Samples were treated with Carrez reagents I and II and 0.3 mL of them was mixed 10 mL of each samples for caffeine. After being centrifuged (10 min at 45,000× g), supernatant was filtered (0.45 µm membrane filter, Acrodisc Syringe Filter, Gelman Sciences, Ann Arbor, MI, USA) and XBridge™ Shield RP18 (5 µm, 4.6 mm × 250 mm, Waters, Milford, MA, USA) was used for analysis. Ten millimolar of citric acid and methanol were used for mobile A and B, respectively and the detector was set at 276 nm. In case of chlorogenic acid, samples were treated in the same way of caffeine except detector was set at 325 nm. For trigonelline, each 10 mL of samples was mixed with 0.3 mL of Carrezz reagents I and II and 0.6 mL of absolute methanol (HPLC-grade, Merck, Damstadt, Frankfurter, Germany). Other processes were identical with caffeine and chlorogenic acid except that a reverse-phase column (XBridge™ Shield RP18; 5 µm, 4.6 mm × 150 mm, Waters, Milford, MA, USA) and 0.5% methanol (HPLC-grade, Merck, Damstadt, Frankfurter, Germany) was used. The flow rate was 0.8 mL/min and the detector was set at 264 nm.
Xbridge shield rp18
Manufactured by Waters Corporation
Sourced in United States
The XBridge Shield RP18 is a reversed-phase high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of organic compounds. The column features a proprietary hybrid silica-based stationary phase that provides good peak shape and reproducibility for a variety of sample types.
Automatically generated - may contain errors
Lab products found in correlation
Chromeleon 7, by Thermo Fisher Scientific (2 mentions)
Plrp s polymeric reversed phase column, by Agilent Technologies (2 mentions)
Hplc grade, by Merck Group (1 mentions)
Millex gs 0.22 μm sterile filters, by Merck Group (1 mentions)
Chromolith performance rp 18e, by Merck Group (1 mentions)
Chromolith semiprep rp 18e, by Merck Group (1 mentions)
Dionex ultimate 3000 hplc, by Thermo Fisher Scientific (1 mentions)
Dionex ultimate 3000, by Thermo Fisher Scientific (1 mentions)
8 protocols using xbridge shield rp18
1
Quantification of Caffeine, Chlorogenic Acid, and Trigonelline
2
Synthesis and Characterization of Stereoisomeric Compounds
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Example 2
To a solution of compound 2-5a (30.0 mg, 41.3 umol) in H2O (10.0 mL) was added Dowex®-50WX8 (Na+ form; 300 mg) and the mixture was stirred at RT for 4 h. The reaction was then filtered, and the filtrate was lyophilized to give Example 2a (RpRp or SpRp; 30.0 mg, 40.6 umol) as a white solid.
MS(ES+) C20H24FN10O9P2S2 requires: 693, found: 693.0 [M+H]+; 1H-NMR (400 MHz, DMSO-d6) δ ppm 8.56 (s, 1H), 8.40 (s, 1H), 8.17 (s, 1H), 8.13 (s, 1H), 7.34 (s, 2H), 7.22 (s, 2H), 6.23 (m, 1H), 6.09 (d, J=8.4 Hz 1H), 5.71 (d, J=52.8 Hz, 1H), 5.54 (s, 1H), 5.15-5.30 (m, 2H), 3.91-4.37 (m, 5H), 3.67-3.70 (m, 1H); 31P NMR (162 MHz, CD3OD) δ ppm 55.97, 53.66; Rt=1.384 minutes [Waters XBridge Shield RP18 2.1*50 mm, 5 um; mobile phase: A: H2O+10 mM NH4HCO3; B: MeCN; A %-B %=0%-30%, 5.2 minutes].
Reaction of compound 2-5b in a similar manner gave Example 2b (SpRp or RpRp, 28.0 mg, 40.6 umol) as a white solid.
