Zorbax sb aq column

Manufactured by Agilent Technologies

Sourced in United States, Japan

The ZORBAX SB-Aq column is a reversed-phase high-performance liquid chromatography (HPLC) column designed for the separation of a wide range of polar and hydrophilic compounds. It features a silica-based stationary phase with an embedded polar group that enhances the retention of polar analytes.

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Zorbax SB-Aq C18 column (30 citations) ZORBAX SB-Aq (24 citations) SB-Aq column (20 citations) SB-Aq (11 citations) Agilent Zorbax-SB-Aq column (3 citations) Zorbax SB-Aq analytical column (3 citations) Zorbax SB-Aq C19 column (2 citations) ZORBAX SB-Aq C18 organic acid column (2 citations)

58 protocols using zorbax sb aq column

Quantification of INH and RIF in Skin and Blood

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Ten microliters of the supernatant from local skin tissues or blood samples was analyzed by HPLC (Agilent 1200, Palo Alto, CA) using Agilent ZORBAX SB-Aq column (4.6 mm × 100 mm, 5 μm) guarded by a ZORBAX SB-Aq column (2.1 mm × 12.5 mm, 5 μm) and detected by a UV2000 ultraviolet detector set at a wavelength of 264 nm for INH and 340 nm for RIF. For INH analysis, the mobile phase consisted of 0.02 M heptanesulfonic acid sodium salt: methanol: acetonitrile (78:17:5, v/v/v) and the column was kept at 25 °C. For RIF analysis, the mobile phase consisted of 0.05 M phosphate buffer: methanol (30:70, v/v) and the column was kept at 25 °C. Standard curves were constructed by dissolving known concentration of INH (1–100 μg/mL) or RIF (2–600 μg/mL) into blank skin homogenates or blank plasma and treating them similar to the experimental samples. Six different concentrations were used for developing a standard curve. The concentration of INH and RIF in each tested sample was calculated on the basis of standard curve. Both of the detection limits of INH and RIF in skin sample or blood sample were 1 μg/mL.

Chen S., Han Y., Yu D., Huo F., Wang F., Li Y., Dong L., Liu Z, & Huang H. (2017). Transdermal delivery of isoniazid and rifampin in guinea pigs by electro-phonophoresis. Drug Delivery, 24(1), 467-470.

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Comprehensive Spectroscopic Characterization

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The NMR spectra were obtained on a Varian 500 spectrometer, with deuterated solvent as reference. The optical rotations were determined using an Autopol^® IV Automatic polarimeter. The FT-IR spectra were measured using a Nicolet NEXUS-470 infrared spectrometer. HR-ESI-MS spectra were recorded on a Xevo-G2 Q-TOF mass spectrometer with an electrospray ionization (ESI) interface (Waters, Milford, MA, USA) in the negative mode. Analytical HPLC was performed on an Agilent 1100 HPLC system, equipped with a diode array detector and an Agilent ZORBAX SB-Aq column (250 mm×4.6 mm, 5 μm). Semi-preparative HPLC was carried out on an Agilent 1200 instrument, using an Agilent ZORBAX SB-Aq column (250 mm×10 mm, 5 μm), detected at a UV wavelength of 230 nm. Column chromatography (CC) was performed on macroporous resin AB-8 (Cangzhou Bon Adsorber Technology Co.), silica gel (100–200 mesh or 200–300 mesh, Qingdao Haiyang Chemical Works, China), Sephadex LH-20 (Pharmacia Co.), and ODS (Merck). Analytical grade solvents were purchased from Beijing Chemical Factory.

Ma X., Guo X., Zhao M., Tu P, & Jiang Y. (2016). Four new phenolic glycosides from Baoyuan decoction. Acta Pharmaceutica Sinica. B, 7(2), 173-178.

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Foliar Hormone Quantification Protocol

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The foliar hormone was measured according to Guo et al. (2017) (link) with some modification. Approximately 300 mg of plant tissue was hand ground in liquid nitrogen and was quickly homogenized in 0.5 mL extraction buffer for 30 min at 4°C with gentle agitation on a shaker. Subsequently, each sample was additionally added 1 mL of CH₂Cl₂, and then agitated for 30 min on a shaker at 4°C. The homogenized sample was centrifuged at 13,000 g for 10 min. After centrifugation, the lower layer was collected, and then was concentrated in a dry machine. The concentrated sample was re-solubilized in 200 μL of MeOH. Next, 1 μL of the sample was injected into an Agilent ZORBAX SB-Aq column (600 bar, 2.1 mm × 100 mm, 1.8 μm) for hormone analysis. The hormone contents were calculated with reference to standard curves.

Guo H., Sun Y., Yan H., Li C, & Ge F. (2018). O3-Induced Leaf Senescence in Tomato Plants Is Ethylene Signaling-Dependent and Enhances the Population Abundance of Bemisia tabaci. Frontiers in Plant Science, 9, 764.

