3200 qtrap mass spectrometer

Drugs were quantified by a Shimadzu HPLC system coupled to a 3200 QTRAP mass spectrometer (Applied Biosystems, Grand Island, NY). The HPLC system consisted of two Shimadzu LC-20A pumps, a DGU-20A5 degasser, and a Shimadzu SIL-20AC HT autosampler. The mass spectrometer was equipped with an electrospray ionization (ESI) TurboIonSpray source. The system was operated with Analyst software, version 1.5.2 (ABSciex, Framingham, MA).
Chromatographic separation of drugs was achieved using a Synergi column (100 × 2.0 mm; 4- μm particle size) with an inline C8 guard column (4.0 × 2.0 mm) (Phenomenex, Torrance, CA). An ammonium acetate buffer/reagent alcohol gradient was used to separate components. Analytes were monitored using multiple-reaction monitoring for positive ions. The following ion transitions were monitored: gemcitabine, m/z 264.066→112.000; paclitaxel, m/z 854.266→286.200; a stable labeled isotope (C₈¹³CH₁₂ClF₂N¹⁵N₂O₄) (m/z 267.067→115.100) was used as an internal standard for gemcitabine; docetaxel (m/z 830.312→549.3) was used as an internal standard for paclitaxel.

For mycotoxin analysis, the samples were injected into an HPLC coupled to a 3200QTRAP mass spectrometer (Applied Biosystems, Foster City, CA, USA). The column used to separate mycotoxins was a Gemini NX C18 column (150 × 2.0 mm I.D, 3.0 mm, Phenomenex, Palo Alto, CA, USA). The mobile phases consisted of water (A) and ACN (B), both with 0.1% formic acid and 5 mM ammonium formate at 0.25 mL/min with a linear gradient. The ions transitions used for the AFB₁ and OTA identification and quantification were m/z 313.1/241.3 and 284.9 (AFB₁) and m/z 404.3/102.1 and 358.1 (OTA) [33 (link)].

Products of 30 enzymatic incubations were purified by HPLC and corresponding peaks were collected as individual fractions. These fractions were directly infused into the mass spectrometer (3200 QTrap mass spectrometer; Applied Biosystems/MDS SCIEX, Darmstadt, Germany), equipped with an electrospray ionization interface (Turbo V), using the integrated syringe pump of the 3200 QTrap instrument (Syringe; 1,000 μl, i.d. 2.3 mm; Hamilton, Nevada, USA) at a flow rate of 10 μl min⁻¹. The MS/MS was operated in the positive mode with a source voltage and declustering potential of 5.5 kV and 76 V, respectively. Nitrogen gas was used for nebulization, with the curtain gas, gas 1, and gas 2 settings at 10, 14 and 0, respectively. Parameters were optimized for benzophenones using standard 2,4,6-trihydroxybenzophenone and for xanthones using 1,3,7-trihydroxyxanthone in methanol at 10 μg ml⁻¹. The molecular ion peaks [M+H]⁺ of the products were further analysed by MS/MS experiments in the enhanced product ion (EPI) mode of the instrument using nitrogen gas for collision-induced dissociation at the high-level setting. The collision energy was 30–50 V. Data acquisition and processing were performed using the Analyst software (version 1.4.2; Applied Biosystems/MDS SCIEX).