Quality Assurance Plan for Virginia In
Contents
1. At the C prompt type cd collectidata and press Enter Type dir to view a list of the files in this directory Fluorometer Laptop Insert a disk into the AA drive At the CA prompt type copy FLMM DD DAT A and press Enter Make a duplicate copy to be stored in a separate location Press the power button to turn off the computer after the file has been copied to the disk GPS Laptop 12 0 12 1 12 2 FLURAU 2 SOP Revision 1 Date 03 10 98 Page 16 of 20 At the C COLLECT DATA prompt type pkzip GPSMM DD zip MONTHGPS ZIP and press Enter Insert a disk into the A drive At the C prompt type copy MONTHGPS ZIP A X and press Enter Make a duplicate copy to be stored in a separate location Press the power button to turn off the computer after the file has been copied to the disk PROCEDURES FOR COLLECTION AND CONCENTRATION OF CALIBRATION SAMPLES Collection To protect the chlorophyll from degradation prior to extraction and analysis collect samples in 1000 mL brown HDPE bottles Rinse the entire inside surface of the sample bottle three times with sample completely discarding the sample before successive rinses Fill the sample bottle with sample leaving approximately one inch of air space in the sample bottle Immediately add 1 mL of magnesium carbonate suspension per liter of sample and invert to mix see definitions section 14 0 Protect from direct sunlight Samples should be concentrated onto filters imme2. Model 10 AU 005 Field Fluorometer User s Manual October 1993 Order Number 998 5313
3. APPARATUS p D E is 4 6 INSTRUMENT MAINTENANCE cssssdiscsccccescotsstessdcssccecctescesvecdocdescedat vsscdseccegesssbessssecdesses 6 Ts ONBOARD PREPARATION sisssissssuessccesossecisastsvsesscossendsnsvscootecssceeanccessvannns eue eure Fee p eU ro ko vn 9 8 VER TICATs SAMPLING itesteesfeekseexsCens beso ebyses ektasc epetpokret VARena Ee Sev pyae eM exa Nd eri een aite d 12 9 HORIZONTAL SAMPLING sees proe ke ec ee VER e PE ePVe ee Coe eva ve vn t uon eh aS reo ea assu eU naga vei ins 13 10 INTERNAL DATA LOGGER SHUT DOWN ee eeeeeeeeee sense esee so oeste toss sete enseee 14 11 LAPTOP COMPUTER SHUT DOWN eee eee ae ee eee oo e etae pn o e ee po eee teen ae eaa ePe e eee a aae 14 12 PROCEDURES FOR COLLECTION AND CONCENTRATION OF CALIBRATION SAMPLE LES inse ee os evo Fent bY oda eyo eHY Fu RESP Fa PY Ue Fa su eS e tareis ERR EN aevo Pe Guo Casu siS 15 13 POST CRUISE ACTIVITIES ie eoe e eco pu bee epp e nee aeu bee cu ea reo Qaa ne tea eeu ba CRe searsncssteeeanees 16 14 DEFINITIONS ostinscesaicovedsnccspccstaceceveseaaceosaieassvosdeonedobessandeseusnandeleasevocddouvestessssustesoetesoness 17 15 FLUOROMETRY FIELD DATA SHEET 4 eee e eee ee eee en eee teen aee eee ese eee e eee 18 16 CHLOROPHYLL FIELD DATA SHEET cce ee ee ee eee ee ee eee en see e eaae eee eno ee 19 17 LITERATURE CITED sicsiscccccsiccsnckodscoccnusecsscutasdoesdibosss conan
4. graduated cylinders and Erlenmeyer flasks and the tops and bottoms of filtration towers with parafilm Store in the appropriate cabinet FLURAU 2 SOP Revision 1 Date 03 10 98 Page 7 of 20 5 2 3 Reagents 6 0 6 1 CHEMICALS USED AND REAGENT PREPARATION Chemicals Used All chemicals must be analytical grade or of a higher purity except as noted Magnesium carbonate MgCO Wash down the counter top or hood with ultrapure water and wipe dry using a paper towel before and after preparing reagents The following information must be recorded on each reagent bottle label Reagent name and concentration Analysis which requires this reagent Date prepared Prepared by initials Expiration date entry is N A if there is no holding time Storage requirements e g room temperature 4EC dark etc NOTE Refer to Safety Precautions and use care when preparing reagents and stock solutions Reagent containers are used only for the intended reagent and are reused Before new reagents are added to the containers the containers are rinsed 9 times with ultrapure water inspected then rinsed 3 times with the fresh reagent that they will contain Magnesium carbonate suspension Measure 100 mL of fresh ultrapure water using a class A volumetric flask Add 1 g of finely powered magnesium carbonate Shake immediately before use until all material is in suspension INSTRUMENT MAINTENANCE AND SERVICE Preventive Maintenance
