Media FAQ

Media for the duckweed test (lemna test)

Steinberg

Stoff	Nährmedium nach Steinberg (modifiziert)
Makroelemente	Molgew.	mg/l	mmol/l
KNO3	101,12	350,00	3,46
Ca(NO3)2*4H2O	236,15	295,00	1,25
KH2PO4	136,09	90,00	0,66
K2HPO4	174,18	12,60	0,072
MgSO4*7H2O	246,37	100,00	0,41
Mikroelemente	Molgew.	µg/l	µmol/l
H3BO3	61,83	120,00	1,94
ZnSO4*7H2O	287,43	180,00	0,63
Na2MoO4*2H2O	241,92	44,00	0,18
MnCl2*4H2O	197,84	180,00	0,91
FeCl3*6H2O	270,21	760,00	2,81
Titriplex III (EDTA)	372,24	1500,00	4,03

Solution 1:

1. Makroelemente (50fach konzentriert)	g/l
Solution 1:
KNO3	17,50
KH2PO4	4,5
K2HPO4	0,63
Solution 2:
MgSO4*7H2O	5,00
Solution 3:
Ca(NO3)2*4H2O	14,75

2. Mikroelemente (1000fach konzentriert)	mg/l
Solution 4:
H3BO3	120,0
ZnSO4*7H2O	180,0
Na2MoO4*2H2O	44,0
MnCl2*4H2O	180,0
Solution 5:
FeCl3*6H2O	760,00
Titriplex III (EDTA)	1500,00

Start Kultivating with:

autoclave at 121°C, for 20 Min. or use 0.2µm sterilfilter

OECD, 1998 – Culture and Test media for Lemna minor (SIS growth medium)

 

Substance	Stock Solution [g/L]	Mediuma [mg/L]	Element	Stock Solution
MgSO4 – 7 H2O	15	75	NIb	II
NaNO3	8,5	85	NI	I
CaCl2 – 2 H2O	7,2	36	NI	III
Na2CO3	4,0	20	NI	IV
KH2PO4	1,34	13,4	NI	I
H3BO3	1,0	1,0	NI	V
MnCl2 – 4 H2O	0,2	0,2	NI	V
Na2MoO4 – 2H2O	0,010	0,010	NI	V
ZnSO4 – 7 H2O	0,050	0,050	NI	V
CuSO4 – 5 H2O	0,005	0,005	NI	V
Co(No3)2 – 6 H2O	0,010	0,010	NI	V
Na2 EDTA	1,4	1,4	NI	VIc
FeCl3 – 6 H2O	0,84	0,84	NI	VIc
MOPS (buffer)d	488	488	NI	VIIc
PH Adjustment	PH adjustment to 6,5 ± 0,2 by addition of NaOH or HCl
Sterilization	Stock solutions I to V are sterilized by autoclaving (120°C, 15 min.) or by membrane filtration (pore diameter 0,2 μm); stock solutions IV (and optional VII) are sterilized by membrane filtration only (i.e. , these should not be autoclaved).
a	Concentration of substance in Medium
b	Not indicated
c	Added after autoclaving
d	MOPS buffer is only required when pH control of the test medium is particularly important (e.g. when testing metals or substances which are hydrolytically unstable)

Zitiert aus: Report EPS 1/RM/37; Dec. 1998,

Biological Test Method: Test for Measuring the Inhibition of Growth using the Freshwater Macrophyte, Lemna minor

Method Development and Application Section  
Environmental Technology Centre
Environment Canada          
Ottawa, Ontario

ASTM, 1991 – 20X-AAP Medium^a for Culturing and Testing Lemna gibba

004

Substance	Stock Solution [g/L]	Medium [mg/L]	Element	Stock Solution
MgSO4 – 7 H2O	14,70	38,22	S	D
NaNO3	25,50	84,00	N	A
CaCl2 – 2 H2O	4,410	24,04	Ca	F
Na2CO3	15,00 ---	220,02 42,86	Na C	B B
KH2PO4	1,044 ---	9,38 3,72	K P	C C
H3BO3	0,18552	0,64920	B	G
MnCl2 – 4 H2O	0,41561	2,30748	Mn	G
Na2MoO4 – 2H2O	0,00726	0,05756	Mo	G
ZnCl2	0,00327	0,0314	Zn	G
CuCl2 – 2 H2O	1,2 x 10^-5	8 x 10^-5	Cu	G
CoCl6– 6 H2O	0,00143	0,00708	Co	G
Na2 EDTA 2 H2O	0,300	---	---	G
FeCl3 – 6 H2O	0,160	0,66102	Fe	G
MgCl2 – 6 H2O	12,164	58,08	Mg	E
PH Adjustment	PH adjustment to 7,4-7,6 by addition 0,1N NaOH or HCl
Sterilization	sterilized by membrane filtration only (pore diameter 0,2 μm)

Zitiert aus: Report EPS 1/RM/37; Dec. 1998,

Biological Test Method: Test for Measuring the Inhibition of Growth using the Freshwater Macrophyte, Lemna minor

Method Development and Application Section  
Environmental Technology Centre
Environment Canada          
Ottawa, Ontario

ASTM, 1991 – Modified Hoagland's Medium^a (no Sucrose or EDTA) for Culturing and Testing Lemna gibba

006

Substance	Stock Solution [g/L]	Mediumb [mg/L]	Element	Stock Solution
MgSO4 – 7 H2O	NIc	75	NI	Ad
KNO3	NI	85	NI	A
Ca(NO3)2 – 4 H2O	NI	36	NI	A
KH2PO4	NI	13,4	NI	A
H3BO3	NI	1,0	NI	Bd
MnCl2 – 4 H2O	NI	0,2	NI	B
Na2MoO4 – 2H2O	NI	0,010	NI	B
ZnSO4 – 7 H2O	NI	0,050	NI	B
CuSO4 – 5 H2O	NI	0,005	NI	B
FeCl3 – 6 H2O	NI	0,010	NI	A
Tartaric Acid	NI	1,4	NI	A
PH Adjustment	PH adjustment to 5,01 ± 0,1 by addition of 1N KOH or HCl
Sterilization	Autoclave 20 min. at 121°C and 1,1 Kg/cm²
a	This medium is the same as Hoagland´s E+ medium exept the sucrose, bacto-tryptone, yeast and EDTA have been excluded
b	Concentration of substance in Medium
c	Not indicated
d	Add each chemical (A) to distilled or deionized water Add 1 ml of micronutrient stock solution

Zitiert aus: Report EPS 1/RM/37; Dec. 1998,

Biological Test Method: Test for Measuring the Inhibition of Growth using the Freshwater Macrophyte, Lemna minor

Method Development and Application Section  
Environmental Technology Centre
Environment Canada          
Ottawa, Ontario