Xbridge amide

Manufactured by Waters Corporation

Sourced in United States

The XBridge Amide is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of polar compounds. It features a proprietary bonded phase that provides excellent retention and selectivity for a wide range of polar analytes, including carbohydrates, glycans, and other polar molecules.

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Lab products found in correlation

Dnapac pa100 4 x 250 mm column, by Thermo Fisher Scientific (2 mentions) Qtrap 6500, by AB Sciex (1 mentions) Lc 30ad, by Shimadzu (1 mentions) Qtrap 6500 mass spectrometer, by Shimadzu (1 mentions) Ultimate 3000 hplc, by Thermo Fisher Scientific (1 mentions) Lcms 8030, by Shimadzu (1 mentions) Api5500 triple quadrupole mass, by AB Sciex (1 mentions) Sil 20a xr system, by Shimadzu (1 mentions) Finnigan surveyor, by Thermo Fisher Scientific (1 mentions) Sucrose, by Yuanye Bio-Technology (1 mentions)

Close spelling variants of this product

Xbridge amide column Xbridge amide 3.5 μm column XBridge BEH Amide XP HILIC column Amide XBridge HILIC column Amide XBridge HPLC column XBridge BEH Amide XP Column XBridge BEH Amide XBridge BEH Amide column XBridge BEH Amide HPLC column XBridge BEH Amide OBD column

18 protocols using xbridge amide

Metabolite Separation by HPLC-MS

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We used two methods as described below to separate the metabolites:
Method A: ESI positive mode was used in method A. The HPLC column was Waters Xbridge Amide 3.5 μm, 4.6 × 100 mm. Mobile phase A and B were 0.1% formic acid in water and acetonitrile, respectively. Gradient: 0 min-85% B; 3–12 min-85 to 30% B, 12–15 min-2% B, 16 min-95% -B, followed by re-equilibration end of the gradient, 23 min to the initial starting condition 85% B. Flow rate: 0.3 ml/min. Injection volume 5 ul.
Method B: ESI negative mode was used in method B. The HPLC column was Waters Xbridge Amide 3.5 μm, 4.6 × 100 mm. Mobile phase A and B were 20 mM ammonium acetate in water with pH 9.0 and 100% acetonitrile, respectively. Gradient: 0 min-85 % B; 0–3 min-85 to 30% B, 3–12 min-30%-2% B, 12–15 min-2% -B, 15–16 min-85% B followed by re-equilibration end of the gradient- the 23 min to the initial starting condition 85% B. Flow rate: 0.3 ml/min. Injection volume 5 ul.

Moye L.M., Liu Y., Coarfa C., Putluri N, & Rhoads J.M. (2019). Plasma Urea Cycle Metabolites May Be Useful Biomarkers in Children With Eosinophilic Esophagitis. Frontiers in Pediatrics, 6, 423.

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Metabolite separation by ESI-LC-MS

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Electrospray ionization (ESI) Positive mode was used to separate the metabolites. The LC column was Waters XBridge Amide 3.5 μm, 4.6 × 100 mm. Mobile phase A and B were 0.1% formic acid in water and acetonitrile respectively. Gradient: 0 min-85% B; 3–12 min- 85% to 10% B, 12–15 min-10% B, 16 min- 85% -B, followed by re-equilibration end of the gradient- the 23 min to the initial starting condition 85% B. Flow rate: 0.3 ml/min. ESI negative mode was also used to separate the metabolites.
The HPLC column was Waters XBridge Amide 3.5 μm, 4.6 × 100 mm. Mobile phase A and B were 20 mM ammonium acetate in water with pH 9.0 and 100% acetonitrile respectively. Gradient: 0 min-85% B; 0–3 min- 85% to 30% B, 3–12 min-30%−2% B, 12–15 min- 2% -B, 15–16 min- 85% B followed by re-equilibration end of the gradient- the 23rd min to the initial starting condition 85% B. Flow rate: 0.3 ml/min.

Vantaku V., Putluri V., Bader D.A., Maity S., Ma J., Arnold J.M., Rajapakshe K., Donepudi S.R., von Rundstedt F.C., Devarakonda V., Dubrulle J., Karanam B., McGuire S.E., Stossi F., Jain A.K., Coarfa C., Cao Q., Sikora A.G., Villanueva H., Kavuri S.M., Lotan Y., Sreekumar A, & Putluri N. (2019). Epigenetic loss of AOX1 expression via EZH2 leads to metabolic deregulations and promotes bladder cancer progression. Oncogene, 39(40), 6265-6285.

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HPLC Analysis of Nucleic Acids

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Analysis of nucleotide samples was performed by running a linear gradient of 90% acetonitrile/10% water to 50% acetonitrile/H₂O for 7 min, followed by isocratic 90% aqueous acetonitrile for 6 min on a Waters XBridge Amide (150 × 2.5 × 3.5 μm³) column. Analytical HPLC analysis of nucleoside samples was performed by running a linear gradient of 85% acetonitrile/15% water to 40% acetonitrile/60% water over 25 min on a Waters XBridge Amide (150 × 2.5 × 3.5 μm³) column.

