LVA-results

Results of trace contaminants in
manure and co-substrates
LVA - Sabrina Geist, MSc; Bernhard Huber; Michael Gartner, PhD; Céline Lesueur, PhD
Introduction
The purpose of this work was to determine trace contaminants in manure and co-substrates for the ManureEcoMine project. Important trace contaminants
in manure are veterinary drugs and heavy metals due to the direct ingestion of these substances. In co-substrates from plant origin, pesticides, mycotoxins
and heavy metals are relevant.
For the extraction of pesticides the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) method was used. The pesticides were detected with GC-MS/MS and HPLC-MS/MS. The veterinary drugs were
extracted with a new developed extraction method (see other poster) and were separated and detected with UHPLC-MS/MS.
The analysis of the heavy metals is based on Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). The samples were treated by microwave assisted digestion and the resulting solutions were analysed by
ICP-MS. To avoid memory effects often experienced with mercury and also for stabilisation, hydrochloric acid was added to all standards and samples.
Results of co-substrates
Results of mycotoxins
Results of heavy metals
Afla b1 Afla b2 Afla g1 Afla g2 DON Fum b1 Fum b2 HT-2 toxin OTA
(µg/kg) (µg/kg) (µg/kg) (µg/kg) (µg/kg) (µg/kg) (µg/kg) (µg/kg) (µg/kg)
Maize corn silage
546
vegetable waste 1
vegetable waste 2
Mais 8/01
521
Greix HP humit
BioBerica Greix HP Humit
Greix HP sec
Bioberica Greix HP sec
Segregats 1
Segregats 2
Co-substrate of corn 1
3010 15709
1881
Co-substrate of corn 2
0,20
2659 18655
2157
Co-substrate of corn 3
0,30
2572 12182
1833
Sample name
T-2 toxin
(µg/kg)
-
ZON
(µg/kg)
756
569
902
608
768
Sample name
Maize corn silage
vegetable waste 1
vegetable waste 2
Mais 8/01
Greix HP humit
BioBerica Greix HP Humit
Greix HP sec
Bioberica Greix HP sec
Segregats 1
Segregats 2
Co-substrate of corn 1
Co-substrate of corn 2
Co-substrate of corn 3
Table 1: Measured values of mycotoxins in co-substrates: Afla b1 (Aflatoxin b1) - LOQ = 0,2 µg/kg, Afla b2
(Aflatoxin b2) - LOQ = 0,2 µg/kg, Afla g1 (Aflatoxin g1) - LOQ = 0,2 µg/kg, Afla g2 (Aflatoxin g2) - LOQ = 0,2
µg/kg, DON (Deoxynivalenol) - LOQ = 40 µg/kg, Fum b1 (Fumonisin b1) - LOQ = 100 µg/kg, Fum b2 (Fumonisin
b2) - LOQ = 100 µg/kg, HT-2 toxin - LOQ = 10 µg/kg, OTA (Ochratoxin-A) - LOQ = 2 µg/kg, T-2 toxin - LOQ =
10 µg/kg, ZON (Zearalenone) - LOQ = 20 µg/kg. (LOQ = Limit of Quantification)
Chromium (Cr) Nickel (Ni)
mg/kg
mg/kg
1,60
1,40
1,70
1,00
-
Copper (Cu)
mg/kg
8,30
8,60
1,40
4,10
4,10
7,40
6,90
0,62
0,59
2,00
2,30
2,50
Results of pesticides
Zinc (Zn)
mg/kg
5,90
37,50
40,60
7,00
21,20
22,10
35,20
34,10
23,60
28,40
30,50
Arsenic (As) Cadmium (Cd) Mercury (Hg)
mg/kg
mg/kg
mg/kg
0,06
0,03
0,02
0,16
0,04
0,19
0,04
0,07
0,02
0,05
0,03
0,06
0,04
0,07
0,05
0,07
0,06