5. Pre and Post Cruise Flush fluorometer with ultrapure water no later than the morning after every cruise 6 2 FLURAU 2 SOP Revision 1 Date 03 10 98 Page 8 of 20 Visually inspect continuous flow cuvette after every cruise Refer to section 6 2 of this SOP for procedure Clean continuous flow cuvette with 4N HCl and rinse with ultrapure water Refer to section 6 2 of this SOP for procedure If cuvette is scratched or cracked replace Refer to section 6 3 of this SOP for procedure Each time the cuvette is removed visually inspect O rings If damaged change O rings Lubricate O rings with silicon grease Cleaning the Cuvette Removal and cleaning of the continuous flow cuvette is an in laboratory procedure only Please refer to the diagram of the fluorometer in the Turner manufacturer s manual on page A7 13 while following these steps Turn off the power and drain all water from the system Remove all connections from the Intake and Exhaust fittings Remove the four small screws on both the Intake and Exhaust fittings with an allen wrench Remove the 3 screws that anchor the Intake fittings Remove the 8 screws which retain the sample compartment cover If the gasket under this cover sticks remember that it is glued to the cover Insert a thin knife gently between the gasket and the sample Compartment Casing to loosen Using an Allen wrench loosen the Upper and Lower Set Screws Pull the exhaust fitting top out of the
6. homogenizing a sample be inverting one inversion is starting with the cap of the sample bottle pointing straight up turn the sample bottle completely upside down so the cap is pointing straight down then turn the sample bottle upright until the cap is pointing straight up again FLURAU 2 SOP Revision Date Page 03 10 98 20 of 20 15 0 FLUOROMETRY FIELD DATA SHEET Light F mol sec n Loran Coord Lat Date Time Event Cruise _CBP FLURAU 2 SOP Revision 1 Date 03 10 98 Page 21 of 20 16 0 CHLOROPHYLL FIELD DATA SHEET CRUISE CBP CRUISEDATE CHLOROPHYLL FILTERED BY DATE CHLOROPHYLL FILTERED EXTRACTED BY DATE CHLOROPHYLL ANALYZED DATE CHLOROPHYLL EXTRACTED CHLOROPHYLL cwsomwuamowS 664 665 To RATIO LOG SAMPLE TIME 750 664 647 630 480 510 750a 665a CAN NOT NUMBER ID FILTERED A A A A A A A A EXCEED 1 7 mna EO a ee a NN ee ee ee Jews 1 TT omes PL ess L e a FLURAU 2 SOP Revision 1 Date 03 10 98 Page 22 of 20 17 0 LITERATURE CITED APHA 1989 Standard Methods for the Examination of Water and Wastewater 17th Edition Method 10200 H Chlorophyll American Public Health Association Washington DC ASTM 1979 Standard Practices for Measurement of Chlorophyll Content of Algae in Surface Waters Method D3731 79 American Society for Testing and Materials Philadelphia PA
7. 1 Date 03 10 98 Page 5 of 20 250 ml 1 MgCO 5 1 3 5 2 5 2 1 5 2 2 FLURAU 2 SOP Revision 1 Date 03 10 98 Page 6of 20 Instruments Turner Model 10 AU series fluorometer Power cable with computer connector to fluorometer 2 Laptop computers with mouse 2 power cables for computers Sampling array 40 lb weight attached to bottom of T frame 2 each 3AG 3AMP SLO BLO and 3AB 1 2AMP SLO BLO spare fuses Set of Allen wrenches Spare cuvette and O rings Silicon grease Connecting hoses 1 green plastic garden hose labeled BB 1 green plastic garden hose labeled C 1 green plastic garden hose labeled AA Plastic Y valve Plastic T valve pipe Laboratory Labware Needed Use only Class A volumetric labware l ea 100 mL volumetric flasks Labware Cleaning To clean volumetric flasks filtration towers flasks graduated cylinders and sample bottles Inspect labware for reside breaks or defects Clean labware by scrubbing with a bottle brush and dilute Liquinox7 soap Thoroughly rinse with tap water Rinse twice with 4N hydrochloric acid HCL then 9 times with fresh ultrapure water ensuring that the entire inside surfaces are rinsed each time Inspect again for reside breaks or defects If not evident dry on a clean drying rack labelled for clean labware only completely covered with clean paper towels When dry store volumetric flasks vials and sample bottles with the caps on Seal the tops of filtration flasks
