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Acquity ultra high performance liquid chromatography

Manufactured by Waters Corporation
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The Acquity ultra-high performance liquid chromatography (UHPLC) system is a laboratory instrument designed for the separation, identification, and quantification of chemical compounds. It utilizes advanced technology to achieve high-resolution separations with increased speed and efficiency compared to traditional HPLC systems.

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Lab products found in correlation

Waterspremier xe triple quadrupole mass spectrometer, by Waters Corporation (2 mentions) Uplc system, by Waters Corporation (2 mentions) Xevo tq s, by Waters Corporation (2 mentions) Acquity uplc beh c18 column, by Waters Corporation (2 mentions) Acquity uplc beh c18 vanguard pre column, by Waters Corporation (2 mentions) Acquity beh c18 pre column, by Waters Corporation (2 mentions) Uplc beh amide column, by Waters Corporation (1 mentions) 1290 hplc, by Agilent Technologies (1 mentions) Tmt reagent, by Thermo Fisher Scientific (1 mentions) 9rkfsg2 ncs 3500r system, by Thermo Fisher Scientific (1 mentions) Q exactive hf x, by Thermo Fisher Scientific (1 mentions) Beh c18 column, by Waters Corporation (1 mentions) Acquity photodiode array detector, by Waters Corporation (1 mentions) Uplc h class system, by Waters Corporation (1 mentions) Inova 400 mhz, by Bruker (1 mentions) Avance 3 hd 850 mhz, by Bruker (1 mentions) Isolera one, by Biotage (1 mentions) Acquity uplc beh c18, by Waters Corporation (1 mentions) Kinetex xb c18 100 column, by Phenomenex (1 mentions) Hplc system, by Waters Corporation (1 mentions)

19 protocols using acquity ultra high performance liquid chromatography

1

Quantitative Phospholipid Profiling in Cells

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Quantitative analysis of PE and PS was carried out using a Waters
Premier XE triple quadrupole mass spectrometer interfaced with an Acquity
ultra-high performance liquid chromatography (UPLC) system (Waters, Milford,
MA). Mass spectrometer was operated in electrospray ionization mode with
negative ion acquisition. Phospholipids were extracted from cells by the
Folch procedure with initial addition of internal standards,
D31-160/181 PE and D31-160/181 PS. Quantification was
based on calibration curves constructed using PE and PS standards with
multiple reactions monitoring function. PE calibration curves include 160
LPE, 181 LPE, 160/181 PE, 160,182 PE, 180,182 PE, 180/204 ES. PE and PS
species without reference compounds were quantified with the standard
sharing the closest structure. PS calibration curves include 160 LPS, 181
LPS, 180 LPS, 160/181 PS, 160,182 PS, 180,182 PS, 180/204 PS. Final
concentration of PE and PS were normalized to DNA contents in the cell and
expressed as ng/mg DNA.
Thomas H.E., Zhang Y., Stefely J.A., Veiga S.R., Thomas G., Kozma S.C, & Mercer C.A. (2018). Mitochondrial Complex I Activity Is Required for Maximal Autophagy. Cell reports, 24(9), 2404-2417.e8.
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2

High-Resolution LC-HRMS Analysis of Metabolites

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LC-HRMS was performed using an Agilent G6540 Quadrupole Time of Flight (QTOF) system consisting of an Agilent 1290 HPLC coupled to a high resolution (QTOF) mass spectrometer. Electrospray ionization (ESI) in both positive and negative ion modes was employed using a dual ESI source under high-resolution exact mass conditions. 2 µL of sample was injected. A Waters Acquity ultra high performance liquid chromatography (UPLC) BEH Amide column with dimensions 2.1×150 mm, 1.7 µM particle size was used for Hydrophilic Interaction Liquid Chromatography (HILIC), and maintained at 40°C. Data was acquired for each sample for 29 minutes at a flow rate of 0.5 mL/minute using a solvent gradient with 0.1% formic acid in water and 0.1% formic acid in acetonitrile. An Agilent Zorbax Eclipse Plus C8 2.1×100 mm, 1.8 µM particle size column was used for C8 chromatography and data was acquired for each sample for 50 minutes at a flow rate of 0.5 mL/minute using a gradient with 0.1% formic acid in water and 0.1% formic acid in acetonitrile and maintained at 40°C.
West P.R., Amaral D.G., Bais P., Smith A.M., Egnash L.A., Ross M.E., Palmer J.A., Fontaine B.R., Conard K.R., Corbett B.A., Cezar G.G., Donley E.L, & Burrier R.E. (2014). Metabolomics as a Tool for Discovery of Biomarkers of Autism Spectrum Disorder in the Blood Plasma of Children. PLoS ONE, 9(11), e112445.
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3

