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6 protocols using lc ms ms grade formic acid

1

HPLC-MS/MS Analytical Procedure

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High-performance liquid chromatography (HPLC)-grade methanol and acetonitrile (ACN) were purchased from Sigma-Aldrich Srl (Milan, Italy). MS-grade water (MilliQ, manufacture, Milan, Italy) was produced with a Milli-DI system coupled with a Synergy 185 system by Millipore (Milan, Italy). Ammonium acetate (ref. 431311), zinc sulfate heptahydrate and LC–MS/MS-grade formic acid (ref. 607001000) were purchased from Sigma (Milan, Italy). All reagents had 98% purity. All solutions were prepared with HPLC-grade water obtained from a Milli-Q Plus water purification system. HPLC mobile phases were filtered using Millipore membrane filters (0.45 µm) (Millipore, Vimodrone, Italy).
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2

HPLC-MS/MS Quantification of Cystic Fibrosis Drugs

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Ivacaftor (ref. I940600), tezacaftor (ref. T321510), and elexacaftor (ref. E201545) were purchased from Toronto Research Chemicals (North York, ON, Canada). Ivacaftor d-9 (ref. I940603), tezacaftor d-9 (T321512), and elexacaftor d-3 (E201547) were purchased from Spectra 2000 (Rome, Italy).
High-performance liquid chromatography (HPLC)-grade methanol and acetonitrile (ACN) and LC–MS/MS-grade formic acid (ref. 607001000) were purchased from Sigma-Aldrich Srl. (Milan, Italy). All reagents had 98% purity.
All solutions were prepared with HPLC-grade water obtained from a Milli-Q Plus water purification system (18.2 MΩ•cm, 25 °C, TOCs < 5 ppb). HPLC mobile phases were filtered by using Millipore membrane filters (0.45 µm) (Millipore, Vimodrone MI, Italy).
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3

Determination of Neonicotinoid Pesticides

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Acetamiprid, flonicamid, thiacloprid, thiamethoxam,
and nitenpyram standard solutions were purchased from Accustandard
(New Haven, CT, USA) with purity ≥99.7%. Imidacloprid and clothianidin
were purchased from Sigma-Aldrich (St. Louis, MO, USA) with purity
of 99.9%. Dinotefuran was purchased from Chem Service (West Chester,
PA, USA) with purity of 99.2%. The isotope-labeled internal standards
(IS) for Imidacloprid-d4 (99.2%), clothianidin-d3 (98.9%), and thiamethoxam-d3 (99.8%) were purchased from C/D/N Isotopes, Inc. (Quebec,
Canada). LC-MS/MS grade formic acid and ammonium formate were purchased
from Sigma-Aldrich. HPLC grade reagents, including acetonitrile and
water, were purchased from J. T. Baker (Center Valley, PA, USA). Vial
(1.5 mL with 0.2 μm nylon filter) were purchased from VWR international
(Radnor, PA, USA). QuEChERS extraction salt packages, which include
4 g of MgSO4, 1 g of NaCl, 1 g of sodium citrate, and 500
mg of of disodium citrate sesquihydrate in each salt pack, and 2 mL
of QuEChERS dispersive SPE containing 50 mg of PSA + 50 mg of C18
+ 150 mg of MgSO4 or 50 mg of PSA + 50 mg of graphitized
carbon black (GCB) + 150 mg of MgSO4 or 25 mg of PSA +
7.5 mg of GCB + 150 mg of MgSO4 ceramic homogenizer were
purchased from Agilent Technologies (Santa Clara, CA, USA).
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4

Quantitative Analysis of Antiviral Drugs

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Aciclovir (ref. A192400) and ganciclovir-d5 (ref. G235002) were purchased from Toronto Research Chemicals (North York, ON, Canada).
Ganciclovir (ref. Y0001129), Ammonium acetate (ref. 431311), LC–MS/MS-grade methanol (ref. 1.06035.2500) and LC–MS/MS-grade formic acid (ref. 607001000) were purchased from Sigma (Milan, Italy). All reagents had 98% purity.
All solutions were prepared with HPLC-grade water obtained from a Milli-Q Plus water purification system. HPLC mobile phases were filtered using Millipore membrane filters (0.45 µm) (Millipore, Vimodrone, Italy).
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5

Characterization of Transporter Substrates

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Ipratropium bromide, IBC, IBC-d6, l-noradrenaline, ritodrine hydrochloride, sumatriptan-d6, thiamine-d3 hydrochloride, and trospium chloride were obtained from Santa Cruz Biotechnology. Ranitidine-d6 and trospium-d8 were obtained from Toronto Research Chemicals, ASP+ was obtained from Life Technologies, and buformin hydrochloride was obtained from Wako Chemicals. Radiolabeled MPP+ (N-[methyl-3H], 80 Ci/mmol) and TEA+ ([ethyl-1-14C]; 55 mCi/mol) were obtained from Hartmann Analytic. All other compounds tested as substrates were obtained from Sigma–Aldrich. All chemicals used in this study were purchased from commercial sources and had purities of 97% or higher.
Dulbecco's modified Eagle's medium, Hank's buffered salt solution (HBSS), and additives used for cell culturing were obtained from Life Technologies. Poly-d-lysine (1–5 kDa), Hepes, bicinchoninic acid, and copper sulfate pentahydrate were obtained from Sigma–Aldrich. Twelve-well plates were obtained from Starlab, and tissue culture flasks were from Sarstedt. Acetonitrile and methanol in LC–MS/MS grade were obtained from LGC Standards, and formic acid (LC–MS/MS grade) and sodium chloride were obtained from Merck. SDS (UltraPure) was obtained from AppliChem.
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6

Quantitative Analysis of Drugs and Metabolites

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Ipratropium bromide and trospium chloride were obtained from Santa Cruz Biotechnology (Heidelberg, Germany). Trospium-d8 was obtained from Toronto Research Chemicals (TRC, Toronto, ON, Canada). Buformin hydrochloride was obtained from Wako Chemicals (Neuss, Germany). Fenoterol hydrochloride, fenoterol-d6, metformin hydrochloride, butylscopolamine bromide, and atropine were obtained from Sigma-Aldrich (Taufkirchen, Germany).
Dulbecco’s Modified Eagle Medium (DMEM), Hank’s Buffered Salt Solution (HBSS), and additives used for cell culturing were obtained from Life Technologies (Darmstadt, Germany). Poly-d-lysine (1-5 kDa), 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES), bicinchoninic acid, and copper sulfate pentahydrate were obtained from Sigma Aldrich. Twelve-well plates were obtained from Starlab (Hamburg, Germany) and tissue culture flasks from Sarstedt (Nümbrecht, Germany). Acetonitrile and methanol in LC-MS/MS grade were obtained from LGC Standards (Wesel, Germany), formic acid (LC-MS/MS grade) and sodium chloride were obtained from Merck (Darmstadt, Germany). Sodium dodecylsulfate (SDS, ultrapure) was obtained from AppliChem (Darmstadt, Germany).
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