JA and JA-IL levels were quantified using an ethyl acetate extraction method in conjunction with HPLC/MS similar to that described in Chung et al. (2008 (link)). Briefly, samples (approximately 150 mg tissue) were frozen in liquid nitrogen and hormones were extracted using 1 mL of extraction solvent (80:20 methanol:water + 0.1% formic acid) for 18 h at −20 C. Extracts were then centrifuged (10,000 × g for 10 min at 4°C) and the supernatant was transferred to autosampler vials. Five μL of each supernatant were injected into a Waters UPLC BEH C18 column (2.1 × 50 mm; 1.7 μm particles) held at 50°C on a Waters (Milford, MA, USA) Acquity ultraperformance liquid chromatography (UPLC) system that was coupled to a Waters Quattro Premier XE tandem quadrupole mass spectrometer. Separation was performed using a linear gradient based upon 0.15% aqueous formic acid (A) and methanol (B) over a 3-min program using a total flow rate of 0.4 mL/min. Quantification of JA and SA was performed using electrospray ionization in negative-ion mode using multiple reaction monitoring (MRM), using m/z 209 ≥ 59 for JA, m/z 322 ≥ 130 for JA-IL (Chung et al., 2008 (link)), and m/z 137 ≥ 93 for SA (Zeng et al., 2011 (link)). Peak areas were integrated, and calibration curves generated, using Waters QuanLynx software.
Quattro premier xe tandem quadrupole mass spectrometer
Manufactured by Waters Corporation
Sourced in United States
The Quattro Premier XE Tandem Quadrupole Mass Spectrometer is a laboratory instrument designed for the detection and analysis of chemical compounds. It uses a combination of quadrupole mass analyzers to separate and identify different molecules based on their mass-to-charge ratio.
Automatically generated - may contain errors
3 protocols using quattro premier xe tandem quadrupole mass spectrometer
1
Quantifying Jasmonates and Salicylates
2
Acylcarnitine Profiling in Muscle Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Two to 4 mg freeze-dried muscle tissue was homogenized in 1 mL 80% acetonitrile containing 50 µL internal standards. After centrifugation (5 min, 16000 × g, 4°C), the resulting supernatant was dried under a stream of nitrogen at 40°C and derivatized by addition of 1-propanol/acetylchloride (4:1, vol:vol) for 15 min at 60°C. After evaporation under nitrogen at 40°C, samples were redissolved in pure acetonitrile. Determination of the propylated acylcarnitines in the medium was performed by MS in an Acquity UPLC System (Waters) coupled to a Quattro Premier XE Tandem Quadrupole Mass Spectrometer (Waters).
3
Skeletal Muscle Acylcarnitines Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We used 2–4 mg of freeze-dried muscle tissue to analyze skeletal muscle acylcarnitines, as previously described (41 (link)). Quantitative determination of the propylated acylcarnitines in the medium was performed using tandem MS in an Acquity UPLC System (Waters) coupled to a Quattro Premier XE Tandem Quadrupole Mass Spectrometer (Waters). The internal standards d3-0, d3-3, d3-6, d3-8, d3-10, and d3-16 carnitine were purchased from Dr. Herman J. ten Brink (Vrije Universiteit Medical Hospital).
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!