MS(ES+) C20H24FN10O9P2S2 requires: 693, found: 693.0 [M+H]+; 1H-NMR (400 MHz, CD3OD) δ ppm 8.86 (br s, 1H), 8.39 (s, 1H), 8.19 (s, 1H), 8.02 (br s, 1H), 6.35-6.41 (m, 2H), 5.70 (d, J=51.8 Hz, 1H), 5.22-5.27 (m, 2H), 4.35-4.60 (m, 5H), 4.05-4.08 (m, 2H); 31P NMR (162 MHz, CD3OD) δ ppm 57.39, 52.28; Rt=1.644 minutes [Waters XBridge Shield RP18 2.1*50 mm, 5 um; mobile phase: A: H2O+10 mM NH4HCO3; B: MeCN; A %-B %=0%-30%, 5.2 minutes].
3
HPLC Quantification of Quercetin-3-O-Glucuronide
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HPLC quantitative analyses were carried out using a Shimadzu LC-2030C 3D HPLC instrument and a photodiode-array (PDA) detector (Shimadzu, Japan). Analysis of Quercetin-3-O-glucuronide was performed using a Waters Xbridge Shield RP18 (150 × 4.6 mm I.D; 3.5 μm) (Waters Corporation, Milford, MA, USA) column.
4
In Vivo Peptide Stability Assay
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The in vivo stability was determined in healthy male Swiss albino mice (12 mice in total, body weights 30 ± 5 g). Briefly, each radiolabeled peptide was injected as a bolus (100 μL, 11–22 MBq, 3 nmol of total peptide in saline/EtOH 9/1 v/v) in the tail vein of mice either untreated (controls) or 20 min after having received per os a slurry of an Entresto® pill (individual 12 mg/200 mL doses of pill per animal; Entresto®-group; Novartis, Basel, Switzerland), as previously described [34 (link),35 (link)]. Blood was collected 5 min post-injection (pi) in a chilled syringe directly from the heart (0.5–1 mL) and samples were processed prior to being analyzed by HPLC for the detection of forming radiometabolites, as previously described [25 (link),35 (link)]. For analyses, an XBridge Shield RP18 (5 μm, 4.6 mm × 20 mm) column (Waters, Vienna, Austria) was eluted at a flow rate of 1 mL/min with 0.1% TFA in H2O (A) and MeCN (B) with the following linear gradient system: 100%A/0%B at 0 min, with B linearly increasing by 1%/min to 60%A/40%B (system 2). The tR of the intact radiopeptide was determined by coinjection with the respective [111In]In-DOTAGA-PEG2-RM26 (tR = 30 min)/[111In]In-AU-RM26-M1 (tR = 29 min) reference in the HPLC.
5
HPLC Analysis of Organic Compounds
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The chromate column was Waters XBridge™ Shield RP18 (4.6 mm × 250 mm, 5 μm); DAD detector detection wavelength: 280 nm; column temperature: 30°C; mobile phase: Acetonitrile (A) 0.1% KH2PO4 (B); flow speed: 1 mL/min; injection volume: 20 μL. Elution procedures are as follows: 0–10 min, V(A) : V(B) = 15 : 85; 10–15 min, V(A) : V(B) = 20 : 80; 15–20 min, V(A) : V(B) = 25 : 75.
6
Synthesis and Purification of OSI-420
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Unless otherwise stated all chemicals were purchased from Sigma-Aldrich Chemie (Schnelldorf, Germany) or Merck (Darmstadt, Germany) at analytical grade and were used without further purification. The Ni catalyst (Shimalilte Ni reduced, 80/100 mesh) was purchased from Shimadzu (Kyoto, Japan). The precursor 6-O-desmethyl-elotinib (OSI-420; GMP grade) was purchased from Syncom B.V. (Groningen, Netherlands) and the reference compound erlotinib (N-(3-ethinylphenyl)-6,7-bis(2-methoxyethoxy)-quinazolin-4-amine) was obtained from Apollo Scientific (Bredbury, UK). Semi-preparative high performance liquid chromatography (HPLC) column (Chromolith® SemiPrep RP-18e, 100–10 mm; guard column: 10–10 mm) and analytical HPLC column (Chromolith Performance RP-18e, 100–4.6 mm; guard column: 10–4.6 mm) were purchased from Merck (Darmstadt, Germany). The analytical HPLC column for the optimized quality control (XBridge® Shield RP-18; 2.5 μm; 3.0–50 mm) and C18plus SepPak® cartridges for solid phase extraction (SPE) were purchased from Waters (Waters® Associates Milford, MA, USA). Low-protein binding Millex GS® 0.22 μm sterile filters were purchased from Millipore® (Bedford, MA, USA). Gas chromatography (GC) capillary column (forte GC Capillary Column ID-BP20; 12 m × 0.22 mm × 0.25 μm) was obtained from SGE Analytical Sciences Pty Ltd. (Victoria, Australia).
