Quattro premier xe

Manufactured by Waters Corporation

Sourced in United States, United Kingdom

The Quattro Premier XE is a high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) system designed for accurate and sensitive quantitative analysis. It features a triple quadrupole mass analyzer that provides reliable performance for a wide range of applications.

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Quattro Premier XE system Micromass-Quattro Premier XE mass spectrometer Quattro Premier XE triple quadrupole mass spectrometer Quattro Premier XE instrument Quattro Premier XE triple Waters Quattro Premier XE Quattro Premier XE LC/MS/MS Micromass Quattro Premier XE tandem quadruple mass spectrometer Quattro Premier XE triple quadrupole MS Quattro Premier XE mass spectrometer

89 protocols using quattro premier xe

Dried Blood Spot Screening for Inborn Errors of Metabolism

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Amino acids, acylcarnitines and succinylacetone were extracted from a single 3.2 mm-diameter punch from each dried blood spot (DBS) using the NeoBase Non-derivatized kit and, from 2019, the NeoBase 2 Non-derivatized kit (PerkinElmer, Turku, Finland) and quantified by flow injection analysis with ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS). These first-tier analyses were performed on two Acquity Xevo-TQS systems, an Acquity Xevo TQS micro or a Quattro Premier XE (Waters, Milford, MA, USA). The first-tier cut-off values used for the 20 IEMs are depicted in Table 1. In the case of an abnormal screening result in the first assessment, two new 3.2 mm diameter DBS punches were re-analyzed.
Biotinidase activity in DBSs was initially analyzed with a Victor Multilabel Plate Reader (PerkinElmer, Turku, Finland) and measured by a semi-quantitative method using biotin-6-amidoquinoline as a substrate [4 (link)]. From 2013, screening for biotinidase deficiency (BD) was performed using the Genetic Screening Processor (GSP^®) and the GSP Neonatal Biotinidase kit, both from PerkinElmer.

Tangeraas T., Sæves I., Klingenberg C., Jørgensen J., Kristensen E., Gunnarsdottir G., Hansen E.V., Strand J., Lundman E., Ferdinandusse S., Salvador C.L., Woldseth B., Bliksrud Y.T., Sagredo C., Olsen Ø.E., Berge M.C., Trømborg A.K., Ziegler A., Zhang J.H., Sørgjerd L.K., Ytre-Arne M., Hogner S., Løvoll S.M., Kløvstad Olavsen M.R., Navarrete D., Gaup H.J., Lilje R., Zetterström R.H., Stray-Pedersen A., Rootwelt T., Rinaldo P., Rowe A.D, & Pettersen R.D. (2020). Performance of Expanded Newborn Screening in Norway Supported by Post-Analytical Bioinformatics Tools and Rapid Second-Tier DNA Analyses. International Journal of Neonatal Screening, 6(3), 51.

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CYP94 and Amidohydrolase Enzyme Assays

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For CYP94 assay, microsome incubations were performed as described in Widemann et al. (2015 (link)) with the following modifications: JA-Ile substrate concentration was 50 μM and the reaction was stopped with 150 μL methanol containing 0.2% acetic acid. The assays were centrifuged at 10000 g and the supernatant was used for LC-MS/MS analysis.
Amidohydrolase assay was performed in 200 μL as described in Widemann et al. (2013 (link)) using 10 μg affinity-purified protein and 30 μM substrates (JA-Ile, 12OH-JA-Ile or IAA). Both types of enzyme activities were analyzed by LC-MS as described in Widemann et al. (2015 (link)) on a Waters Quattro Premier XE (Waters, Mildorf, MA USA) instrument, using the following detection parameters: in negative mode: JA 209 > 59; 12OH-JA 225 > 59; 12OH-JA-Ile 338 > 130; 12COOH-JA-Ile 352 > 130; in positive mode: JA-Ile 324 > 151; IAA-Ala 247 > 130; IAA 176 > 130.

Hazman M., Sühnel M., Schäfer S., Zumsteg J., Lesot A., Beltran F., Marquis V., Herrgott L., Miesch L., Riemann M, & Heitz T. (2019). Characterization of Jasmonoyl-Isoleucine (JA-Ile) Hormonal Catabolic Pathways in Rice upon Wounding and Salt Stress. Rice, 12, 45.

