LuminoTox - a tool for rapid toxicity testing by JasoRobinson


     Technological Innovation
ABSTRACT                                                                   HIGHLIGHTS
LuminoTox, a new easy to use tool for     INNOVATIVE TOOLS AND PROCESSES   Technology
                                                                           • Portable fluorescent biosensor
toxicity assessment, has been tested
on various types of wastewaters to                                           that uses photosynthetic enzyme
validate its effectiveness and scope                                         complexes
of use.                                                                    • In situ screening test

LuminoTox proved effective for rapid
                                              LUMINOTOX :                  • Toxicity detection in 15 minutes

toxicity detection (15 minutes) in
both treated and untreated effluents,
including municipal wastewaters,
                                               A TOOL FOR                  • Detection of a wide variety
                                                                             of toxic molecules
                                                                           • Validation of the efficacy of
leachates from landfill sites, pulp and
paper mill effluents and mining             RAPID TOXICITY                   wastewater treatment systems
                                                                           • Monitoring of sewage overflows,
industry effluents. It was also found
                                                                             industrial spills and water that rises
to be an effective means of assessing
the level of toxicity reduction in                TESTING                    to the surface on contaminated sites
                                                                             and sanitary landfill sites
these types of effluents following                                         • Rapid detection of toxic shocks
treatment in various systems, such                                           associated with inflows to biological
as those using activated sludge,                                             treatment systems
aerated ponds or sequential
biological reactors.                                                       Cost-effectiveness
                                                                           • Optimization of wastewater
LuminoTox is a tool that can comple-                                         treatment processes
ment routine physico-chemical                                              • Reduction in the amount of time
analyses and standardized bioassays                                          that must be spent in the field
by rapidly providing an index of                                           • Faster and more cost-effective
the toxic potential of effluents.                                            than most conventional bioassays
PROJECT                       BACKGROUND                                  TECHNOLOGY
OBJECTIVES /                  Conventional physico-                       LuminoTox, a patented                         generate a concentration-
                              chemical analyses cannot                    process, has two key                          response curve, allowing the
PHASES                        be used to evaluate the
                              toxic potential of
                                                                          components: stabilized
                                                                          photosynthetic enzyme
                                                                                                                        user to derive conventional
                                                                                                                        endpoints such as the IC50
The aim of this project
was to demonstrate            wastewater streams as they                  complexes (PECs) and a                        value (the concentration
the effectiveness of a        enter treatment plants or                   specialized fluorometer for                   that inhibits 50% of the
rapid, sensitive toxicity
test for potable water        the toxicity reduction in                   measuring photosynthetic                      fluorescence reaction)
and various effluents
(municipal, agricultu-        treated effluents that are                  activity based on chloro-                     or toxic units.
ral, agri-food or indus-      discharged into receiving                   phyll fluorescence, the
trial) that can also be
used to assess the            waters. The bioassays that                  LuminoTox Analyzer. As                        LuminoTox uses multi-
toxicity reduction that       are customarily used for this               PECs naturally fluoresce in                   enzyme complexes that
occurs after various
treatment processes.          purpose produce results after               response to light stimula-                    have been isolated from
The main stages in            a variable time period (up to               tion, this device measures                    plant extracts, providing an
the development of            96 hours). Environmental                    the inhibition of the fluores-                easy to use and rapid
LuminoTox were as
follows:                      decision-makers therefore                   cence reaction that occurs                    technique that can be
• December 2002:              have few tools available for                in the presence of toxic                      employed in the field. It is
  LuminoTox test kit          rapid, on-site toxicity                     substances.                                   therefore a practical and
  for isolated bioactive
  substances                  testing.                                                                                  cost effective tool for the
• December 2003:                                                          The conditions of fluores-                    environmental management
  LuminoTox test kit
  for toxicity                                                            cence measurement and                         of municipal, agricultural,
  (municipal and
  industrial effluents)                                                   photosynthetic efficiency                     agri-food and industrial
• April 2004:                                                             are pre-programmed into                       effluents. LuminoTox is
  LuminoTox test kit
  for herbicides                                                          the apparatus. Percent                        also an effective means of
  (triazines and urea)
• September 2004:                                                         inhibition is automatically                   monitoring the quality and
  LuminoTox test kit                                                      calculated and displayed on                   toxicity of drinking, surface
  for nitrogen
  ammonia                                                                 screen. These data can be                     and underground water
• September 2004:                                                         used directly or can be                       supplies.
  Read-out for
  polycyclic aromatic                                                     processed statistically to
  hydrocarbons (PAH)
• December 2004: Fully
  automated Robot
  LuminoTox, for                                     PECS AND INHIBITION SITES OF TOXIC SUBSTANCES
  continuous on-line
  monitoring of water
  quality (surface or
  underground) and
  industrial effluents

