Uplc hss t3

Manufactured by Waters Corporation

Sourced in United States

The UPLC HSS T3 is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of compounds. It utilizes a sub-2 micron particle size and proprietary hybrid silica technology to provide efficient and reproducible separations.

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Lab products found in correlation

Exionlc system, by AB Sciex (1 mentions) Qtrap 6500, by AB Sciex (1 mentions) Waters xevo g2 qtof, by Waters Corporation (1 mentions) Masslynx 4, by Waters Corporation (1 mentions) Ultra performance liquid chromatography (uplc), by Waters Corporation (1 mentions) Compound discoverer 3, by Thermo Fisher Scientific (1 mentions) Q exactive, by Waters Corporation (1 mentions) Compound discover, by Thermo Fisher Scientific (1 mentions) 6530 accurate mass q tof, by Agilent Technologies (1 mentions) 1290 infinity binary lc system, by Agilent Technologies (1 mentions) Waters acquity uplc hss t3, by Waters Corporation (1 mentions)

Spelling variants of this product

ACQUITY UPLC HSS T3 C18 (132 citations) HSS T3 (52 citations) ACQUITY UPLC HSS T3 1.8 μm column (7 citations) ACQUITY UPLC HSS T3 1.8 μm (2.1 × 100 mm) column (3 citations) Waters Acquity UPLC HSS T3 (2.1×50 mm) column (3 citations) ACQUITY UPLC M-Class HSS T3 column (2 citations) ACQUITY UPLC HSS T3 C18 (1.8 μm (2 citations) ACQUITY UPLC® HSS T3 130 Å column (2 citations) Acquity UPLC HSS T3 1.8-mm column (2 citations) ACQUITY UPLC BEH HSS T3 Column, (2 citations)

12 protocols using uplc hss t3

Targeted Metabolite Analysis by UPLC-MS/MS

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Column: Waters UPLC HSS T3. Mobile phase: methanol, water + 0.1% formic acid. Flow rate: 0.3 ml/min. Sample injection volume: 10 µl. The mass spectrometer was a quadrupole orbital ion trap mass spectrometer (containing a thermospray ion source, Q Active ™) (Table 2).Table 2

Elution gradient table

Time (min)	Mobile phase
Time (min)	A (v%)	B (v%)
0	2	98
1.0	2	98
41.0	100	0
50.0	100	0
50.1	2	98
52.0	2	98

Fu Y., Gao C., Sun X., Zhao Y, & Zhang H. (2024). Study on the mechanism of action of Wu Mei Pill in inhibiting rheumatoid arthritis through TLR4-NF-κB pathway. Journal of Orthopaedic Surgery and Research, 19, 65.

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UPLC Gradient Elution for Compound Analysis

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Chromatographic separation was conducted on a WATERS ACQUITY UPLC HSS T3 (1.7 μm, 2.1 mm × 100 mm) at 40°C. The mobile phase contained 0.1% formic acid aqueous solution A) and acetonitrile B), at a flow rate of 0.3 ml min^-1. The gradient elution program was applied as follows: 2% B–11% B (0–10 min); 11% B–20% B (10–15 min); 20% B–30% B (15–25 min); 30% B–40% B (25–28 min); 40% B–98% B (28–47 min); 98% B (47–51 min); 98%–2% B (51–51.5 min); 2%–2% B (51.5–55 min). The injection volume was 2 μL.

Jiang Y., Chen M., Gang H., Li X., Zhai C., Feng Z., Luo G, & Gao X. (2023). A funnel-type stepwise filtering strategy for identification of potential Q-markers of traditional Chinese medicine formulas. Frontiers in Pharmacology, 14, 1143768.

