C18 reverse phase hplc column

Manufactured by Agilent Technologies

Sourced in United States

The C18 reverse phase HPLC column is a commonly used analytical tool in High-Performance Liquid Chromatography (HPLC) applications. It is designed to separate and analyze a wide range of organic compounds based on their hydrophobic interactions with the stationary phase.

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Lab products found in correlation

Ctc pal chilled autosampler, by Agilent Technologies (2 mentions) 1200 hplc, by Agilent Technologies (2 mentions) Masshunter software, by Agilent Technologies (2 mentions) High performance liquid chromatography (hplc), by Shimadzu (1 mentions) Hplc ms ms system, by Agilent Technologies (1 mentions) 6410 triple quad, by Agilent Technologies (1 mentions) 6410b triple quadrupole mass spectrometer, by Agilent Technologies (1 mentions) Agilent 1200 series hplc, by Agilent Technologies (1 mentions)

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C18 column (380 citations) Reverse-phase HPLC (16 citations) C18 HPLC column (16 citations) Reverse HPLC (3 citations)

7 protocols using c18 reverse phase hplc column

Bortezomib Quantification in Spinal Cord

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Flash frozen spinal cord tissue samples were mixed with PBS (pH 7.2, 1:1 ratio) and homogenized with a pestle-type homogenizer. The homogenate was treated with 2 volumes of acetonitrile to precipitate the proteins, and the supernatant was analyzed by LCMS/MS. Samples were created for calibration and quality control with a working dilution of bortezomib in warfarin at 50 times the final concentration, and this was serially diluted to make the standard samples. These samples were diluted 31-fold into blank tissue extract and processed as above.
The signal was optimized for bortezomib by electrospray negative ionization (ESI) mode. A MS2 targeted SIM scan was used to optimize the precursor ion, and a product ion analysis was used to identify the best fragment for analysis and to optimize the collision energy.
Samples were analyzed with an ABI3000 mass spectrometer coupled with a Shimadzu HPLC and a Sil-HTc chilled autosampler, all controlled by Analyst software (ABI). After separation on a C18 reverse phase HPLC column (Agilent, Waters, or equivalent) mobile phase A was 0.1% formic acid in water and mobile phase B was 0.1% formic acid in acetonitrile.

Foran E., Kwon D.Y., Nofziger J.H., Arnold E.S., Hall M.D., Fischbeck K.H, & Burnett B.G. (2016). CNS uptake of bortezomib is enhanced by P-glycoprotein inhibition: implications for spinal muscular atrophy. Neurobiology of disease, 88, 118-124.

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Extraction and Purification of DSF Signaling Molecules

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The protocol for extraction and purification of DSF family components was described previously48 (link). In brief, Xcc strains were cultured in liquid medium for 24–48 hours and 50 mL of bacterial supernatant was collected by centrifugation at 3,800 × g for 30 minutes at 4 °C. The pH of the supernatants was adjusted to 4.0 by adding hydrochloric acid prior to two extractions with an equal volume of ethyl acetate. The ethyl acetate fractions were collected and the solvent was removed by rotary evaporation to dryness at 40 °C. The residue was dissolved in 1 mL of methanol. The crude extract was subjected to a 0.45 μm Minisart filter unit and the collected filtrate was concentrated to 0.5 mL. Three microliters of the extract was injected into a C18 reverse-phase HPLC column (4.6 × 150 mm, Agilent Technologies), eluted with water in methanol (23:77, v/v, respectively; 0.1% formic acid) at a flow rate of 1 mL/minute in an Agilent Technologies 1260 Infinity system with a DAD G1315D VL detector.

Yu Y.H., Hu Z., Dong H.J., Ma J.C, & Wang H.H. (2016). Xanthomonas campestris FabH is required for branched-chain fatty acid and DSF-family quorum sensing signal biosynthesis. Scientific Reports, 6, 32811.

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Serum L-Arginine Depletion in Mice

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Example 5

Serum Depletion of L-Arg in the Mouse Model

Balb/c mice were treated by single IP injection with 500 μg of pharmacologically prepared, pegylated Co-hArgI or an equal volume of PBS. Mice were sacrificed by cardiac veni-puncture for blood collection at the time points of 0, 48, 72, and 96 hrs. Blood samples were immediately mixed 50:50 (v/v) with a 400 mM sodium citrate buffer pH 4, allowed to clot for 30 minutes and centrifuged for serum separation. The resulting serum was then filtered on a 10,000 MWCO device (Amicon) for the removal of large proteins and precipitates and the flow-through was collected for analysis. L-arginine standards, control mouse serum and experimental samples were derivatized with OPA (Agilent) and separated on a C18 reverse phase HPLC column (Agilent) (5 μm, 4.6×150 mm) essentially as described by Agilent Technologies (Publication Number: 5980-3088) except for modification of the separation protocol slightly by reducing the flow rate by ½ and doubling the acquisition time to get better peak separation. An L-arginine standard curve was constructed by plotting L-Arg peak area versus concentration in order to quantify serum L-Arg levels. A single dose of pharmacologically prepared Co-hArgI was sufficient to keep L-Arg at or below detection limits for over 3 days (FIG. 1).

US10729752B2. Compositions and methods for treating cancer with arginine depletion and immuno oncology agents (2020-08-04). AERASE, INC. [US]. Inventors: David Lowe [US], Scott W. Rowlinson [US], Susan Alters [US], Giulia Agnello [US].