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Quantification of UDP-GlcNAc in Activated T Cells

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Purified splenic CD4⁺ T cells activated for 72 hr in the presence or absence of T_H17-inducing conditions were washed twice with ice-cold 1x PBS and counted. 15 × 10⁶ cells were pelleted in 1.5 ml Eppendorf tubes by centrifuging at 350 g for 7 min at 4°C. Metabolites were extracted from the pellets by the addition of 1 ml of ice-cold solution of 40% acetonitrile, 40% methanol and 20% water. The tubes were vortexed for 1 hr at 4°C and spun down at 14,000 rpm for 10 min at 4°C (Eppendorf, Germany). The supernatant was transferred to fresh tubes and evaporated to dryness in an Eppendorf Vacufuge at 30°C (Eppendorf, Germany). The dry samples were stored at −80°C until analyzed, at which point they were reconstituted in 100 μl of LC-MS grade water. The samples were run on a Waters (Micromass) Quattro Premier XE LC-MS/MS machine in negative mode with Water:Acetonitrile (+0.2% Acetic Acid and 5 mM Ammonium Acetate) solvent. UPLC with C18 reverse phase column was employed. Area was converted to UDP-GlcNAc concentration using a standard curve simultaneously generated with purified UDP-GlcNAc (Sigma).

Araujo L., Khim P., Mkhikian H., Mortales C.L, & Demetriou M. (2017). Glycolysis and glutaminolysis cooperatively control T cell function by limiting metabolite supply to N-glycosylation. eLife, 6, e21330.

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Analytical Characterization of Chemical Compounds

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Analytical grade chemical reagents were purchased from Chengdu Changzheng Chemical Factory (Sichuan, People’s Republic of China). Thin-layer chromatography was performed on 0.20 mm silica gel 60 F₂₅₄ plates (Qingdao Ocean Chemical Factory, QingDao, People’s Republic of China). Proton nuclear magnetic resonance (¹H NMR) spectra were recorded at 400 MHz on a Varian spectrometer model Gemini 400 and reported in parts per million (ppm). Chemical shifts (δ) are quoted in ppm relative to tetramethylsilane as internal standard, where (δ) tetramethylsilane =0.00 ppm. The multiplicity of the signal is indicated as s, singlet; d, doublet; t, triplet; q, quartet; and m, multiplet, defined as all multipeak signals where overlap or complex coupling of signals makes definitive descriptions of peaks difficult. Mass spectra were obtained using a Q-TOF Premier mass spectrometer utilizing electrospray ionization (Waters Quattro Premier XE, Milford, MA, USA). The purity of compounds was determined to be ≥97% by high-performance liquid chromatography analysis with a photodiode array detector (Waters, Milford, MA, USA) and an Atlantis C₁₈ chromatographic column (150×4.6 mm², id 5 μm; Waters).

Ma L., Wang T., Shi M, & Ye H. (2016). Synthesis, activity, and docking study of phenylthiazole acids as potential agonists of PPARγ. Drug Design, Development and Therapy, 10, 1807-1815.

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Quantitative Analysis of Imatinib in Plasma

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In house liquid chromatography-tandem mass spectrometry method was developed and validated for quantitative analysis of Imatinib in plasma samples. The developed method was validated for various validation parameters (Table 2) according to the guidelines [23 ]. Analytical procedure was performed at Lambda Therapeutics Research Limited (Gujrat, India) using an instrument equipped with a MS/MS component (Quattro Premier XE, Waters, Manchester, UK), sample manager with column oven (Aquity SM, Waters), and binary solvent manager (Acquity BSM, Waters). The mobile phase consisted of methanol and ammonium formate buffer (5 mM, pH 7.0) at a ratio of 75:25 v/v. Chromatographic separation was achieved using a C18 column (ACE, 5μ, 150x4.6 mm) at the flow rate of 1 mL/min and injection volume (10 μL) in full loop mode. Mass-spectrometric detection was performed using an electrospray-ionization (ESI) technique and the ESI source was kept in positive ionization mode. MRM transitions for imatinib (analyte) and Imatinib-d8 (internal standard) were m/z 494.3 > 394.16 and 502.44 > 394.21, respect-ively. Data acquisition and evaluation was performed using Mass Lynx software ver. 4.1 (Waters).