8. FLURAU 2 SOP Revision 1 Date 03 10 98 Page 1 of 20 STANDARD OPERATING PROCEDURE FOR THE MEASUREMENT OF FLUOROMETRY IN WATER AND SEAWATER USING A MODEL 10 AU TURNER FLUOROMETER PREPARED BY Shannon Lee FOR Water Quality Laboratory at NAUTICUS Applied Marine Research Laboratory College of Sciences Old Dominion University Norfolk Virginia 23529 0456 DOCUMENT FILE Quality Assurance Office LOCATION AMRL EFFECTIVE DATE April 1997 TERMINATION DATE APPROVED BY Dr Alan W Messing Date Associate Director AMRL Joseph G Winfield Date Quality Assurance Officer AMRL DISCLAIMER This SOP applies to the analysis of marine and estuarine water samples in support of the USEPA Chesapeake Bay Water Quality Monitoring Program for samples collected in the Chesapeake Bay and Elizabeth River This SOP may not be applicable to any other studies FLURAU 2 SOP Revision 1 Date 03 10 98 Page 2 of 20 TABLE OF CONTENTS MUNG PASE eMe toD br mde net Meneame E SEIN 1 Table of Contents mI EEES Esi ie 2 1 EOCATIONS iiic tab Rte E AS A EEEN 3 2 PURPOSE saseicesiessscastesncths ctacdtaiecsuiccacvssreaneconddoaveas epiac soistes E teess oek Essa e sE Lee epa Eseese siia 3 3 APPLICABUATY uio cescednsee Qe ecescttseexude so ssdori saa cteas seosest dU e coissa peux Pepe p Sodos he ia ded 3 4 OVERVIEW OF METHODBD s ovekc ster ee eeoo re sese poe eee esae cep epo exul eun suu en kane a e ES CUM enun n 3 5
9. diately after collection section 12 2 If this is not possible store samples at 4EC by refrigerating or packing the sample bottles in ice up to the level of the sample but below the bottom of the sample bottle cap Keep container lids closed as much as possible Storage temperature must be measured using a thermometer which has been calibrated against an NIST traceable thermometer within the last six months Storage temperature must be documented Concentration Labware needed 6 ea filtration towers 2 ea 500 mL graduated cylinders 2 ea 1000 mL graduated cylinders 1 ea 4000 mL filtration flasks FLURAU 2 SOP Revision 1 Date 03 10 98 Page 17 of 20 2 ea Forceps with rounded tips 2 ea 2000 mL or 4000 mL filtration flasks 4 25 cm Whatman7 GFF glass fiber filters box of 100 Ultrapure water squirt bottles Labels for aluminum foil Labels for sample bottles Aluminum foil for chlorophyll filter pads Notebook with fluorometry field data sheets Black ink pens See section 5 2 2 for the appropriate labware cleaning procedures Using forceps gripping only the filter edge transfer a 4 25 cm Whatman7 GF F glass fiber filter onto the base of a filtration tower Replace the top of the filtration tower onto the base Moisten the filter with ultrapure water Rinse the entire inside surface of a 500 mL graduated cylinder three times with ultrapure water Mix the sample thoroughly by inverting see definitions the
10. e bottom opening of the fluorometer by holding the Intake fitting Insert the Exhaust fitting into the top opening of the fluorometer Rotate the Intake and Exhaust fittings slowly back and forth until the cuvette is properly aligned with the fittings CAUTION Excess force can easily break the fragile cuvette Tighten the Upper and Lower Set Screws Wipe off any fingerprints on the cuvette To make a visual inspection for leakage move the instrument to the dish washing section and place it on the side of the sink near the ultra pure system Connect the tubing from the end of the ultra pure system to the bottom connection of the flow through cell and turn on the flow of water 7 0 7 1 T2 FLURAU 2 SOP Revision 1 Date 03 10 98 Page 10 of 20 Make sure that the over flow from the top fitting runs in the sink If leakage occurs the installation process must be repeated Replace the front cover and the Intake and Exhaust fittings and screws ONBOARD PREPARATION Fluorometer Position the fluorometer so that the instrument panel faces you Uncap the power connector at the bottom right front of the fluorometer Line up the power cable with the connector and screw in the cable The cable must be pushed in and screwed tightly in stages to be fully connected Plug the power cable into the nearest electrical outlet Connect the computer cable to the laptop computer that is designated for use with the fluorometer Hose Arrangem
