CSF metabolite concentrations of the entire cohort were measured in 2015 in one batch using a triple-quadrupole mass spectrometer (API4000, Sciex, Framingham, MA, USA) with an electrospray-ionization ion source coupled to a high-performance liquid chromatography system (SIL-HTc, Shimadzu, Japan) and the AbsoluteIDQ™ p180 kit and MetIDQ™ software (Biocrates Life Sciences, Innsbruck, Austria) as described in detail in [14 (link)]. The following lipid nomenclature is used. Sphingomyelin (SM) and hydroxysphingomyelin (SM[OH]): Cx:y, where x = total number of carbon atoms and y = total number of double bonds in the amide bond. Phosphatidylcholine (PC): aa: both side chains are fatty acids linked to the glycerol backbone by ester bonds, ae: one of the side chains is a fatty alcohol linked to the glycerol backbone by an ether bond; Cx:y: x = total number of carbon atoms and y = total number of double bonds in both fatty acid chains.
Sil htc
Manufactured by Shimadzu
Sourced in Japan
The SIL-HTc is a high-throughput autosampler designed for liquid chromatography (LC) systems. It features a compact size, high sample capacity, and precise sample handling capabilities. The core function of the SIL-HTc is to automatically introduce liquid samples into an LC system for analysis.
Automatically generated - may contain errors
Lab products found in correlation
Api 4000, by AB Sciex (3 mentions)
Absoluteidq p180 kit, by Biocrates (2 mentions)
Metidq software, by Biocrates (2 mentions)
Lc 20ad, by Shimadzu (2 mentions)
Lc 10ad, by Shimadzu (2 mentions)
Cto 10as vp, by Shimadzu (1 mentions)
Dgu 20a3, by Shimadzu (1 mentions)
Api 5000 mass spectrometer, by AB Sciex (1 mentions)
Qtrap 5500, by AB Sciex (1 mentions)
Mxp quant 500 kit, by Biocrates (1 mentions)
Quantstudio 6 flex rt pcr system, by Thermo Fisher Scientific (1 mentions)
Odyssey imager, by LI COR (1 mentions)
7 protocols using sil htc
1
Comprehensive CSF Metabolite Profiling
2
HPLC Analysis of Carbocisteine and Rosiglitazone
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Chromatography separation was carried out by using the Symmetry Shield RP8, 150 × 3.9 mm, 5 μm. The retention times of carbocisteine and rosiglitazone were approximately 2.20 and 3.01 min, respectively. The overall run time was 4.50 min. An HPLC system of Shimadzu (Japan) containing an autosampler (SIL-HTc), pumps (LC-20AD), column oven (CTO-10AS VP), and degasser (DGU-20A3) was used for the separation of the analytes.
The injection volume was 5 μL at the autosampler temperature of 10°C. The mobile phase consisting of methanol: 0.5% and formic acid (40:60, v/v) was used for the analysis.
The injection volume was 5 μL at the autosampler temperature of 10°C. The mobile phase consisting of methanol: 0.5% and formic acid (40:60, v/v) was used for the analysis.
3
Quantification of Compound in Rat Brain
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rats were terminally anesthetized and perfused transcardially with heparinized saline (0.1% heparin in 0.9% saline) and whole brains were removed and weighed. Brains were homogenized in distilled water at a volume (μl) that was 2x brain weight. A 5μL aliquot of sample was injected onto a Phenomenex Luna C8 (50x2.0 mm) 5 mm HPLC column with a Shimadzu SILHTC auto sampler and an integrated HPLC pumping system Shimadzu LC10AD. The compound was detected by an Sciex API 5000 Mass Spectrometer with a positive ESI ionization mode. Mobile phase A was 95% acetonitrile in water, mobile phase B was 10 mM ammonium acetate buffer in water, pH 7.0 with a flow rate of 500 μL/min. The starting condition for HPLC gradient was 25:75 (A/B) at time 0 min, 100:0 (A/B) from 0.1 to 1.9 min and 25:75 (A/B) from 2 to 3 min. Multiple reaction monitoring (MRM) was used to monitor the compound with m/z transitions 363.2–347.2 and a retention time of 1.25 min.
4
Urine Sample Preparation for HPLC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Urine samples were diluted 1:10 with 80% acetonitrile aqueous solution (dilution solution) and centrifuged at 4000×g for 5 min. The supernatant was then diluted 1:5, and the centrifugation step was repeated. Obtained samples were additionally diluted 1:100 (for lactulose quantitation) and 1:500 (for mannitol quantitation). From these samples, 120 μL were transferred to 96-well plates, and the plates were transferred to the autosampler (Model: SIL-HTc, Shimadzu Co., Tokyo, Japan), which was connected to a high-performance liquid chromatography pump (Model: LC-10AD VP Shimadzu Co., Tokyo, Japan).
5
Targeted Metabolomics of CSF and Serum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CSF metabolite concentrations were measured on a triple-quadrupole mass spectrometer (API4000, Sciex, Framingham, MA, USA) with an electrospray-ionization ion source coupled to a high-performance liquid chromatography system (SIL-HTc, Shimadzu, Japan) and the AbsoluteIDQ™ p180 kit and MetIDQ™ software (Biocrates Life Sciences, Innsbruck, Austria), as described in detail in [17 (link)]. Serum concentrations of phosphatidylcholines were measured on an AB SCIEX 5500 QTrap™ mass spectrometer (AB SCIEX, Darmstadt, Germany) using the MxP™ Quant 500 kit (Biocrates), following the manufacturer’s protocols (https://biocrates.com/mxp-quant-500-kit , accessed on 15 December 2020). Details about internal standards and quality controls (QC) are given in the manufacturer’s application note [22 ]. The Quant 500 kit measures the same glycerophospholipids as the p180 kit and is downward compatible with it, ensuring comparability of the data.
6
Multimodal Analysis of Signaling Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plates for the total STAT4, phosphorylated STAT4, and IFNγ assay were analyzed using the Sector S 600 (Meso Scale Discovery Technologies). Plates for the phosphorylated STAT2 Assay were analyzed using the SpectraMax-M2 plate reader. Dot blots for the phosphorylated PLCγ1 assay were analyzed using the Odyssey Imager (LiCor). qRT-PCR was performed on the Applied Biosystems QuantStudio 6 Flex RT-PCR system. Mass spectrometry was conducted on the API-4000 (Sciex, Framingham, MA). Liquid chromatography was assessed on a Shimadzu Scientific Instruments (Columbia, MD) SIL-HTc.
7
HPLC Separation and Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The compounds separation was performed by an HPLC Shimadzu prominence with a C18 column Shim-Pac GIST Shimadzu (150 mm, 4.6 mm, 5 μm) supported by a binary pump (LC-20AD) and an auto-sampler (SIL-HTC, Shimadzu, Japan). The diode array detector (DAD SPD M-20A) was set for compound identification using a threedimensional (3D) scan mode in the wavelength range from 190 to 350 nm. While for compounds quantification, a channel of 260, 280, and 320 nm was selected. This chromatographic system was managed using LabSolutions software. The mobile phases consisted of phase A containing 2% acetic acid, 5% methanol in water, and mobile phase B containing 2% acetic acid, 88% methanol in water. Both phases were filtered using a 45 µm nylon filter then were sonicated in an ultrasound bath for 20 min before use.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!