Serum, cecum content and colon content reversed-phase HPLC analyses were performed on a Shimadzu HPLC equipped with a LC-10ADvp pump, SCL 10Avp system controller and a SPD-M10Avp PDA detector. All the operations were controlled by Shimadzu EZ start 7.2.1 software in Windows XP system. The PDA detector was set between 190 to 800 nm and analysis was done at 341 nm. HPLC solvents consisted of 65% Acetonitrile and 35% Distilled water. An isocratic method with flow rate 0.18 ml/min used with total run time 40 min (serum samples), and 20 min (cecum and colon content samples). The injection volume was 70 µL for serum samples and 5 or 10 µl for cecum content and colon content samples. A XBridgeTM C18 (Waters, BEH 130, 3.5 µm, 2.1×150 mm) column with a Precolumn XBridge (BEH C18 Sentry Guard Cartridge, 130Å, 3.5 µm, 2.1 mm × 10 mm) was used. Retention times for DQ (IS) is 9.6 min, and SHetA2 is 11.7 min.
Lc 10advp pump
Manufactured by Shimadzu
Sourced in Japan, United Kingdom, Germany
The LC-10ADvp is a high-performance liquid chromatography (HPLC) pump designed for reliable and precise solvent delivery. It features a dual-plunger design and advanced flow control technology to ensure accurate and stable flow rates. The pump is capable of operating at a maximum pressure of 40 MPa (400 bar, 5,800 psi) and can deliver a flow rate range of 0.001 to 10.000 mL/min, making it suitable for a variety of HPLC applications.
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Lab products found in correlation
Sil 10advp auto injector, by Shimadzu (6 mentions)
Spd 10a vp, by Shimadzu (5 mentions)
Spd m10avp, by Shimadzu (4 mentions)
Hplc system, by Shimadzu (4 mentions)
Sil htc autosampler, by Shimadzu (3 mentions)
Sil 20a ht, by Shimadzu (3 mentions)
High performance liquid chromatography (hplc), by Shimadzu (3 mentions)
Scl 10avp controller, by Shimadzu (3 mentions)
Sil 10advp autosampler, by Shimadzu (3 mentions)
Spd 10a vp uv vis detector, by Shimadzu (3 mentions)
Scl 10a vp system controller, by Shimadzu (3 mentions)
Ez start 7, by Shimadzu (2 mentions)
Class vp software, by Shimadzu (2 mentions)
Eclipse xdb c18, by Agilent Technologies (2 mentions)
Microtof spectrometer, by Bruker (2 mentions)
Spd m10avp uv vis photodiode array detector, by Shimadzu (2 mentions)
Inertsil ods 3 octadecylsilane column, by GL Sciences (2 mentions)
Tsq quantum discovery max, by Thermo Fisher Scientific (2 mentions)
Xcalibur software, by Thermo Fisher Scientific (2 mentions)
Xbridgetm c18, by Waters Corporation (1 mentions)
54 protocols using lc 10advp pump
1
Serum and Gut Content HPLC Analysis
2
HPLC Analysis of Glutamate and Aspartate
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Analysis of glutamate and aspartate in the dialysate was performed by reversed-phase high performance liquid chromatography (HPLC) with precolumn derivatization with o-phthaldialdehyde (OPA) and fluorescence detection, as reported elsewhere with some modifications (31 (link)). Briefly, 15 μl of each dialysate sample were mixed with 15 μl OPA solution (40 mM, pH 9.3) and allowed to react for 1 min at room temperature. After the reaction was completed, 20 μl of the resulting mixture were injected into an HPLC system (Shimadzu Corp.) consisting of a solvent delivery system (LC-10 ADVP pump; Shimadzu Corp.), an octadecyldisulfonate (ODS) C18 (25×4.6 mm i.d., 5 μm particle size; Shimpak; Shimadzu Corp.) column and a fluorescent detector (RF-10 AXL; Shimadzu Corp.) with excitement (Ex) set at 350 λ and emission (Em) set at 470 λ. The mobile phase consisted of 92.5% sodium acetate buffer (0.1 M, pH 6.96), 5% methanol and 2.5% tetrahydrofuran (flow rate 1.2 ml/min).