0,01
0,01
0,02
0,03
0,01
Lead (Pb)
mg/kg
0,23
0,47
0,57
0,21
0,04
0,04
0,02
0,02
0,02
Table 2: Measured values of heavy metals in co-substrates: Chromium - LOQ 1 mg/kg, Nickel - LOQ 0,5 mg/kg,
Copper - LOQ 0,5 mg/kg, Zinc - LOQ 1 mg/kg, Arsenic - LOQ 0,01 mg/kg, Cadmium - LOQ 0,01 mg/kg, Mercury
- LOQ 0,01 mg/kg, Lead – LOQ 0,01 mg/kg (LOQ = Limit of Quantification)
Sample name
Results
LOQ
mg/kg
(mg/kg)
Maize corn silage
0,01
vegetable waste 1
0.71 mg/kg Ethoxyquin, 0.050 mg/kg Isoprocarb, 0.010 mg/kg Pirimiphos-methyl 0,01
vegetable waste 2
0.33 mg/kg Ethoxyquin, 0.045 mg/kg Isoprocarb
0,01
Mais 8/01
0,01
Greix HP humit
0,01
BioBerica Greix HP Humit
0,01
Greix HP sec
0,01
Bioberica Greix HP sec
0,01
Segregats 1
0,01
Segregats 2
0,01
Co-substrate of corn 1 0.13 mg/kg Deltamethrin, 1.60 mg/kg Piperonylbutoxide
0,01
Co-substrate of corn 2 0.22 mg/kg Deltamethrin, 2.4 mg/kg Piperonylbutoxide
0,01
Co-substrate of corn 3 0.19 mg/kg Deltamethrin, 2.2 mg/kg Piperonylbutoxide
0,01
Table 3: Measured values of pesticides in co-substrates, around 450 pesticides were
analysed. (LOQ = Limit of Quantification)
Results of manure
Results of veterinary drugs in liquid manure
Results of heavy metals in liquid manure
Chromium (Cr) Nickel (Ni)
mg/kg
mg/kg
Copper (Cu)
mg/kg
Zinc (Zn)
mg/kg
Lead (Pb)
mg/kg
dry
matter (%)
0,66
19,5
78,9
0,09
0,04
-
0,25
7,8
0,45
0,53
18,2
74,9
0,08
0,03
-
0,23
6,7
Manure 4.1 (NL)
0,42
0,65
16,7
68,9
0,08
0,03
-
0,21
6,8
6,5
Manure 4.2 (NL)
0,46
0,55
17,7
71,9
0,08
0,03
-
0,23
6,5
-
12
Manure 9.1 (NL)
0,64
0,89
26,1
108
0,12
0,05
-
0,27
12
-
-
11,7
Manure 9.2 (NL)
0,67
0,81
24,9
105
0,12
0,046
-
0,27
11,7
-
59,164
-
3,9
Manure from Balsa glass (ESP)
0,2
0,22
3
17
0,05
0,01
-
0,56
3,9
-
-
63,374
-
4
Manure from Balsa plastic 1 (ESP)
0,2
0,23
2,9
16,8
0,05
0,01
-
0,53
4
-
-
60,84
detected < LOQ
3,9
Manure from Balsa plastic 2 (ESP)
0,19
0,22
2,9
16,2
0,05
0,01
0,02
0,52
3,9
Lead (Pb)
mg/kg DM
dry
matter (%)
Oxytetracycline
(µg/kg)
Tylosin
(µg/kg)
Lincomycin
(µg/kg)
Enrofloxacin
(µg/kg)
dry
matter (%)
Manure 1.1 (NL)
18,1
5,8
-
-
7,8
Manure 1.1 (NL)
0,48
Manure 1.2 (NL)
17,8
6,5
-
-
6,7
Manure 1.2 (NL)
Manure 4.1 (NL)
22,7
7,8
-
-
6,8
Manure 4.2 (NL)
18,4
5,5
-
-
Manure 9.1 (NL)
12,6
detected < LOQ
-
Manure 9.2 (NL)
12,8
detected < LOQ
Manure from Balsa glass (ESP)
-
Manure from Balsa plastic 1 (ESP)
Manure from Balsa plastic 2 (ESP)
Sample name
Table 4: Measured values of veterinary drugs in liquid manure: Oxytetracycline - LOQ 1 mg/kg,
LOD = 0,5 mg/kg; Tylosin - LOQ 5 mg/kg, LOD = 1 mg/kg; Lincomycin - LOQ 1 mg/kg, LOD =
0,5 mg/kg; Enrofloxacin - LOQ 1 mg/kg, LOD = 0,5 mg/kg (LOQ = Limit of Quantification; LOD =
Limit of Detection)
Results of veterinary drugs in manure (dry matter)
Sample name
Table 5: Measured values of heavy metals referring to the liquid sample. The values include the water content of manure.