Cafferty B.J., Fialho D.M., Khanam J., Krishnamurthy R, & Hud N.V. (2016). Spontaneous formation and base pairing of plausible prebiotic nucleotides in water. Nature Communications, 7, 11328.

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Enzymatic Synthesis of Glucan Polymers

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All reactions were performed in PBS buffer (pH 6.0, 50 mM). A mixture of the purified CBS (100 μg), 0.5 mM ATP and glucose (20 mg) or UDPG (20 mg) was suspended in 1 mL of PBS buffer and the mixture was stirring at 30 °C for 1 day. After reaction, the mixture was centrifuged and treated with filter membrane (pore size 0.45 μm). HPLC was utilized to analyze the composition of the samples. Measurement conditions: column, XBridge Amide (Waters); column temperature, 30 °C; mobile phase, 82% acetonitrile, and 18% water; flow rate, 0.4 mL/min; injection volume, 20 μL. Standard configuration: standard mixture was prepared by 1% mobile phase.

Zhang Y., Tang B, & Du G. (2017). Self-induction system for cellulase production by cellobiose produced from glucose in Rhizopus stolonifer. Scientific Reports, 7, 10161.

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Targeted Metabolite Quantification by LC-MS/MS

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Targeted LC-MS/MS analysis
was performed using a Shimadzu LC system coupled to a triple quadrupole
mass spectrometer (QTRAP 6500+, ABSciex). Liquid chromatography was
performed with 20 mM ammonium acetate and 0.1% ammonium hydroxide
in 95:5 water/ACN as phase A and ACN as phase B. LC-method-1 uses
a HILIC column (iHILIC-(P) Classic column, 5 μm, 150 ×
2.1 mm, 200 A, made in Sweden). Mobile phases A and B are the same
as method 1. The flow rate is 0.2 mL/min, the total runtime is 23
min, and the gradient is: 0 min, 85% B; 2 min, 85% B; 7 min, 60% B;
12 min, 35% B; 12.1 min, 20% B; 15.9 min, 20% B; 16 min, 85% B; 23
min, 85% B. ESI parameters are GS1, 60; GS2, 60; CUR, 35; temperature,
500; ISVF, 5000 or −4500 in positive or negative modes, respectively.
LC-method-2 uses a different HILIC column (Waters XBridge Amide, 3.5
μm, 4.6 × 100 mm, made in Ireland). The flow rate is 0.4
mL/min, the total run time is 26 min, and the gradient is 0 min, 85%
B; 0.1 min, 85% B; 3.5 min, 32% B; 12 min, 2% B; 16.5 min, 2% B; 17
min, 85% B; 25.5 min, 85% B; 26 min, 85% B. ESI parameters: GS1, 33;
GS2, 33; CUR, 25; temperature, 475; ISVF, 4500 or −4500 in
positive or negative modes, respectively. All results were obtained
using LC-method-1, unless specified. All metabolite standards’
retention time and MRM transition were recorded in Supporting Information 3.

Jia Z., Qiu Q., He R., Zhou T, & Chen L. (2023). Identification of Metabolite Interference Is Necessary for Accurate LC-MS Targeted Metabolomics Analysis. Analytical Chemistry, 95(20), 7985-7992.

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Metabolite Profiling via QTRAP LC-MS

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The SCIEXSCIEX QTRAP 6500+ mass spectrometer and a Shimadzu LC30AD liquid chromatography system was used to analyze the supernatant. The Waters XBridge Amide (100 mm × 4.6 mm i.d., 3.5 μm) was used for LC separation. A 5μL sample was needed. The electrospray ionization mass spectra were acquired in positive ion mode (4850 V ion spray voltage) and negative ion mode (4500 V ion spray voltage), respectively. The multiple reaction monitoring (MRM) acquisition methods were used to collect MS information simultaneously. The heated capillary temperature was maintained at 475 °C. The curtain gas flow, nebulizer, and heater gas were set to 25, 33, and 33 arbitrary units, respectively.

Wang R., Wu X., Lin K., Guo S., Hou Y., Ma R., Wang Q, & Wang R. (2022). Plasma Metabolomics Reveals β-Glucan Improves Muscle Strength and Exercise Capacity in Athletes. Metabolites, 12(10), 988.

