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for Alga Growth Inhibition Tests
OECD Medium, Stock Solution I, II, III & IV



OECD Medium, Stock Solution I, II, III & IV are intended for preparation of OECD medium described in OECD Guidelines for the Testing of Chemicals 201.
This Solution is for Alga Growth Inhibition Tests that is one of WET (Whole Effluent Toxicity) methods. WET tests measure wastewater' s effects on specific test organisms’ ability to survive, grow and reproduce.



Pseudokirchneriella subcapitata
OECD Medium, Stock Solution I-IV


INDEX

  1. Features
  2. Preparation of OECD Culture Medium based on OECD Test Guidelines 201
  3. Composition List of OECD Stock Solution I ~ IV
  4. Example Analysis
  5. Product List
  6. Related Products



  7. Leaflet   (393KB/2P)


1. Features

  • No weighing and dissolution necessary
  • Mycoplasma tested
  • Endotoxin tested
  • Sterilized by 0.2 μm filtration
  • Based on OECD Test Guidelines 201

2. Preparation of OECD Culture Medium based on OECD Test Guidelines 201

For preparation of 1 L of OECD Culture Medium based on OECD Test Guidelines 201, take the OECD Medium, Stock solution I ~ IV as shown below,
mix together and dilute with ion-exchanged water, etc. to make the 1 L solution.

Product Name   Amount used  
  · OECD Medium, Stock Solution   I       ( x 100)
10 mL
  · OECD Medium, Stock Solution   II   ( x 1,000)
1 mL
  · OECD Medium, Stock Solution   III   ( x 1,000)
1 mL
  · OECD Medium, Stock Solution   IV   ( x 1,000)
1 mL
Prepare 1 L of OECD Culture Medium by adding an appropriate amount of ion-exchanged water, etc.    
1 L

3. Components of OECD Stock Solution I ~ IV



Product Components Concentration
(Stock Soln.; mg/L )
Concentration
(after dilution; mg/L)
OECD Medium, Stock Solution I    ( x 100)     [Wako Cat. #158-03315; 500 mL]
x 100
x 1
      NH4Cl
1,500.00
15.0000
    MgCl2 · 6H2O
1,200.00
12.0000
    CaCl2 · 2H2O
1,800.00
18.0000
    MgSO4 · 7H2O
1,500.00
15.0000
    KH2PO4
160.00
1.6000
OECD Medium, Stock Solution II (x 1,000)     [Wako Cat. #153-03321; 50 mL]
x 1,000
x 1
      FeCl3 · 6H2O
64.00
0.0640
    EDTA-Na2 · 2H2O
100.00
0.1000
OECD Medium, Stock Solution III ( x 1,000)     [Wako Cat. #150-03331; 50 mL]
x 1,000
x 1
      H3BO3
185.00
0.1850
    MnCl2 · 4H2O
415.00
0.4150
    ZnCl2
3.00
0.0030
    CoCl2 · 6H2O
1.50
0.0015
    CuCl2 · 2H2O
0.01
0.00001
    Na2MoO4 · 2H2O
7.00
0.0070
OECD Medium, Stock Solution IV (x 1,000)     [Wako Cat. #157-03341; 50 mL]
x 1,000
x 1
      NaHCO3
50,000.00
50.0000

 


4. Example Analysis

Data was provided by Chemicals Evaluation and Research Institue, Japan.
Target Item Pseudokirchneriella subcapitata
Exposure Condition
· Term 72 hours
· Test type Incubation with shaking (approx. 100 rpm)
· Concentration 1.6; 0.80; 0.40; 0.20 mg/L as potassium dichromate and the blank
Preparation of test solution
  Prepare OECD medium using OECD Medium, Stock Solution I ~ IV, followed by adding an appropriate volume of 100 mg/L potassium dichromate solution.
Environmental Condition
· Temperature in an incubator
21 ~ 24°C (fluctuation range: within ±2°C)
· Initial cell concentration
  0.75 x 103 cells/mL
· Volume of Test Solution
  600 mL/test level
    (100 mL/test vessel - 6 replicates/test level)
300 mL/test level
    (100 mL/test vessel - 3 replicates/test level)
· Test apparatus
  A sterile 500 mL-glass Conical flask with a stopper having regular pores such as SILICOSDEN®
Lighting Set point: 90 μE/m2/s
(Fluctuation range: within ±20% of the set point; within ± 15% of the mean)
Measurement of cell growth:    Cell concentration
Test Results
  ⇒ Effective concentration (EC50) (72 hours) = 1.1 mg/L
  This value was within the stipulated range of the background data at the test facility.
  ⇒ Efficacy of tests
  1. Growth rate in the target area at the end of exposure: 187-fold
    (Efficacy criterion: Growth rate of more than 16-fold)
  2. Mean of the variation coefficient from day to day of the growth rate in the target area: 10 %
    (Efficacy criterion: 35 % or less)
  3. Reproducibility coefficient of the growth rate in the target area: 1.1 %
    (Efficacy criterion: 7 % or less)
Conclusion
According to the test results, OECD Medium prepared with OECD Medium, Stock Solution I ~ IV is decided that it is useful as medium for alga growth inhibition tests.



Growth speed in test level Growth curve in test level

5. Product List

Product Name Pkg. Size Wako Cat. No. Grade Storage Condition
OECD Medium, Stock Solution I     ( x 100)
500 mL
for Plant Culture
Keep at 2-10°C
OECD Medium, Stock Solution II     ( x 1,000)
50 mL
OECD Medium, Stock Solution III     ( x 1,000)
50 mL
OECD Medium, Stock Solution IV     ( x 1,000)
50 mL

6. Related Products

Product Name Pkg. Size Wako Cat. No. Grade Storage Condition
Potassium Dichromate, 99.5+% (Titration)
25 g
JIS Special Grade
Keep at
room temperature
100 g
500 g
Sterile Water, Endotoxin Free
500 mL
for Cell Culture
Distilled Water, Deionized, Sterile
100 mL
Manufactured
by Nippon Gene
100 mL x 6
500 mL
Agar (Powder)
25 g
for Plant Culture
500 g


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