Sequant zic hilic

Manufactured by Merck Group

Sourced in Germany, United Kingdom

The SeQuant ZIC-HILIC is a hydrophilic interaction liquid chromatography (HILIC) column for the separation and analysis of polar and hydrophilic analytes. It utilizes a zwitterionic stationary phase to provide high selectivity and resolution for a wide range of polar compounds.

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Lab products found in correlation

4 hydroxyproline, by Avantor (2 mentions) Api 4000, by AB Sciex (2 mentions) Xcalibur quan software, by Thermo Fisher Scientific (2 mentions) Q exactive orbitrap, by Thermo Fisher Scientific (2 mentions) Agilent 1260 infinity, by Agilent Technologies (1 mentions) Tmao d9, by LGC (1 mentions) 4000 qtrap, by AB Sciex (1 mentions) Agilent 1200 hplc system, by Agilent Technologies (1 mentions) Xevo tq xs, by Waters Corporation (1 mentions) Acquity uplc, by Waters Corporation (1 mentions) Agilent 1290 infinity 2 lc system, by Agilent Technologies (1 mentions) Acquity uplc glycan beh amide column, by Waters Corporation (1 mentions) Maxis q tof mass spectrometer, by Bruker (1 mentions) Q exactive plus, by Thermo Fisher Scientific (1 mentions) Tsq quantum ultra, by Thermo Fisher Scientific (1 mentions) Prominence lc system, by Shimadzu (1 mentions) Htc pal autosampler, by CTC Analytics (1 mentions) 1200 series hplc, by Agilent Technologies (1 mentions) Lc 10ad vp instrument, by Shimadzu (1 mentions) Uv spd m20a diode array detector, by Shimadzu (1 mentions)

16 protocols using sequant zic hilic

Quantification of Uremic Toxins by LC-MS/MS

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Reference standards, i.e., ADMA, SDMA, and TMAO, as well as internal standards ADMA-d6 and TMAO-d9 were purchased from Toronto Research Chemicals (Toronto, Canada).
Uremic toxins were determined with validated high-performance liquid chromatography coupled to mass spectrometry (LC-MS/MS) using multiple-reaction-monitoring (MRM) mode on Agilent 1260 Infinity (Agilent Technologies, Santa Clara, CA, USA) coupled to QTRAP 4000 (AB Sciex, Framingham, MA, USA). MRM transitions, declustering potential (DP), and collision energy (CE) were: ADMA, m/z 203 > 46 (DP = 61V, CE = 41V); SDMA, m/z 203 > 172 (DP = 61V, CE = 19V); ADMA-d6, m/z 209 > 77 (DP = 66V, CE = 45V); TMAO, m/z 76 > 42 (DP = 66V, CE = 53V); and TMAO-d9, m/z 85 > 46 (DP = 61V, CE = 59V), respectively. Chromatographic separation was achieved using a SeQuant^® ZIC^®-HILIC (50 × 2.1 mm, 5 μm, Merck (Darmstadt, Germany)) column. The column was maintained at 25 °C at a flow rate of 0.5 mL min⁻¹. The mobile phases consisted of 20 mM ammonium acetate as eluent A and acetonitrile with 0.2% formic acid as eluent B. The gradient (%B) was as follows: 0 min, 95%; 1 min, 95%; 7 min, 50%; and 8 min, 50%. The injection volume was 5 μL. Urine samples (0.1 mL) prior to injection with LC were mixed with internal standards (0.1 mL, 6 μg/mL) and acetonitrile (0.6 mL), vortexed at high speed (3 min), and centrifuged (5 min at 10,000 g).

Osredkar J., Baškovič B.Ž., Finderle P., Bobrowska-Korczak B., Gątarek P., Rosiak A., Giebułtowicz J., Vrhovšek M.J, & Kałużna-Czaplińska J. (2023). Relationship between Excreted Uremic Toxins and Degree of Disorder of Children with ASD. International Journal of Molecular Sciences, 24(8), 7078.

