Uplc beh amide column

Manufactured by Waters Corporation

Sourced in United States

The UPLC BEH Amide column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of polar compounds. It features a bridged ethylene hybrid (BEH) technology and an amide-based stationary phase, which provides enhanced selectivity and resolution for the separation of polar analytes. The column is compatible with ultra-high performance liquid chromatography (UPLC) systems and can be used for a variety of applications in analytical chemistry and life sciences.

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Lab products found in correlation

Analyst tf 1, by AB Sciex (17 mentions) Tripletof 5600 q tof, by AB Sciex (10 mentions) Tripletof 6600 q tof, by AB Sciex (10 mentions) 1290 uhplc system, by Agilent Technologies (8 mentions) Uhplc system, by Agilent Technologies (8 mentions) 1290 infinity series uhplc system, by Agilent Technologies (5 mentions) Tripletof 6600 mass spectrometry, by AB Sciex (5 mentions) Qtof 6550, by Agilent Technologies (4 mentions) 1290 infinity lc, by Agilent Technologies (3 mentions) Ammonia hydroxide, by CNW Technologies (2 mentions) Ammonium acetate, by CNW Technologies (2 mentions) 1290 infinity, by Agilent Technologies (2 mentions) Tripletof 5600, by AB Sciex (2 mentions) Tripletof 6600, by AB Sciex (2 mentions) Analyst tf v 1, by AB Sciex (1 mentions) Uhplc q tof ms, by Agilent Technologies (1 mentions) Scaa 104, by ANPEL (1 mentions) Acquity uplc 1 class, by Waters Corporation (1 mentions) Uhplc system, by Waters Corporation (1 mentions) Triple tof 5600 system, by AB Sciex (1 mentions)

Spelling variants of this product

ACQUITY UPLC BEH Amide column (127 citations) BEH Amide column (63 citations) ACQUITY UPLC Glycan BEH Amide column (17 citations) Waters ACQUITY UPLC BEH Amide column (7 citations) UPLC BEH Amide (3 citations) Aqcuity UPLC BEH amide column (3 citations) ACQUIY UPLC BEH Amide 1.7 μm column (3 citations) ACQUIY UPLC BEH Amide column (2 citations) ACQUITY UPLC BEH Amide 2.1 mm × 100 mm column (2 citations) AQUITY UPLC BEH Amide Column (2 citations)

57 protocols using uplc beh amide column

UHPLC-MS/MS Analysis of Metabolites

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The UHPLC separation was conducted using a 1,290 Infinity series UHPLC System (Agilent Technologies, MA, United States) coupled to a UPLC BEH amide column (1.7 µm, 2.1 × 100 mm; Waters, Milford, United States). The mobile phase was composed of ammonium acetate (25 mmol/L, CNW Technologies, Shanghai, China) and ammonia hydroxide (25 mmol/L, CNW Technologies), which were added to water (pH = 9.75) (A) and acetonitrile (B), respectively. The following elution gradient was used for analysis: 0–0.5 min, 95% B; 0.5–7.0 min, 95%–65% B; 7.0–8.0 min, 65%–40% B; 8.0–9.0 min, 40% B; 9.0–9.1 min, 40%–95% B, and 9.1–12.0 min, 95% B. The injection volume, auto-sampler temperature, and column temperature were 2 µl (both positive and negative modes), 4°C, and 25°C, respectively (19 (link)).
MS/MS data acquisition and analysis were conducted using the Analyst TF v1.7 (AB Sciex, MA, United States) according to a preset standard. In each cycle, a collision energy (CE) of 30 eV and cycle time of 0.56 s was used to select the densest 12 precursor ions with an intensity >100 for MS/MS. The following were set as the electrospray ionization (ESI) source conditions: curtain gas (35 psi), gas 1 (60 psi), gas 2 (60 psi), ion spray voltage floating (5,000 V in the positive mode and −4,000 V in the negative mode), declustering potential (60 V), and source temperature (600°C) (19 (link)).

Tan R., Ou S., Kang T., Wu W., Xiong L., Zhu T, & Zhang L. (2023). Altered serum metabolome associated with vascular calcification developed from CKD and the critical pathways. Frontiers in Cardiovascular Medicine, 10, 1114528.

