Male or female Sprague-Dawley rats (8–12 weeks old) were purchased from Damul Sciences (Daejeon-si, Korea). This study was conducted in accordance with the Guidelines for the Care and Use of Laboratory Animals and approved by the Institutional Animal Care and Use Committee (IACUC, No: IACUC140047). Over the acclimatization period and during the study, the range of temperatures and humidity were recorded daily; ambient temperature was maintained in the 25 ± 1 °C range; humidity level was measured as 50 ± 5% RH; ventilation was 10–15 th/h. The room had an entirely artificial fluorescent lighting with a controlled cycle of 12 h light and 12 h dark. The measurement of body weight was conducted just before the administration. Fasting was done for
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Blood samples were collected from the jugular vein at 0.17, 0.33, 0.5, 0.75, 1, 2, 4, 6, 8, 12, and 24 h after the administration. The female rats (n = 5/group) were given a single dose 2 mg/kg of PFOS by oral and IV injection. Blood samples were collected from the jugular vein at 0.25, 0.5, 1, 2, 4, 8, 12, 18, 24 h and 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70 day after IV administration and at 0.25, 0.5, 1, 3, 5, 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70 day after the oral administration. In the case of PFHxS, female rats (n = 5/group) were administered a single dose of PFHxS at 4 mg/kg by oral and IV. Blood samples were collected from the jugular vein at 0.5, 1, 2, 4, 6, 12, 24, 48, 72, 120, 168, 240, and 336 h after the administration. Each PFC solution was prepared in saline or distilled water. To obtain plasma sample, all the blood samples were collected in heparinized tubes and immediately centrifuged (5,000 × g, 10 min). Following the centrifugation, plasma was stored at –80 °C until analysis. Tissue samples of the liver, kidney, heart, lung, and spleen were dissected and taken immediately after taking the blood sample at the terminal time (PFOA, 24 h; PFOS, 70 day; PFHxS, 336 h) and stored frozen (−80 °C) until …show more content…
The liquid chromatography was performed using an Acquity UPLC® system (Waters Corp., Milford, MA, USA) coupled to a mass spectrometer (Xevo TQ-S, Waters Corp.). The bioanalytical method for the simultaneous quantitation of the three PFCs was optimized through the various mobile phase conditions and sample preparation processes. A 3 µL aliquot of each sample was injected into a Kinetex® 2.6 µm C8 100 Å column (2.1 mm × 100 mm, 2.7 µm particle size, Phenomenex, Inc., USA) with a column temperature of 40 ± 0.5 °C. Gradient elution was used for chromatographic separation using 5 mM ammonium acetate in water (solution A) and methanol (solution B) at a flow rate of 0.3 mL/min. The gradient elution program used is as follows: the initial mobile phase was maintained at 40% B for 1.0 min, linearly increased to 90% B over 3.0 min, and held constant for until 4.0 min, and finally returned to the initial condition for 1.0 min. The mass spectrometer was operated with electrospray ionization (ESI) interface in the negative ion mode. Multiple reaction monitoring (MRM) transitions, m/z 413.1→169.0, 499.0→79.9, 399.0→80.0, 417.0→172.0, 503.0→99.0, and 403.0→103.0 for PFOA, PFOS, PFHxS, MPFOA, MPFOS, and MPFHxS for IS, respectively, were used for the
Blood samples were collected from the jugular vein at 0.17, 0.33, 0.5, 0.75, 1, 2, 4, 6, 8, 12, and 24 h after the administration. The female rats (n = 5/group) were given a single dose 2 mg/kg of PFOS by oral and IV injection. Blood samples were collected from the jugular vein at 0.25, 0.5, 1, 2, 4, 8, 12, 18, 24 h and 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70 day after IV administration and at 0.25, 0.5, 1, 3, 5, 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70 day after the oral administration. In the case of PFHxS, female rats (n = 5/group) were administered a single dose of PFHxS at 4 mg/kg by oral and IV. Blood samples were collected from the jugular vein at 0.5, 1, 2, 4, 6, 12, 24, 48, 72, 120, 168, 240, and 336 h after the administration. Each PFC solution was prepared in saline or distilled water. To obtain plasma sample, all the blood samples were collected in heparinized tubes and immediately centrifuged (5,000 × g, 10 min). Following the centrifugation, plasma was stored at –80 °C until analysis. Tissue samples of the liver, kidney, heart, lung, and spleen were dissected and taken immediately after taking the blood sample at the terminal time (PFOA, 24 h; PFOS, 70 day; PFHxS, 336 h) and stored frozen (−80 °C) until …show more content…
The liquid chromatography was performed using an Acquity UPLC® system (Waters Corp., Milford, MA, USA) coupled to a mass spectrometer (Xevo TQ-S, Waters Corp.). The bioanalytical method for the simultaneous quantitation of the three PFCs was optimized through the various mobile phase conditions and sample preparation processes. A 3 µL aliquot of each sample was injected into a Kinetex® 2.6 µm C8 100 Å column (2.1 mm × 100 mm, 2.7 µm particle size, Phenomenex, Inc., USA) with a column temperature of 40 ± 0.5 °C. Gradient elution was used for chromatographic separation using 5 mM ammonium acetate in water (solution A) and methanol (solution B) at a flow rate of 0.3 mL/min. The gradient elution program used is as follows: the initial mobile phase was maintained at 40% B for 1.0 min, linearly increased to 90% B over 3.0 min, and held constant for until 4.0 min, and finally returned to the initial condition for 1.0 min. The mass spectrometer was operated with electrospray ionization (ESI) interface in the negative ion mode. Multiple reaction monitoring (MRM) transitions, m/z 413.1→169.0, 499.0→79.9, 399.0→80.0, 417.0→172.0, 503.0→99.0, and 403.0→103.0 for PFOA, PFOS, PFHxS, MPFOA, MPFOS, and MPFHxS for IS, respectively, were used for the