Analytical Workflow for High-Resolution Mass Spectrometry

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ACQUITY Ultra-High Performance Liquid Chromatography (Waters, USA), SYNAPT G2-Si Q-TOF/MS Quadrupole Time-of-flight Mass Spectrometer (Waters, USA), Ultra-High-Speed Low-Temperature refrigerated centrifuge (Shanghai Anting Scientific Instrument Factory), ME155DU electronic balance (Shanghai Mettler-Toledo Instrument Co., Ltd.), KQ-300DB CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.).
Kui H., Lei Y., Jia C., Xin Q., Tursun R., Zhong M., Liu C, & Yuan R. (2024). Antithrombotic pharmacodynamics and metabolomics study in raw and processed products of Whitmania pigra Whitman. Heliyon, 10(7), e27828.
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4

Bile Acid Profiling in Mouse Serum and Liver

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BAs were extracted from mouse serum and liver tissues then analyzed by LC-MS. An aliquot of 25 μl of serum was deproteinated with 100 μl of acetonitrile containing 1 μM d5-TCA (internal standard). After centrifugation for 10 min at 15,000 ×g, 100 μl of the supernatant was further diluted with 100 μl of water containing 0.1% formic acid. The liver samples were homogenized with 1/10 (w/v) acetonitrile containing 1 μM d5-TCA. After centrifugation for 10 min at 15,000 ×g, 40 μl of supernatant was diluted with 360 μl of water containing 0.1% formic acid. A 5 μl aliquot of the supernatants was injected into an Acquity ultra-high-performance liquid chromatography/Synapt G2Si quadrupole time-of-flight mass spectrometry system (UPLC-Q/TOF MS, Waters Corporation, Milford, MA).
Xie C., Takahashi S., Brocker C.N., He S., Chen L., Xie G., Jang K., Gao X., Krausz K.W., Qu A., Levi M, & Gonzalez F.J. (2019). Hepatocyte peroxisome proliferator-activated receptor α regulates bile acid synthesis and transport. Biochimica et biophysica acta. Molecular and cell biology of lipids, 1864(10), 1396-1411.
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5

Porphyrin Isomers Analysis by LC-MS

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The analyses of isomers of porphyrins and of the crude extracts obtained under acidic conditions were conducted on a LC-MS in a Xevo TQ-S (Waters Corporation) Mass Spectrometer coupled with Acquity Ultra High-Performance Liquid Chromatography (UPLC, Waters). Samples were eluted using a gradient of 0.1% formic acid (phase A) and methanol/0.1% formic acid (phase B). The gradient started with 40% phase B and increased linearly to 95% within 5 min, with an additional hold for 2 min in 95% phase B. The flow rate was 0.4 mL.min−1. The mass spectrometer operated with electrospray ionization in the positive mode (ESI+). The capillary voltage and spray voltage were set at 3.0 kV and 40 V, respectively; with desolvation temperature 300 °C, source temperature 120 °C, argon used as the collision gas, and collision gas flow 0.2 mL.min−1.
Martinez A.F., de Almeida L.G., Moraes L.A, & Cônsoli F.L. (2017). Tapping the biotechnological potential of insect microbial symbionts: new insecticidal porphyrins. BMC Microbiology, 17, 143.
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6

Quantification of Sterols and Carotenoids

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The sterols content was determined as the methodology substantive in Stuper-Szablewska et al. [51 (link)]. The carotenoids were determined utilizing the Acquity ultra-high performance liquid chromatography (Waters, Milford, MA, USA) as the same methodology and conditions substantive in Kurasiak-Popowska et al. [52 (link)].
Abdel-Razek A.G., Badr A.N., Alharthi S.S, & Selim K.A. (2021). Efficacy of Bottle Gourd Seeds’ Extracts in Chemical Hazard Reduction Secreted as Toxigenic Fungi Metabolites. Toxins, 13(11), 789.
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7

