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717 plus autosampler

Manufactured by Waters Corporation
Sourced in United States, United Kingdom, Italy, France, Germany, Japan, Ireland

The 717 plus autosampler is a laboratory equipment product from Waters Corporation. It is designed to automate the sample injection process for analytical instrumentation, such as liquid chromatography systems. The 717 plus autosampler features programmable sample handling and injection capabilities to improve efficiency and throughput in analytical workflows.

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169 protocols using 717 plus autosampler

1

Flavanone Disaccharide Extraction and Separation

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The extraction and separation of the flavanone disaccharides were conducted as described previously [17 (link)]. Flavonoids were separated using a gradient elution program with a flow rate of 1 ml/min, a column temperature of 35°C and a Waters 1525 binary system equipped with a 2996 photodiode array detector, a 717 Plus autosampler (Waters Corp., Milford, MA, USA) and a C18 column (250 mm × 4.6 mm, 5 μm, Thermo Scientific, Waltham, MA, USA). Subsequently, 0.15% formic acid diluted in water was prepared as mobile phase A, and 0.15% formic acid-acetonitrile was prepared as mobile phase B. The data were analyzed using Empower Chromatography Manager software (Waters Co., Milford, MA, USA).
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2

FP Quantification by Reverse-Phase HPLC

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Samples obtained from the diffusion and dissolution experiments were diluted 3:7 with methanol:water (50:50), while the donor wash mixture was diluted 1:1 with methanol:water (50:50). FP concentrations were quantified by a reverse-phase HPLC method using a Waters® HPLC system (Waters Co, Milford, MA, USA) with a 717 plus Autosampler (Waters Co., Milford, MA, USA), UV detection at 254 nm (Waters 2487 Dual λ Absorbance Detector (UV/Vis)), a Waters C18 Symmetry column, (100Å, 3.5 µm, 4.6 mm × 30 mm) and an injection volume of 100 µL. A mixture of acetonitrile:water (70:30), at a 1 mL/min flow rate, was selected as the mobile phase. The method, providing a limit of quantification of 0.2 µg/mL, was validated with intra-day and inter-day variabilities of less than 15% and an R2 larger than 0.998 for all the calibration curves.
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3

Quantification of Short-Chain Fatty Acids

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Short-chain fatty acids in samples at all time points were measured using a modified method of Ahmadi et al. [52 (link)]. Samples were thawed and centrifuged at 12,000× g for 5 min. The supernatant (300 μL) was mixed with 10 μL of 1 mM of 4-methyl valeric acid as an internal standard. The sample was transferred into a syringe fitted with a 4 mm 45-micron nylon filter and analyzed immediately using HPLC. HPLC was equipped with a Bio-Rad HPLC Organic Acid Analysis Aminex HPX-87H ion exclusion column (300 × 7.8 mm). The column was maintained at 65 °C (0.1 °C by a temperature control unit). The mobile phase consisted of water acidified with sulfuric acid (pH 2.28) with a flow rate of 0.65 mL/min. The solvent delivery system was a Waters 515 HPLC pump equipped with a Waters 717 plus autosampler. A Waters 996 photodiode array detector monitored the eluting compounds at 210 nm. As reference standards, individual and an equimolar mixture of acetic, propionic and butyric acid were used to quantify individual SCFA content.
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4

HPLC-UV Analysis of CIP Content

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Samples were analyzed for CIP content using an HPLC-UV method. The system comprised of Waters 717 plus Autosampler, Waters 2487 Dual λ Absorbance detector, Water 600 controller pump, and Agilent 3395 Integrator. A Phenomenex Luna® C18 4.6 mm × 250 mm column was used under isocratic elution for chromatographic analysis. The mobile phase used was mixture of acetonitrile and triethanolamine buffer (150:850 v/v) with pH adjusted to 2.36 using orthophosphoric acid. Triethanolamine buffer is made up of water, triethanolamine and 25 mM phosphoric acid in the ratio of (996:1.6:1.57 v/v). The flow rate was set at 1 mL/min with λmax (detection wavelength) of 299 nm during the analysis (27 ).
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5

Polymer Characterization by NMR and GPC

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1H NMR spectrum was examined on a Varian-400 FT-NMR spectrometer at 400.0 MHz with d6-DMSO as the solvent. Molecular weight (Mn and Mw) and distribution (Mw/Mn) of the synthesized polymers were measured by gel permeation chromatography (GPC) performed on a Waters 515 HPLC pump and a Waters 717 Plus Autosampler equipped with a Waters 2414 refractive index detector. Tetrahydrofuran (THF) was used as the eluent with a flowing rate of 1.0 mL/min at 35°C. A series of commercial polystyrene standards with narrow molecular weight distribution were applied to calibrate the GPC elution traces.
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6

