6495 qqq ms

Manufactured by Agilent Technologies

Sourced in United States

The 6495 QQQ-MS is a triple quadrupole mass spectrometer designed for high-performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography (UHPLC) applications. It features a high-sensitivity, high-speed quadrupole mass analyzer that enables accurate quantification and identification of target compounds in complex samples.

Automatically generated - may contain errors

Lab products found in correlation

1290 infinity 2 uhplc, by Agilent Technologies (4 mentions) Masshunter software, by Agilent Technologies (2 mentions) Luna propylamine column, by Phenomenex (1 mentions) Masshunter b 07, by Agilent Technologies (1 mentions) 1290 hplc system, by Agilent Technologies (1 mentions) Advancebio peptide map column, by Agilent Technologies (1 mentions) 1290 infinity 2, by Agilent Technologies (1 mentions) Nucleoside digestion mix, by New England Biolabs (1 mentions) Nitrocellulose, by Bio-Rad (1 mentions)

Spelling variants of this product

6495 Triple Quad QQQ-MS (6 citations)

8 protocols using 6495 qqq ms

Extracellular Nucleotide Quantification

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Supernatants of HUVEC and PMNs previously incubated with ABC (20 µmol/L, 37°C) were collected (3 and 8 min and 4 h), mixed with MeOH (10 min, 4°C) and centrifuged (10 min, 15.000 rpm). 100 µL of the respective samples were dried under N₂ flow prior to the addition of H₂O and subsequent analysis of the extracellular nucleotides by means of a 1200 Series uHPLC coupled to a 6495 QqQ/MS (Agilent Technologies) (Chen et al., 2009 (link); Zhang et al., 2014 (link)).
To determine the levels of CBV-TP, supernatants of HUVEC and PMNs previously incubated with ABC (20 µmol/L, 37°C) were collected (at 4 h) and treated with phosphatase to degrade the CBV-TP to CBV, and CBV-TP was measured as total CBV (CBV-TP + CBV) by uHPLC coupled to a 6495 QqQ/MS (Agilent Technologies).

Collado-Díaz V., Martinez-Cuesta M.Á., Blanch-Ruiz M.A., Sánchez-López A., García-Martínez P., Peris J.E., Usach I., Ivorra M.D., Lacetera A., Martín-Santamaría S., Esplugues J.V, & Alvarez A. (2021). Abacavir Increases Purinergic P2X7 Receptor Activation by ATP: Does a Pro-inflammatory Synergism Underlie Its Cardiovascular Toxicity?. Frontiers in Pharmacology, 12, 613449.

+ Open protocol

+ Expand

Targeted Metabolite Quantification by LC-MS

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were centrifuged for 2 min at 500g at 4 °C. The pellet was washed with ice-cold PBS and centrifuged for 2 min at 500g at 4 °C. The supernatant was discarded. Samples were extracted in 750 μl 50:30:20 v/v/v methanol/acetonitrile/water, and samples were centrifuged for 10 min at maximum speed at 4 °C. The supernatant was stored at −80 °C. Targeted metabolite quantification by LC–MS was carried out using an Agilent 1290 Infinity II UHPLC in-line with an Agilent 6495 QQQ-MS operating in multiple reaction monitoring (MRM) mode. MRM settings were optimized separately for all compounds using pure standards. Liquid chromatography separation was on a Phenomenex Luna propylamine column (50 × 2 mm, 3-μm particles) using a solvent gradient of 100% buffer B (5 mM ammonium carbonate in 90% acetonitrile) to 90% buffer A (10 mM NH₄ in water). Flow rate was from 1,000 to 750 μl min⁻¹. Autosampler temperature was 5 °C and injection volume was 2 μl. Data processing was performed by an R script developed in-house.

Kelly B., Carrizo G.E., Edwards-Hicks J., Sanin D.E., Stanczak M.A., Priesnitz C., Flachsmann L.J., Curtis J.D., Mittler G., Musa Y., Becker T., Buescher J.M, & Pearce E.L. (2021). Sulfur sequestration promotes multicellularity during nutrient limitation. Nature, 591(7850), 471-476.