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HPLC-DAD Analysis of Pharmaceutical Compound

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Chromatographic analyses were performed using the Agilent 1200 HPLC system (Agilent Technologies, Santa Clara, California, USA) equipped with ChemStation revision B.04.01, Degasser G1379B, Binary Pump G1312A, Autosampler (ALS) G1329A, Thermostatted Column Compartment (TCC) G1316A and Diode-Array Detector (DAD) G1315D. Merck SeQuant® ZIC®-HILIC (3.5 µm, 200 Å) 150 × 4.6 mm column was used for separation. The column temperature was set at 25°C. The mobile phase that consisted of water and acetonitrile (40/60, v/v) adjusted to pH 4.8 with ammonium acetate buffer (20 mM) was used for isocratic elution at a flow rate of 1 ml/min. The injection volume was 1 μl. The eluent was monitored at 267 nm for drug detection over a period of 7 min. The pH measurement of the CaLev was performed with the pH meter XS Instruments pH 7+ DHS.

Sanogo S., Silimbani P., Gaggeri R, & Masini C. (2020). Stability of calcium levofolinate reconstituted in syringes and diluted in NaCl 0.9% and glucose 5% polyolefin/polyamide infusion bags. Journal of Oncology Pharmacy Practice, 27(2), 288-296.

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HILIC-MS/MS Protocol for Metabolite Quantification

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LC-MS/MS: LC-MS/MS was performed on ACQUITY UPLC (Waters, Milford, MA) with SeQuant ZIC-HILIC, 150 × 2.1 mm, 3.5 μm, 100 Å analytical column (Merck, Kenilworth, NJ) and Xevo TQ-XS (Waters, Milford, MA) equipped with electrospray ionization source. Column oven was set to 40 °C and an autosampler to 20 °C. The injection volume was 1 µl. Elution was performed using mobile phases A (20 mM Ammonium acetate buffer pH 5) and B (100% acetonitrile (ACN)). Chromatographic and MS parameters are described in Tables 123.
Table 1. Chromatographic conditions.
Loading [Contrib]/a11y/accessibility-menu.js Download CSVDisplay Table Table 2. Specific mass spectrometric parameters and retention times (RT) for the analytes and internal standards (IS). Download CSVDisplay Table Table 3. General mass spectrometric parameters.

Svistounov D., Solbu M.D., Jenssen T.G., Mathisen U.D., Hansen T., Elgstøen K.B.P, & Zykova S.N. (2022). Development of quantitative assay for simultaneous measurement of purine metabolites and creatinine in biobanked urine by liquid chromatography-tandem mass spectrometry. Scandinavian journal of clinical and laboratory investigation, 82(1).

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HILIC-Based Glycopeptide Separation and Analysis

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The measurements were performed on an Agilent 1290 Infinity II LC System with a binary pump (Agilent Technologies, Inc., Waldbronn, Germany) coupled with maXis™ Q-TOF mass spectrometer (Bruker Daltonics, Bremen, Germany). The glycopeptide separation was tested in three different HILIC columns: HALO^® penta-HILIC (2.1 × 150 mm; 2.7 µm; Advanced Materials Technology, Wilmington, Delaware, USA), ACQUITY UPLC Glycan BEH Amide Column (2.1 × 150 mm; 1.7 µm; Waters Corporation, Milford, MA, USA) and SeQuant ZIC-HILIC (2.1 × 150 mm; 3.5 µm; Merck, Darmstadt, Germany). The mobile phase consisted of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The gradient program was optimized to enhance, as much as possible, the resolution of all glycopeptides analyzed in this study. In the case of HALO^® penta-HILIC and ACQUITY UPLC Glycan BEH Amide, the following gradient ((min)/% B) was used: 0/80, 10/80, 25/65, 35/40, 40/40, 43/80, 55/80. For the SeQuant ZIC-HILIC column, a shallower gradient program was used: 0/80, 10/80, 35/65, 45/40, 55/40, 58/40, 70/80. The flow rate was 0.3 mL min⁻¹ for every column, and the column temperature was maintained at 40 °C (if not stated otherwise).

Molnarova K, & Kozlík P. (2020). Comparison of Different HILIC Stationary Phases in the Separation of Hemopexin and Immunoglobulin G Glycopeptides and Their Isomers. Molecules, 25(20), 4655.

