not available. 12

Chemical and physicochemical testing is carried out to drinking water, irrigation water or other waters coming from industrial use. Also, biological and microbiological analyses can be performed as the necessary equipment is available. Equipment like the Varian atomic absorption flame and graphite spectrometers for metal analysis or the Axio Observer A1 Zeiss microscope that uses fluorescence technology for phytoplankton analysis can be characterized as the state of the art equipment of the laboratory. All the necessary benches, fitments, extractors, refrigerators, air or liquid supplies, electrical and water systems are constructed or bought for the good operation of the laboratory. Also, other basic laboratory equipment or means as volume glasses, pipettes, thermometers, washing appliances and every other needed for the good operation of the laboratory are afforded. The equipment of the Laboratory of Monitor & Certification of Hygiene & Safety of Shellfish and the type/use of each of them can be seen in the following table 1. Table 1: Laboratory equipment 1a, 1b instrument pcs type / use a) Distilled & bis-distilled water, 1 Water distillation GFL 2108, sn:10315109j b) Deionized water, Zalion 1 Water deionization 2 Rotavapor, Heidolph, Vacuum filtration & 2 Laborota 4001, sn:591-24000-00-0 Vacuum solvent condensation 3 Radwag, PS750/Y, sn:288108/10 1 Balance 4 HMC, HiClave, HV-50L, sn:30309060912 1 Sterilizer clave 5 Fiocchetti 1 Refrigerator 6 Consort C861 1 PH, EC, Salinity, TDS analyzer 7 Candy CCF200 1 Freezer 8a, 8b a) Gorenje, ST-5B50-FOT8, sn:s053f09204 1 Incubator cabinet 9 b) Lovibond Oxidirect Radwag, XA60/220, sn:242689/2008 1 B.O.D analyzer 2 Balance 13

10 Hettich Rotanda 460R 1 Centrifuge 11 WTW CR4200 1 Thermoreactor 12a, 12b 13a, 13b a) Spectroflex 6100 WTW, ULSTD6100-1 b) WTW kits a) Varian AA240 FS flame b) Varian AA240 Z graphite 1 1 1 1 Spectrophotometer Kits Spectrometer Spectrometer 14 Primostar Zeiss 1 Microscope 15a. 15b a) POL-ECO CLN53STD, sn:cn5sd08895 1 Incubator b) POL_ECO ILW115-T TOP, sn:iw1te08894 1 Incubator 16a, 16b a) Schuett Phoenix 2 Bunsen burner b) Stuart Vortex Mixer 3 Mixer 17 GFL 1103, sn:11548510k 1 Waterbath 18 Stomacher 400 Circulator Seward, sn:46623 1 Blender 19 Stuart Heat Stir SB162 2 Stirrer 20 Axio Observer A1 Zeiss 1022198127, sn:3832000763 1 Microscope 21 HMC, HiClave, HV-110L 1 Sterilizer clave 22 Sanyo MLS 3781L 1 Sterilizer clave 23 POL-ECO CLN115STD, sn:sn1sd08903 1 Incubator A laboratory plan with the points that equipment is placed can be seen in picture 7. 14

Picture 7: Laboratory plan A detailed description of the equipment of the Laboratory of Monitor & Certification of Hygiene & Safety of Shellfish is following. 1a: Distilled & bis-distilled water GFL 2108, sn:10315109j (picture 8). It is a material combination of stainless steel / glass, operating fully automatic. Distillation involves boiling the water and then condensing the steam into a clean container. It produces eight liters of ultra-pure, low-gas, bacteria and pyrogen free distillate with a very low conductivity. 15

1b: Deionized water, Zalion (picture 8). This product works by exposing water to electrically charged resins, which bind to the mineral ions or salts in the water, including calcium, iron, copper, sodium, bromide, chloride and other and remove them. It is contained by a special resistance of 3.000.000 Ω/cm and produces 1-10 liters/min of deionized water. Resins need replacing when saturated. Picture 8: Distilled, bis-distilled water and deionized water 2: Rotavapor, Heidolph, Laborota 4001, sn:591-24000-00-0 (picture 9). This product is designed for simple and complex distillations of volumes from 50 ml to 3l. It is contained by a long lasting graphite-filled PTFE vacuum seal between drive and vapour tube that provides leak-free, greaseless operation. Drive motor is quiet, brush-less, sparkless and maintenance free. Rotary drive is electronically controlled, range from 20 to 270 rpm. 16