Arora R., Sharma M., Monif T, & Iyer S. (2016). A Multi-centric Bioequivalence Trial in Ph+ Chronic Myeloid Leukemia Patients to Assess Bioequivalence and Safety Evaluation of Generic Imatinib Mesylate 400 mg Tablets. Cancer Research and Treatment : Official Journal of Korean Cancer Association, 48(3), 1120-1129.

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UPLC-MS/MS Acylcarnitine and LysoPC Profiling

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The ACQUITY UPLC system (Waters, Milford, MA, USA) consisted of a binary solvent manager, a vacuum degasser, a column heater and sample manager. The column temperature was maintained at 50°C. The samples were injected onto a Kinetex C8 column, 50 × 2.1 mm, 2.6 μm particle diameter (Phenomenex, Torrance, CA, USA). The acylcarnitines were separated by a linear gradient between solution A (0.1% formic acid in H₂O) and solution B (0.1% formic acid in methanol). The gradient was as follows: at T = 0 min: 36% A, 64% B, flow 0.4 mL/min towards T = 6 min: 0% A, 100% B, flow 0.4 mL/min; T = 6–11 min: 0% A, 100% B, flow 0.4 mL/min, and T = 11–11.1 back to 36% A, 64% B, flow 0.4 mL/min. A Quattro Premier XE (Waters, Milford, MA, USA) was used in the positive electrospray ionization mode. Nitrogen was used as desolvation gas (900 L/h) and cone gas (50 L/h). Desolvation temperature was 350°C, capillary voltage was 3.5 kV and the source temperature was 130°C. Argon was used as collision gas (2.5 x 10e-3 mbar). For the very long-chain acylcarnitines and lysophosphatidylcholines (lysoPC) multiple reaction monitoring (MRM) traces were acquired with optimized cone voltage and collision energy for each transition with a dwell time of 0.01 s (Table 1).

van de Beek M.C., Dijkstra I.M., van Lenthe H., Ofman R., Goldhaber-Pasillas D., Schauer N., Schackmann M., Engelen-Lee J.Y., Vaz F.M., Kulik W., Wanders R.J., Engelen M, & Kemp S. (2016). C26:0-Carnitine Is a New Biomarker for X-Linked Adrenoleukodystrophy in Mice and Man. PLoS ONE, 11(4), e0154597.

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Acylcarnitine Analysis by LC-MS/MS

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Acylcarnitine analysis was performed according to a published procedure (Liu and Pasquali, 2005 )) (Figure 1E, S1A–B). A Waters Acquity Ultra Performance LC solvent manager system was used for sample and mobile phase delivery. MS/MS analysis was performed on a Waters Quattro Premier XE tandem mass spectrometer operated in positive ion electrospray mode using MassLynx software. Acylcarnitines extracts were derivatised with butanolic HCl, dried, and resuspended in mobile phase for MS/MS analysis. Free carnitine and acetylcarnitine were monitored in selective reaction monitoring mode (SRM), while all other acylcarnitines were monitored using parent ion scan (m/z 85). A list of butylated acylcarnitine species are shown in Table S2. Acylcarnitines were quantified using stable isotope dilution method and NeoLynx software.

Simcox J., Geoghegan G., Maschek J.A., Bensard C.L., Pasquali M., Miao R., Lee S., Jiang L., Huck I., Kershaw E.E., Donato A.J., Apte U., Longo N., Rutter J., Schreiber R., Zechner R., Cox J, & Villanueva C.J. (2017). Global analysis of plasma lipids identifies liver-derived acylcarnitines as a fuel source for brown fat thermogenesis. Cell metabolism, 26(3), 509-522.e6.

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Quantitative Analysis of ABA in Soybean Seeds

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For analysis of ABA content in soybean seeds, the previously protocol we used (Shu et al., 2013 (link)) were employed in this study. Firstly, the seeds were ground to powder in liquid nitrogen, and 300 mg of seeds powder was homogenized and extracted for 24 h in methanol containing D6-ABA (OIChemIm Co. Ltd.) as an internal standard. Purification was performed with an Oasis Max solid phase extract cartridge (Waters) and eluted with 5% formic acid in methanol. Subsequently, the elution was dried and reconstituted, and it was then injected into a liquid chromatography–tandem mass spectrometry system consisting of an Acquity ultra performance liquid chromatograph (Acquity UPLC; Waters) and a triple quadruple tandem mass spectrometer (Quattro Premier XE; Waters). Three biological replications were performed.