            E N V I R O N M E N T   •   T E C H N O L O G I C A L   I N N O V A T I O N   –   I N N O V A T I V E   T O O L S   A N D   P R O C E S S E S
Effects of physico-chemical                                                              sensitivity was found to increase with                                     incubation time after the effluents had
parameters                                                                               incubation time to a maximum of                                            gone through the first settling tank or
LuminoTox can be used under a wide                                                       60 minutes. The toxicity results for                                       the first aerated pond. LuminoTox is
range of physico-chemical conditions                                                     treated effluents showed a low level                                       therefore sensitive to waste matter
(no pre-treatment necessary) without                                                     of toxicity after 15 minutes of                                            that has not been completely
any risk of interference with the IC20                                                   incubation (inhibition < 25% in                                            decomposed during the initial
and IC50 values for atrazine and                                                         all effluents tested).                                                     treatment stage. Nonetheless, these
copper, used as reference toxicants.                                                                                                                                treatment processes were found to be
                                                                                         Effluents treated in a sequential                                          effective after the effluents had gone
Assays of industrial                                                                     biological reactor showed no toxicity                                      through the secondary settling tank
and municipal effluents                                                                  even after an incubation period of 60                                      or the second or third aerated pond,
LuminoTox was used to assess the                                                         minutes. Wastewaters treated in                                            since in these cases no toxicity was
toxicity of a number of wastewater                                                       activated sludge and aerated pond                                          detected even after 60 minutes of
samples and to derive an IC50 value for                                                  systems, on the other hand, showed a                                       incubation.
them, in just 15 minutes. Detection                                                      rise in toxicity with increasing

                                              PHYSICO-CHEMICAL CONDITIONS FOR THE EFFECTIVE USE OF LUMINOTOX1
                                                                                                                            Stability range of IC20 and IC50s(± 2 )
                           Parameters tested                 2
                                                                                                 Analyses of organic compounds                                          Analyses of inorganic compounds
                                                                                                      (reference: atrazine)                                                   (reference: copper)
                           Conductivity ( S)                                                                   0-3000                                                                     0-1000
                           Colour (Pt. Co. CU.)                                                                 0-450                                                                       0-450
                           Turbidité (NTUs)                                                                     0-350                                                                        0-503
                           Turbidity (CaCO3 ppm)                                                                0-320                                                                       0-180
                           Suspended solids (mg/L)                                                              0-500                                                                       0-500
1. Statistical validation by the Centre national en électrochimie et en technologies environnementales (CNETE) and Shawinigan College.
2. Tested at 22°C and pH 7.5. Testing can be done at temperatures between 4°C and 25°C, provided the blank and the test sample are at the same temperature.
   The pH must be adjusted if it is not a toxicity parameter and it does not fall in the 6.5-7.8 range.
3. The turbidity effect may be due to copper chelation or adsorption in response to the standards used

                                                   RESULTS OBTAINED FOR EFFLUENTS (BEFORE AND AFTER TREATMENT)
                                            Municipal wastewater                                                                           Pulp and paper mill

                                            100                                                                                           100

                                            80                                                                                            80
                                                                                                                       Inhibition (%)
                           Inhibition (%)

                                            60                                                                                            60
                                                                                                     15 min                                                                                          15 min
                                             40                                                      30 min                               40                                                         30 min
                                                                                                     60 min                                                                                          60 min
                                             20                                                                                           20

                                              00    20       40       60    80     100     120                                              0      20          40        60        80     100
                                            -20                                                                                           -20
                                                                  Effluent dilution (% v/v)                                                                                   Effluent dilution (% v/v)

                                            100                                                                                           100

                                                                                           Étang 1, 15 min
                                                                                           Étang 2, 15 min
                           Inhibition (%)

                                                                                                                        Inhibition (%)

                                            60                                             Étang 3, 15 min                                 60
                                                                                            Étang 1, 60 min                                                                                          15 min
                                            40                                              Étang 2, 60 min                                40                                                        30 min
                                                                                            Étang 3, 60 min                                                                                          60 min
                                            20                                                                                             20