RP-HPLC Analysis of DNA and RNA Nucleosides

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RP-HPLC analysis was performed using the Waters 625 LC System (encompassing: Waters 625 Pump, Waters 600 Controller, Waters 717plus Autosampler, Waters Degasser and Waters 996 PDA detector) Synergy Max-RP C12 (250 × 4.6 mm, 4u, Phenomenex) column, combined with Synergy Max-RP C12 pre-column according to an adapted procedure [74 (link), 75 (link)]. Separation of nucleosides was conducted in the presence of ‘A’ buffer (0.5 % v/v methanol in 10 mM KH2PO4, pH 3.7) and ‘B’ buffer (10 % v/v methanol in 10 mM KH2PO4, pH 3.7). The pH of the buffers was adjusted with phosphoric acid. The linear gradient used for separation consisted of 100 % ‘A’ buffer to 100 % of ‘B’ buffer for 10 min, next 100 % of ‘B’ buffer for 10–25 min and then, at the end of 25 min program 100 % ‘A’ buffer was pumped for 5 min. Flow rate was 1 ml per min and column temperature was set at 30 °C. UV-detection was used at the wavelength of 280 nm. The external standard consisted of major DNA (0.5–50 µM) and RNA nucleosides (1.5–150 µM) and 5-methyl-2′-deoxycytidine (5mdC) dissolved in deionized water. Peaks corresponding to 2′-deoxycytidine (dC) and 5mdC had retention time equal to 6.5 and 9.3 min, respectively. The contribution of 5mdC was calculated based on Millennium 32 v. 4.0 software (Waters Corporation, Milford, Massachusetts, USA).
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7

Spectroscopic Analysis and Chromatography

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UV and IR spectra were obtained employing Perkin-Elmer Lambda 40 and Perkin-Elmer Spectrum BX instruments (PerkinElmer, Inc., Waltham, USA), respectively. VLC grade (Macherey-Nagel, Polygoprep 60-50C18) was used for vacuum liquid chromatography. All organic solvents were distilled prior to use. HPLC was carried out using a Waters system, controlled by Waters Millenium software, consisting of a 600E pump, a 996 PDA, and a 717 plus autosampler (Waters Corporation, Milford, MA, USA). All NMR spectra were recorded on a Bruker Avance 300 and 500 DRX spectrometers (Bruker Corporation, Rheinstetten, Germany). Spectra were referenced to the residual solvent signals with resonances at δH/C 2.04/29.8 (acetone-d6) and δH/C 7.26/77.0 (CDCl3). ESI mass spectra were obtained on an Applied Biosystems/MDS Sciex API 2000 MS spectrometer (Applera Corporation and MDS Inc., Foster, CA, USA). HRESIMS were recorded on a Bruker Daltonik micrOTOF-Q Time-of-Flight mass spectrometer with ESI source, and UPLC-Synapt G2 HDMS mass spectrometer (Bruker CorporationCompany, Rheinstetten, Germany).
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8

HPLC Analysis of Flavonoids in SBTE

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HPLC analysis of SBTE was performed using Waters HPLC system (Waters Corporation, Milford, MA) equipped with Waters 515 HPLC pump, Waters 717 plus autosampler and Waters 2487 PDA detector. Separation was performed in a symmetry C18 250 mm ×4.7 mm ID; 5 μm column by maintaining a flow rate of 1 mL/min for the mobile phase (1% acetic acid (A):methanol (B) as linear gradient run consisting of (A) 90–85% in 0–2 min, 85–70% in 3–20 min, 70–60% in 21–45 min, 60–90% in 46–60 min). Each run was followed by a 10 min equilibrium period. Two flavonoids, kaempferol and isorhamnetin, were quantified in the similar conditions. Peaks were assigned by spiking the samples with standard compounds and comparison with the retention times and spectral matching.
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9

Instrumentation Setup for Natural Product Library

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The instrumentation setup for our natural product peak library generation has been recently published.21 (link) The HPLC stage utilized two Waters 510 pumps and a Waters 717plus autosampler, both controlled with Empower 2 software. Separation was performed on a 250 × 10 mm 5 μm Luna C18 column (Phenomenex). Spectra from three detectors were acquired during peak library fractionation: Waters 996 photo diode array, SEDEX 55 ELSD, and Mariner 5054 ESI-TOF-MS. The mobile phase parameters are CH3CN (A) and H2O (B) with a flow rate of 2 mL/min and the following elution conditions: 30 min gradient from 10:90, 10 min isocratic 100:0, 1 min gradient from 100:0 to 10:90, 9 min isocratic 10:90. The injection amount was 15 mg/150 μL. Sample collection was performed using a Gilson 215 liquid handler controlled with Gilson Unipoint LC software. Samples were collected into BD Biosciences 96-deep-well plates, with a working volume of 2 mL (part number: 353966). Fractions were collected every minute. After the LC-MS-UV-ELSD library is collected, a duplicate archive plate is generated for analytical reference using a 12-channel pipet, creating an exact copy and counter balance for centrifugal drying. Plates were dried and concentrated using a Savant AES2010 SpeedVac.
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10

Polymer Characterization by NMR and GPC

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1H and 13C NMR spectra were recorded on a Bruker AVANCE 500 spectrometer. The chemical shifts reported were referenced to internal tetramethylsilane (0.00 ppm) or to the solvent resonance at the appropriate frequency. The number and weight average molecular weights (Mn and Mw, respectively) of polymers were measured by gel permeation chromatography (GPC) using a Waters HPLC system equipped with a model 1515 isocratic pump, a 717 plus autosampler, and a 2414 refractive index (RI) detector with Waters Styragel columns HT6E and HT2 in series. Empower II GPC software was used for running the GPC instrument and for calculations. Both the Styragel columns and the RI detector were heated and maintained at 40 °C temperature during sample analysis. Chloroform was used as the eluent at a flow rate of 1.0 mL min−1. Sample concentrations of 2 mg mL−1 and injection volumes of 100 µL were used. Polymer molecular weights were determined based on a conventional calibration curve generated by narrow polydispersity polystyrene standards from Aldrich Chemical Co.
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