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Plasma Pharmacokinetics of Test Agent

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CD-1 female mice were injected intravenously with a single dose of 20 mg per kg in water and showed no adverse effects. Plasma samples were taken from 3 mice per time point (5, 15, 30 min; 1, 2, 4, 8 and 24 h post-dose). An aliquot of plasma sample or calibration sample was mixed with three volumes of methanol containing internal standard, incubated on ice for 5 min, and centrifuged. The protein-free supernatant was analysed by LC/MS/MS using an Agilent 6410 mass spectrometer coupled with an Agilent 1200 HPLC and a CTC PAL chilled autosampler, all controlled by MassHunter software (Agilent). After separation on a C18 reverse phase HPLC column (Agilent) using an acetonitrile-water gradient system, peaks were analysed by mass spectrometry using ESI ionization in MRM mode. The product m/z analysed was 134.1D, which provided a low limit of quantification of 1 ng ml⁻¹. The mean plasma concentration and the standard deviation from all 3 animals within each time point were calculated. PK parameters of test agent were calculated with a non-compartmental analysis model basedon WinNonlin. The mean plasma concentrations fromall3 mice at each time point were used in the calculation.

Ling L.L., Schneider T., Peoples A.J., Spoering A.L., Engels I., Conlon B.P., Mueller A., Schäberle T.F., Hughes D.E., Epstein S., Jones M., Lazarides L., Steadman V.A., Cohen D.R., Felix C.R., Fetterman K.A., Millett W.P., Nitti A.G., Zullo A.M., Chen C, & Lewis K. (2015). A new antibiotic kills pathogens without detectable resistance. Nature, 517(7535), 455-459.

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ADME In Vitro Analytical Workflow

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ADME in vitro studies were performed by APREDICA (Watertown, MA). Samples were analyzed by LC/MS/MS using an Agilent 6410 mass spectrometer coupled with an Agilent 1200 HPLC and a CTC PAL chilled autosampler, all controlled by MassHunter software (Agilent). After separation on a C18 reverse phase HPLC column (Agilent, Waters, or equivalent) using an acetonitrile-water gradient system, peaks were analyzed by mass spectrometry (MS) using ESI ionization in MRM mode.

Papadopoulou M.V., Bloomer W.D., Lepesheva G.I., Rosenzweig H.S., Kaiser M., Aguilera-Venegas B., Wilkinson S.R., Chatelain E, & Ioset J.R. (2015). Novel 3-nitrotriazole-based amides and carbinols as bifunctional anti-Chagasic agents. Journal of medicinal chemistry, 58(3), 1307-1319.

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Multiresidue Pesticide and Antibiotic Analysis

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The HPLC-MS/MS system (Agilent technologies, USA) consisted of a 1200 Series liquid chromatographer coupled to a triple quadrupole mass spectrometer (6410 Triple Quad) equipped with an electrospray ionization interface (ESI). All target compounds were separated using a HPLC reverse-phase C18 column (50 mm × 2.1 mm × 3.5 μm, Agilent technologies, USA). The gradient mobile phase consisted of methanol (phase A) and 0.1% formic acid (phase B). The gradient elution program for the 47 pesticides and 10 antibiotics was shown in ESI (Tables S1 and S2†). The injection volume was 5 μL and the column temperature was maintained at 30 °C. The total chromatographic run times for the 47 pesticides and 10 antibiotics were 25 min and 13.5 min, respectively. The HPLC–MS/MS was performed in multiple-reaction monitoring (MRM) mode and positive ESI mode. The desolvation gas (N₂) temperature was maintained at 350 °C with the gas flow being maintained at 8.0 L min⁻¹, and the nebulizer pressure being maintained at 35 psi. The optimized parameters of the 47 pesticides and 10 antibiotics are provided in Tables S3 and S4.†

Pan L., Feng X., Cao M., Zhang S., Huang Y., Xu T., Jing J, & Zhang H. (2019). Determination and distribution of pesticides and antibiotics in agricultural soils from northern China. RSC Advances, 9(28), 15686-15693.

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HPLC-MS/MS Analysis of Pesticides

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The HPLC–MS/MS analysis was achieved using an Agilent 1200 HPLC series (Agilent Technologies, Santa Clara, CA, USA) and an Agilent 6410B triple-quadrupole mass spectrometer equipped with an electrospray ionization interface (ESI±). A HPLC reverse-phase C₁₈ column (50 mm × 2.1 mm × 3.5 μm, Agilent, Santa Clara, CA, USA) was employed for the separation of dichlorprop-P, bentazone, 6-hydroxy-bentazone and 8-hydroxy-bentazone at 30 °C. The mobile phase was acetonitrile and 0.1% formic acid water (v/v = 90/10) at a flow rate of 0.25 mL/min, and the injection volume was 5 μL. The total run time was 2.5 min. The HPLC–MS/MS was performed in negative multiple-reaction monitoring (MRM) mode. The desolvation gas (N₂) temperature was set at 350 °C with the gas flow at 8.0 L/min, and the nebulizer pressure at 35 psi. The parameters were optimized individually for each target compound (Table 2).

Feng X., Yu J., Pan L., Song G, & Zhang H. (2016). Dissipation and Residues of Dichlorprop-P and Bentazone in Wheat-Field Ecosystem. International Journal of Environmental Research and Public Health, 13(6), 534.

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