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Rice Metabolite Extraction and Analysis

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The metabolites of rice were extracted and analyzed by the method described previously (Zhang Y. et al., 2017 (link)). Briefly, a total of 50 mg of fresh leaves was ground in liquid nitrogen, and transferred into the new EP tube containing 1 mL of an internal target substance (V_methanol:V_acetonitrile:V_water = 2:2:1, which was kept at -20°C in advance), and homogenized by ball mill at 45 Hz for 4 min, then treated on ice bath by continuous three times of ultrasound for 5 min, then incubated at -20°C for 1 h to precipitate proteins, then centrifuged by 14500 × g at 4°C for 15 min. A total of 500 μL of supernatants were transferred into the new EP tubes, and dried in a vacuum concentrator without heating, and was supplemented by a reconstitution with 100 μL of extraction solution (V_acetonitrile:V_water = 1:1), and mixed for 30 sec, and sonicated in a water bath at 4°C for 10 min, and centrifuged by 14500 × g at 4°C for 15 min. The supernatants were filtered through the PTFE membrane of 0.22 mm (Sigma, United States), and subjected to analyses on the UHPLC-QTOF-MS (1290, Agilent Technologies) equipped with a UPLC BEH Amide column (1.7 μm, 2.1 mm × 100 mm, Waters Corporation, United States) using optimized mobile phases. The extraction and analysis of metabolites were assisted by the Allwegene Technology Co., Ltd. (Beijing, China).

Wang B., Xie G., Liu Z., He R., Han J., Huang S., Liu L, & Cheng X. (2019). Mutagenesis Reveals That the OsPPa6 Gene Is Required for Enhancing the Alkaline Tolerance in Rice. Frontiers in Plant Science, 10, 759.

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Metabolite Extraction and LC-MS/MS Analysis

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A mixer mill (MM400, Retsch) was used to ground the samples into fine powders (1.5 min) at 30 Hz. Lyophilized powder (100 mg) was used to extract metabolites at 4°C using 0.8 ml of 70% aqueous methanol (methanol: H₂O₂, 70:30, v/v). After adding pure methanol, centrifugation was conducted for 10 min (10000 g). We gathered, homogenized, and sieved the supernatants accordingly (SCAA-104, 0.22 mm pore size; ANPEL Shanghai, China, www.anpel.com.cn/). Samples were mixed into tissue-specific samples, namely root tissue and rhizosphere soil, to assess the inter-tissue variations in the different compounds using LC-MS/MS analyses. Biomarker biocloud platform (www.biocloud.net) was used to carry out instrument stability after combining the samples. After that, UHPLC system (1290, Agilent Technologies) containing of a TripleTOF 5600 (Q-TOF, AB Sciex) and UPLC BEH Amide column (1.7 μm 2.1*100 mm, Waters) was used to carry out LC-MS/MS analyses. More details regarding LC-MS/MS analyses, metabolites processing, and annotation were documented in previous studies (Fallah et al., 2022 (link); Yuan et al., 2022 (link))

Fallah N., Pang Z., Lin Z., Nyimbo W.J., Lin W., Mbuya S.N., Ishimwe C, & Zhang H. (2023). Sustained organic amendments utilization enhances ratoon crop growth and soil quality by enriching beneficial metabolites and suppressing pathogenic bacteria. Frontiers in Plant Science, 14, 1273546.

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Quantifying Trehalose in Arabidopsis

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To measure trehalose contents, 10‐d‐old Arabidopsis seedlings (100 mg) were harvested and rapidly frozen in liquid nitrogen. After being ground to powder, each sample was evenly mixed with 500 μL chloroform: acetonitrile (3:7, v‐v) and incubated in a ‐10°C freezer with occasional shaking. The organic matter was extracted with 400 μL water at 4°C and centrifuged for 4 min at 10,000 g; this step was repeated twice. The supernatants were combined and dried in a centrifugal vacuum dryer. The dry samples were dissolved in 500 μL methanol: water (1:1, v‐v) and filtered through a 0.22 μm membrane at room temperature.
ACQUITY UPLC I‐Class (Waters) coupled with a Xevo G2 Q‐TOF high‐resolution mass spectrometer (Waters) was used to measure trehalose content. Chromatographic separation was conducted using a UPLC BEH Amide column (1.7 μm, 2.1 mm × 100 mm, Waters). Elution was performed with mobile 90% phase A (water, 0.1% ammonia) and 10% phase B (acetonitrile: water = 95:5, 0.1% ammonia, 2 mM ammonium acetate) at a flow rate of 0.3 mL/min.

Wang W., Chen Q., Xu S., Liu W., Zhu X, & Song C. (2020). Trehalose‐6‐phosphate phosphatase E modulates ABA‐controlled root growth and stomatal movement in Arabidopsis. Journal of Integrative Plant Biology, 62(10), 1518-1534.