+ Open protocol

+ Expand

Plasma S1P Quantification Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Plasma S1P was determined as previously described (Burla et al., 2018 (link)). Briefly, lipids were extracted by mixing 20 µL of plasma with 200 µL butanol/methanol (1:1, v/v) solution containing an internal S1P d18:1 ¹³C2D2 standard (Toronto Research Chemicals, S681502), followed by derivatization of the lipids by a trimethylsilyl-diazomethane derivatization step (Narayanaswamy et al., 2014 (link)). Lipids were then separated on an Agilent 1290 UHPLC, using a Waters ACQUITY UPLC BEH HILIC, C18 (130 Å, 2.1 × 100 mm, 1.7 μm) column, and measured in an Agilent 6495 QQQ MS. The mobile phase A consisted of acetonitrile and 25 mM ammonium formate buffer (50:50, v/v) and mobile phase B was composed of acetonitrile and 25 mM ammonium formate buffer (95:5, v/v).
The Agilent MassHunter software was used to analyze the data, with lipid peaks being identified based on the specific precursor and product ion transitions, as well as the retention time. Finally, normalization to the internal standards was carried out as previously discussed (Burla et al., 2018 (link)).

Bisgaard L.S., Christensen P.M., Oh J., Torta F., Füchtbauer E.M., Nielsen L.B, & Christoffersen C. (2024). Kidney derived apolipoprotein M and its role in acute kidney injury. Frontiers in Pharmacology, 15, 1328259.

+ Open protocol

+ Expand

Comprehensive Metabolite Quantification by LC-MS

Check if the same lab product or an alternative is used in the 5 most similar protocols

Three
different chromatographic methods were
used. All targeted metabolite quantifications by LC-MS were carried
out using an Agilent 1290 Infinity II UHPLC in line with an Agilent
6495 QQQ-MS operating in MRM mode (HILIC method 1) or DMRM mode (HILIC
method 2 and reversed-phase method). MRM transitions were optimized
separately for all compounds using pure standards or inferred from
closely related compounds. Both data sets HILIC 1a and HILIC 1b were
recorded using HILIC method 1 but with different sets of target compounds.

Eilertz D., Mitterer M, & Buescher J.M. (2022). automRm: An R Package for Fully Automatic LC-QQQ-MS Data Preprocessing Powered by Machine Learning. Analytical Chemistry, 94(16), 6163-6171.

+ Open protocol

+ Expand

Targeted Metabolite Quantification by LC-MS

Check if the same lab product or an alternative is used in the 5 most similar protocols

Targeted metabolite quantification by LC–MS was carried out using an Agilent 1290 Infinity II UHPLC in line with an Agilent 6495 QQQ-MS operating in MRM mode. MRM settings were optimized separately for all compounds using pure standards. LC separation was on a Phenomenex Luna propylamine column (50 × 2 mm, 3 μm particles) using a solvent gradient of 100% buffer B (5 mM ammonium carbonate in 90% acetonitrile) to 90% buffer A (10 mM NH₄ in water). Flow rate was from 1,000 to 750 μl min⁻¹. Autosampler temperature was 5 °C and injection volume 2 μl. Peak areas were measured using MassHunter B.07.01 (Agilent).

Baixauli F., Piletic K., Puleston D.J., Villa M., Field C.S., Flachsmann L.J., Quintana A., Rana N., Edwards-Hicks J., Matsushita M., Stanczak M.A., Grzes K.M., Kabat A.M., Fabri M., Caputa G., Kelly B., Corrado M., Musa Y., Duda K.J., Mittler G., O’Sullivan D., Sesaki H., Jenuwein T., Buescher J.M., Pearce E.J., Sanin D.E, & Pearce E.L. (2022). An LKB1–mitochondria axis controls TH17 effector function. Nature, 610(7932), 555-561.

