Transcript boars2018.com

Federal Department of Economic Affairs FDEA
Agroscope Liebefeld-Posieux Research Station ALP
Analysis of Androstenone,
Skatole and Indole in Biopsy
Boar Samples
Ampuero Kragten1, S., Robatel1, A., Mattei2, S., Bee1,
G., Hofer3, A.
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Agroscope Liebefeld-Posieux ALP, 2 University of Zurich, 3 SUISAG
Boars heading for 2018, Amsterdam, 2. December 2011
The Project
• Developed at ALP as part of a breeding project from Suisag,
Switzerland
• Funded by the Swiss Commission of Innovation and
Technology
• Boars of Swiss Large White breed
• Samples taken at the neck region
Boars heading for 2018, Amsterdam, 2. December 2011
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Biopsy / Liquid Fat
120
Liquid Fat Mass [mg]
100
80
60
R² = 0.2789
40
20
0
100
150
200
250
Biopsy Sample Mass [mg]
Heavy boars
Light boars
R2 = 0.014
N=6
R2 = 0.317
N = 14
Boars heading for 2018, Amsterdam, 2. December 2011
300
350
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Biopsy / Liquid Fat
300
Liquid Fat Mass [mg]
250
200
150
R² = 0.8316
100
50
0
250
350
450
550
Biopsy Sample Mass [mg]
Boars heading for 2018, Amsterdam, 2. December 2011
650
750
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Biopsy / Liquid Fat
Optimum microwave heating cycle: 9min ( 3x3min; 600 W)
Biopsy mass set to 300 mg
150
Liquid Fat Mass [mg]
140
130
120
110
100
90
80
70
60
50
100
200
300
400
500
Biopsy Sample Mass [mg]
Boars heading for 2018, Amsterdam, 2. December 2011
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Biopsy / Liquid Fat
Sample Mass for Analysis [mg]
140
"1st aliquot"
120
"2nd aliquot"
100
80
60
40
20
0
0
100
200
300
400
Sample Number
Life Weight, mean (STD):
138 (61) kg
Liquid Fat, mean (STD):
91.3 (40.1) mg
Boars heading for 2018, Amsterdam, 2. December 2011
500
600
700
583 samples
58 replicates
2 samples ~0 LF
5 samples < 20 mg LF
42 samples < 40 mg LF
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Extraction procedure
200 ml MeOH:H2O (95:5) +
Std int (androstanone + 2-methyl indole)
(150 ml)
70 – 100 mg Liquid Fat
(40 – 70 mg LF)
Boars heading for 2018, Amsterdam, 2. December 2011
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HPLC procedure
Derivatization in-situ:
3 ml
Dansylhydrazine
2 ml
H2O
+
+
+
+
+
2 ml
BF3
20 ml
extract
2 min
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HPLC procedure
Injection:
Derivat.:
Column:
Eluent:
Flow rate:
pH:
Temp.:
FLD:
10 ml
Androstenone
Skatol, Indole
yes
SB C18
THF/phosphate buffer (50:50)
1.3 ml/min
6
40°C
346/521 nm
no
Eclipse XDB C18
MeOH/phosphate buffer (50:50)
1.3 ml/min
6
40°C
285/340 nm
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Biopsy vs. Normal Analysis
Biopsy Method, A conc. [mg/kg]
1.00
y = 0.8511x - 0.0828
R² = 0.825
0.90
y = 1.0138x - 0.1367
R² = 0.9193
0.80
0.70
y = 0.9029x - 0.0571
R² = 0.8636
0.60
0.50
0.40
H1-biopsie
0.30
H2-biopsie
0.20
0.10
0.00
0.00
R-biopsie
0.20
0.40
0.60
0.80
1.00
Normal Method, A conc. [mg/kg]
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Biopsy vs. Normal Analysis
1.80
y = 0.6484x + 0.016
R² = 0.8836
Biopsy Method, A conc. [mg/kg]
1.60
y = 0.6952x + 0.0186
R² = 0.889
1.40
y = 0.8489x - 0.0308
R² = 0.9536
1.20
1.00
0.80
H1-biopsie
0.60
H2-biopsie
0.40
R-biopsie
0.20
0.00
0.00
0.50
1.00
1.50
Normal Method, A conc. [mg/kg]
Boars heading for 2018, Amsterdam, 2. December 2011
2.00
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Conclusions
• Biopsy method is adapted to breeding programs
• A good compromise between biopsy size and critical
analytical mass was found (0.9 % n.d.)
• Analysis of skatole and indole is less problematic
• Deviation in accuracy: <3% at 2.0 mg/kg A (70-100 mg LF)
• Deviation in accuracy: <12% at 0.3 mg/kg A
or for analysis with < 70 mg LF
• Good correlation with normal method
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