We quantified 16 bile acids hypothesized to guide adult lake trout to spawning reefs (Zhang et al., 2001 ) to determine their presence as odorants during the spawning season (Li et al., 2015 ). We added an internal standard of 10 ng deuterated cholic acid and taurocholic acid (CA‐d4 and TCA‐d4) to a 10 ml subsample of the 1 L samples collected from aquaria over 16 weeks, as described above. Each sample was freeze‐dried using a CentriVap Cold Trap with CentriVap Concentrator (Labconco, MO) and reconstituted in 100 μl of methanol and water (1:1, v:v). Reconstituted samples were subjected to UHPLC‐MS/MS (Waters Acquity ultra‐performance liquid chromatography system, Waters, Milford, MA; Micromass Quattro Premier XE tandem quadruple mass spectrometer, Waters, Manchester, UK) using described methods (Li et al., 2015 ).
Micromass quattro premier xe tandem quadruple mass spectrometer
Manufactured by Waters Corporation
Sourced in United Kingdom
The Micromass Quattro Premier XE is a tandem quadrupole mass spectrometer designed for high-performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography (UHPLC) applications. It features a mass range up to 2,000 m/z and can operate in positive and negative ion modes. The instrument is capable of performing multiple reaction monitoring (MRM) and selected reaction monitoring (SRM) experiments for sensitive and selective quantitative analysis.
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3 protocols using micromass quattro premier xe tandem quadruple mass spectrometer
1
Quantifying Bile Acids in Lake Trout Spawning
2
Comprehensive Water Quality Analysis
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Water quality analyses were firstly conducted by measuring chemical oxygen demand (COD), UV absorbance at 254 nm (UV 254 ), total organic carbon (TOC), total phosphorus (TP), NH 4 + -N, total nitrogen (TN), electric conductivity, turbidity and color according to the standard methods [11] . Specific ultraviolet absorbance (SUVA) was calculated as the ratio of UV 254 over TOC.
For trace organic chemicals, instrumental methods were applied targeting 59 chemicals including 13 polycyclic aromatic hydrocarbons (PAHs), 3 pesticides, 16 phenols, and 27 pharmaceuticals. PAHs, pesticides and phenols were analyzed on an Agilent 6890 GC coupled to a 5975 MS (GC/MS), equipped with a DB-5MS (30 m × 0.25 mm × 0.25 μm) capillary column. Detailed instrument conditions for determining PAHs, pesticides and phenols are described in Table SI-1. Pharmaceuticals were analyzed on a UPLC coupled with a Waters Micromass Quattro Premier XE tandem quadruple mass spectrometer (UPLC-MS/MS), equipped with a reversed-phase BEH C18 column (100 m × 2.1 mm × 1.7 μm). Instrument conditions for analyzing different categories of pharmaceuticals were described in detail in previous studies [12] .
For trace organic chemicals, instrumental methods were applied targeting 59 chemicals including 13 polycyclic aromatic hydrocarbons (PAHs), 3 pesticides, 16 phenols, and 27 pharmaceuticals. PAHs, pesticides and phenols were analyzed on an Agilent 6890 GC coupled to a 5975 MS (GC/MS), equipped with a DB-5MS (30 m × 0.25 mm × 0.25 μm) capillary column. Detailed instrument conditions for determining PAHs, pesticides and phenols are described in Table SI-1. Pharmaceuticals were analyzed on a UPLC coupled with a Waters Micromass Quattro Premier XE tandem quadruple mass spectrometer (UPLC-MS/MS), equipped with a reversed-phase BEH C18 column (100 m × 2.1 mm × 1.7 μm). Instrument conditions for analyzing different categories of pharmaceuticals were described in detail in previous studies [12] .
3
Extraction and Quantification of PPCPs in Water Samples
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Before extraction, 12.5 mL of 20 g/L EDTA-2Na was added to 500 mL water
samples individually, and the pH was adjusted to 6-8. Samples were then extracted using series-connected Waters WAX (200 mg, 6 mL) and Waters Oasis HLB (500 mg, 6 mL) cartridges. The cartridges were individually preconditioned with 15 mL of methanol followed by 15 mL of milli-Q water. The elution process followed Sun et al. (2015) (link). Finally, 1 mL extracts were prepared for quantifying PPCPs in the water samples using UPLC-MS/MS analysis. The UPLC-MS/MS was a UPLC coupled with a Waters Micromass Quattro Premier XE tandem quadruple mass spectrometer, equipped with a reverse-phase BEH C18 column (100 m × 2.1 mm × 1.7 m). The gradient conditions and instrument parameters for analyzing different categories of PPCPs were listed in Table S-2 and Table S-3 in SI 1.
samples individually, and the pH was adjusted to 6-8. Samples were then extracted using series-connected Waters WAX (200 mg, 6 mL) and Waters Oasis HLB (500 mg, 6 mL) cartridges. The cartridges were individually preconditioned with 15 mL of methanol followed by 15 mL of milli-Q water. The elution process followed Sun et al. (2015) (link). Finally, 1 mL extracts were prepared for quantifying PPCPs in the water samples using UPLC-MS/MS analysis. The UPLC-MS/MS was a UPLC coupled with a Waters Micromass Quattro Premier XE tandem quadruple mass spectrometer, equipped with a reverse-phase BEH C18 column (100 m × 2.1 mm × 1.7 m). The gradient conditions and instrument parameters for analyzing different categories of PPCPs were listed in Table S-2 and Table S-3 in SI 1.
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