3: Radwag, PS750/Y, sn:288108/10 (picture 9) This is a simple operated and maximum automated of the weighing process. Measurement reliability and accuracy is assured by internal adjustment/calibration system triggered by time flow or temperature conditions. It has a maximum capacity of 750g, readability of 1mg and a stabilization time of 2sec. Picture 9: Vacuum filtration, vacuum solvent condensation and balance 4: HMC, HiClave, HV-50L, sn:30309060912 (picture 10) A steam sterilizer with a chamber volume of 50l, temperature 105-135 C and 2.2 bar maximum pressure. The model has model includes an easy slide quick lock, a flexible temperature sensor thermo lock, steam condenser and single key programming. 5: Fiocchetti (picture 10) This refrigerator designed and produced to preserve sanitary products and/or heat-sensitive and thermo labile biological materials in the best way, as an accurate control of the storage temperature is requested for them. 17

Picture 10: Sterilizer clave and refrigerator 6: Consort C861 (picture 11) A basic multi-parameter analyser for ph, mv, conductivity, Salinity, TDS and temperature. It can read ph 0-14 (res. 0.01), conductivity 0-1000mS/cm (res. 0.01mS/cm), ±1000mV (res. 1mV), salinity 0 70ppt (res.0.01ppt), TDS 0-100g/l (res. 0.1mg/l) and temperature 0-100 C (res. 0.1 C). Picture 11: PH, ev, EC, Salinity, TDS analyzer 18

7: Candy CCF200 (picture 12) This is a chest freezer with a total capacity of 192 litres and a freezing capacity of 10kg/24h. 8a: Gorenje, ST-5B50-FOT8, sn:s053f09204 (picture 12) This thermostatically controlled cabinet is used for continuous temperature control over a range of 2 C to 40 C. This makes it ideal for the temperaturecontrolled storage of samples or BOD determinations in effluent analysis work. The temperature can be set in steps of 0.1 C and an LED display shows both the set temperature and the current temperature in the cabinets. Picture 12: Freezer and incubator for BOD determination 8b: Lovibond Oxidirect (picture 13) Respirometric methods provide direct measurement of the oxygen consumed by microorganisms from an air or oxygen-enriched environment in a closed vessel under conditions of constant temperature and agitation. Carbon dioxide produced metabolically by the bacteria is chemically bound by the potassium hydroxide 19

solution contained in the seal cup in the bottle. The result is a pressure drop in the system, which is directly proportional to the BOD value and is measured by the Lovibond BOD sensor. The BOD level is then displayed directly in mg/l. Instrument includes sample bottles, measuring sensors, non-wearing inductive stirring system, overflowed measuring flasks for metering of sample volumes, nitrification inhibitor and potassium hydroxide as an absorbent. Picture 13: BOD analyzer 9: Radwag, XA60/220, sn:242689/2008 (picture 14) This is a simple operated and maximum automated of the weighing process. Measurement reliability and accuracy is assured by internal adjustment/calibration system triggered by time flow or temperature conditions. It has a maximum capacity of 220g (minimal load 10mg) and readability of 0.1mg. 20

Picture 14: Balance 10: Hettich Rotanda 460R (picture 15) A centrifuge with brushless induction motor, refrigeration, high capacity 4x750ml, max RPM/RCF:15,000/24,400 (fixed angle) that is available to accommodate the most varied types of tubes. Picture 15: Centrifuge 21

11: WTW CR4200 (picture 16) This thermoreactor is a dry temperature control device for that facilitates and secures the digestion using reaction cells. It ensures complete digestion of the sample, as it maintains the necessary high reaction temperature throughout the defined period. It takes 24 reaction cells with an outer diameter of 16 mm and has two independent thermoblocks that enable two temperature programs to run simultaneously. 12a: Spectroflex 6100 WTW, ULSTD6100-1 (picture 16) This instrument is developed for measurements in the range of 320-1100 nm. It is used for precise measurements of test kits up to complete scans for COD, TOC, nitrogen, phosphorous, heavy metals and much other. It can also scan functions for absorption measurement and kinetics. Picture 16: Thermoreactor and spectrophotometer 12b: WTW kits (picture 17) The kits are used for the spectrophotometer analysis of the instrument above. NO - 3, NO - 2, PO 3-4, NH + 2-4, SiO 4 and COD kits are kits used by the laboratory in present. 22