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Synthesis and Characterization of HBV Inhibitors

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All the organic solvents used for the synthesis were of analytical grade. Unless otherwise stated, all chemicals were purchased from Aladdin (Shanghai, China). Melting points were measured on a WRX-4 micro melting point instrument (Shanghai YiCe Equipment, China) and were uncorrected. ¹H NMR and ¹³C NMR spectra were recorded on BRUKER DPX-400 and DPX-600 spectrometers (Bruker Company, Germany), using TMS as an internal standard and CDCl3, MeOD-d4, Pyridine-d5 and DMSO-d6 as solvents. Chemical shifts (δ values) and coupling constants (J values) are given in ppm and Hz, respectively. All the solvents and reactants were of analytical grade and were used without further purification unless noted. Column chromatography was accomplished on silica gel (100–200 or 200–300 mesh, Qingdao, China). The purity of compounds was confirmed to be over 95% and was determined by high performance liquid chromatography (HPLC) with a ZORBAX SB-Aq column (250 mm × 4.6 mm, 5 μm, Agilent) using ACN/water (10%, v/v) as the mobile phase (1.0 mL/min).HepG2.2.15 cells were provided by Beijing 302 Hospital and presented by Prof.Su of Guilin Medical College. HepG2.2.15 cells are hepatoma cell lines capable of expressing HBV antigen and secreting complete HBV particles.

Ye Z., Zhao T.S., Li S.B., Zhou X.L., Luo Q., Qin J.K., Liang C.Q., Wang P, & Ge G.B. (2023). Synthesis and biological evaluation of esculetin derivatives as potential anti-HBV agents. Medicinal Chemistry Research, 32(5), 899-909.

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HPLC Quantification of Phenolic Compounds in Barley

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The individual phenolic compounds in the above extracts were quantified using a Shimazu LC-20AD HPLC system. A ZORBAX SB-Aq column (5 m, 4.6 mm × 250 mm ID, Agilent) was used at a column temperature of 30°C. The mobile phase consisted of a 0.1% aqueous solution of formic acid (solution A) and acetonitrile (solution B) with the following gradient program: 0–10 min, solution B 3–3%; 10–20 min, solution B 3–80%; 20–30 min, solution B 80–3%; 30–35 min, solution B 3–3%. Other chromatographic conditions included a constant flow rate of 0.8 ml/min, an injection volume of 20 μL, a run time of 35 min, and a detection wavelength of 254 and 320 nm. Prior to analysis, all samples were filtered through a 0.25–l m membrane filter. The identification of each peak was based on the retention time and the chromatography of authentic standards. The concentrations of each compound were calculated according to a standard curve, and the results were expressed as μg/g DW of barley powder.

Xiao X., Li J., Xiong H., Tui W., Zhu Y, & Zhang J. (2022). Effect of Extrusion or Fermentation on Physicochemical and Digestive Properties of Barley Powder. Frontiers in Nutrition, 8, 794355.

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HPLC Analysis of Organic Compounds

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Quantitative analysis was performed using an Agilent 1100 system (Agilent, USA). Chromatographic separations were carried out on a Zorbax SB-Aq column (250 mm × 4.6 mm, 5 μm; Agilent, USA), maintained at 35 °C. The mobile phases were acetonitrile (A) and 0.1% (v/v) phosphoric acid (B) (with a gradient elution program of 0–15 min, 12% A; 15–25 min, 13–14% A; 25–30 min, 14% A; 30–40 min, 14%–12% A), at a flow rate of 0.8 mL/min. The injection volume was 5–20 μL each time. The detection wavelength was set to 340 nm.

Wang Y., Liang Z., Liao X., Zhou C., Xie Z., Zhu S., Wei G, & Huang Y. (2019). Identification of C-glycosyl flavones by high performance liquid chromatography electrospray ionization mass spectrometry and quantification of five main C-glycosyl flavones in Flickingeria fimbriata. BMC Chemistry, 13(1), 94.

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Comprehensive Metabolic Profiling Using HILIC and RPLC

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Metabolic extracts were analyzed four times using hydrophilic liquid chromatography (HILIC) and reverse phase liquid chromatography (RPLC) separation in both positive and negative ionization modes as previously described^{124 (link)}. Data were acquired on a Thermo Q Exactive plus mass spectrometer equipped with a HESI-II probe and operated in full MS scan mode. MS/MS data were acquired on pool samples consisting of an equimolar mixture of all the samples in the study. HILIC experiments were performed using a ZIC-HILIC column 2.1×100 mm, 3.5μm, 200Å (Merck Millipore) and mobile phase solvents consisting of 10mM ammonium acetate in 50/50 acetonitrile/water (A) and 10 mM ammonium acetate in 95/5 acetonitrile/water (B). RPLC experiments were performed using a Zorbax SBaq column 2.1 × 50 mm, 1.7 μm, 100Å (Agilent Technologies) and mobile phase solvents consisting of 0.06% acetic acid in water (A) and 0.06% acetic acid in methanol (B). Data quality was ensured by (i) injecting 6 and 12-pool samples to equilibrate the LC-MS system prior to run the sequence for RPLC and HILIC, respectively, (ii) sample randomization for metabolite extraction and data acquisition, and (iii) checking mass accuracy, retention time and peak shape of internal standards in every samples.