11. el and allow it to warm up for a minimum of 10 minutes The HOME screen should be visible on the fluorometer Press ENT to access the Main Menu Press 5 to choose the Internal Data Logger menu Press 3 to choose Status Press ENT to toggle between stop and logging The internal data logger is now collecting data Press lt ESC gt 3 times to get back to the HOME screen Logging data should be displayed on the screen and raw data will appear While data is logging numbers on the keypad can be pressed to mark events 1 start Vertical Fluorometry profile 0 5m 3 start Horizontal Fluorometry profile 4 end Horizontal Fluorometry profile 5 Horizontal grab 6 Vertical grab 8 each depth during the Vertical profile 1m 2m 3m 6m 9m This is not pressed if a vertical grab is taken and 6 has been pressed Laptop Computer for Fluorometer Ensure that the fluorometer is connected to the laptop 7 5 FLURAU 2 SOP Revision 1 Date 03 10 98 Page 12 of 20 Connect the mouse and power cables to the appropriate ports on the computer Plug the power cable into the nearest electrical outlet Turn the computer on by pressing the green power button At the C prompt type win Click on OK if a New Mail message appears The computer should be in the Collect W Jr software If not double click on this icon Click on Instruments from the main menu Choose Open Instrument from the drop menu Choose F
12. ent Screw the end of the hose labelled A to the intake bottom fitting of the fluorometer Screw the end of the hose labelled AA to the end of the T valve labelled AA Screw the end of the hose labelled B to the exhaust top fitting This is a short piece of hose with a Y valve on one end Screw the end of the hose labelled BB to the fittings on the V valve labelled BB this valve is left open parallel to the direction of flow at all times so water can flow freely through the cuvette The end of this hose is laid out on the deck of the ship so the water goes overboard Screw the end of the hose labelled C hose to the end of the T valve labelled C Screw the other end of the hose labelled C to the valve on the hull pump When doing a vertical profile the end of the hose from the exhaust hose of the submersible pump is connected to the T valve labelled D As quickly as possible turn off the water flow from the 12 7 4 FLURAU 2 SOP Revision 1 Date 03 10 98 Page 11 of 20 hose labelled C and turn on the valve to the hose labelled D Water should flow from the submersible pump into D connection through the T valve then into the end of the hose labelled AA From this point the water flows through the fluorometer and out the exhaust of the hose labelled BB Internal Data Logger After attaching the hoses to the fluorometer and ensuring that there are no leaks Turn on the fluorometer by pressing the red button on the front pan
13. esvscabsavessduusveseiveasbesesseavecasessneees 20 FLURAU 2 SOP Revision 1 Date 03 10 98 Page 3 of 20 1 0 LOCATION This analytical procedure will be used by Water Quality Laboratory staff at NAUTICUS located at One Waterside Drive Norfolk Virginia The procedure will be performed at the field preparation section and in the field for in situ monitoring 2 0 PURPOSE This procedure is designed to use a fluorometric technique to estimate chlorophyll a in estuarine and coastal waters as an indicator of water quality and to provide estimates of chlorophyll a concentrations throughout the study area for comparison to remote sensing data The intended user groups are the subcommittees and work groups of the USEPA Chesapeake Bay Monitoring Program CBP These data will be used to assess the effectiveness of the multijurisdictional action plan for restoring the water quality of the Chesapeake Bay This assessment will include various analyses of the data such as transectional data compilations and comparison with photosynthetic pigment data collected using discrete grab samples at selected locations These data may also be used for ground truthing of remote sensing e g satellite image data These data may also be used by State and Federal government officials to implement management actions designed to help improve the quality of the water in Chesapeake 3 0 APPLICABILITY This procedure is applicable to the continuous in vivo measureme