Picture 17: Kits 13a: Varian AA240 FS flame (picture 18) A fully automated PC-controlled true double-beam spectrometer with fast sequential operation for fast multi-element flame AA determinations. It features 4 lamp positions and automatic lamp selection. 13b: Varian AA240 Z graphite (picture 18) A fully automated PC-controlled true double-beam spectrometer dedicated to graphite furnace AA determinations with graphite tube atomizer and programmable sample dispenser. It features 4 lamp positions, automated lamp selection, automatic wavelength and slit selection, deuterium background correction and a wide range photomultiplier tube. 23

Picture 18: Spectrometers 14: Primostar Zeiss (picture 19) An easy to operate microscope, equipped with good optics. It contains three illuminations, a 30 Watt halogen light, and LED with stable color temperature and higher efficiency for long-term usage. Picture 19: Microscope 24

15a: POL-ECO CLN53STD, 5-100 C, sn:cn5sd08895 (picture 20) A laboratory incubator with natural air convection and 56lt capacity. It has stainless steel interior and microprocessor time and temperature control with a temperature range of 5 to 100 C. 15b: POL_ECO ILW115-T TOP, -10-100 C, sn:iw1te08894 (picture 20) A cooled incubator with forced air convection and 112lt capacity. It has stainless steel interior and microprocessor time and temperature control with a temperature range of -10 to 100 C. Picture 20: Incubators 16a: Schuett Phoenix (picture 21) This unit features user-adjustable controlled burn-time from 1 s to 120 min and automatic safety shutdown when the unit is not in use. Sensors measure the temperature of the burner head and report a warning if overheating occurs. The flame is stable even in strong air current situations. The Bunsen burners are suitable for use with natural gas (central gas supply in the lab) or propane/butane gas. 25

16b: Stuart Vortex Mixer (picture 21) A mixer with speed that can be selected from 200rpm for very gentle mixing to 2500rpm for vigorous agitation. The robust die-cast body avoids unnecessary movement during use while integral retort rod fixing allows vessels to be secured above the vortex action for long-term mixing. Picture 21: Bunsen burners and mixers 17: GFL 1103, sn:11548510k (picture 22) This water bath is successfully used for incubations and inactivation of cultures. It has a 14lt capacity and microprocessor-controlled temperature regulation. Heating element, bath interior, cover, lid and perforated tray is made of stainless steel. 18: Stomacher 400 Circulator Seward, sn:46623 (picture 22) This blender is used for organism recovery by a unique paddle action that extracts more organisms into suspension than any other blending process. The sample and diluents are retained in an irradiated sterile bag and there is no direct contact between the blender mechanism and the sample. It uses a combination of mechanical forces like crushing and stirring to produce homogenization. 26

19: Stuart Heat Stir SB162 (picture 22) A stirrer with a glass ceramic top which has excellent chemical and temperature resistance. Powerful magnets and motor give stirring speed up to 1500rpm and volumes up to 15 litres. Picture 22: Waterbath, blender and stirrer 20: Axio Observer A1 Zeiss 1022198127, sn:3832000763 (picture 23) This microscope is developed for the observation, manipulation and analysis of living cells. It also uses fluorescence technology to produce better contrast quality, and blends negative and positive phase contrast into one objective lens. 27

Picture 23: Microscope 21: HMC, HiClave, HV-110L (picture 24) A steam sterilizer with a chamber volume of 110l, temperature 105-123 C and 1.2 bar maximum pressure. The model has model includes an easy slide quick lock, a flexible temperature sensor thermo lock, steam condenser and single key programming. 22: Sanyo MLS 3781L (picture 24) A steam sterilizer with a chamber volume of 75l, temperature 115-135 C and 2.3 bar maximum pressure. It includes a microprocessor temperature control. 28