3
Monosaccharide Analysis of Polysaccharide Sample
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The polysaccharide sample (5 mg) was dissolved in 2 mL of 2 M trifluoracetic acid (TFA, Sigma, Egypt) and hydrolyzed for 3 h at 110 °C in a sealed tube. Under nitrogen flow, the soluble portion was evaporated to dryness. The samples and monosaccharide standards (Bio-Rad, USA) were injected onto a platinum amino column (5 µm, 250 mm 4.6 mm i.d.; Grace, Lokeren, Belgium), which was kept at 20 °C in a column oven following the removal of TFA (CTO-10 ASvp, Shimadzu). The analysis used an LC-10 ADvp pump from HPLC (Shimadzu, Japan) and ELSD (Sedex 55, SEDERE, Olivet, France) at 60 °C and 230 kPa of nitrogen pressure. The chromatograms were obtained, and the data were processed with Class-VP software (Shimadzu, version 6.1, Tokyo, Japan) [47 (link)].
4
Quantifying Sugars in Wastewater
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The wastewater samples were suitably diluted with distilled water and filtered through a 0.45 μm PTFE filter (Waters, Milford, MA). Then, the effluent filtrates were subjected to high-performance liquid chromatography (HPLC) analysis for the determination of glucose and fructose. The HPLC system was equipped with a LC-10Advp pump (Shimadzu, Kyoto, Japan) and a RID-6A refractive index detector (Shimadzu). The separation was achieved on an Agilent HI-plex H (Agilent Technologies, Santa Clara, CA, USA) column by isocratic elution with ultrapure water at 65 °C. The flow rate was 0.5 mL/min and the injection volume 10 μL. Each sample was analyzed in triplicate. Quantification was performed by linear regression analysis using external calibration curves for glucose (g/L) and fructose (g/L). Total soluble sugar content (g/L) in the effluents was determined spectrophotometrically by the phenol-sulfuric acid method and expressed as glucose equivalents (g/L) [21 (link)].
5
Quantitative Analysis of Aflatoxins
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Aflatoxins analyses were performed using the methodology described by Bragulat et al. (2001 (link)). The strains were incubated in Petri dishes containing yeast extract sucrose (YES agar, Katsurayama et al. 2018 (link)) at 25ºC for 14 days in the dark. Then, 3 plugs from each Petri dish were transferred to an Eppendorf tube and 1 ml of methanol was added. After 1 hour, the solution was filtered with a Millipore filter (Ø 0.22 mm), an aliquot (200 µl) was derivatised with 700 µl trifluoroacetic acid: acetic acid: water (20:10:70, v/v/v). The derivatised solution was analysed using a reverse phase HPLC consisting of a Shimadzu LC-10ADvp pump, a RF-10Axl fluorescence detector (Shimadzu; excitation and emission wavelength of 360 nm and 440 nm, respectively), and a C18 reversed-phase column (150 mm x 4.6 mm i.d., 5 µm particle size; Nucleodur®, Macherey-Nagel, Düren, Germany) connected to a pre-column Security Guard (8 mm x 4 mm i.d., 5 µm particle size; Nucleodur®, Macherey-Nagel, Düren, Germany). The mobile phase was water: methanol: acetonitrile (4:1:1, v/v/v) at a flow rate of 1.5 ml min-1. The injection volume was 20 µl. Aflatoxins production was measured in ng g-1 of culture medium. The limit of detection was 1 ng g-1 of AFB1 and AFG1, and 0.8 ng g-1 of AFB2 and AFG2.
6
HPLC Quantification of Temozolomide Release
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The HPLC system consisted of a LC-10ADvp pump and a PD-10vp UV/VIS absorbance detector (Shimadzu, Kyoto, Japan) set at 316 nm. It featured a 150 × 4.6-mm, 5-μm ZORBAX ODS column (Agilent Technologies, Santa Clara, CA, USA). The mobile phase consisted of 0.1% aqueous acetic acid-acetonitrile (90:10, v/v) and was delivered at 1.0 mL/min. The temperature was 40°C. Peak data were recorded with a chromatography management system (Shimadzu). A calibration curve was constructed using eight calibration standards prepared at concentrations of 0–2 mM TMZ. The amount of TMZ released from FG was based on the calibration curve standards.