14. he fluorometer into the port at the back of the computer where the modem is located The port is labelled A Ensure that the fluorometer is plugged in Turn the fluorometer on by pressing the red power button on the front panel On the computer Log in to the computer by typing your login name and password Press Enter Select 6 from the Main Menu to return to the DOS prompt Type H and press Enter At the DOS prompt type IDL_1B1 Press Enter Select lt 6 gt from the Main Menu This option allows you to download data and convert the file to ASCII in one step On the fluorometer Using the fluorometer keypad press lt ENT gt to exit the HOME screen Press lt 5 gt to choose Internal Data Logger from the Main Menu Press lt 4 gt to access the downloading sequence Press lt 8 gt five times to start downloading On the computer When downloading is complete the program will prompt you to name the file The filename should be CBP BIN Where cruise number The program will then prompt you to name the ACSII file The ASCII filename should be CBP PRN Where cruise number Press lt 9 gt to exit the program 13 2 Maintenance See section 6 0 of this SOP Should be performed after every cruise 13 3 Storage The fluorometer is stored in the cruise prep area until the next cruise FLURAU 2 SOP Revision 1 Date 03 10 98 Page 19 of 20 14 0 DEFINITIONS HPDE High density polyethylene invert In reference to
15. holder rotating it back and forth to free it BE CAREFUL The cuvette may come out with the exhaust fitting When this happens remove it with the exhaust fitting Remove the lower intake fitting in the same manner as the upper fitting If the cuvette did not come out with the Exhaust fitting it should come out with the Intake fitting Wipe up any spilled liquids in the instrument and dry thoroughly to remove any saltwater that is highly corrosive to metal components 6 3 FLURAU 2 SOP Revision 1 Date 03 10 98 Page 9 of 20 Clean the cuvette with 4N HCL and rinse with ultrapure water If this will not clean the cuvette soak it in dilute acetic acid then rinse in Milli Q water Dry with lens tissue or Kimwipes Inspect the Intake and Exhaust fittings for any salt residue Clean with Lime Away and rinse with Milli Q if necessary Carefully inspect the two O rings on the exhaust fitting and on the intake fitting If there are any signs of nicks tears or deterioration replace with new O rings The O rings are Nitrile Bunda N rubber therefore chemical resistance is good Lubricate the O rings with silicon grease using lubricant in the O ring replacement kit The O rings must be lubricated before installation Installation of the continuous flow cuvette Ensure that the cuvette is clean and that the O rings have been lubricated Position the cuvette securely onto the Intake fitting Insert the cuvette into th
16. les For vertical profiles try to collect one at each sampling station covering the range of fluorometry values observed on the digital read out on the fluorometer Collect 10 vertical grabs per cruise day A grab sample is collected by opening the valve to the hose labelled BB and filling a 1 L brown HDPE chlorophyll bottle to the shoulder following the procedures in Section 12 1 of this SOP Press 6 on the fluorometer keypad to mark this event After collecting the sample close the valve Add 1 mL of magnesium carbonate suspension to the bottle The sample should be filtered immediately After the profile is completed the T valve labelled D is closed the valve labelled C is opened and the end of the submersible pump is removed to allow the bottom sample to be collected in the appropriate carboy 9 0 HORIZONTAL PROFILE SAMPLING 9 1 Horizontal Profile A horizontal profile is taken between each station The profile is started as soon as the boat begins to move toward the next station Press 3 on the fluorometer keypad to mark this event Check to make sure the valve for the hull pump is turned on Open the T valve to the hose labelled C and close the valve to the hose labelled D Water should now be flowing through the hose labeled CC into the T valve then into the hose labeled AA through the fluorometer and out the exhaust hose labelled BB During the horizontal profile GPS coordinates are collected on the laptop computer Ho
