Table 1.
Quantification limit using LC-MS/MS for acetamiprid, azoxystrobin, estriol, estrone, progesterone, and testosterone in rat liver.
Fig 1.
Linearity coefficients of calibration curves for all six compounds (acetamiprid, azoxystrobin, estrone, estriol, progesterone, and testosterone).
Fig 2.
Experiment design to study the toxicity of acetamiprid and azoxystrobin on rats.
Fig 3.
A. Column comparison (poroshel, supelco, and synergi) with three compounds (estriol, estrone, and progesterone). HPLC columns comparison (n = 5). Poreshel 120 EC-C18 50X 3 mm, 2.7 μm (Agilent), Titan C18 50 X 2.1 mm, 1.9 μm (Supelco), and Synergi Fusion 50 X 2 mm, 4 μm (Phenomenex). B. Column comparison (poroshel, supelco, and synergi) with three compounds (testosterone, acetamiprid, and azoxystrobin). HPLC columns comparison (n = 5). Poreshel 120 EC-C18 50X 3 mm, 2.7 μm (Agilent), Titan C18 50 X 2.1 mm, 1.9 μm (Supelco), and Synergi Fusion 50 X 2 mm, 4 μm (Phenomenex).
Fig 4.
Extracted ion chromatogram of azoxystrobin 50 ppb, injection volume of 2, 5, and 25 μL.
a 5 μL injection volume was chosen to achieve maximum detection and quantification limitations.
Fig 5.
LC-MS/MS chromatograms of a spiked liver sample on LOQ level.
Table 2.
Mass parameters of parent and daughter ions of pesticides and hormones on LC-MS/MS (n = 5).
Table 3.
Extraction solvent comparison by using different solvents in the liver tissue (n = 5).
Table 4.
Effect of acetamiprid and azoxystrobin on the body weight (g) of rats during the two months of exposure.
Fig 6.
A. Serum testosterone levels and acetamiprid residues levels after a month of different acetamiprid doses treatment of the male rats. Data presented as mean S.E. a Significant difference as compared to 1/40 LD50, and b significant difference as compared to 1/20 of LD50 (n = 4, P≤ 0.05). B. Serum testosterone and progesterone levels as well as acetamiprid residues after 2 months of different acetamiprid doses treatment of male rats. Data presented as mean S.E. a Significant difference as compared to 1/40 LD50, and b significant difference as compared to 1/20 of LD50 (n = 4, P≤ 0.05).
Fig 7.
A. Serum testosterone levels and azoxystrobin residues after a month of different azoxystrobin doses treatment of male rats. Data presented as mean S.E. a Significant difference as compared to 1/40 LD50, and b significant difference as compared to 1/20 of LD50 (n = 4, P≤ 0.05). B. Serum testosterone and progesterone levels as well as azoxystrobin residues after 2 months of different azoxystrobin doses treatment on male rats. Data presented as mean S.E. a Significant difference as compared to 1/40 LD50, and b significant difference as compared to 1/20 of LD50 (n = 4, P≤ 0.05).
Fig 8.
A. Testis testosterone and progesterone levels as well as acetamiprid residues after a month of different treatment of acetamiprid doses on male rats. Data presented as mean S.E. a Significant difference as compared to 1/40 LD50, and b significant difference as compared to 1/20 of LD50 (n = 4, P≤ 0.05). B. Testis testosterone and progesterone levels as well as acetamiprid residues after 2 months of different treatment of acetamiprid doses on male rats. Data presented as mean S.E. a Significant difference as compared to 1/40 LD50, and b significant difference as compared to 1/20 of LD50 (n = 4, P≤ 0.05).
Fig 9.
A. Testis testosterone and progesterone levels as well as azoxystrobin residues after a month treatment of different azoxystrobin doses on male rats. Data presented as mean S.E. a Significant difference as compared to 1/40 LD50, and b significant difference as compared to 1/20 of LD50 (n = 4, P≤ 0.05). B: Testis testosterone and progesterone levels as well as azoxystrobin residues after 2 months treatment of different azoxystrobin doses on male rats. Data presented as mean S.E. a Significant difference as compared to 1/40 LD50, and b significant difference as compared to 1/20 of LD50 (n = 4, P≤ 0.05).
Fig 10.
Liver acetamiprid residues after 1 and 2 months treatment of with different doses of acetamiprid on male rats.
Data presented as mean S.E. a Significant difference as compared to 1/40 LD50, and b significant difference as compared to 1/20 of LD50 (n = 4, P≤ 0.05).
Fig 11.
Liver azoxystrobin residues after 1 and 2 months treatment with different doses of azoxystrobin on male rats.
Data presented as mean S.E. a Significant difference as compared to 1/40 LD50, and b significant (n = 4, P≤ 0.05).
Table 5.
Mean values of testosterone and progesterone in control samples (n = 4/each month).