A triple quadrupole mass spectrometer (AB SCIEX Triple QuadTM 5500, AB SCIEX Corp., Framingham, Massachusetts, USA) was used to qualitatively and quantitatively analyze the target compounds, and Analyst version 1.6.1 software (AB SCIEX Pte. Ltd., Concord, Ontario, Canada) was used for the analysis. Tandem mass spectrometry was used to determine the precursor and product ions of the target compounds in positive ionization and multiple reaction detection (MRM) modes. The typical MS parameters were as follows: electrospray ionization voltage, 5500 V; ion source temperature, 550 °C; spray gas, auxiliary gas (nitrogen), curtain gas, and collision gas pressure, 45, 50, 35, and 8 psi, respectively; collision chamber outlet voltage, 12 V; and injection voltage, 10 V. The precursor ions, product ions, the corresponding cone voltage, and collision energy of the eight coccidiostats are listed in Table 1 .
Triple quad 5500
Manufactured by AB Sciex
Sourced in United States, Japan
The Triple Quad 5500 is a triple quadrupole mass spectrometer designed for quantitative and qualitative analysis. It features high sensitivity and selectivity, enabling precise and reliable detection and measurement of target analytes.
Automatically generated - may contain errors
Lab products found in correlation
Xterra ms c18 column, by Waters Corporation (2 mentions)
Eclipse plus c18 column, by Agilent Technologies (2 mentions)
Analyst 1, by AB Sciex (2 mentions)
Exion lc, by AB Sciex (2 mentions)
Nexera ultra high performance liquid chromatography system, by Shimadzu (2 mentions)
Zorbax sb aq column, by Agilent Technologies (2 mentions)
Ultra performance liquid chromatography (uplc), by Shimadzu (2 mentions)
Analyst version 1, by AB Sciex (1 mentions)
Silica gel 60, by Merck Group (1 mentions)
Behamidexp column, by Waters Corporation (1 mentions)
Phoenix winnonlin, by Certara (1 mentions)
Acquity uplc system, by Waters Corporation (1 mentions)
Nexera x2 uhplc system, by Shimadzu (1 mentions)
Acquity uplc beh c18, by Shimadzu (1 mentions)
Lc ac system, by AB Sciex (1 mentions)
Agilent 1200 infinity series, by Agilent Technologies (1 mentions)
Florisil cartridge, by Waters Corporation (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Acquity uplc classic system, by Waters Corporation (1 mentions)
Cd 1 mice, by Charles River Laboratories (1 mentions)
56 protocols using triple quad 5500
1
Quantitative Analysis of Coccidiostats
2
Glycoglycerolipid Analysis by TLC and HPLC-MS
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The GT cp -synthesized glycoglycerolipids were analysed by TLC using a Camag Automatic TLC Sampler III (Camag, Muttenz, Switzerland) for spotting. Glycoglycerolipids were separated on silica gel 60 (Merck, Darmstadt, Germany) with chloroform/methanol/water (65:35:4 vol/vol), and stained with sulfuric acid/methanol/water (45:45:10 vol/vol) for visualization. The resulting solutions were further analysed by high-performance liquid chromatography using a 1290 Infinity II UPLC instrument (Agilent Corp., Santa Clara, CA, USA) coupled with an AB SCIEX Triple Quad TM 5500 (AB SCIEX, Framingham, MA, USA) equipped with an electrospray-ion (ESI) detector. A BEH Amide XP column (2.5-µm inner diameter, 3 mm by 150 mm, Waters, Milford, MA, USA) was used for chromatographic separation. The mobile phase consisted of acetonitrile (solvent A) and 10 mM ammonium acetate, pH 9.2 (solvent B). The column was equilibrated for 10 min with 95% A: 5% B prior to sample injection. The separation was conducted using a stepwise gradient starting from 95% A: 5% B to 70% A: 30% B after 15 min with a constant flow rate of 150 µl min -1 . Mass spectrometric analysis was performed in the ESI positive ion mode with the ion spray voltage at 3500 V and temperature at 350°C. The nebulizer gas and heater gas were set at 40 psi. The analytical data were processed by Analyst software (version 1.6.3).
3
Ubiquinol Quantification in Serum
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Serum concentrations of ubiquinol were measured using LC/MS/MS (commissioned at Kaneka Techno Research Corporation). Specifically, 0.1 mL of serum was fractionated immediately after collection, mixed with 0.7 mL of isopropanol to prevent oxidation of ubiquinol and stored at −80°C until measurement. At the time of measurement, samples were centrifuged at 12,000 rpm for 5 min; supernatants were filtered through a membrane filter and subjected to LC/MS/MS analysis using the AB SCIEX Triple Quad5500 instrument.