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Non-Structural Carbohydrate Profiling in Plants

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The contents of NSCs, including sucrose, glucose, fructose, and raffinose, in the leaves of allodiploid and allotetraploid plants at five developmental stages during development and flowering were measured using the hydrophilic interaction liquid-chromatography-charged aerosol detection (HILIC-CAD) method according to a previous study, with modifications [48 (link)]. Two milliliters of 80% acetonitrile solution was added to 0.5 g of ground sample, after which the mixture was fully shaken. Then, ultrasound was conducted in a low-temperature water bath for 30 min using a PS40A ultrasonic cleaner (Shen Hua Tai, Shen Zhen, China). After centrifugation at 4 °C at 12,000 × g for 5 min, the supernatant was collected. The supernatants from the two repeated procedures were collected and brought to a volume of 5 mL. One milliliter of supernatant was filtered and used to measure the NSC contents using an Ultimate 3000 HPLC (Thermo, Germering, Germany) coupled with a Corona CAD Ultra instrument (Thermo, Chelmsford, MA, USA). The column was a Waters XBridge Amide type (250 mm × 4.6 mm, 3.5 µm).

Zhang Q., Hu H., Jiang Y., Wang L., Kong X., Huang Y, & Jia G. (2022). Effects of Polyploidization on Morphology, Photosynthetic Parameters and Sucrose Metabolism in Lily. Plants, 11(16), 2112.

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Cellobiose Quantification in R. stolonifer

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An LC/MS/MS method was developed for the measurement of cellobiose produced by R. stolonifer TP-02 from glucose in vivo using a Shimadzu LCMS-8030 liquid chromatography mass spectrometer. The LC column used was an XBridge Amide (Waters). Mobile phase was 82% acetonitrile and 18% water. Measurement conditions: ESI; DL 250 °C; nebulizing gas 3 L/min; heat block 400 °C; drying gas 15 L/min.

Zhang Y., Tang B, & Du G. (2017). Self-induction system for cellulase production by cellobiose produced from glucose in Rhizopus stolonifer. Scientific Reports, 7, 10161.

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Quantification of Tranexamic Acid in Plasma

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To quantify trough TXA concentrations, we developed and validated a liquid chromatography tandem mass spectrometric assay. A 1-step protein precipitation extraction using acidified acetonitrile was optimized for preparation of plasma samples to avoid the logistical complications and analytical variability/dilution introduced from derivatization reactions commonly used to quantify TXA (eg, perchloric acid precipitation with NaOH neutralization). Chromatographic separation was achieved using Waters XBridge Amide column (3.5 μm × 2.1 mm × 50 mm). TXA and an isotopically labeled TXA (internal standard) were detected by multiple reaction monitoring using electrospray ionization in positive mode. The final validated assay was linear over the concentration range of 0.2 to 400 μg/mL (linearity [R²] = 0.9977; lower limit of detection = 0.12 μg/mL; lower limit of quantification ∼ 0.2 μg/mL). Analytical robustness was evaluated across multiple independent preparations by analyzing replicate samples (n = 6) at 4 concentrations within the linear concentration range (including lower limit of detection of 0.12 μg/mL). The intra- and interday precision values were below 10%, and accuracy was better than 4% (range, 97%-104%).

Ilich A., Gernsheimer T.B., Triulzi D.J., Herren H., Brown S.P., Holle L.A., Lucas A.T., de Laat B., El Kassar N., Wolberg A.S., May S, & Key N.S. (2022). Absence of hyperfibrinolysis may explain lack of efficacy of tranexamic acid in hypoproliferative thrombocytopenia. Blood Advances, 7(6), 900-908.

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LCMS Analysis of Organic Compounds

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Chromatographic analysis was performed on a Shimadzu SIL-20A XR system (Shimadzu, Kyoto, Japan), consisting of a binary solvent delivery system and an autosampler. Separation was performed on a Waters XBridge Amide (2.1 mm × 100 mm, 3.5 μm) column. The mobile phase was composed of water with 0.2% formic acid (A) and acetonitrile with 0.2% formic acid (B) using a gradient elution of 15% A at 0–2.5 min, 15%–50% A at 2.5–5 min, 50% A at 5–7 min, 50%–15% A at 7–8 min, 15% A at 8–11 min. The flow rate was 0.6 mL/min, and the column temperature was set at 30 °C.
Mass spectrometry detection was performed using an API5500 triple quadrupole mass (AB SCIEX, Framingham, MA, USA) equipped with an electrospray ionization (ESI) source operating in the negative ion mode. The ESI-MS spectra were acquired in the multiple reaction monitoring (MRM). The parameters in the source were set as follows: GS1 flow 55 L/min, GS2 flow 55 L/min, CUR flow 40 L/min; gas temperature 550 °C; pressure of nebulizer of MS −4500 V. All MS data were acquired using the Analyst 1.6.2 software to ensure mass accuracy and reproducibility.

Hua Y., Wang S., Chai C., Liu Z., Liu X., Zou L., Wu Q., Zhao H, & Ying Y. (2016). Quality Evaluation of Pseudostellariae Radix Based on Simultaneous Determination of Multiple Bioactive Components Combined with Grey Relational Analysis. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 22(1), 13.

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