Monitoring Algae Growth in Marine Water Culture Solutions Using TOC/TN ACS - NERM - Dan Davis Shimadzu Scientific Instruments

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Monitoring Algae Growth in Marine Water Culture Solutions Using TOC/TN ACS - NERM - Dan Davis Shimadzu Scientific Instruments October 7, 2016

Monitoring Algae Growth in Marine Water Culture Solutions Using TOC/TN The culturing of microalgae in various types of nonedible biomass has been receiving considerable attention in recent years. Microalgae that inhabit seawater are favored due to the low cost of the culture solution and the wide choice of culture locations. Various nutrients, such as nitrogen compounds, are added to these types of microalgae cultures. Nutrient concentrations must be managed within an appropriate range depending on the type of microalgae and the culture conditions.

Monitoring Algae Growth in Marine Water Culture Solutions Using TOC/TN TOC for Microalgal biomass measurement permits accurate measurement of total organic carbon even if the sample is in the form of a suspension, such as microalgae, and with minimal impact due to sedimentation. The following types of measurements can be conducted with a TOC for microalgal biomass measurement. Measurement of total carbon and nitrogen in water, consisting of dissolved and suspended carbon and nitrogen Measurement of total carbon, organic carbon, and inorganic carbon in water Measurement of dissolved CO 2 content in water

Monitoring Algae Growth in Marine Water Culture Solutions Using TOC/TN TOC/TN can be utilized for researching the following purposes for microalgal biomass : Obtaining information related to the nature and physiological state of microalgae Grasping the time-course changes in the culture, and the material changes in cells in a light and dark environment Grasping the time-course changes in the carbon / nitrogen ratio in the culture system TOC supports measurements using small sample volumes of 10 to 20 ml, it is also suitable for laboratory scale research.

Monitoring Algae Growth in Marine Water Culture Solutions Using TOC/TN TC (Total Carbon) : Consists of 2 types of carbon (in water), Total Organic Carbon (TOC) : Organic carbon bonding with hydrogen or oxygen forming organic compounds. Inorganic carbon (IC) : Carbonates, bicarbonates, and carbon dioxide i.e. TC = TOC + IC

Monitoring Algae Growth in Marine Water Culture Solutions Using TOC/TN IC (Inorganic Carbon): Carbonates, bicarbonates and CO 2, dissolved in solution Samples with ph >3 will absorb IC from the surroundings IC Analysis: Samples are acidified to a ph of ~ 2 using phosphoric acid The dissolved carbonates are converted to CO 2 Sample is sparged to remove dissolved CO 2 out of solution

Monitoring Algae Growth in Marine Water Culture Solutions Using TOC/TN TOC (Total Organic Carbon): Organic carbon bonding with hydrogen or oxygen to form organic compounds Consists of: Purgeable Organic Carbon (POC) Non-purgeable Organic Carbon (NPOC) i.e. TOC = POC + NPOC Analysis: There are 2 methods, Difference method : TOC = (TC IC) Direct method : TOC NPOC (if no POC)

Total Organic Carbon Measurement Measure the sample (dilute) Inorganic Carbon Removal Hydrochloric Acid Solution Agitate Removal of CO 2 Decomposition : 680 with Pt catalyst Formation of CO 2 Obtain the TOC Result

Catalytic Combustion Oxidation 680 o C Sample + O 2 CO 2 + H 2 O* Platinum Catalyst Movie

Total Nitrogen TN (Total Nitrogen): Sum of Both Organic Nitrogen and Inorganic Nitrogen: Organic Nitrogen (R-N) (Inorganic) Nitrate (NO3) (Inorganic) Nitrite (NO2) (Inorganic) Ammonia (NH3) Does not include: Nitrogen Gas (N2) Chemical unreactive and not very reactive biologically Not detected by TN Method

Total Nitrogen Measurement Measure the sample (dilute) Air (20% O 2 ) Decomposition : 720 with Pt catalyst Electricity Formation of NO Formation of NO 2*, -> NO 2, Photon Emission Ozone Obtain the TN Result Relaxation of NO 2* to NO 2 by emitting light

