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Evaluation of the effect of surgical and immunological castration of male pigs on boar taint compounds in oral fluid and fat tissue by LC-MS/MS method

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Fig. 1

Chromatographic separation of tested compounds on a column: a) ZORBAX SB-C18 (50 mm × 2.1 mm, 1.8 μm) and b) Poroshell 120-EC C18 (150 × 2.1 mm, 2.7 μm)
Chromatographic separation of tested compounds on a column: a) ZORBAX SB-C18 (50 mm × 2.1 mm, 1.8 μm) and b) Poroshell 120-EC C18 (150 × 2.1 mm, 2.7 μm)

Fig. 2

LC-MS/MS chromatograms of oral fluid samples in which were detected: a) skatole at a concentration of 136 μg L−1 (gilts); b) indole with a concentration of 86.1 μg L−1 (SC boars)
LC-MS/MS chromatograms of oral fluid samples in which were detected: a) skatole at a concentration of 136 μg L−1 (gilts); b) indole with a concentration of 86.1 μg L−1 (SC boars)

Fig. 3

LC-MS/MS chromatogram of a fat sample taken from an immunologically castrated boar in which an indole concentration of 21 μg L−1 was determined
LC-MS/MS chromatogram of a fat sample taken from an immunologically castrated boar in which an indole concentration of 21 μg L−1 was determined

HPLC gradient

Time (min)Flow (mL min−1)Eluent A (%)Eluent B (%)
0.010.354060
2.000.356040
3.000.359010
3.990.359010
4.000.459010
6.000.451000
7.000.451000
7.010.351000
7.100.354060
10.000.354060

Results of studies on compounds responsible for boar taint in fat samples taken from gilts, surgically and immunologically castrated boars

GiltsSkatoleIndoleAndrostenone
Number of samples tested (n)151515
Number of analyte-containing samples7110
C average (μg L−1)65.225.3
SD (μg L−1)8.040.9
C min-max (μg L−1)57.7–78.35.4–146.5
IMC boars
Number of samples tested (n)151515
Number of analyte-containing samples10110
C average (μg L−1)60.315.6
SD (μg L−1)5.88.8
C min-max (μg L−1)51.3–69.25.5–30.0
SC boars
Number of samples tested (n)151515
Number of analyte-containing samples790
C average (μg L−1)60.318.4
SD (μg L−1)5.19.0
C min-max (μg L−1)54.0–67.55.2–35.2

LC-MS/MS parameters (MRM) used for identification of indole, skatole and androstenone

AnalyteStructureMRM (mass-to-charge (m/z))Collision energy (eV)
Indole (2,3-benzopyrrole)118.0 > 91.123
118.0 > 65.033
118.0 > 39.147
Skatole (3-methylindole)132.5 > 117.122
132.5 > 89.041
132.5 > 90.233
2-methylindole (IS)132.5 > 117.223
132.5 > 89.141
132.5 > 90.034
Androstenone273.5 > 255.313
273.5 > 93.226
273.5 > 77.225
Testosterone-d2 (IS)291.2 > 111.126
291.2 > 99.224
IS – internal standard

LC-MS/MS method validation parameters for indole, skatole and androstenone in swine oral fluid and fat

ParameterIndoleSkatoleAndrostenone
Oral fluid
Calibration curvescorrelation coefficient0.99900.99880.9967
Matrix calibration curvescorrelation coefficient0.99800.99640.9859
Limit of quantificationLOQ (μg L−1)55050
RepeatabilityC.V. (%)28.2030.024.4
Recovery(%)75.470.095.3
Matrix effect%101130
Fat
Calibration curvescorrelation coefficient0.99880.99890.9953
Matrix calibration curvescorrelation coefficient0.99680.98750.9840
Limit of quantificationLOQ (μg L−1)550100
RepeatabilityC.V. (%)5.032.926.3
Recovery(%)121.070.0102.8
Matrix effect%75.68227

Results of tests of pooled oral fluid samples taken from animals from four experimental groups each consisting of 14 animals

Sampling timeSkatoleIndoleAndrostenone
Gilts (control groups for IMC boars)(μg L−1)(μg L−1)(μg L−1)
Before giving Improvac to the boars---
Two weeks after the first boar immunisation with Improvac (pen 1)---
Two weeks after the first boar immunisation with Improvac (pen 2)---
Two weeks after the second immunisation with Improvac before slaughter
Pen 1309.4--
Pen 2---
Pen 3106.3--
Pen 4136.438.6-
Immunologically castrated boars (IMC)
Before giving Improvac82.5--
Two weeks after the first immunisation with Improvac (pen 1)---
Two weeks after the first immunisation with Improvac (pen 2)---
Two weeks after the second immunisation with Improvac, before slaughter---
Surgically castrated boars (SC) – samples taken before slaughter
Pen 1-23.8-
Pen 269.712.2-
Pen 3---
Pen 4-86.1-
Gilts and boars (SC), 50/50 – samples taken before slaughter
Pen 1153.148.9-
Pen 2---
Pen 3-125.4-
Pen 4---
eISSN:
2450-8608
Langue:
Anglais
Périodicité:
4 fois par an
Sujets de la revue:
Life Sciences, Molecular Biology, Microbiology and Virology, other, Medicine, Veterinary Medicine