4
LC-MS/MS Quantification of BB and THP
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The plasma concentrations of BB were determined using an AB Sciex Triple Quad 5500 (AB Sciex, Framingham, MA, USA) LC–MS/MS system with an ESI probe. A Waters Acquity UPLC HSS T3 (2.1 × 100 mm, 1.8 μm) column (Milford, MA, USA) was used for separation at 35 °C. The mobile phase consisted of 0.2% formic acid in water and 100% acetonitrile. The flow rate was 0.2 mL/min. The gradient program was 40% acetonitrile for 0–3.0 min, from 40% to 90% acetonitrile for 3.0–4.0 min, and returned to initial conditions at 4.1 min to re-equilibrate for 2.0 min. Only data from 0 to 4.0 min were acquired through MS. Analytes were quantified using the multiple-reaction monitoring mode. The precursor-to-product ion transition for BB and THP were the same as those aforementioned. For BB, they were as follows: CE, 39 V; declustering potential (DP), 100 V; and cell exit potential, (CEP) 18 V. For THP, they were as follows: CE, 27 V; DP, 119 V; and CEP, 16 V. Data were analyzed using the Analyst v1.5.2 software (Version 1.5.2, AB Sciex, Redwood City, CA, USA, 2011).
These assays were fully validated with reference to the US Food and Drug Administration Guidance for Industry Bioanalytical Method Validation, including selectivity, accuracy, precision, recovery, linearity, the matrix effect, and stability.
These assays were fully validated with reference to the US Food and Drug Administration Guidance for Industry Bioanalytical Method Validation, including selectivity, accuracy, precision, recovery, linearity, the matrix effect, and stability.
5
Pharmacokinetics of LY3502970 in Rats and Monkeys
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LY3502970 was administered orally at doses of 0.05, 0.15, or 0.45 mg/kg or i.v. at 0.15 mg/kg to 8-wk-old male rats (RccHan: WIST, Japan SLC; n = 4 rats/group) or oral doses of 0.04, 0.12, or 0.36 mg/kg or i.v. at 0.12 mg/kg to 3-y-old male cynomolgus monkeys (Hamri Co., Ltd.; n = 4 monkeys/group). Blood was collected predose and 30 min and 1, 2, 3, 4, 6, 8, 12, 16, and 24 h after administration in orally dosing group. Blood samples were also collected predose and 2, 10, and 30 min and 1, 2, 4, 8, 12, 16, and 24 h after i.v. administration. Compound concentrations were determined by liquid chromatography–tandem mass spectrometry (AB SCIEX Triple Quad 5500, AB SCIEX), which had a lower limit of quantification of 0.1 ng/mL. Pharmacokinetic parameters were calculated by noncompartmental analysis (linear/log trapezoidal rule) in Phoenix WinNonlin (Version 6.4, Certara). Oral bioavailability (BA) was calculated with area under the concentration-time curve from zero to infinity (AUCinf) after oral and i.v. administration by BA (%) = AUCinf, by mouth, orally (p.o.)/AUCinf, i.v. × 100.
6
Determining EFV Concentrations in Plasma and Vitreous
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EFV concentrations in the plasma and vitreous samples were assessed using a validated ultra-high-performance liquid chromatography/tandem mass spectrometry method. In brief, the ultra-high-performance liquid chromatography/tandem mass spectrometry system consisted of a Waters Acquity UPLC system (Waters Corporation, Milford, MA) and an AB Sciex Triple Quad 5500 mass spectrometer (AB SCIEX, Framingham, MA). The compounds were separated using a Venusil ASB C18 column (2.1 × 50 mm; 5 µm; Agela, Beijing, China) with a mobile phase consisting of 5 mmol/L ammonium formate in water and acetonitrile by gradient elution. The autosampler temperature was set at 4°C, and the injection volume was 10 μL. Compounds were detected by multiple reaction monitoring in the positive ion mode. The ion pairs for EFV and its internal standard (EFV-d5) were monitored at 316.1/244.0 and 321.1/173.1, respectively. The quantification range of EFV in the plasma and vitreous samples was 12.5-5000 ng/mL. The compounds were quantified using the Analyst software (version 1.6.3) by comparing the peak area of the analyte to that of the internal standard with a weighted regression of 1/X 2 .