+ Open protocol

+ Expand

Quantification of OATP1B1 Protein Expression

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

The LC–MS/MS-based
QTAP approach was used to quantify the absolute protein expression
of OATP1B1 in the crude membrane preparations. The method for the
protein sample preparation and quantitation using targeted LC–MS
is described earlier.^{14 (link),30 (link),31 (link)} OATP1B1 and Na⁺/K⁺ ATPase signature peptides
were quantified from 50 μg of crude membrane fractions. After
denaturation and breakdown of the tertiary structure of the proteins,
the crude membrane preparations were digested first with 1/100 LysC
endopeptidase and after that with 1/100 TPCK-treated trypsin. A previously
used isotope-labeled peptide mixture (3 fmol/μg protein) served
as an internal standard.^{14 (link)} Quantification
was conducted on a 6495 QQQ MS with a 1290 HPLC system and an AdvanceBio
peptide Map Column, 2.7 μm, 2.1 × 250 mm (Agilent Technologies,
Santa Clara, CA, USA) as described previously.^{14 (link),31 (link)} The SNVs did not alter the amino acids in the analyzed peptide sequences.
The peak area ratios of the analyte peptides and their respective
internal standards were compared using the Skyline application (MacCoss
Lab Software, Seattle, WA). The results of OATP1B1 expression are
presented as relative to the Na⁺/K⁺–ATPase
expression level and normalized to the reference OATP1B1 protein.
The absolute amount of OATP1B1 in proteomics samples is presented
in Supporting Information Table S2 and
Figure S1.

Häkkinen K., Kiander W., Kidron H., Lähteenvuo M., Urpa L., Lintunen J., Vellonen K.S., Auriola S., Holm M., Lahdensuo K., Kampman O., Isometsä E., Kieseppä T., Lönnqvist J., Suvisaari J., Hietala J., Tiihonen J., Palotie A., Ahola-Olli A.V, & Niemi M. (2023). Functional Characterization of Six SLCO1B1 (OATP1B1) Variants Observed in Finnish Individuals with a Psychotic Disorder. Molecular Pharmaceutics, 20(3), 1500-1508.

+ Open protocol

+ Expand

Quantitative Cellular Metabolomics

Check if the same lab product or an alternative is used in the 5 most similar protocols

A total of 250 000 cells were washed in 3% glycerol solution and extracted in 10 μl of 80% acetonitrile or 100 μL of 80% cold methanol containing 13C yeast extract as an internal standard. The samples in methanol were vacuum concentrated. Metabolite quantification by liquid chromatography–tandem mass spectrometry was carried out using an Agilent 1290 Infinity II ultra-high-performance liquid chromatography in line with an Aglient 6495 QQQ-MS operating in MRM mode.

Zhang Y.W., Velasco-Hernandez T., Mess J., Lalioti M.E., Romero-Mulero M.C., Obier N., Karantzelis N., Rettkowski J., Schönberger K., Karabacz N., Jäcklein K., Morishima T., Trincado J.L., Romecin P., Martinez A., Takizawa H., Shoumariyeh K., Renders S., Zeiser R., Pahl H.L., Béliveau F., Hébert J., Lehnertz B., Sauvageau G., Menendez P, & Cabezas-Wallscheid N. (2023). GPRC5C drives branched-chain amino acid metabolism in leukemogenesis. Blood Advances, 7(24), 7525-7538.

+ Open protocol

+ Expand

DNA Methylation Analysis by Dot Blot and LC-MS

Check if the same lab product or an alternative is used in the 5 most similar protocols

Genomic DNA was purified by phenol/chloroform extraction and ethanol precipitation according to standard procedures. DNA dot blot for 5mC was performed according to standard procedures^{51 (link)}. DNA was spotted on nitrocellulose (Biorad) in 2× SSC and UV-cross-linked at 254 nm, 120 mJ/cm². Membranes were probed with anti-5-methylcytosine antibody (Zymo research, clone 10G4, diluted 1:3000 in 3% BSA/PBST). For MS analysis of 5mC and 5hmC levels, genomic DNA was digested to nucleosides using Nucleoside Digestion mix (NEB, M0649S). Targeted nucleoside quantification by LC-MS was carried out using an Agilent 1290 Infinity II UHPLC inline with an Agilent 6495 QQQ-MS operating in MRM mode. MRM settings were optimized separately for unmodified nucleosides using pure standards and adapted for methylated nucleosides. LC separation was on a Waters T3 column (100 × 2.1 mm, 1.8-μm particles) using a solvent gradient of 100% buffer A (0.1% formic acid in water) to 90% buffer B (50:50 acetonitrile:methanol). Flow rate was 400 μL/min. Autosampler temperature was 4 °C, and the injection volume was 2 μL. Data processing was performed using Agilent MassHunter Software (version B.08).

Montavon T., Shukeir N., Erikson G., Engist B., Onishi-Seebacher M., Ryan D., Musa Y., Mittler G., Meyer A.G., Genoud C, & Jenuwein T. (2021). Complete loss of H3K9 methylation dissolves mouse heterochromatin organization. Nature Communications, 12, 4359.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!