Shu K., Qi Y., Chen F., Meng Y., Luo X., Shuai H., Zhou W., Ding J., Du J., Liu J., Yang F., Wang Q., Liu W., Yong T., Wang X., Feng Y, & Yang W. (2017). Salt Stress Represses Soybean Seed Germination by Negatively Regulating GA Biosynthesis While Positively Mediating ABA Biosynthesis. Frontiers in Plant Science, 8, 1372.

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Pharmacokinetics of Compound 15 in Mice

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Test compound 15 was dosed intravenously
or orally by gavage as a solution at 3 or 10 mg free base/kg respectively
(dose volume, 5 or 10 mL/kg respectively; dose vehicle, 10% (v/v)
dimethyl sulfoxide (DMSO), 60% polyethylene glycol 400, and 30% deionized
water) to female Balb/c mice (n = 3). Blood samples
were taken from each mouse at 0.08, 0.25, 0.5, 1, 2, 4, 6, 8, and
24 h post dose and mixed with two volumes of distilled water. After
suitable sample preparation, the concentration of test compound in
blood was determined by UPLC-MS/MS using a Quattro Premier XE (Waters,
USA). Pharmacokinetic parameters were derived from the mean blood
concentration time curve using PK solutions software v 2.0 (Summit
Research Services, USA).

Thomas M.G., De Rycker M., Ajakane M., Albrecht S., Álvarez-Pedraglio A.I., Boesche M., Brand S., Campbell L., Cantizani-Perez J., Cleghorn L.A., Copley R.C., Crouch S.D., Daugan A., Drewes G., Ferrer S., Ghidelli-Disse S., Gonzalez S., Gresham S.L., Hill A.P., Hindley S.J., Lowe R.M., MacKenzie C.J., MacLean L., Manthri S., Martin F., Miguel-Siles J., Nguyen V.L., Norval S., Osuna-Cabello M., Woodland A., Patterson S., Pena I., Quesada-Campos M.T., Reid I.H., Revill C., Riley J., Ruiz-Gomez J.R., Shishikura Y., Simeons F.R., Smith A., Smith V.C., Spinks D., Stojanovski L., Thomas J., Thompson S., Underwood T., Gray D.W., Fiandor J.M., Gilbert I.H., Wyatt P.G., Read K.D, & Miles T.J. (2018). Identification of GSK3186899/DDD853651 as a Preclinical Development Candidate for the Treatment of Visceral Leishmaniasis. Journal of Medicinal Chemistry, 62(3), 1180-1202.

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Quantification of Metyrapone and Metabolites

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Metyrapone and metyrapol were measured with Ultra Performance Liquid Chromatography (BEH C₁₈ 2.1x50 mm column at 45°C) coupled to a Quattro Premier XE mass spectrometer (Waters, Milford, MA). Acetonitrile (150 µL) with metyrapone-d6 (Alsachim, France) internal standard was added to 50 µL of sample. Ten microliters of supernatant was chromatographically resolved over 2 minutes with 2 mmol/L ammonium acetate plus 0.1% formic acid and a linear methanol gradient. Mass transitions (mz) were monitored in positive ionization mode: metyrapol 229.3 > 121; metyrapone 227.2 > 120.9 0; metyrapone d6 233.1 > 126.9. The detection limits for metyrapone and metyrapol were 0.2 and 0.5 µg/L and imprecision at 25 and 250 µg/L were < 7%. Cortisol, 11-deoxycortisol, and aldosterone were measured with tandem mass spectrometry [3 ]. ACTH was assayed on an Immulite 2000 analyzer (Siemens, Munich, Germany).

Duke M.E., Britten F.L., Pretorius C.J., McWhinney B.C., Ungerer J.P., Barrett H.L, & Donovan P. (2019). Maternal Metyrapone Use During Breastfeeding: Safe for the Breastfed Infant. Journal of the Endocrine Society, 3(5), 973-978.

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