17. luorometer from the menu Click on Open In Instrument Data Destination click on File An X should appear in the box beside File Name the file C COLLECT DATA FLMM DD DAT Where MM is the month DD is the day Click on OK The laptop should now be collecting data Laptop Computer for GPS Plug in the GPS and turn on the power to the GPS and to the differential beacon Allow the GPS to warm up for approximately 15 min Ensure that the GPS is connected to the appropriate laptop computer Connect the mouse and power cables to the appropriate ports on the computer Plug the power cable into the nearest electrical outlet Turn the computer on by pressing the green power button At the C prompt type win FLURAU 2 SOP Revision 1 Date 03 10 98 Page 13 of 20 Click on OK if a New Mail message appears The computer should be in the Collect W Jr software If not double click on this icon Click on Instruments from the main menu Choose Open Instrument from the drop menu Choose GPS from the menu Click on Open In Instrument Data Destination click on File An X should appear in the box beside File Name the file CACOLLECTIDATAAGPSMM DD DAT Where MM is the month DD is the day Click on OK The laptop should now be collecting data from the GPS 8 0 VERTICAL PROFILE SAMPLING 8 1 8 2 Vertical profile At Im depth fill the surface carboy with water from the submersible pump hose Place the exhaust hose of the submersible pum
18. n system The data collected from the fluorometer are calibrated using spectrophotometry Discrete water samples are collected from the outlet port of the fluorometer flow through system and analyzed using spectrophotometric analysis for chlorophyll a concentration These values are used to create a regression equation that is used to interpolate chlorophyll a concentrations from fluorometric measurements Samples for fluorometry calibration are collected in brown plastic high density polyethylene HDPE sample containers Samples are buffered using 1 mL of saturated 1 magnesium carbonate per 1000 mL of sample then filtered on board immediately The procedures for filtration sample handling preservation and analysis are described in the Chlorophyll Spectrophotometric Method SOP 5 0 APPARATUS 5 1 Field 5 1 1 Labware Use only class A glassware For vertical and horizontal grab samples 20 ea 1000 mL brown HDPE High Density Polyethylene chlorophyll sample bottles for each sampling day 6 ea filtration towers 2 ea 500 mL graduated cylinders 2 ea 1000 mL graduated cylinders 1 ea 4000 mL filtration flasks 2 ea Forceps with rounded tips 2 ea 2000 mL or 4000 mL filtration flasks Ultrapure water squirt bottles Labels for aluminum foil Labels for sample bottles Aluminum foil for chlorophyll filter pads Notebook with fluorometry field data sheets Black ink pens 5 1 2 Reagents FLURAU 2 SOP Revision
19. nt of chlorophyll a in estuarine and coastal waters It can also be used for monitoring chlorophyll a in fresh water The salinity ranges encountered in the Chesapeake Bay are 0 36 parts per thousand The procedure assumes that spectrophotometric chlorophyll a data obtained from grab samples for calibration purposes are accurate and that fluorescence of chlorophyll a using the excitation and emission wavelengths specified in the procedure is proportional to the pigment concentration of the grab samples Chlorophyll a concentration is reported in units of F g L 4 0 OVERVIEW OF METHOD The procedure described in this SOP is based on the fluorescence of chlorophyll molecules at specific wavelengths of light At these wavelengths the fluorescence of chlorophyll is proportional to ambient chlorophyll concentration A submersible sampling pump is used to pump ambient water through a flow through cuvette fitted to the fluorometer to provide in vivo measurement of chlorophyll concentrations Fluorometry data are collected using a laptop computer and internal data logger that captures data every FLURAU 2 SOP Revision 1 Date 03 10 98 Page 4 of 20 30 seconds The position latitude and longitude and time date and time stamp for each discrete sample or for continuous flow through measurements are recorded at 2 second intervals using a laptop computer coupled to the fluorometer and Global Positioning System GPS with a differential beacon correctio