7
Quantification of Domperidone in Plasma
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Venous blood samples of 4 mL were taken at 10 minutes before dosing and at 8 predefined timepoints after dosing (within 5 minutes after the last of the triplicate 12-lead ECG recording) on Day 1 and Day 4 of each period for the determination of plasma concentrations of domperidone. The whole blood samples for PK evaluation were centrifuged at room temperature for 10 minutes, beginning within 2 hours of collection; the resulting ∼1.7 mL plasma samples were frozen within 2 hours of whole blood sample collection and stored at approximately −20°C until transferred to the bioanalytical facility (PRA Early Development Services, Assen, The Netherlands) for analysis.
Concentrations of domperidone in EDTA plasma were determined using a validated,9 ,10 (link) sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) method. The method consisted of a solid-phase extraction in a micro-elution plate with Oasis MCX, followed by reversed phase chromatography using a Waters XBridge C18 column (Waters, Milford, MA) using a HTC PAL autosampler (CTC Analytics, Zwingen, Switzerland) with LC-10Advp pump (Shimadzu, Columbia, MD) coupled with a Applied Biosystems/MDS SCIEX API3000 triple quadrupole mass spectrometer. The quantification range was 1–500 ng/mL.
PK parameters were calculated for domperidone using standard non-compartmental analysis methods.
Concentrations of domperidone in EDTA plasma were determined using a validated,9 ,10 (link) sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) method. The method consisted of a solid-phase extraction in a micro-elution plate with Oasis MCX, followed by reversed phase chromatography using a Waters XBridge C18 column (Waters, Milford, MA) using a HTC PAL autosampler (CTC Analytics, Zwingen, Switzerland) with LC-10Advp pump (Shimadzu, Columbia, MD) coupled with a Applied Biosystems/MDS SCIEX API3000 triple quadrupole mass spectrometer. The quantification range was 1–500 ng/mL.
PK parameters were calculated for domperidone using standard non-compartmental analysis methods.
8
3D-printed Device Characterization
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For constructing the devices, the Form-2 3D-printer (Formlabs) and Form Cure chamber (405 nm wavelength; Formlabs) were used. Packing of the devices was performed using a Shimadzu LC-10AD VP pump (Shimadzu‘s Hertogenbosch, The Netherlands). The flow measurements were performed with an Agilent 1100 Series Pump (Agilent, Waldbronn, Germany). The LC–MS measurements were performed with Waters ACQUITY UPLC and Waters Synapt G2 (Waters Corporation, Milford, US).
9
Size-Exclusion HPLC Analysis of Protein Isolates
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The SE-HPLC separation of the protein isolate and its hydrolysates was carried out according to a previously described method [10 (link)]. The Shimadzu HPLC system composed of a LC-10ADVp pump, a SPD-M10AVp photo-diode array detector, and a SCL-10AVp system controller (Kyoto, Japan) was applied. A 20 μL sample at a concentration of 5 mg/mL was injected onto a TSKgel G2000SWXL column (7.8 × 300 mm, 5 μm, Tosoh Bioscience, Tokyo, Japan) and eluted with acetonitrile:water:trifluoroacetic acid (45:55:0.1 v/v/v) at a flow rate of 0.5 mL/min. Six standard molecular markers, albumin from chicken egg white (45 kDa), cytochrome C (12.4 kDa), aprotinin (6.5 kDa), angiotensin II acetate (1.046 kDa), leucine enkephalin (556 Da) and Val-Tyr-Val (379 Da), were used for column calibration.
10
HPLC Analysis of Brain Amino Acids
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The brain levels of the amino acids were determined using HPLC method, coupled to an electrochemical detection, as described previously (Szyndler et al. 2008 (link)). Briefly, the compounds were eluted isocratically with the mobile phase delivered at 0.7 ml/min, using a Shimadzu Class LC-10ADvp pump. An electrochemical detector with a flow-through cell (Intro-Antec Leyden) linked to the Shimadzu Class VP Integrator SCL-10 Avp was used. A high-density glass carbon-working electrode (Antec) was used at + 0.85 V. A Rheodyne injection valve with 20-µl sample loops was used to manually inject the samples. The preparation of the mobile phase and the derivatizing agents was based on the slightly modified method of Rowley et al. (1995 (link); Szyndler et al. 2008 (link)). The concentrations of GABA, alanine, taurine, glutamine and glutamate were calculated in µmol/g of tissue. Similarly as in case of monoamines, ratios of the glutamate concentration to both glutamine and GABA were also computed and investigated.
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