Quantitative Phospholipid Profiling in Cells

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Quantitative analysis of PE and PS was carried out using a Waters
Premier XE triple quadrupole mass spectrometer interfaced with an Acquity
ultra-high performance liquid chromatography (UPLC) system (Waters, Milford,
MA). Mass spectrometer was operated in electrospray ionization mode with
negative ion acquisition. Phospholipids were extracted from cells by the
Folch procedure with initial addition of internal standards,
D31-160/181 PE and D31-160/181 PS. Quantification was
based on calibration curves constructed using PE and PS standards with
multiple reactions monitoring function. PE calibration curves include 160
LPE, 181 LPE, 160/181 PE, 160,182 PE, 180,182 PE, 180/204 ES. PE and PS
species without reference compounds were quantified with the standard
sharing the closest structure. PS calibration curves include 160 LPS, 181
LPS, 180 LPS, 160/181 PS, 160,182 PS, 180,182 PS, 180/204 PS. Final
concentration of PE and PS were normalized to DNA contents in the cell and
expressed as ng/mg DNA.
Thomas H.E., Zhang Y., Stefely J.A., Veiga S.R., Thomas G., Kozma S.C, & Mercer C.A. (2018). Mitochondrial Complex I Activity Is Required for Maximal Autophagy. Cell reports, 24(9), 2404-2417.e8.
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8

Quantitative Proteomics Analysis of Bone Tissues

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Proteomics analyses were performed at Sinotech Genomics Inc. (Shanghai, China) according to a standard procedure. Briefly, bone tissues were ground into powder in liquid nitrogen and suspended in lysis buffer (1% sodium deoxycholate (SDS) and 8 M urea) to extract total protein. Then, the extracted protein was quantified, qualified, and digested. The resulting peptide mixture was labeled using 10-plex tandem mass tag (TMT) reagent (Thermo Fisher, USA). Next, the samples were pooled and fractionated by ACQUITY ultrahigh-performance liquid chromatography (Waters, USA) on an ACQUITY UPLC BEH C18 column (1.7 µm, 2.1 mm × 150 mm, Waters, USA) to increase the proteomic depth. Labeled peptides were analyzed by online nanoflow liquid chromatography with tandem mass spectrometry on a 9RKFSG2_NCS-3500R system (Thermo Fisher, USA) connected to a Q Exactive HF-X (Thermo Fisher, USA) with a nanoelectrospray ion source. Finally, the raw data were analyzed to identify differentially expressed proteins at a false discovery rate (FDR) cutoff of 0.05 (95% confidence interval).
Li S.D., Xing W., Wang S.C., Li Y.B., Jiang H., Zheng H.X., Li X.M., Yang J., Guo D.B., Xie X.Y., Jiang R.Q., Fan C., Li L., Xu X, & Fei J. (2023). Fibulin2: a negative regulator of BMSC osteogenic differentiation in infected bone fracture healing. Experimental & Molecular Medicine, 55(2), 443-456.
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9

UPLC-Triple-TOF-MS/MS Analysis of Compounds

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UPLC-Triple-TOF-MS/MS analysis was performed using an ACQUITY ultra-high performance liquid chromatography (Waters, MA, USA) coupled with an ABSCIEX-Triple TOF 5600 triple time-of-flight mass spectrometry (AB Sciex, MA, USA). The chromatographic conditions were as follows: BEH C18 column (100 mm × 2.1 mm × 1.7 μm; Waters, Milford, MA), aqueous solution of 0.1% formic acid as mobile phase A, acetonitrile/isopropanol solution (1 : 1) containing 0.1% formic acid as mobile phase B, gradient elution at 0.4 mL min−1, injection volume of 10 μL, and column temperature of 40 °C. For mass spectroscopy, the scanning modes of positive ion and negative ion were both adopted for collecting the mass spectrum signals. The ionspray voltage at anode (ESI+) was +5000 V, ionspray voltage at cathode (ESI−) was −4000 V, source temperature was 500 °C, MS/MS collusion energy was 20–60 V, and mass range was 50–1000 m/z.
Sun J., Liu J., Ren G., Chen X., Cai H., Hong J., Kan J., Jin C., Niu F, & Zhang W. (2022). Impact of purple sweet potato (Ipomoea batatas L.) polysaccharides on the fecal metabolome in a murine colitis model. RSC Advances, 12(18), 11376-11390.
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10

Sterol Oil Content and Carotenoid Analysis

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The oil content of sterols was measured due to the methodology described by Stuper-Szablewska et al. [59 (link)]. While, carotenoids separated and their quantity in samples was evaluated using Acquity ultra-high performance liquid chromatography (Waters, Milford, MA, USA) according to a method and conditions described by Kurasiak-Popowska et al. [60 (link)].
Badr A.N., Ali H.S., Abdel-Razek A.G., Shehata M.G, & Albaridi N.A. (2020). Bioactive Components of Pomegranate Oil and Their Influence on Mycotoxin Secretion. Toxins, 12(12), 748.
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