Picture 24: Sterilizer claves 23: POL-ECO CLN115STD, sn:sn1sd08903 A laboratory incubator with natural air convection and 112lt capacity. It has stainless steel interior and microprocessor time and temperature control with a temperature range of 5 to 100 C. The analyses conducted by the Laboratory of Monitor & Certification of Hygiene & Safety of Shellfish, the method and the equipment used for each of them are summarized in the table 2 below: physicochemical parameters Table 2: Analyses performed parameters units method equipment temperature o C ph 4-10 APHA-AWWA- WEF, 2005-4500H + b conductivity μs/cm APHA-AWWA- WEF, 2005-2510b salinity ppt APHA-AWWA- WEF, 2005-2520b 6 29

organic load biochemical oxygen demand (BOD 5 ) chemical oxygen demand (COD) mg/l mg/l APHA-AWWA- WEF, 2005-5210b APHA-AWWA- WEF, 2005-5220c 8a, 8b 11, 12a, 12b solids total suspended solids (TSS) total dissolved solids (TDS) mg/l mg/l APHA-AWWA- WEF, 2005-2540d APHA-AWWA- WEF, 2005-2540c 2, 9, 15a or 15b 6 nitrates (NO 3 - ) mg/l nitrites (NO 2 - ) mg/l nutrients phosphates (PO 3-4 ) ammonium (NH + 4 ) mg/l mg/l APHA-AWWA- WEF, 2005-4500b 11, 12a, 12b silicates (SiO 2-4 ) mg/l potassium mg/l ions sodium calcium chloride ions sulphate mg/l mg/l mg/l mg/l APHA-AWWA- WEF, 2005-4110 11, 12a, 12b metals microbiological parameters hardness ο F APHA-AWWA- WEF, 2005-2340 lead Pb cadmium Cd mercury Ηg mg/l, μg/l mg/l, μg/l mg/l, μg/l APHA-AWWA- WEF, 2005-3111b,d APHA-AWWAe.coli cfu/100 ml WEF, 2005-9221 APHA-AWWAtotal coliforms cfu/100 ml WEF, 2005-9225 13a or 13b 14, 15a or 15b, 16a, 16b, 17, 18, 19 30

phytoplankton analysis biological parameters heterotrophic plate count APHA-AWWA- 37 o C cfu/100 ml WEF, 2005- heterotrophic 9215 plate count 22 o C APHA-AWWAintestinal cfu/100 ml WEF, 2005- enterococci 9230 clostridim cfu/250 ml perfringens pseudomonas cfu/250 ml aeroginosa quality determination 20 quantitative cells/ml chlorophyll α μg/l 2, 7, 10, 12a, 12b 31

3.1.3 Reports and communication Results of every test performed by the Laboratory of Monitor & Certification of Hygiene & Safety of Shellfish is reported with accuracy, clarity, objectiveness, without doubts and according to the special instructions of the methods used. All the necessary information, required by the customer, is reported for the method and the interpretation of the results. When opinions or special interpretations are included, the laboratory documents the basis that they have been corroborated. The form of the testing report is designed so as to include types of testing performed by the laboratory and minimize the possibility of misunderstanding or bad use. Testing reports include the following information: The title Testing Report. The one single identity of the report with page numbers so as to define easily the size and the end of the report. The sampling date or the reception date of sample at the laboratory and the time that testing was performed. Testing results. The customer s name, manufacturer s name and the sampling area. The methods used for testing. A note for testing performed by the laboratory and is out of the Hellenic Accreditation System certification. A note for testing results performed by a subcontractor with the names and signatures of the persons that license. A laboratories declaration that report cannot be reproduced without the written approval of the laboratory. The testing report form can be seen below: 32

Testing report Certification Number: Sample Code: Customer Code: Customer Name: Customer Address: Sampling Date: Sample Reception Date: Date of Testing Performance: Responsible for Sampling: Sample was handed by: Sample Condition: Temperature of Reception: Possible Deviations or Non- Conformities according to the method requirements specifications: Results of Analysis Sample Characterization Parameter Method Result Units Limit Uncertainty n.d.: Not determined at the reporting limit of the method Address: e-mail : Reproduction of the analysis certification is not allowed except as a whole after written approval by the lab The present certification does not contain opinions and interpretations Measurement uncertainty is expressed by the same units as the measurement units. Measurement value is applied by: Result value: +/- Uncertainty value Thessaloniki date: For the laboratory The laboratory manager Wherever necessary for the testing results interpretation, the following information could also be contained: Deviations due to additions or exceptions of the testing method and information for special conditions of testing e.g. environmental conditions A conformity or non-conformity declaration according to the method requirements / specifications 33