All samples were analyzed on HPLC/MS/MS systems which consist of Shimadzu LC‐30AD pumps (Shimadzu), a rack changer II auto‐sampler (Shimadzu) and either a Shimadzu 8060 (Shimadzu) or an AB Sciex API 4000/5500 (AB Sciex) mass spectrometer. 20 μL of plasma samples was mixed with 200 μL of acetonitrile containing internal standards and centrifuged at 4000g for 15 minutes (4°C). The mixture was then thoroughly mixed with 200 μL of acetonitrile containing internal standard and centrifuged at 4000g for 15 minutes (4°C). 10 μL of dialysates and CSF samples were mixed with 100 μL of acetonitrile containing internal standard. Supernatants were diluted with appropriate volumes of water before analysis on HPLC/MS/MS.
Lc 30ad pump
Manufactured by Shimadzu
Sourced in Japan
The Shimadzu LC-30AD pump is a high-performance liquid chromatography (HPLC) pump designed for reliable and precise solvent delivery. It features a dual-piston design and can operate at a maximum pressure of 60 MPa (600 bar, 8,700 psi). The pump is capable of delivering flow rates from 0.0001 to 10 mL/min, with a flow rate precision of ±0.1%.
Automatically generated - may contain errors
Lab products found in correlation
Sil 30ac autosampler, by Shimadzu (17 mentions)
Cto 20ac column oven, by Shimadzu (5 mentions)
Cbm 20a, by Shimadzu (4 mentions)
Dgu 20a5r degasser, by Shimadzu (4 mentions)
Sil 30ac, by Shimadzu (4 mentions)
Nexera hplc system, by Shimadzu (4 mentions)
Nexera x2, by Shimadzu (3 mentions)
Cbm 20a system controller, by Shimadzu (3 mentions)
Cbm 20a controller, by Shimadzu (3 mentions)
Api 4000 tandem mass spectrometer, by AB Sciex (3 mentions)
Analyst software, by AB Sciex (3 mentions)
Rack changer 2 auto sampler, by Shimadzu (2 mentions)
Api 4000 5500, by AB Sciex (2 mentions)
Lcms 8050, by Shimadzu (2 mentions)
Uhplc ms ms, by Shimadzu (2 mentions)
Dgu 20a5 degasser, by Shimadzu (2 mentions)
Cto 30a column oven, by Shimadzu (2 mentions)
Nexera uhplc, by Shimadzu (2 mentions)
Cto 20ac, by Shimadzu (2 mentions)
Sil30 acmp autosampler, by Shimadzu (2 mentions)
41 protocols using lc 30ad pump
1
Bioanalytical Assay for Plasma, Dialysate, and CSF
2
HPLC/MS/MS Analysis of Dialysate Samples
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All samples were analyzed on HPLC/MS/MS systems, which consist of Shimadzu LC‐30AD pumps (Shimadzu), a rack changer II autosampler (Shimadzu), and either a Shimadzu 8060 (Shimadzu) or an AB Sciex API 4000/5500 (AB Sciex) mass spectrometer. Ten μl of dialysates samples were mixed with 100 μl or 200 μl of acetonitrile containing internal standard. The supernatants were diluted with appropriate volumes of water before analysis on HPLC/MS/MS.
3
HPLC-MS/MS Metabolite Quantification
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The HPLC system consisting of two LC-30AD pumps and an SIL-30AC autosampler (Shimadzu, Kyoto, Japan) was coupled to an AB Sciex Qtrap 5500 System (Applied Biosystems, Foster City, USA) equipped with a TurboIonSpray ionization interface. Analyst Version 1.5.2 (Applied Biosystems) was used for data acquisition. Centrifugation employed a CT 15 RE high-speed desktop centrifuge (Hitachi, Tokyo, Japan).
4
UPLC-MS/MS for Compound Identification
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The Shimadzu Nexera UPLC system consisted of two LC‐30 AD pumps, a CTO‐30AS column oven and a SIL‐30 AC autosampler, together with an LC–MS/MS 8060 system (Shimadzu Scientific Instruments, Columbia, MD, USA). An electrospray ionization source in positive ionization mode was used to ionize samples. LabSolutions LCMS software version 5.80 (Shimadzu Scientific, Inc., Columbia, MD, USA) was used for data acquisition.