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Plasma Metabolomics Analysis Protocol

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Metabolomics analyses were performed by Profilomic SA (Huningue, France) on plasma from blood collected at the end of the first experiment (i.e., 9 weeks after surgery—Figure 1). In brief, 50 μL of plasma were homogenized in 200 μL methanol cooled at −20 °C. The samples were vortexed for 10 s and then incubated on ice for 90 min. After centrifugation (20,000 g, 10 min, 4 °C), precipitates were dried using a stream of nitrogen and then solubilized in 150 μL ammonium carbonate (10 mM, pH 10.5/can 40:60, v/v), before storage at −80 °C until the LC-MS analysis. The metabolomic profile of the samples was determined using a high-performance liquid chromatograph adapted to polar compounds (SeQuant ZIC-HILIC, Merck, Germany) coupled to a high-resolution mass spectrometer (Q-Exactive Plus, Thermo Fisher Scientific, Illkirsh, France). The mass spectra were collected using a high resolving power (70,000 Full Width at Half Maximum, FWHM) by alternating positive and negative ionization modes. The identification of metabolites in humans and mice was performed using their respective mass and retention time according to the Cribiom and Profilomic databases, respectively.

Bernard A., Le May C., Dastugue A., Ayer A., Blanchard C., Martin J.C., Pais de Barros J.P., Delaby P., Le Bourgot C., Ledoux S, & Besnard P. (2021). The Tryptophan/Kynurenine Pathway: A Novel Cross-Talk between Nutritional Obesity, Bariatric Surgery and Taste of Fat. Nutrients, 13(4), 1366.

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Quantitative LC-MS/MS Analysis of Bilirubin

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Quantitative analysis of IS using LC-MS/MS was performed using a Prominence LC system (Shimadzu) coupled to a TSQ Quantum-Ultra (Thermo Fisher Scientific, San Jose, CA, USA) and operated in the negative mode. Each sample (4 μL) was injected onto a 100 × 2.0 mm SeQuant ZIC-HILIC (Merck Schuchardt) with a flow rate of 0.4 mL/min. For gradient elution, mobile phase A was H₂O/acetonitrile/HCOOH = 98/2/0.1 and mobile phase B was H₂O/acetonitrile/HCOOH = 2/98/0.1. Linear and stepwise gradients were programmed as follows: 0–1 min: 10% solvent B; 1.1–2 min: 10–55% solvent B; 2.1–4 min: 55–90% solvent B; 4.1–6 min: 90–100% solvent B; 6.1–11 min: 100% solvent B; 11.1–15 min: 10% solvent B. Bilirubin and biliverdin-d₄ were detected in selected reaction monitoring mode by monitoring the transitions of m/z 212 to 80 and 216 to 80, respectively. Spray voltage was 3 500 V, vaporizer temperature was 275 °C, and ion transfer tube temperature was 350 °C. To validate each assay, the followings were performed. Intra and inter assay coefficients of precision were 4.03% and 0.049% at 0.2 μg/mL, respectively, and intra and inter assay coefficients of accuracy were 8.62% and 3.1% at 0.2 μg/mL, respectively. Calibration curve was prepared in every batch, and the correlation coefficient was over 0.999.

Sato E., Mori T., Mishima E., Suzuki A., Sugawara S., Kurasawa N., Saigusa D., Miura D., Morikawa-Ichinose T., Saito R., Oba-Yabana I., Oe Y., Kisu K., Naganuma E., Koizumi K., Mokudai T., Niwano Y., Kudo T., Suzuki C., Takahashi N., Sato H., Abe T., Niwa T, & Ito S. (2016). Metabolic alterations by indoxyl sulfate in skeletal muscle induce uremic sarcopenia in chronic kidney disease. Scientific Reports, 6, 36618.

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Quantifying GAG and Hydroxyproline in Samples

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GAG was measured with the DMMB (dimethyl methylene blue) assay as previously described.4 The HPro measurement was performed using a high‐performance liquid chromatography‐mass spectrometry/mass spectrometry (HPLC‐MS/MS) assay, using 4‐hydroxyproline (VWR International, Radnor, PA) 0.1–50.0 µg/ml as calibration standards. Then, 5 µl of the samples were mixed with 10 µl of internal standard (1.2 µg/ml 4‐hydroxyproline [²H₃] from C/D/N‐Isotopes) and 200 µl of 25% (v/v) hydrochloric acid and hydrolyzed overnight at approximately 110°C. After centrifugation, samples were evaporated at 55°C/10 Torr, resuspended in 1 ml water and mixed 1:5 in acetonitrile. The HPLC separation (1200 Series HPLC, Agilent, Santa Clara, CA; HTC PAL Autosampler, CTC Analytics, Zwingen Switzerland) was achieved on a hydrophilic interaction chromatography column (Sequant ZIC‐HILIC, 50‐2.1 mM, 3.5 µM, 200 Å column; Merck KGaA) using a mobile phase gradient (eluent A: 0.1% [v/v] formic acid in water, eluent B: acetonitrile). Detection was performed on a tandem MS (API4000; Sciex, Darmstadt, Germany) with a turbo ion spray interface operating in positive ion mode.