                                              00    20       40       60    80     100     120                                              0      20          40        60         80     100
                                            -20                                                                                           -20
                                                                  Effluent dilution (% v/v)                                                                                   Effluent dilution (% v/v)
Note :
-Sampling was carried out in summer
-The toxicity of nitrogen ammonia was not assessed in these samples as the tests were performed using PECs insensitive to Nitrogen Ammonia
-The results for other treatments and industries are available upon request.
     E N V I R O N M E N T                               •       T E C H N O L O G I C A L       I N N O V A T I O N      –              I N N O V A T I V E        T O O L S     A N D   P R O C E S S E S
                                                                                                                                    Technological Innovation
                                                                                                                                    Technological Innovation Data
POTENTIAL AND LIMITATIONS                                                                                                           Sheets, produced by Environ-
                                                                                                                                    ment Canada, are intended for
                                                                                                                                    all firms, industries, organiza-
Potential                                                                                                                           tions and individuals interested
                                                                                                                                    in new environmental technolo-
• Useful to detect a wide                • Can reduce the number                       • Sensitivity of PECs to                     gies. Their purpose is to dissemi-
  variety of toxic molecules:              of samples sent to the                        light, except to green light               nate the results of technology
  phenols, sulphites, herbi-               laboratory for subsequent                     (regeneration occurs after                 development and demonstration
                                                                                                                                    projects carried out in the
  cides, hydrocarbons,                     analysis                                      30 minutes’ incubation in                  following sectors: wastewater,
  polycyclic aromatic                    • Toxicity detection limits                     the dark)                                  atmospheric emissions, contami-
  hydrocarbons (PAHs),                     are generally lower than                    • Industrial plants that                     nated soil, waste management,
  organic solvents and                     those required by regula-                     conduct Daphnia or trout                   hazardous waste, agri-environ-
                                                                                                                                    ment and innovative tools and
  heavy metals                             tory standards and other                      tests will have to correlate               processes.
• The use of PECs and algae                bioassays                                     their toxicity responses
  allows a larger number of                                                              with those obtained using                  Data sheets can be obtained
                                                                                                                                    from: Environment Canada
  molecules to be detected,                                                              LuminoTox                                  Innovation, Monitoring and
  including nitrogen                      Limitations                                  • Approval process                           Industrial Sectors
  ammonia                                                                                is ongoing                                 105 rue McGill, 4th Floor
                                         • PECs can be conserved for                                                                Montreal, Quebec H2Y 2E7
• Automation enables                                                                   • Does not replace
                                           12 days at 20°C, 2 months                                                                Tel.: 1 800 463-4311
  on-line water toxicity                                                                 the tests prescribed
                                           at 4°C and 4 months
  monitoring (Robot                                                                      by the regulations                         Publications are available on the
                                           at -20°C                                                                                 Environment Canada Web site in
                                                                                                                                    the “Publications” section:


INFORMATION                                                                                                                         Julie Leduc
                                                                                                                                    Writing Team:
                                                                                                                                    François Bellemare
In developing this technology,           For more information, please contact:                                                      Nathalie Boucher
Lab_Bell inc. received                                                                                                              Reviewers:
technical support from                                                                                                              Jean-René Michaud
                                          Lab_Bell inc.                                 Environment Canada
Environment Canada and                                                                                                              Christian Blaise
                                          François Bellemare, Ph.D.,                    Innovation, Monitoring and
financial support from the                                                                                                          Graphic Design:
                                          President and Scientific                      Industrial Sectors                          Lacroix O’Connor Lacroix
Canada Revenue Agency
                                          Director                                      Jean-René Michaud, Eng.,
(tax credits), Revenue Québec                                                                                                       Printing:
                                          Tel.: (819) 539-8508, ext. 107                M.Sc.A.                                     Les Impressions IntraMédia
(tax credits) and National
                                          Fax: (819) 539-8880                           Tel.: (514)283-9207
Research Council                                                                                                                    Published under the authority of
                                          E-mail:                                       Fax: (514) 496-2901
of Canada (NRC).                                                                                                                    the Minister of the Environment
                                                       E-mail:                                     (c) Her Majesty the Queen in
The demonstration of the
                                          Web Site:                                             Right of Canada, 2005
effectiveness of this technology                                                                                                    Cat. No.: En153-113/56-2005E
in the different test media was                                                                                                     ISSN: 1712-0209
funded under the Quebec                                                                                                             ISBN: 0-662-39710-X
Department of Education’s
                                                                                                                                    March 2005
PART program. Lab_Bell inc.
is grateful to the cities,                                                                                                          Cette fiche est également
municipalities and industrial                                                                                                       disponible en français sous
                                                                                                                                    le titre : LuminoTox : Outil de
plants that provided effluent                                                                                                       dépistage rapide de la toxicité

     E N V I R O N M E N T   •   T E C H N O L O G I C A L   I N N O V A T I O N   –   I N N O V A T I V E   T O O L S   A N D   P R O C E S S E S

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