Agilent eclipse plus c18

Manufactured by Agilent Technologies

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The Agilent Eclipse Plus C18 is a reversed-phase high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of compounds. The column features a stationary phase of high-purity, endcapped silica particles with a C18 bonded phase, providing efficient and reproducible chromatographic performance.

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Eclipse Plus C18 (181 citations) Eclipse Plus (30 citations) Agilent Eclipse Plus C18 column (24 citations) Agilent Zorbax Eclipse Plus C18 column (21 citations) Agilent Zorbax Eclipse Plus C18 (8 citations) Agilent Eclipse plus C18 RRHD (2 citations) Agilent Zorbax Eclipse Plus C18 guard column (2 citations) Agilent Eclipse Plus C18 RRHD column (2 citations)

10 protocols using agilent eclipse plus c18

HPLC Characterization of Compound with m/z 699

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Example 8

HPLC: Retention time=5.93 min (Agilent Eclipse Plus C18 Column 3.5 μm, 3×150 mm, 3.5 μm particles; Mobile Phase A: water with 20 mM ammonium acetate; Mobile Phase B: methanol. Gradient: 0% B to 50% B over 15 min, then 50% B to 95% B over 2 min; Flow: 0.5 mL/min; Detection: MS and UV (260 nm)) MS (ES): m/z=699 [M+H]⁺.

HPLC: Retention time=6.18 min (Agilent Eclipse Plus C18 Column 3.5 μm, 3×150 mm, 3.5 μm particles; Mobile Phase A: water with 20 mM ammonium acetate; Mobile Phase B: methanol. Gradient: 0% B to 50% B over 15 min, then 50% B to 95% B over 2 min; Flow: 0.5 mL/min; Detection: MS and UV (260 nm)) MS (ES): m/z=699 [M+H]⁺.

HPLC: Retention time=6.81 min (Agilent Eclipse Plus C18 Column 3.5 μm, 3×150 mm, 3.5 μm particles; Mobile Phase A: water with 20 mM ammonium acetate; Mobile Phase B: methanol. Gradient: 0% B to 50% B over 15 min, then 50% B to 95% B over 2 min; Flow: 0.5 mL/min; Detection: MS and UV (260 nm)) MS (ES): m/z=699 [M+H]⁺.

HPLC: Retention time=7.88 min (Agilent Eclipse Plus C18 Column 3.5 μm, 3×150 mm, 3.5 μm particles; Mobile Phase A: water with 20 mM ammonium acetate; Mobile Phase B: methanol. Gradient: 0% B to 50% B over 15 min, then 50% B to 95% B over 2 min; Flow: 0.5 mL/min; Detection: MS and UV (260 nm)) MS (ES): m/z=699 [M+H]⁺.

US10519187B2. Cyclic dinucleotides as anticancer agents (2019-12-31). BRISTOL-MYERS SQUIBB COMPANY [US]. Inventors: Brian E. Fink [US], Dharmpal S. Dodd [US], Steven J. Walker [US], Libing Chen [US], Yufen Zhao [US], Zheming Ruan [US], Lan-Ying Qin [US], Peter Kinam Park [US], Muthoni G. Kamau [US], Lalgudi S. Harikrishnan [US].

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HPLC Characterization of Compound with m/z 699

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Example 8

HPLC: Retention time=5.93 min (Agilent Eclipse Plus C18 Column 3.5 μm, 3×150 mm, 3.5 m particles; Mobile Phase A: water with 20 mM ammonium acetate; Mobile Phase B: methanol. Gradient: 0% B to 50% B over 15 min, then 50% B to 95% B over 2 min; Flow: 0.5 mL/min; Detection: MS and UV (260 nm)) MS (ES): m/z=699 [M+H]⁺.

HPLC: Retention time=6.18 min (Agilent Eclipse Plus C18 Column 3.5 μm, 3×150 mm, 3.5 m particles; Mobile Phase A: water with 20 mM ammonium acetate; Mobile Phase B: methanol. Gradient: 0% B to 50% B over 15 min, then 50% B to 95% B over 2 min; Flow: 0.5 mL/min; Detection: MS and UV (260 nm)) MS (ES): m/z=699 [M+H]⁺.