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Quantification of Ferulic Acid via HRMS

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Verification of ferulic acid in the samples was performed on an Agilent 6550 high-resolution mass spectrometry (HRMS) coupled to an Agilent Infinity 1290 II UHPLC system. Metabolites were separated on an UPLC HSS T3 (1.8 μm, 2.1 × 100 mm, Waters) column with a water-MeOH gradient solvent system containing 0.04% formic acid. The gradient started at 5% MeOH with formic acid (MPB) and ramped to 100% MPB over 6 min and held for 4.50 min at 100% MPB. Column temperature was set to 45 °C and the flow at 0.4 ml/min. Mass spectra were acquired using a Jetstream ESI source in negative ionization mode scanning from 50 to 1700 m/z at 1.67 spectra/s. The capillary voltage was set at 3500 V. The source parameters were as follows: gas temperature at 175 °C, drying gas flow at 12 l/min, nebulizer at 45 psig, sheath gas temperature at 375 °C, and flow at 11 l/min and fragmentor at 300 V. Ferulic acid was identified based on its accurate mass (193.4546 m/z) and retention time (2.8 min). MS/MS data was acquired for the selected precursor at 237.3767 m/z identified in reactions containing DmCE1A and for a standard of 3,4,5-trimethoxycinnamic acid (molecular weight 238.24 g/mol) by using three different collision energies (10, 20 and 40 eV) with a duty cycle of 2.032 s/cycle. MS data was acquired with MassHunter Workstation Data Acquisition.

Kmezik C., Mazurkewich S., Meents T., McKee L.S., Idström A., Armeni M., Savolainen O., Brändén G, & Larsbrink J. (2021). A polysaccharide utilization locus from the gut bacterium Dysgonomonas mossii encodes functionally distinct carbohydrate esterases. The Journal of Biological Chemistry, 296, 100500.

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LC-MS/MS Metabolite Profiling Protocol

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Samples were injected into an ExionLC
System interfaced with a QTRAP 6500+ (Sciex, USA). The temperatures
of the autosampler were maintained at 10 °C. The injection volume
of samples was 5 μL. Eluents consisted of 10 mM ammonium formate
and 0.1% formic acid in 6:4 acetonitrile/water (eluent A) and 1:9
acetonitrile/methanol contained 10 mM ammonium formate and 0.1% formic
acid (eluent B). The flow rate was set at 0.35 mL/min. A 20 min elution
gradient with an UPLC HSS T3 (1.8 μm, 2.1 × 100 mm, Waters)
column was carried out as follows: During the first 1.5 min, the eluent
composition was set at 0% B, which was linearly increased to 55% B
at 5 min, then 60% B at 10 min, 70% B at 13 min, and 90% B at 15 min;
in the next 1 min, B was changed to 100% and held for 2 min. Mass
data were acquired in the positive/negative ion mode using the time-scheduled
multiple reaction monitoring (MRM) method, respectively. The source
conditions were as follows: curtain gas was 40 psi, collision gas
was medium, ion spray voltage was −4500 V/+5500 V, and gas
1 and gas 2 were 50 and 55 psi, respectively.

Song Q., Liu H., Zhang Y., Qiao C, & Ge S. (2022). Lipidomics Revealed Alteration of the Sphingolipid Metabolism in the Liver of Nonalcoholic Steatohepatitis Mice Treated with Scoparone. ACS Omega, 7(16), 14121-14127.

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UPLC-ESI-MS/MS Analysis of Phenolic Metabolites

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For the purpose of qualitatively and quantitatively detecting target metabolites, UPLC-ESI-MS/MS analysis was employed.
An ultrahigh-performance liquid chromatograph from AB Company (Framingham, Massachusetts, USA) was used as the chromatographic system. To separate phenolic substances, a Waters UPLC HSS T3 (100 × 2.1 mm, 1.8 m) liquid chromatographic column was utilized (Milford, Massachusetts, USA). The following chromatographic conditions were used: injection volume of 5 μL, flow rate of 0.35 mL/min, mobile phase comprising A (0.1% formic acid water solution) and B (acetonitrile). The gradient elution procedure was set as follows: 0 min A/B (95:5, V/V), 0.8 min A/B (95:5, V/V), 3 min A/B (75:25, V/V), 12 min A/B (56.2:43.8, V/V), 13 min A/B (1:99, V/V), 14.4 min A/B (1:99, V/V), 14.41 min A/B (95:5, V/V), and 15 min A/B (95:5, V/V).
For the mass spectrum conditions, the following settings were employed: Positive ion mode with CUR at 35 psi, EP at 10, IS at 5500, CXP at 10, TEM at 500 ℃, Gas1 at 60 psi, and Gas2 at 50 psi. Negative ion mode with CUR at 35 psi, EP at −10, IS at −4500, CXP at −20, TEM at 500 ℃, Gas1 at 60 psi, and Gas2 at 50 psi. The column temperature was maintained at 40 ℃.