5
Steroid Profiling Using UHPLC-MS/MS
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The analyses were performed using a Shimadzu Nexera UHPLC (Kyoto, Japan) consisting of two LC-30AD pumps, a SIL-30AC autosampler, a CTO-20AC column oven and a CBM-20A controller. The chromatographic system was coupled online to a triple quadrupole LCMS-8050 (Shimadzu, Kyoto, Japan) equipped with an Electrospray Ionization (ESI) source. A Kinetex ® Biphenyl Column 100 × 2.1 mm, 2.6 μm (Phenomenex ® , Bologna Italy) was used for steroid separation. Column oven temperature was set at 45 °C; flow rate was set to 0.5 mL/min; mobile phases composition was as follows: (A) H 2 O + 0.2 mM NH 4 F, (B) MeOH and the following gradient has been employed: 0 min, 20% B, 0.01-4.00 min, 20-60% B, 4.01-6.00 min, 60-70% B, 6.0.1-6. Interface temperature, Desolvation line temperature, Heat Block temperature were set, respectively to 300 °C, 250 °C and 400 °C. Nebulizing gas, drying (N 2 ) and heating gas (air) were set, respectively, to 3, 10 and 10 L/min.
6
UHPLC-MS/MS Analysis of Metabolites
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Methanol, acetonitrile, ammonium acetate (NH4Ac), and aqueous ammonia (NH4OH) were all high-performance liquid chromatography (HPLC)-grade and were purchased from Thermo Fisher Scientific (Waltham, MA, United States). All the experiments were conducted on an ultra-high performance liquid chromatography (UHPLC) system including two Shimadzu LC-30AD pumps, a SIL-30AC auto-sampler, and a CTO-20AC column oven (Shimadzu Corporation, Kyoto, Japan), and coupled with QTRAP 4500 mass spectrometer (AB SCIEX, CA, United States). The PLRP-S column (3.0 µm, 150 mm × 2.1 mm) was purchased from Agilent Technologies (Santa Clara, CA, United States).
7
Urinary BPA and Phthalate Quantification
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BPA and phthalate (MEP, MIbP, MEHHP, MBzP, MEHP) levels were measured in mid-stream clean urine samples instead of in first or 24 h urine samples, according to Lee et al. [45 (link)], and creatinine concentrations were also assessed.
Briefly, urinary samples were collected into polypropylene cups for urine culture, and then women were asked to store the cups in a cool place until transportation. The urine samples were transported under refrigeration, aliquoted into tubes made from phthalate-free materials and stored at −80 °C until analysis.
BPA and phthalates levels were measured by UHPLC-MS/MS (Shimadzu, Milan, Italy). The UHPLC system consisted of two LC 30 AD pumps, a SIL 30 AC autosampler, a CTO 20 AC column oven and a CBM 20 A controller, and the system was coupled online to a triple quadrupole LCMS 8050 (Shimadzu, Kyoto, Japan) equipped with an electrospray ionization (ESI) source. Results were compared to previous data (Table S1, Supplementary Materials ).
Creatinine levels were measured by routine methods on an Atellica® CH Analyzer (Siemens Healthcare Diagnostics Inc., Tarrytown, NY, USA) and expressed as μmol/L.
All urinary samples were considered acceptable because the creatinine concentrations were within the range of 0.3–3.0 g/L [46 ].
Briefly, urinary samples were collected into polypropylene cups for urine culture, and then women were asked to store the cups in a cool place until transportation. The urine samples were transported under refrigeration, aliquoted into tubes made from phthalate-free materials and stored at −80 °C until analysis.
BPA and phthalates levels were measured by UHPLC-MS/MS (Shimadzu, Milan, Italy). The UHPLC system consisted of two LC 30 AD pumps, a SIL 30 AC autosampler, a CTO 20 AC column oven and a CBM 20 A controller, and the system was coupled online to a triple quadrupole LCMS 8050 (Shimadzu, Kyoto, Japan) equipped with an electrospray ionization (ESI) source. Results were compared to previous data (
Creatinine levels were measured by routine methods on an Atellica® CH Analyzer (Siemens Healthcare Diagnostics Inc., Tarrytown, NY, USA) and expressed as μmol/L.
All urinary samples were considered acceptable because the creatinine concentrations were within the range of 0.3–3.0 g/L [46 ].