HPLC: Retention time=6.81 min (Agilent Eclipse Plus C18 Column 3.5 μm, 3×150 mm, 3.5 m particles; Mobile Phase A: water with 20 mM ammonium acetate; Mobile Phase B: methanol. Gradient: 0% B to 50% B over 15 min, then 50% B to 95% B over 2 min; Flow: 0.5 mL/min; Detection: MS and UV (260 nm)) MS (ES): m/z=699 [M+H]⁺.

HPLC: Retention time=7.88 min (Agilent Eclipse Plus C18 Column 3.5 μm, 3×150 mm, 3.5 m particles; Mobile Phase A: water with 20 mM ammonium acetate; Mobile Phase B: methanol. Gradient: 0% B to 50% B over 15 min, then 50% B to 95% B over 2 min; Flow: 0.5 mL/min; Detection: MS and UV (260 nm)) MS (ES): m/z=699 [M+H]⁺.

US11339185B2. Cyclic dinucleotides as anticancer agents (2022-05-24). Bristol-Myers Squibb Company [US]. Inventors: Brian E. Fink [US], Dharmpal S. Dodd [US], Steven J. Walker [US], Libing Chen [US], Yufen Zhao [US], Zheming Ruan [US], Lan-Ying Qin [US], Peter Kinam Park [US], Muthoni G. Kamau [US], Lalgudi S. Harikrishnan [US].

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Quantifying Balanol Production in Cultures

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The concentration of balanol was determined according to the method described by He et al. (2018) [10 (link)]. Culture broth was sampled for analysis of balanol by HPLC using a reverse-phase C18 column (Agilent Eclipse Plus C18, 4.6 × 250 mm, 5 μm) (1260 Infinity, Agilent Technologies, Santa Clara, CA, USA). Chromatographic conditions were composed of solvent A and solvent B. Solvent A comprised water with 0.001 M trifluoroacetic acid (TFA), and solvent B comprised acetonitrile-0.001 M TFA; the solvent gradient was 5% B in the first 5 min and increased to 58% at 35 min and to 95% B at 36 min, followed by 4 min with 95% B, with a flow rate of 1 mL/min and UV detection at 254 nm. The structure-identified balanol was used as the standard control. Through analysis, the peak area of balanol with different concentrations was determined by HPLC, and the standard curve of balanol concentration was established (Figure 1). The concentration of balanol production in culture broth was determined with the regression equation from the standard curve: Y = 31.764X − 203.51 (R² = 0.9993), where Y indicates the concentration of balanol (mg/L), and X is the peak area of balanol.

Li R.Q., Liu X., Zhang M., Xu W.Q., Li Y.Q, & Chen X.A. (2022). Gram-Level Production of Balanol through Regulatory Pathway and Medium Optimization in Herb Fungus Tolypocladium ophioglossoides. Journal of Fungi, 8(5), 510.

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Characterizing Drug Release from Hydrogel

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The release behavior of VDA and EPI in the hydrogel was investigated by High-Performance Liquid Chromatography (HPLC) and UV-Vis spectrometer. An amount of 1 mL V+E@Gel was immersed in PBS buffer (pH 7.4) at 37 °C, during which 1 mL sample was collected at specific time intervals. The concentration of VDA in the collected samples was determined by HPLC at 305 nm using Agilent 1260 Infinity II liquid chromatography system with an Agilent Eclipse Plus C18 column (250 mm × 4.6 mm, inside diameter (i.d.), particle size 5 μm) (Agilent Technologies, Santa Clara, CA, USA). The mobile phase was set as 50% A: 0.05mol/L potassium dihydrogen phosphate; 50% B: methanol + acetonitrile (v/v: 80:20) at a flow rate of 1 mL/min. The concentration of EPI in the collected samples was determined by UV-Vis spectrometer at 480 nm.

Li F., Shao X., Liu D., Jiao X., Yang X., Yang W, & Liu X. (2022). Vascular Disruptive Hydrogel Platform for Enhanced Chemotherapy and Anti-Angiogenesis through Alleviation of Immune Surveillance. Pharmaceutics, 14(9), 1809.