20. p onto the T valve labelled D and using the valves on the T valve open the valve to the hose labelled D and close the valve to the hose labelled C Water should now be flowing through the valve labelled D into the T valve then into the hose labelled AA through the fluorometer and out the exhaust hose To begin the vertical profile bring the array up to 0 5m let the water flush through the hoses and fluorometer for 1 5 min this includes the 45 seconds flush time for the submersible pump and the additional hoses Record this as the start of the vertical profile by pressing 1 on the fluorometer keypad Record the event number time the event was made and event depth on the fluorometry field sheet Appendix 1 After 0 5m repeat the previous steps at 1m 2m 3m and every 3m intervals thereafter to the bottom Press 8 on the fluorometer keypad to mark these events After the last vertical profile is completed at the last station to be sampled the recording session can be ended Vertical Grab Samples FLURAU 2 SOP Revision 1 Date 03 10 98 Page 14 of 20 A grab is sample water that has passed through the fluorometer filtered and analyzed for chlorophyll a using the SOP for Chlorophyll Spectrophotometric Method and then correlated to its corresponding value on the fluorometer for a calibration curve Two sets of grab samples 10 each are taken for two calibrations one is for vertical profiles and the other is for horizontal profi
21. rizontal fluorometry is terminated upon arrival at the next station Press 4 on the fluorometer keypad to mark this event After the last vertical profile is completed at the last station to be sampled the recording session can be ended 9 2 10 0 11 0 FLURAU 2 SOP Revision 1 Date 03 10 98 Page 15 of 20 Horizontal Grab Samples A grab sample is taken by opening the valve to the hose labelled BB filling a 1 0 L brown HDPE chlorophyll bottle to the shoulder following the procedures in Section 12 1 of this SOP Press 5 on the fluorometer keypad to mark this event After collecting the sample close the valve The samples are processed the same way as the vertical profile samples For horizontal profiles 10 grab samples are collected to cover all ranges observed on the digital display on the fluorometer Collect not less than 10 horizontal grabs per cruise day INTERNAL DATA LOGGER SHUT DOWN Eliminate the water supply by turning off both the hull pump and the submersible flows to the fluorometer On the fluorometer keypad Press lt ENT gt on the fluorometer keypad to access the Main Menu Press lt 5 gt to choose the Internal Datalogger menu Press lt 3 gt to choose Status Press ENT to toggle from Logging to Stop Turn off the power to the fluorometer LAPTOP COMPUTER SHUT DOWN Close the Fluorometer or GPS window Click on OK Close the Collect W Jr software Close the Program Manager window Click on OK
22. sample bottle 20 times Immediately rinse the graduated cylinder twice with the sample to be concentrated Mix the sample thoroughly by inverting the sample bottle 20 times Immediately pour a pre determined volume of sample into the rinsed graduated cylinder Record the filtration volume in the appropriate space on the chlorophyll field data sheet Concentrate the phytoplankton by filtering the sample water through the filter using 12 KPa vacuum pressure To avoid cell damage during filtration do not exceed this vacuum pressure and limit filtration duration to 5 minutes or less If it takes longer than 5 minutes to filter the selected sample volume discard filter and remaining sample rinse the filtration tower and refilter using a smaller sample volume Fold the filter in half using forceps being careful not to contact or disturb the filtrate Place the folded filter on a labelled square of aluminum foil and seal tightly Freeze as soon as possible at 20EC to 70EC If freezing immediately is not possible refrigerate or store in a water and light tight container packed in ice and freeze as soon as possible Completely rinse with the vacuum applied the empty filtration tower three times with ultrapure water before seating a new filter for the next sample Repeat the above steps for each sample FLURAU 2 SOP Revision 1 Date 03 10 98 Page 18 of 20 13 0 POST CRUISE ACTIVITIES 13 1 Downloading Fluorometry Data Plug the cord from t
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