8
Quantification of HsTX1[R14A] in Samples
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Before analysis, samples were diluted to an appropriate concentration of HsTX1[R14A] within the linear range of the standard curve. Samples were then injected onto an Ascentis ® Express C 18 column (2.7 mm particle size, 2.1 Â 50 mm internal diameter) with a Phenomenex SecurityGuard TM C 18 guard column (2.0 Â 4.0 mm). LC was performed on a Shimadzu HPLC system consisting of 2 LC-30AD pumps, a SIL-30AC autoinjector, and a DGU-20A 5 degasser (Shimadzu, Kyoto, Japan). Mobile phase A consisted of 0.1% (vol/vol) formic acid in MilliQ water and mobile phase B was methanol. Concentrations of HsTX1 [R14A] were determined using the following gradient profile: 0-1 min, 90% A; 1-1.5 min, 75% A; 1.5-2 min, 70% A; 2-2.5 min, 60% A; 2.5-3.5 min, 60% A; and 3.5-4.5 min, 90% A followed by a 1.5 min equilibration period at the initial conditions. Mass spectrometry was performed on a Shimadzu LCMS-8030 quadrupole mass spectrometer (Shimadzu) using electrospray ionization in the positive mode. Mass detection was performed by multiple reaction monitoring (m/z 747.0 to 84.10). Nitrogen was used as the nebulizing gas and drying gas with flow rates set at 3.0 and 15.0 L/min, respectively. The desolvation line and heat block temperatures were 250 C and 400 C, respectively, and the dwell time was set at 500 ms.
9
UPLC-DAD-ESI-IT-TOF Metabolite Analysis
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The UPLC-DAD-ESI-IT-TOF analysis of the metabolites was performed using Shimadzu LC-30AD instrument equipped with two LC-30AD pumps, an SIL-30AC autosampler, SPD-M20A, CTO-20AC column oven, and CBM-20A system controller, and coupled to an IT-TOF mass spectrometer with an ESI interface.
The chromatography separations were performed on a Shim-pack HR-ODS column (150 mm × 2.1 mm, 3 μm). The column was eluted with a gradient mobile phase A of water-formic acid (100: 0.1, v/v) and B of acetonitrile, The elution gradient was 0–2 min, 5–10% B, 2–4 min, 10–12% B, 4–7 min, 12–18% B, 7–13 min, 18–27% B, 13–32 min, 27–56% B, 32–37 min, 56–100% B, and 37–47 min 100% B. The flow rate was 0.2 mL/min; the injection volume was 5 μL and the column oven temperature was kept at 30°C.
The tandem mass spectrometry analyses were carried out on an IT-TOF (Shimadzu, Japan) with the full scan over m/z 100–900 (MS1) and m/z 50–900 (MS2 and MS3) in the ion model of negative(NI) and positive(PI). The parameters were as follows: heat block and curved desolvation line temperature, 200°C; nebulizing nitrogen gas flow, 1.5 L/min; interface voltage: (+), 4.5 kv; (−), 3.5 kv; detector voltage, 1.56 kv; relative collision-induced dissociation energy, 50%.
The chromatography separations were performed on a Shim-pack HR-ODS column (150 mm × 2.1 mm, 3 μm). The column was eluted with a gradient mobile phase A of water-formic acid (100: 0.1, v/v) and B of acetonitrile, The elution gradient was 0–2 min, 5–10% B, 2–4 min, 10–12% B, 4–7 min, 12–18% B, 7–13 min, 18–27% B, 13–32 min, 27–56% B, 32–37 min, 56–100% B, and 37–47 min 100% B. The flow rate was 0.2 mL/min; the injection volume was 5 μL and the column oven temperature was kept at 30°C.
The tandem mass spectrometry analyses were carried out on an IT-TOF (Shimadzu, Japan) with the full scan over m/z 100–900 (MS1) and m/z 50–900 (MS2 and MS3) in the ion model of negative(NI) and positive(PI). The parameters were as follows: heat block and curved desolvation line temperature, 200°C; nebulizing nitrogen gas flow, 1.5 L/min; interface voltage: (+), 4.5 kv; (−), 3.5 kv; detector voltage, 1.56 kv; relative collision-induced dissociation energy, 50%.
10
Chiral Separation by Liquid Chromatography
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Liquid chromatography was performed using a Shimadzu HPLC-IT-TOF/MS system consisting of a controller (CBM-20A), two LC-30AD pumps, a SIL-30AC auto-sampler and a CTO-20AC column heater. The separation was operated on Chiralpak AD-H column (250 mm × 4.6 mm, 5 µm) using n-hexane (containing 0.1% acetic acid):ethanol (75:25, v/v) as mobile phase. The column temperature was maintained at 30 °C; the flow rate was 0.6 mL/min; the injection volume was 5 µL; the auto-sampler temperature was 10 °C. The LCMSsolutionVer3 LC-MS software (Shimadzu, Kyoto, Japan) was used to control instruments and process the data.
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