Monitoring Algae Growth in Marine Water Culture Solutions Using TOC/TN

Monitoring Algae Growth in Marine Water Culture Solutions Using TOC/TN

Salt A Challenge 14

Salt A Challenge 15 500 Injection of 150uL (1500 Injection of 50 ul as Experiment)

Salt A Challenge High Salts Kit Increases amount of salt that can buildup, uses new combination of Catalysts and H2SO4 Combustion Tube life extended 3-5x 16

Extend Life of Combustion Tube As salts melts it breaks down quartz combustion tube. Why Change from HCl to H2SO4? NaCl (801C)+ H2SO4 Na2SO4 (881C) MgCl (714C) + H2SO4 MgSO4 (1124C) CaCl (772C) + H2SO4 CaSO4 (1450C) As melting point rises combustion tube becomes more stable. 17

Concentration[mg/L] Area TOC-L with 220 measurements High Salt Conbutiuon tube 28%NaCl in 5mg/LKHP[Inj:50uL Acid:15%] 10 8 6 4 2 0 Concentratoin Area 0 50 100 150 200 250 Injections 50 40 30 20 10 0 Check Standards at 1, 110, & 220 50 ul Injection, 15% H2SO4 18

Workflow Manual Technique Collection Analysis Reporting Sampling Physical Treatment Report Generation Preserving Chemical Treatment Review/React Storage Batch Measurement

Autosampler ASI-L Vials 9mL -93 24mL -93 40mL -68 Options Magnetic stirrer (for 24mL/40mL) Septum and Cap (for 24mL/40mL)

Autosampler - OCT-L Low cost alternative to ASI-L Fits almost any container The container that the customer uses daily can be use directly Maximum 2 units can be installed (16 samples) Magnetic stirrers are available

Workflow Manual Technique Collection Analysis Reporting Sampling Physical Treatment Report Generation Preserving Chemical Treatment Review/React Storage Batch Measurement

Workflow Automated Technique Collection Analysis Reporting Sampling Physical Treatment Report Generation Preserving Chemical Treatment Review/React Storage Measurement

On-Line Sampling Considerations When selecting a sampling device Consider: Particulates Are there a lot of particulates? Do I need to include them in my reading? Concentration of Analyte Are there special precautions based on level? Do I need to consider environmental influences? Number of Sampling Points Can I monitor all necessary points?

On-Line Sampling Considerations Particulates can add to the complexities of collecting and analyzing samples. Do you have particulates? If yes will they interfere with the analysis Do I need to consider them in my analysis? Do they contribute to your analysis?

On-Line Sampling Considerations Performing low level analysis can be influenced by environmental influences such as contact with air or water used in the process. Concentration of Analyte Are there special precautions based on level? Do I need to consider environmental influences?

Multi-Stream Suspended Solids Unit Sample flows in the top and drains out the bottom Actuators move the sampling arm into the stream Sample flows down into the sampling chamber through the strainer removing larger particles Mixing blades homogenizes the sample System back flushes then washes Movie

Multi-Stream Sample Switching Unit Sample flows in the bottom and out the top Some sample flows in lower chamber on the outside of the sample collection cup Actuators move the stream to the sample collection chamber. Reduces Sample-Air Contact Eliminates need for washing

Backwash Strainer Sampling Unit Sample flows in the bottom and out the top Air pressure prevents sample from entering inner chamber until sampling Air is turned off and sample is filtered as it enters chamber Sample flow dynamics self clean the sample mechanism Movie

Monitoring Algae Growth in Marine Water Culture Solutions Using TOC/TN TOC/TN can be utilized for researching the following purposes for microalgal biomass : Obtaining information related to the nature and physiological state of microalgae Grasping the time-course changes in the culture, and the material changes in cells in a light and dark environment Grasping the time-course changes in the carbon / nitrogen ratio in the culture system quantitatively Analysis can done manually in the laboratory or automatically with on-line analyzers.

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