Xiong Q., Zhang J., Sun C., Wang R., Wei H., He H., Zhou D., Zhang H, & Zhu J. (2023). Metabolomics revealed metabolite biomarkers of antioxidant properties and flavonoid metabolite accumulation in purple rice after grain filling. Food Chemistry: X, 18, 100720.

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Ultra-high-performance LC-QTOF analysis of metabolites

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Data were collected using an ultra-high-performance liquid phase (Waters Acquity UPLCTM I-class system, Waters Corp, Milford, MA, USA) coupled with a high-resolution time-of-flight mass spectrometer (Waters Xevo G2 QTof, Waters Corp., Milford, MA, USA). Chromatography was performed using Waters UPLC HSS T3 at 40°C. The data were processed using Waters MassLynx 4.1. The mobile phase was water (A) and acetonitrile (B) containing 0.1% formic acid, the flow rate was 0.3 mL/min, and the elution conditions were as follows: 0–2 min, 0–5% B; 2–10 min, 5%–15% B; 10–15 min, 15%–25% B; 15–18 min, 25%–50% B; 18–23 min, 50%–100% B; 23–25 min, 100%–2% B; and 25–30 min, 2% B. The injection volume was 2 μL. The mass spectra were obtained in Fast DDA and positive ion mode, with an ESI ion source, and the following settings were used: electrospray ionization ion scan range: 50–2000 m/z; capillary voltage: 3.0 kV; ion source temperature: 100°C; desolvation gas (N₂) temperature: 500°C; desolvation gas flow rate: 1000 L/h; cone gas flow rate (N₂): 100 L/h; and collision gas: argon.

Li Z., Liu Q., Zhu Y., Wu L., Liu W., Li J., Zhang Z, & Tao F. (2022). Network Pharmacology, Molecular Docking, and Experimental Validation to Unveil the Molecular Targets and Mechanisms of Compound Fuling Granule to Treat Ovarian Cancer. Oxidative Medicine and Cellular Longevity, 2022, 2896049.

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LC-MS Metabolomics Profiling Protocol

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ACQUITY UPLC HSS T3 (2.1 × 100 mm 1.8 m columns) was used for liquid chromatography–mass spectrometry (LC-MS) (Waters, UPLC; Thermo, Q Exactive). The following chromatographic separation conditions were used: column temperature, 40°C; flow rate, 0.30 mL/min; mobile phase A, water + 0.05 percent formic acid; mobile phase B, acetonitrile; injection volume, 2 μL; automatic injector temperature, 4°C. The MS parameters are as follow: ESI+: spray voltage, 3.0 (ESI+) or 3.2 (ESI-) kV; S-Lens RF level, 30% (ESI+) or 60% (ESI-); heater temperature, 300 °C; sheath gas flow rate, 45 arb; auxiliary gas flow rate, 15 arb; sweep gas flow rate, 1arb; capillary temperature, 350°C. Scan duration is 100 ms, interscan time is 50 ms, and the scan range is 70-1050 m/z. Data was analyzed using Compound Discoverer 3.1 software (Thermo Fisher Scientific,USA), normalized, and converted into a two-dimensional matrix using Excel 2010 software, containing retention time, compound molecular weight, observations (samples), and peak intensity (39 (link), 40 (link)).