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Untargeted Metabolomics Analysis Protocol

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Metabonomic profiles of included samples were detected with 290 Infinity series ultrahigh-performance liquid chromatography (UHPLC) System (Waters Corporation, Milford, MA, United States), equipped with a UPLC BEH Amide column (2.1 mm×100 mm, 1.7 μm). Ultrahigh-performance liquid chromatography (UHPLC) (Waters, Milford, United States) system equipped with AB SCIEX Triple TOF 5600 System (AB SCIEX, Framingham, United States). The mobile phase consisted of 25 mmol/L ammonium acetate and 25 mmol/L ammonia hydroxide in water (A) and acetonitrile (B). All analyzed samples were kept at 4°C, and the temperature of the column was kept at 25°C. The analysis was performed with an elution gradient as previously described (Darwish et al., 2021 (link); Wu et al., 2022b (link)). The Triple TOF 6600 mass spectrometry (AB Sciex, Boston, MA, United States) was applied to catch MS/MS spectra on an information-dependent basis (IDA) during the LC/MS experiment.

Wang X., Gao Y, & Jiang R. (2022). Diagnostic and predictive values of serum metabolic profiles in sudden sensorineural hearing loss patients. Frontiers in Molecular Biosciences, 9, 982561.

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LC-MS/MS Analysis of Metabolites

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The LC-MS/MS analyses were performed using an UHPLC system (1290, Agilent Technologies) with a UPLC BEH Amide column (1.7 μm 2.1 mm × 100 mm, Waters) coupled to Triple TOF 5600 (Q-TOF, AB Sciex). The mobile phase consisted of 25 mmol/L NH₄OAc and 25 mmol/L NH₄OH in water (pH = 9.75) (A) and acetonitrile (B) and was used with an elution gradient as follows: 0 min, 95% B; 7 min, 65% B; 9 min, 40% B; 9.1 min, 95% B; and 12 min, 95% B, which was delivered at 0.5 mL min⁻¹. The injection volume was 2 μL. The Triple TOF mass spectrometer was used for its ability to acquire MS/MS spectra on an information-dependent basis (IDA) during an LC/MS experiment. In this mode, the acquisition software (Analyst TF 1.7, AB Sciex) continuously evaluates the full-scan survey MS data as it collects and triggers the acquisition of the MS/MS spectra depending on preselected criteria. In each cycle, 12 precursor ions with an intensity greater than 100 were chosen for fragmentation at a collision energy (CE) of 30 V (15 MS/MS events with a product ion accumulation time of 50 ms each). The ESI source conditions were set as follows: ion source gas 1 at 60 Psi, ion source gas 2 at 60 Psi, curtain gas at 35 Psi, source temperature at 650°C, and ion spray voltage floating (ISVF) at 5000 V or −4000 V in positive or negative modes, respectively.

Wang B., Ma M.P., Diao Q.Y, & Tu Y. (2019). Saponin-Induced Shifts in the Rumen Microbiome and Metabolome of Young Cattle. Frontiers in Microbiology, 10, 356.

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UHPLC-MS Analysis of Metabolites

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UHPLC-MS analysis was performed on an Agilent 1290 UHPLC system (Agilent Technologies) which was equipped with TripleTOF 6600 mass spectrometry (AB Sciex). Sample was separated on a UPLC BEH Amide column (2.1 × 100 mm, 1.7 μm, Waters) with column temperature at 25 °C. The injection volume was 2 μL for each sample. Mobile phase A was acetonitrile. Mobile phase B consisted of ammonium acetate and ammonia hydroxide in water (25 mmol/L, respectively, pH = 9.75). Gradient elution was applied (0–0.5 min, 95% A; 0.5–7.0 min, 95%-65% A; 7.0–8.0 min, 65%-40% A; 8.0–9.0 min, 40% A; 9.0–9.1 min, 40%-95% A; 9.1–12.0 min, 95% A). The mass spectrometry was in tandem with UHPLC via an electrospray ion (ESI) source to acquire MS and MS/MS spectra under IDA mode. In this mode, the top 12 precursor ions from each MS scan (m/z 60–1200) were chosen for MS/MS scan (m/z 25–1200) at collision energy of 30 eV. The cycle time was 0.56 s. Gas 1, gas 2, and curtain gas of the ESI sourse was 60, 60, and 35 psi, respectively. The source temperature was 600 °C. The ion spray voltage was 5000 V and -4000 V in positive and negative ion modes, respectively.

Song W., Zhang Z., Jiang Y., Cao Y., Zhang B., Wang Y., Shi H, & Zhu L. (2022). Integrative metabolomic profiling reveals aberrations in myometrium associated with adenomyosis: a pilot study. Reproductive Biology and Endocrinology : RB&E, 20, 49.