7
HPLC Chromatographic Characterization Protocol
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The HPLC DIONEX Ultimate 3000 (Thermo Scientific
Inc.) coupled to an RS diode array and the Chromeleon 7.2.10 software
(www.thermofisher.com )
was used for all chromatographic measurements. The variety of experimental
descriptors required specific chromatographic columns. Three different
columns were used: IAM.PC.DD2 (300 Å, 10 μm, 10 cm ×
4.6 mm) from REGIS, PLRP-S polymeric reversed-phase column (100 Å,
5 μm, 50 × 4.6 mm) from Agilent (www.agilent.com ), and XBridge Shield RP18 (130 Å, 5 μm,
5 cm × 4.6 mm) from Waters (www.waters.com ). High-performance ergonomic single-channel
variable volume pipettors, 1.5 mL HPLC vials, and 9 mm PP screw caps
were purchased from VWR Signature. The pH of each buffer and sample
was controlled using a Eutech pH Meter 2700 (www.fishersci.com ).
Inc.) coupled to an RS diode array and the Chromeleon 7.2.10 software
(www.
was used for all chromatographic measurements. The variety of experimental
descriptors required specific chromatographic columns. Three different
columns were used: IAM.PC.DD2 (300 Å, 10 μm, 10 cm ×
4.6 mm) from REGIS, PLRP-S polymeric reversed-phase column (100 Å,
5 μm, 50 × 4.6 mm) from Agilent (
5 cm × 4.6 mm) from Waters (
variable volume pipettors, 1.5 mL HPLC vials, and 9 mm PP screw caps
were purchased from VWR Signature. The pH of each buffer and sample
was controlled using a Eutech pH Meter 2700 (
8
HPLC Analysis of Biomolecular Compounds
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Chromatographic measurements were performed
using instrument DIONEX Ultimate 3000, Thermo Scientific Inc. coupled
to RS Diode Array and Chromeleon 7.2.10 software (www.thermofisher.com ) (HPLC).
HPLC columns IAM.PC.DD2 (300 Å, 10 μm, 10 cm × 4.6
mm) from REGIS, XBridge Shield RP18 (130 Å, 5 μm, 5 cm
× 4.6 mm) from Waters (www.waters.com ) and PLRP-S polymeric reversed phase column (100 Å, 5 μm,
50 mm × 4.6 mm) from Agilent were used. Ergonomic high-performance
single-channel variable volume pipettors, HPLC 1.5 mL vials, 0.1 mL
microinsert, and PP screw 9 mm caps were purchased from VWR Signature.
pH was controlled with Eutech pH Meter 2700 (www.fishersci.com ).
using instrument DIONEX Ultimate 3000, Thermo Scientific Inc. coupled
to RS Diode Array and Chromeleon 7.2.10 software (
HPLC columns IAM.PC.DD2 (300 Å, 10 μm, 10 cm × 4.6
mm) from REGIS, XBridge Shield RP18 (130 Å, 5 μm, 5 cm
× 4.6 mm) from Waters (
50 mm × 4.6 mm) from Agilent were used. Ergonomic high-performance
single-channel variable volume pipettors, HPLC 1.5 mL vials, 0.1 mL
microinsert, and PP screw 9 mm caps were purchased from VWR Signature.
pH was controlled with Eutech pH Meter 2700 (
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