Chromatography was performed on a modular HPLC system from Shimadzu (Kyoto, Japan); it contained a system controller (CBM-20A), two Nexera X2 pumps, a degasser (DGU-20ASR), and a column oven (CTO-20AC). Automated extractions were carried out with a DBS-MS 500 (CAMAG, Muttenz, Switzerland). Analytes were separated on a Shim-pack GIST (4.6 × 50 mm, 5 μm STEAROYL, 227-30017-3) analytical column (Shimadzu, Kyoto, Japan). A filter frit (KrudKatcher Ultra, Phenomenex, Torrance, CA, USA) was connected upstream to the analytical column. Mobile phase A consisted of water plus 0.1% formic acid and 2 mM ammonia fluoride, while methanol supplemented with 0.1% formic acid and 2 mM ammonia fluoride was used as mobile phase B. The following stepwise gradient was applied: 40% A (0–1.0 min), 40–90% A (1.0–2.0 min), 90% A (2.0–3.0 min), and 40% A (3.01–4.0 min). The flow rate was set at 1.0 mL/min at 40 °C. The HPLC liquid stream was connected to an 8060 tandem mass spectrometer (Shimadzu, Kyoto, Japan). The mass transitions and compound specific settings were included in Table S1 .
Krudkatcher ultra
Manufactured by Phenomenex
Sourced in United States, Germany
The KrudKatcher Ultra is a filtration device designed to remove particulates and contaminants from liquid samples prior to analysis. It features a durable construction and efficient filtration capabilities to help maintain the integrity of analytical instruments and samples.
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3 protocols using krudkatcher ultra
1
Quantitative Analysis of Compounds by HPLC-MS/MS
2
UHPLC-MS Dust Sample Extraction Protocol
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Prior
to mass spectrometric
sample analysis, chromatographic separation was realized using Elute
HPG 1300 UHPLC-systems (Bruker Daltonics, Bremen, Germany). In QTOF-HRMS
and TQMS analysis, 30 μL of the diluted dust extract was injected
using PAL HTC-xt and RSI autosamplers (CTC Analytics AG, Zwingen,
Switzerland). In IM-QTOF-HRMS analysis, the maximum injection volume
was limited to 27 μL due to technical limitations of the applied
Elute autosampler (Bruker Daltonics, Bremen, Germany). Chromatographic
separation was performed on Nucleodur C18 Gravity-SB columns (75 ×
2 mm, 1.8 μm, Macherey-Nagel GmbH & Co. KG, Düren,
Germany) equipped with a KrudKatcher Ultra (Phenomenex, Aschaffenburg,
Germany) or a Nucleodur C18 Gravity-SB pre-column (1.8 μm, Macherey-Nagel
GmbH & Co. KG, Düren, Germany) at 40 °C in Elute column
oven systems (Bruker Daltonics, Bremen, Germany). Gradient elution
conditions using MeCN + 0.1% FA (A) and H2O + 0.1% FA (B)
were applied as follows: 0.0 min 5% A, 2.0 min 5% A, 3.9 min 25% A,
6.5 min 70% A, 7.5 min 70% A, 9.0 min 95% A, 12.0 min 95% A, 12.2
min 5% A, and 15.0 min 5% A. Additionally, a flow gradient of 350
μL/min respective 450 μL/min (3.9–12.2 min) was
included. The first 2 min of each run were directed into waste. Software
Compass HyStar (versions 4.1, 5.1, and 6.0) (Bruker Daltonics, Bremen,
Germany) was used for the operation of the UHPLC systems.
to mass spectrometric
sample analysis, chromatographic separation was realized using Elute
HPG 1300 UHPLC-systems (Bruker Daltonics, Bremen, Germany). In QTOF-HRMS
and TQMS analysis, 30 μL of the diluted dust extract was injected
using PAL HTC-xt and RSI autosamplers (CTC Analytics AG, Zwingen,
Switzerland). In IM-QTOF-HRMS analysis, the maximum injection volume
was limited to 27 μL due to technical limitations of the applied
Elute autosampler (Bruker Daltonics, Bremen, Germany). Chromatographic
separation was performed on Nucleodur C18 Gravity-SB columns (75 ×
2 mm, 1.8 μm, Macherey-Nagel GmbH & Co. KG, Düren,
Germany) equipped with a KrudKatcher Ultra (Phenomenex, Aschaffenburg,
Germany) or a Nucleodur C18 Gravity-SB pre-column (1.8 μm, Macherey-Nagel
GmbH & Co. KG, Düren, Germany) at 40 °C in Elute column
oven systems (Bruker Daltonics, Bremen, Germany). Gradient elution
conditions using MeCN + 0.1% FA (A) and H2O + 0.1% FA (B)
were applied as follows: 0.0 min 5% A, 2.0 min 5% A, 3.9 min 25% A,
6.5 min 70% A, 7.5 min 70% A, 9.0 min 95% A, 12.0 min 95% A, 12.2
min 5% A, and 15.0 min 5% A. Additionally, a flow gradient of 350
μL/min respective 450 μL/min (3.9–12.2 min) was
included. The first 2 min of each run were directed into waste. Software
Compass HyStar (versions 4.1, 5.1, and 6.0) (Bruker Daltonics, Bremen,
Germany) was used for the operation of the UHPLC systems.
3
HPLC Analysis of Compound PP
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High performance liquid chromatography (HPLC: Alliance 2695, Waters) was employed to monitor the concentration of PP. Separation was achieved on a Kinetex XB-C18 100A column (2.6 μm, 2.1 × 50 mm) and a 0.5 μm inline filter (KrudKatcher Ultra) both purchased from Phenomenex. The
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