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UHPLC-MS/MS Analysis of Metabolites

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The supernatant in the sampler bottle was analyzed using a 1,290 Infinity series UHPLC System equipped with an UPLC BEH Amide column (2.1 × 100 mm, 1.7 μm, Waters) (Agilent Technologies) with a mobile phase comprising 25 mmol L⁻¹ ammonium acetate and 25 mmol L⁻¹ ammonia hydroxide in water (pH = 9.75) (A) and acetonitrile (B). The analysis was carried out with an elution gradient as follows: 0 ∼ 0.5 min, 95% B; 0.5 ∼ 7.0 min, 95 ∼ 65% B; 7.0 ∼ 8.0 min, 65 ∼ 40% B; 8.0 ∼ 9.0 min, 40% B; 9.0 ∼ 1 min, 40 ∼ 95% B; and 9.1 ∼ 12.0 min, 95% B. The column temperature was 25°C, and the injection volume was 2 μL (positive) or 2 μL (negative).
A triple TOF 6600 mass spectrometer (AB Sciex) was used to acquire MS/MS spectra on an information-dependent basis (IDA). The acquisition software (Analyst TF1.7, AB Sciex) continuously evaluates the full-scan survey MS data as it collects and triggers the acquisition of MS/MS spectra depending on preselected criteria. The ESI source conditions were as follows: Gas 1 was 60 psi, Gas 2 was 60 psi, curtain gas was 35 psi, source temperature was 600°C, declustering potential was 60 V, and ion spray voltage floating (ISVF) was 5000 V or -4000 V in positive or negative modes, respectively.

Chen Y., Ji H., Guo J., Chen Y., Li W., Wang S, & Zhen L. (2022). Non-targeted Metabolomics Analysis Based on LC–MS to Assess the Effects of Different Cold Exposure Times on Piglets. Frontiers in Physiology, 13, 853995.

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UPLC-Q-TOF/MS Quantification Protocol

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Liquid phase conditions were shown as follows: UPLC BEH Amide Column (2.1 mm ×100 mm, 1.7 μm, Waters, USA); injection volume 5 μL; column temperature 55°C; mobile phase A-100% H₂O, B-100% ACN. Gradient elution conditions were as follows: at 0-1 min, 85% B; at 1-12 min, 65% B; at 12-12.1 min, 40% B; at 12.1-15 min, 40% B; at 15-15.1 min, 85% B; at 15.1-20 min, 85% B. The flow rate was 0.3 mL/min. Mass spectrometry conditions were as follows: UPLC-Q-TOF/MS (ESI) electrospray ionization source (X500R, AB SCIEX, USA); ion source temperature 600°C; ion source voltage -4500 V or 5500 V; curtain gas 20 psi, atomized gas and auxiliary gas both 60 psi. Multiple responses monitoring (MRM) was used for scanning.

Zhang B., Chen K., Liu L., Li X., Wu E., Han L., Shi Z, & Deng X. (2022). Effects of Lycium Barbarum Polysaccharides on the Metabolism of Dendritic Cells: An In Vitro Study. Journal of Immunology Research, 2022, 5882136.

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UHPLC-MS/MS for Metabolite Profiling

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UHPLC separation was performed using a 1290 Infinity series UHPLC System (Agilent Technologies) with a UPLC BEH amide column (2.1*100 mm, 1.7 μm, Waters). The column temperature was maintained at 25°C. The mobile phase was A, containing 25 mmol/L ammonium acetate and 25 mmol/L ammonia hydroxide (pH=9.75) in water, and B was acetonitrile. The gradient conditions were as follows: 95% B at 0–0.5 min, 95–65% B at 0.5–7.0 min, 65–40% B at 7.0–8.0 min, 40% B at 8.0–9.0 min, 40–95% B at 9.0–9.1 min, and 95% B at 9.1–12.0 min. The flow rate of the mobile phase was 0.5 L/min, and the autosampler temperature was set at 4°C. The injection volume was 1 μL for both negative mode and positive mode.
During the LC/MS experiment, TripleTOF 6600 mass spectrometry (AB Sciex) and information-dependent acquisition (IDA) were adapted to acquire MS/MS spectra. In IDA mode, Analyst TF (version 1.7, AB Sciex), the acquisition software, evaluated the full scan survey MS data and collected the MS/MS spectra depending on the preselected criteria. The 12 most intensive precursor ions with intensities greater than 100 were selected for MS/MS in each cycle; the collision energy was 30 eV, and the cycle time was 0.56 s. The ESI source parameters were set as follows: gas 1:60 psi; gas 2: 60 psi; CUR: 35 psi; TEM:600°C; DP: 60 V; ISVF: 5000 V in positive mode and −4000 V in negative mode.

Pan X., Chen W., Nie M., Liu Y., Xiao Z., Zhang Y., Zhang W, & Zou X. (2021). A Serum Metabolomic Study Reveals Changes in Metabolites During the Treatment of Lung Cancer-Bearing Mice with Anlotinib. Cancer Management and Research, 13, 6055-6063.

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