FlavonQ-Data Analysis
Profiling flavonoids in natural products poses great challenges due to the diversity of flavonoids, the lack of commercially available standards, and the complexity of food matrices. Data-mining of ultra-high-performance liquid chromatography diode array detection high-resolution accurate-mass mass spectrometry (UHPLC HRAM-MS) for the analysis of flavonoids is very daunting, labor-intensive, and expertise-dependent. FlavonQ is an automated data processing tool that transfers field-acquired expertise into flavonoid data analysis.
You can analyze your acquired data by FlavonQ. We are in the process of populating our databases, if you want your analysis to be included in the databases, please contact the administrator. Your support for FlavonQ is greatly appreciated.
Instructions
Start data analysis, please click to link.
Download
test data (note: test data are about 655 mb)Data format:
MS data format must be .raw or any other formats that can be imported into ProteoWizard (Note: set Binary precision for 32-bit and uncheck the 'zlib compression' option). MSGet was used as a tool that exports PDA data in text files from the binary .raw data of Xcalibur software (ThermoFisher). Xcalibur is the control and data acquisition software for the mass spectrometers of ThermoFisher. To convert .mzXML and text files to mat. files, please use data conversion tools here. If version 9.10 (R2021a) of the MATLAB Runtime is installed on the computer, please download and use the UV2MAT/MS2MAT executable directly (about 1.4 mb each); otherwise, please download the installer files (about 830 mb each). Both MS and PDA data are required and MS must be collected from a high resolution MS.Example instrument settings:
A standard UHPLC condition is used to develop FLavonQ The separation was carried out on a UHPLC column (200 mm 2.1 mm i.d., 1.9 um, Hypersil Gold AQ RP-C18) (Thermo-Scientific) with an HPLC/UHPLC precolumn filter (UltraShield Analytical Scientific Instruments, Richmond, CA) at a flow rate of 0.3 mL/min. The mobile phase consisted of a combination of A (0.1% formic acid in water, v/v) and B (0.1% formic acid in acetonitrile, v/v). The linear gradient was from 4 to 20% B (v/v) at 40 min, to 35% B at 60 min, and to 100% B at 61 min and held at 100% B to 65 min.The PDA was set at 520, 330, and 280 nm to record the peaks, and UV/vis spectra were recorded from 200 to 700 nm.
Both positive and negative ionization modes were used, and the conditions for the LTQ-Orbitrap were set as follows: sheath gas at 70 (arbitrary units), auxiliary and sweep gas at 15 (arbitrary units), spray voltage at 4.8 kV, the capillary temperature at 300 C, the capillary voltage at 15 V, and tube lens at 70 V. The mass range was from m/z 200 to 2000 with a resolution of 15000, FTMS AGC target at 2e5, FT-MS/MS AGC target at 1e5, isolation width of 1.5 amu, and maximum ion injection time of 500 ms. The most intense ion was selected for the data-dependent scan to offer their MS2 , MS3 , and MS4 product ions with a normalization collision energy at 35%. The conditions were given as references only. However, the closer your instrument setting is to the condition mentioned above, the more accurate the results will be.
The workflow of FlavonQ and some tips for how to use FlavonQ are listed below:
Smoothing is important for reliable peak detection. Several smoothing filters are available in FlavonQ including un-weighted sliding average filter, weighted sliding average filter, pseudo-Gaussian smooth and the Savitzky-Golay filter.
Among the various classes of flavonoids, some subclasses share a characteristic absorption band in UV, e.g. flavones and flavonols contain the cinnamoyl structure in the B and C ring has UV absorption in the range from 305 to 390 nm; proanthocyanidins have characteristic UV absorbance at 274-280 nm. The similarity of the UV-vis spectra from 200 to 600 nm between all the other peaks and the reference peak can be evaluated, and flavonoids peaks can be automatically extracted by FlavonQ.