7
Quantification of Elvitegravir by LC-MS/MS
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A 50 µL of media or cell lysate samples were mixed with 3-volumes of cold acetonitrile containing internal standard (IS) ritonavir (50 ng/mL). The mixed solution was vortexed and centrifuged at 8000 rpm for 10 min for protein precipitation. A clear supernatant was collected and analyzed using our established LC-MS/MS method66 (link). Briefly, a Shimadzu liquid chromatographic system (Kyoto, Japan) coupled with an AB SCIEX Triple Quad 5500 tandem mass spectrometer (Framingham, MA) was used for the analysis. Chromatographic separation was performed on an Xterra® MS C18 column (125 Å, 3.5 μm, 4.6 mm × 50 mm; Waters, Milford, MA). The mobile phase used for EVG separation consisted of (A) water with 0.1% formic acid and (B) acetonitrile with 0.1% formic acid (v/v) at a flow rate of 1 mL/min. The gradient elution was as follows: 0–1.5 min, 50% B; 1.5–5.1 min, 60% B (v/v). The quantification of the validated assay of EVG was 1 to 500 ng/mL. EVG and IS were eluted separately at approximately 3.27 and 2.72 min, respectively. The multiple reactions monitoring (MRM) transitions (m/z) Q1/Q3 selected for quantitative analyses were 447.9/343.8 for EVG and 721.3/296.1 for the IS. In order to reduce matrix effects, calibration curves were prepared with blank media or cell lysate based on the sample types.
8
Quantitative Analysis of Clofazimine in Biological Samples
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Clofazimine was extracted by protein precipitation from dog plasma. Clofazimine from tissues was first extracted by homogenization of tissues with a Bead Rupter during the extraction process. Reversed-phase high-pressure liquid chromatography (HPLC) separation was achieved with a Waters Acquity UPLC BEH C18 (2.1 × 50 mm, 1.7 μm) column on a Shimadzu Nexera X2 UHPLC system. Subsequently, MS/MS detection (Sciex Triple Quad 5500) was set at mass transitions of m/z 473.2→431.1 for clofazimine, and 480.2→432.1 for clofazimine-d7, respectively, in positive mode. Retention time and peak area were determined by Analyst Data Acquisition/Processing software (version 1.6.3). Analyte concentrations were obtained from a calibration curve constructed by plotting the peak area versus the nominal concentration using Analyst.
9
UHPLC-MS/MS Analysis of A. tenuissima
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The UHPLC–ESI–MS/MS analysis of the ethyl acetate extract of A. tenuissima was performed on an ExionLC AC system coupled with a SCIEX Triple Quad 5500 + MS/MS system equipped with an electrospray ionization (ESI) system. An Ascentis C18 Column (4.6 × 150 mm, 3 μm) was employed as the stationary phase, and the sample was eluted with mobile phases consisting of eluent A (0.1% formic acid) and eluent B (acetonitrile, LC grade) with following mobile phase gradient: 10% B at 0–1 min, 10%–90% B at 1–21 min, 90% B at 21–25 min, and 10% at 25.01–28 min. The flow rate was 0.5 mL/min, and the injection volume was 10 μL. MS/MS analysis used positive and negative ionization modes with a scan (EMS-IDA-EPI) [35 (link)]. The compounds were identified by using MS-DIAL version 4.70, Natural Products Atlas, and CFM-ID version 4.0 software [39 (link),40 (link),41 (link),42 (link)].
10
Polyphenol Quantification by LC-MS/MS
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The sample was analyzed with liquid chromatography−electrospray ionization−tandem mass spectrometry (LC–ESI–MS/MS) using an Exion LC AC system for separation and a SCIEX Triple Quad 5500 + MS/MS system with ESI for detection. For the separation, a ZORBAX Eclipse Plus C18 Column (4.6100 mm, 1.8 m) was used. The mobile phase was 0.1% formic acid in water (solvent A) and acetonitrile (solvent B) with the following gradient: 2% B from 0–1 min, 2–60% B from 1–8 min, 60% B from 8–12 min, and 2% B from 12.01–15 min. The flow rate was 0.8 mL/min and the injection volume was 3 µL. The selected polyphenols were analyzed using the negative ionization mode using Multiple Reaction Monitoring (MRM). The curtain gas was 25 psi, the ion spray voltage was 4500, the source temperature was 400 °C, the ion source gases 1 and 2 were 55 psi with a declustering potential of 50, the collision energy was 25 eV and the collision energy spread was 10 eV.
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