Su Y.N., Wang M.J., Yang J.P., Wu X.L., Xia M., Bao M.H., Ding Y.B., Feng Q, & Fu L.J. (2023). Effects of Yulin Tong Bu formula on modulating gut microbiota and fecal metabolite interactions in mice with polycystic ovary syndrome. Frontiers in Endocrinology, 14, 1122709.

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Untargeted Metabolomics of Mouse Feces

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Untargeted metabolomics was used to analyze the metabolome in mouse feces. One-hundred microliter feces was mixed with 300 μl methanol and 20 μl internal standard. Samples were extracted with ultrasound for 5 min in an ice bath and then stood at −20°C for 2 h. After centrifuging at 13,000 rpm at 4°C for 15 min, 200 μl supernatant was taken in a 2 ml vial for LC-MS analysis. The instrument platform for LC-MS analysis consisted of Agilent 1290 ULTRA performance liquid chromatography in tandem with Thermo Fisher Scientific Q Exactive Orbitrap High-resolution mass spectrometer. The chromatographic column was UPLC HSS T3 (1.7 μm 2.1 × 100 mm, Waters). Compound Discover (Version 2.0, Thermo) and OSI-SMMS (Version 1.0, Dalian Dasuo Information Technology Co., LTD.) were used in conjunction with mzCloud database and self-built database for substance identification.

Rong Z.J., Cai H.H., Wang H., Liu G.H., Zhang Z.W., Chen M, & Huang Y.L. (2022). Ursolic Acid Ameliorates Spinal Cord Injury in Mice by Regulating Gut Microbiota and Metabolic Changes. Frontiers in Cellular Neuroscience, 16, 872935.

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UPLC-MS/MS Analytical Protocol

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Chromatography was performed on 1290 Infinity Binary LC System from Agilent together with Waters Acquity UPLC HSST3 1.8μm 2.1 × 100 mm column in connection with a Water Acquity UPLC HSS T3 1.8μm VanGuard Pre-column. Mass spectrometry was performed using Agilent Technologies 6530 Accurate-Mass Q-T of with a dual ASJ ESI ion source.

Sarkar M.K., Kaplan N., Tsoi L.C., Xing X., Liang Y., Swindell W.R., Hoover P., Aravind M., Baida G., Clark M., Voorhees J.J., Nair R.P., Elder J.T., Budinova I., Getsios S, & Gudjonsson J.E. (2017). Endogenous glucocorticoid deficiency in psoriasis promotes inflammation and abnormal differentiation. The Journal of investigative dermatology, 137(7), 1474-1483.

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Analytical Techniques for Areca Nut Compounds

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Arecoline was quantified using high-performance liquid chromatography (HPLC), as described previously, with some modifications (25 (link)). For HPLC, we used a Waters UPLC HSS T3 chromatographic column (100 mm × 2.1 mm × 1.8 μm), a 215-nm wavelength, a column temperature of 30°C, a flow rate of 0.15 mL/min, a sample volume of 10 μL, and a mobile phase comprising 0.1% phosphoric acid-acetonitrile in a 65:35 ratio. Furthermore, areca nut polyphenols were detected using enzyme-linked immunosorbent assay (ELISA). The ELISA kit was purchased from Suzhou Michy Biology Technology Co., Ltd. The HPLC analysis and ELISA showed that the extracted ANE contained 8.75 μg/g and 8.86 mg/g of arecoline and phenol, respectively. The secondary metabolites in ANE were detected using liquid chromatography-mass spectrometry (LC–MS) (Supplementary Table S1).

Wang S., Wang H., Jiang Q., Dai J., Dai W., Kang X., Xu T., Zheng X., Fu A., Xing Z., Chen Y., He Z., Lu L, & Gu L. (2023). Supplementation of dietary areca nut extract modulates the growth performance, cecal microbiota composition, and immune function in Wenchang chickens. Frontiers in Veterinary Science, 10, 1278312.

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