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Fermentation and LC-MS Analysis of Youssoufene A

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The ΔdtlA mutant strain was fermented for 50 mL in the medium (1% soluble starch, 2% glucose, 0.4% corn syrup, 1% yeast extract, 0.3% beef extract, 0.05% MgSO₄·7H₂O, 0.05% KH₂PO₄, 0.2% CaCO₃, and 3.3% sea salt, pH = 7.0) at 30 °C, 220 rpm for 7 days. The fermentation supernatant was extracted twice with an equal volume of EtOAc. The resulting EtOAc extract was subjected to HPLC-HRESIMS analysis, eluting with a linear gradient of 20–100% B/A (phase B: 100% ACN + 0.1% HCOOH; phase A: H₂O + 0.1% HCOOH; flow rate: 0.2 mL/min; wavelength: 300 nm) using an Agilent Eclipse Plus C18 (100 × 2.1 mm, 3.5 μm) (Agilent, Santa Clara, CA, USA) column to trace the youssoufene A analogs.

Liu J., Li H., Liu Z., Li T., Xiao F, & Li W. (2022). Youssoufenes A2 and A3, Antibiotic Dimeric Cinnamoyl Lipids from the ΔdtlA Mutant of a Marine-Derived Streptomyces Strain. Marine Drugs, 20(6), 394.

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UPLC-QTOF Analysis of Derivatized Nucleosides

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LC experiments were performed on Agilent 1290 UPLC (Agilent, America). Two reverse phase separation columns (Agilent Eclipse Plus C18, 100 × 2.1 mm, 3.5 μm; Agilent UPLC SB C18, 50 × 2.1 mm, 1.8 μm; Agilent Technologies) were picked to compare the separation of the derivatization products. The column temperature was set at 30°C. Water containing variable proportion of formic acid and acetic acid (v/v, solvent A) and ACN (solvent B) were investigated to confirm the mobile phase. A gradient of 15–35% ACN for 4 min and 35–40% ACN for 5 min was used. The flow rate of mobile phase was set at 0.3 mL min^-1. The injection volume was 1 μL.
High resolution mass spectrometric experiments were performed on Agilent 6550 Q-TOF mass spectrometer (Agilent, America). Agilent Data Analysis software version 5.0 was used for the data processing. The detection was performed under positive electrospray ionization (ESI) mode. The nucleosides and derivatives were monitored by MS analysis mode. Solutions were infused from the ESI source at 0.3 mL min^-1 with parameters: capillary 4000 V, drying gas 12 L min^-1, drying gas temperature 280°C, Sheath gas temperature 400°C. Nitrogen was used as the nebulizing and drying gas. All MS conditions were optimized to achieve maximal detection sensitivity.

Guo M., Li X., Zhang L., Liu D., Du W., Yin D., Lyu N., Zhao G., Guo C, & Tang D. (2017). Accurate quantification of 5-Methylcytosine, 5-Hydroxymethylcytosine, 5-Formylcytosine, and 5-Carboxylcytosine in genomic DNA from breast cancer by chemical derivatization coupled with ultra performance liquid chromatography- electrospray quadrupole time of flight mass spectrometry analysis. Oncotarget, 8(53), 91248-91257.

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Stability Analysis of ABN401 by HPLC

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An HPLC system (Shimadzu LC-20, Shimadzu, Kyoto, Japan) was used to analyze the solubility, dissolution profile, and degradation products as well as to determine the storage stability of ABN401. The wavelength of the UV detector was set at 282 nm. For quantification analysis, the Agilent Eclipse Plus C18 (5 μm, 4.6 × 150 mm) (Agilent technologies, Santa Clara, CA, USA) was used and maintained at 30 °C. The mobile phase was a mixture of acetonitrile and 50 mM acetate buffer at pH 5.0 at a volume ratio of 50 to 50 (v/v %). The flow rate of the mobile phase was 0.5 mL/min and the injection volume was 10 μL. A Kromasil C8 (5 μm, 4.6 × 250 mm) column was used for qualification analysis to observe the impurity profile during the storage stability test. The following gradient was applied: 0–20 min, 80% acetonitrile and 20% acetate buffer to 20% acetonitrile and 80% acetate buffer, maintained up to 25 min; 25– 28 min, back to 80% acetonitrile and 20% acetate buffer, maintained up to 35 min. The flow rate of the mobile phase was 1 mL/min and the injection volume was 10 μL.