Müller S., Lindemann S, & Gigout A. (2019). Effects of Sprifermin, IGF1, IGF2, BMP7, or CNP on Bovine Chondrocytes in Monolayer and 3D Culture. Journal of Orthopaedic Research, 38(3), 653-662.

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HPLC Analysis of AFA Components

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HPLC analyses were performed with a HPLC Shimadzu LC-10AD VP instrument (Tokyo, Japan) equipped with a binary pump LC-10AD VP, a UV SPD-M20A Diode Array detector, a 20 μL injector and a computer integrating apparatus (EZ Start 7.4 software, Shimadzu Scientific Instruments, Inc., Columbia, MD, USA). Chromatographic separation was achieved on a reversed-phase column SeQuant^® Zic^®-Hilic, (5 μm, 200 Å, 150 × 2.1 mm, Merck, Germany), a mobile phase consisted of acetate buffer 5 mM pH 6.5 (A) and Acetonitrile (B). For separation of AFA components the gradient method was developed as follows: A:B (0.5:99.5→0.01–5.00 min, 40:60→5.00–11.00 min; 40:60→11.00–30.00 min). The flow rate was set at 0.3 mL/min, the UV wavelength range 200–700 nm and set at 260, 334, 407, 665 nm to identification.

De Caro V., Murgia D., Seidita F., Bologna E., Alotta G., Zingales M, & Campisi G. (2019). Enhanced In Situ Availability of Aphanizomenon Flos-Aquae Constituents Entrapped in Buccal Films for the Treatment of Oxidative Stress-Related Oral Diseases: Biomechanical Characterization and In Vitro/Ex Vivo Evaluation. Pharmaceutics, 11(1), 35.

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Quantification of 4-Hydroxyproline by HPLC-MS/MS

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The HPro concentration was determined by using high‐performance liquid chromatography‐mass spectrometry/mass spectrometry (HPLC‐MS/MS) assays. 4‐hydroxyproline (0.1‐200 µg/mL, diluted in medium; VWR) was used as calibration standard. A mixture of 5 µL sample, 10 µL 4‐hydroxyproline (1.2 µg/mL) and 200 µL HCL (25%) was hydrolysed overnight at 110°C. The mixture was centrifuged, evaporated at 55°C/10 Torr, resuspended in 1 mL MilliQ‐water and mixed 1/5 in acetonitrile (Merck) for injection into the HPLC‐MS/MS system. High‐performance liquid chromatography separation was performed on a hydrophilic interaction chromatography (HILIC) column (Sequant ZIC‐HILIC, 50‐2.1 mM, 3.5 µM, 200 Å column; Merck) with a mobile phase gradient (eluent A: 0.1% formic acid from Merck, eluent B: acetonitrile from Merck). Detection was performed with a Tandem MS (API4000, Sciex) with a turbo ion spray interface operating in positive ion mode.

Mang T., Kleinschmidt‐Dörr K., Ploeger F., Lindemann S, & Gigout A. (2020). The GDF‐5 mutant M1673 exerts robust anabolic and anti‐catabolic effects in chondrocytes. Journal of Cellular and Molecular Medicine, 24(13), 7141-7150.

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Zwitterion HILIC Separation and Detection

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A chromatographic separation was performed using a Zwitterion hydrophilic interaction liquid chromatography (ZIC-HILIC) column with a 200-Å pore size (SeQuant ZIC-HILIC, 150 mm × 4.6 mm × 5 μm, Merck Millipore, VIC, Australia).
For Corona CAD, nebuliser temperature was set at 25°C. Nitrogen gas was used as carrier gas at 2 L/minute flow rate to evaporate mobile phase and to produce particles of analytes. The data collection rate, power function and filter time constant were kept at 100 Hertz, 1 and 3 seconds, respectively. The temperature of the sample compartment was set at 10°C and the column compartment was either 25°C or 40°C. The injection volume was 5 μL.

Asthana C., Peterson G.M., Shastri M, & Patel R.P. (2019). Development and validation of a novel high performance liquid chromatography-coupled with Corona charged aerosol detector method for quantification of glucosamine in dietary supplements. PLoS ONE, 14(5), e0216039.

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