Lu R.J., Taylor S., Contrepois K., Kim M., Bravo J.I., Ellenberger M., Sampathkumar N.K, & Benayoun B.A. (2021). Multi-omic profiling of primary mouse neutrophils predicts a pattern of sex and age-related functional regulation. Nature aging, 1(8), 715-733.

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Metabolomic Profiling of Serum and Cecal Samples

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A well-developed metabolomic platform based on the GC/TOF-MS technique was used to profile the metabolites in the serum samples from each group, as previously reported, with a few modifications ^{26 (link)}. The amount of serum was reduced to 30 μL and extracted with 120 μL of methanol. The supernatants were dried and derivatized for GC/TOF-MS analysis. For extraction and analysis of metabolites in cecal content, cecal content (0.5 g) were added with 1000 μL purified water and vigorously vortexed for 3 min (1:2, w). An aliquots of 50 μL mixture was transferred to an Eppendorf tube, and then 200μL methanol (containing internal standard [¹³C₂]-myristic acid 5μg/mL) was added and vortexed for 3 min. An aliquot of 100 μL supernatant was dried and derivatized for GC/TOF-MS analysis in the same way as that of serum samples. The raw data acquired with GC/TOF-MS were processed, and the metabolites were identified as previously reported ^{26 (link)}. The multivariate data were evaluated using the SIMCA P-13 software (Umetrics, Umeå, Sweden). Bile acids, such as cholic acid(CA), glycocholic acid (GCA), taurocholic acid (TCA), deoxycholic acid(DCA), ursodeoxycholic acid(UDCA), lithocholic acid(LCA) and chenodeoxycholic acid (CDCA) were quantitative analysed in Shimadzu LC/MS 2010, equipped with an Aglient ZORBAX SB-Aq column (2.1 × 150 mm, 3.5 μm), Method S1.

Gu S., Cao B., Sun R., Tang Y., Paletta J.L., Wu X.L., Liu L., Zha W., Zhao C., Li Y., Radlon J.M., Hylemon P.B., Zhou H., Aa J, & Wang G. (2014). Metabolomic and pharmacokinetic study on the mechanism underlying the lipid-lowering effect of oral-administrated berberine. Molecular bioSystems, 11(2), 463-474.

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Quantifying mRNA m6A Modification

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Nuclease P1 (1 U; Sigma) in 20 μl of buffer, which contained 10 mM of NH₄Ac (pH 5.3), was used to digest mRNA at 42 °C for 4 h. Then, 100 mM NH₄HCO₃ and alkaline phosphatase (0.5 U) were then added to about 50 to 100 ng purified mRNA for another incubation at 37 °C for 4 h. Next, the supernatant of digested sample was collected by centrifugation (4 °C, 13,000 rpm, 20 min) and then injected into Ultraperformance liquid chromatography-MS/MS. Ultraperformance liquid chromatography (SHIMADZU) equipped with ZORBAX SB-Aq column (Agilent) was used to separate the nucleosides. Then, Triple Quad 5500 (AB SCIEX) in positive ion multiple reaction-monitoring mode was used to detect the nucleosides. According to nucleoside-to-base ion mass transitions, the modifications were quantified: m/z 268.0 to 136.0 for A and m/z 282.0 to 150.1 for m6A. Pure nucleosides were used to generate standard curves. Then, the concentrations of A and m6A in the sample were calculated. Finally, the percentage of total unmodified A represents the level of m6A.

Xie F., Zhu X., Liu X., Chen H, & Wang J. (2022). N6-methyladenosine (m6A) RNA methylation mediated by methyltransferase complex subunit WTAP regulates amelogenesis. The Journal of Biological Chemistry, 298(12), 102715.

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Quantitative Analysis of CHR and ANT

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Cultured broth was centrifuged at 15,000 ×g for 10 min. The supernatant was filtered using a Nylaflo nylon membrane filter and stored at -20°C. The concentrations of CHR and ANT were determined by high-performance liquid chromatography (HPLC) using a Zorbax SB-Aq column (4.6 x 250 mm, Agilent, USA). The mobile phase was 0.1% trifluoroacetic acid (TFA) in 40% MeOH and the flow rate was 0.5 ml/min. CHR and ANT were detected at 273 nm and 330 nm, respectively.

Kim H.J., Choi S.S, & Kim E.S. (2023). CRISPR-Driven Genome Engineering for Chorismate- and Anthranilate-Accumulating Corynebacterium Cell Factories. Journal of Microbiology and Biotechnology, 33(10), 1370-1375.

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