Kim N.A., Hong S., Kim K.H., Choi D.H., Kim J.S., Park K.E., Choi J.Y., Shin Y.K, & Jeong S.H. (2020). New Preclinical Development of a c-Met Inhibitor and Its Combined Anti-Tumor Effect in c-Met-Amplified NSCLC. Pharmaceutics, 12(2), 121.

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Glycan Mapping by HPLC-Chip Q-TOF

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The glycan mapping was performed on an Agilent 1200 series HPLC-Chip system coupled to an Agilent 6520 Q-TOF (Agilent Technologies, Santa Clara, CA). Agilent Zorbax C18 stationary phase (300 Å, 5 μm) and porous graphitized carbon (250 Å, 5 μm) stationary phase were used to separate the trypsin digest and pronase digest, respectively. The microfluidic C18 chip consists of two columns: one for enrichment (4 mm, 40 nL) and one for separation (150 mm × 75 μm). The microfluidic PGC chip also consists of one enrichment column (4 mm, 40 nL) and one separation column (43 mm × 75 μm).
The quantitative analyses were performed on an Agilent 1290 infinity LC system coupled to an Agilent 6490 triple quadrupole (QqQ) mass spectrometer (Agilent Technologies, Santa Clara, CA). An Agilent Eclipse plus C18 (RRHD 1.8 μm, 2.1 × 100 mm) coupled with an Agilent Eclipse plus C18 pre-column (RRHD 1.8 μm, 2.1 × 5 mm) was used for UPLC separation.

Hong Q., Ruhaak L.R., Stroble C., Parker E., Huang J., Maverakis E, & Lebrilla C.B. (2015). A method for comprehensive glycosite-mapping and direct quantitation of plasma glycoproteins. Journal of proteome research, 14(12), 5179-5192.

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In Vitro Methyltransferase Assay for Alkaloid Detection

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The in vitro methyltransferase reactions were performed as previously described50 (link) at 50 μl scale containing 100 mM Gly-NaOH buffer (pH 9.0) in the presence of 25 mM sodium ascorbate, 2 mM DTT, 10% glycerol, 300 μM S-Adenosyl methionine, 10 μM purified 9 and 50 μg purified enzyme or no enzyme as a control. The reactions were performed overnight and quenched with 150 μl of 99.9% MeOH/0.1% formic acid. The quenched reaction mixture was analysed on an Agilent 6420 triple quad LC–MS (Agilent EclipsePlus C18, 2.1 × 50 mm, 1.8 μm) using positive ionization. Noscapine and narcotoline were detected with MRM using the highest characteristic precursor ion/product ion transition (Supplementary Table 8).

Li Y, & Smolke C.D. (2016). Engineering biosynthesis of the anticancer alkaloid noscapine in yeast. Nature Communications, 7, 12137.

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E. coli Metabolite Extraction and LC-MS Analysis

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For production analysis, E. coli BL21I(DE3)/pHW4 was grown and induced as described above. The pellet was extracted with methanol and the supernatant with an equal volume of ethyl acetate three times. The extracts were combined and analyzed on an Agilent 1200 HPLC instrument (Agilent Eclipse Plus C18, 5 μm, 4.6 mm × 250 mm) connected with an Agilent 6130 Single Quadrupole mass spectrometer, eluted with a linear gradient of 75 to 90 % (v/v) acetonitrile-water (containing 0.1% trifluoroacetic acid) over 45 min at a flow rate of 1 mL/min. The products were detected at 210 nm.

Yan H., Sun L., Huang J., Qiu Y., Xu F., Yan R., Zhu D., Wang W, & Zhan J. (2018). Identification and heterologous reconstitution of a 5-alk(en)ylresorcinol synthase from endophytic fungus Shiraia sp. Slf14. Journal of microbiology (Seoul, Korea), 56(11).

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