Results of heavy metals in manure (dry matter)
Oxytetracycline
(µg/kg) DM
Tylosin
(µg/kg) DM
Lincomycin
(µg/kg) DM
Enrofloxacin
(µg/kg) DM
dry
matter (%)
Manure 1.1 (NL)
231,5
74,6
-
-
7,8
Manure 1.1 (NL)
6,2
Manure 1.2 (NL)
265,3
97,2
-
-
6,7
Manure 1.2 (NL)
Manure 4.1 (NL)
334,4
114,0
-
-
6,8
Manure 4.2 (NL)
283,8
84,9
-
-
Manure 9.1 (NL)
104,9
detected < LOQ
-
Manure 9.2 (NL)
109,5
detected < LOQ
Manure from Balsa glass (ESP)
-
Manure from Balsa plastic 1 (ESP)
Manure from Balsa plastic 2 (ESP)
Sample name
Arsenic (As) Cadmium (Cd) Mercury (Hg)
mg/kg
mg/kg
mg/kg
Copper (Cu)
mg/kg DM
Zinc (Zn)
mg/kg DM
8,5
250,0
1011,5
1,2
0,5
-
3,2
12
6,7
7,9
271,6
1117,9
1,2
0,4
-
3,4
11,7
Manure 4.1 (NL)
6,2
9,6
245,6
1013,2
1,2
0,4
-
3,1
7,8
6,5
Manure 4.2 (NL)
7,1
8,5
272,3
1106,2
1,2
0,5
-
3,5
6,7
-
12
Manure 9.1 (NL)
5,3
7,4
217,5
900,0
1,0
0,4
-
2,3
6,8
-
-
11,7
Manure 9.2 (NL)
5,7
6,9
212,8
897,4
1,0
0,4
-
2,3
6,5
-
1517,0
-
3,9
Manure from Balsa glass (ESP)
5,1
5,6
76,9
435,9
1,3
0,3
-
14,4
3,9
-
-
1584,4
-
4
Manure from Balsa plastic 1 (ESP)
5,0
5,8
72,5
420,0
1,3
0,3
-
13,3
4
-
-
1560,0
detected < LOQ
3,9
Manure from Balsa plastic 2 (ESP)
4,9
5,6
74,4
415,4
1,3
0,3
0,5
13,3
3,9
Table 6: Measured values of veterinary drugs referring to the dry matter. The values consider
the water content of manure.
Results & Discussion
Sample name
Chromium (Cr) Nickel (Ni)
mg/kg DM
mg/kg DM
Arsenic (As) Cadmium (Cd) Mercury (Hg)
mg/kg DM
mg/kg DM
mg/kg DM
Table 7 Measured values of heavy metals in liquid manure: Chromium - LOQ 2 mg/kg dm, Nickel - LOQ 1 mg/kg dm, Copper - LOQ 1 mg/kg
dm, Zinc - LOQ 4 mg/kg dm, Arsenic - LOQ 0,4 mg/kg dm, Cadmium - LOQ 0,03 mg/kg dm, Mercury - LOQ 0,02 mg/kg dm, Lead – LOQ 2
mg/kg dm (LOQ = Limit of Quantification, dm = dry matter)
Heavy metals are naturally occuring substances in the earth crust and some of them are essential trace elements in feed for health and physical working capacity of animals. Especially zinc and copper are heavy
metals which are added to animal feed. This fact is reflected by the results of the manure samples. The values of zinc and copper were definitely higher than the other analysed heavy metals.
Veterinary drugs were analysed after 15 months since the new extraction method was developed. Nevertheless some veterinary drugs (oxytetracyclin, tylosin and lincomycin) could still be quantified after the
samples were stored in a freezer. Pesicides were found in vegetable waste and maize. These results are not unusual because only these co-substrates could be treated directly with plant protection products.
The detected mycotoxins were only found in maize based co-substrates and are common for maize products.
Conclusion
For the risk evaluation it is very important to monitor the substances which could effect the properties of the end products, thus all inputs and outputs have to be analysed. As heavy metals can not be degraded,
they will be very relevant for the risk assessment, especially since they were found in the manure and co-substrate samples. Veterinary drugs, mycotoxins and pesticides have the property to degrade with time
(depending on the substance), but since these substances were found in the samples, they should and will also be tested in the end products.
Images from http://pinstake.com/kuh-jpg/ and http://diepresse.com/home/panorama/oesterreich/532887/Lawinenexperiment-mit-Schweinen-vorerst-gestoppt