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2
Sample preparation / Re: Mice brain preparation for HPLC
Last post by Alexandra -
Hi! Your question has been tweeted by the MetSoc EMN (https://twitter.com/EMN_MetSoc/status/1382607928599965696)and received the following answers:

CAROLINA GONZALEZ: we use an extraction method for multiplataform analyses. We add MeOH:H2O 50% (1mg tissue:10uL solvent) for Mouse face Brain homogenization using a TissueLyser. Then, we mix 100uL of brain homogenate with 320uL MeOH + 80uL MTBE (for lipids) DOI: 10.1021/acs.jproteome.8b00816.

WENYUN LU: For tissue samples our preferred extraction method is first grind into powder then add 40:40:20 methanol:acetonitrile:h2o+0.5% formic acid then neutralize with 15% NH4 HCO3 followed by two rounds of centrifugation.
https://doi.org/10.1089/ars.2017.7014

Feel free to follow up with further Qs here or there!
3
Other / Progenesis QI and HMDB spectra
Last post by gregopim -
Hello everyone,
By any chance, does anyone have found a way to import the HMDB spectra library in Progenesis?
Progenesis only support .msp files for spectra libraries, but HMDB only provides .xml or .txt versions (https://hmdb.ca/downloads).
Is there a way to convert .xml files to .msp?
Grégory
4
MS-DIAL / Re: Not all my internal standards are detected
Last post by BBonnefille -
Hi Hiroshi,

I did all my samples data processing following the exact parameters you gave me, but I still have problems with my IS:
- One of my IS is still not found by MS-DIAL (one of those that were missing previously)
- The other detected by MS-DIAL, which is better. But the identification using a .txt file as in-house library did not work, and the feature in the ion table present a high mass deviation compared to my IS mass (approx 13 ppm instead of < 5 ppm when I am checking with XCalibur).

One of the group I am processing a few samples were not spiked with the IS(3 vs 28 samples spiked). Do you think it can be related (even if it should not)?

Thank you for your help,
Bénilde
6
MS-DIAL / Re: Not all my internal standards are detected
Last post by Hiroshi Tsugawa -
Hi,

I just checked your positive ion mode data. It works fine.
Of course, several IS compounds were not detected owing to the structure property.
However, at least, I could detect the peaks that you wrote as "Not detected by MS DIAL in ESI pos" in the excel.

I also pasted the parameters that I used. Thanks,

Hiroshi
MS-DIAL ver. 4.60-dev

#Project
MS1 Data type   Profile
MS2 Data type   Profile
Ion mode   Positive
Target   Metablomics
Mode   ddMSMS

#Data collection parameters
Retention time begin   0
Retention time end   100
Mass range begin   0
Mass range end   2000
MS2 mass range begin   0
MS2 mass range end   2000

#Centroid parameters
MS1 tolerance   0.01
MS2 tolerance   0.025

#Isotope recognition
Maximum charged number   2

#Data processing
Number of threads   1

#Peak detection parameters
Smoothing method   LinearWeightedMovingAverage
Smoothing level   3
Minimum peak width   5
Minimum peak height   10000

#Peak spotting parameters
Mass slice width   0.1
Exclusion mass list (mass & tolerance)

#Deconvolution parameters
Sigma window value   0.5
MS2Dec amplitude cut off   0
Exclude after precursor   True
Keep isotope until   0.5
Keep original precursor isotopes   False

#MSP file and MS/MS identification setting
MSP file   
Retention time tolerance   100
Accurate mass tolerance (MS1)   0.01
Accurate mass tolerance (MS2)   0.05
Identification score cut off   80
Using retention time for scoring   False
Using retention time for filtering   False

#Text file and post identification (retention time and accurate mass based) setting
Text file   E:\0_SourceCode\BugReports\20210414_GrosFich\pos_mzrt_lib.txt
Retention time tolerance   0.5
Accurate mass tolerance   0.01
Identification score cut off   85

#Advanced setting for identification
Relative abundance cut off   0
Top candidate report   False

#Adduct ion setting
[M+H]+

#Alignment parameters setting
Reference file   E:\0_SourceCode\BugReports\20210414_GrosFich\210322_SWS_QC4_ESIpos_14.raw
Retention time tolerance   0.05
MS1 tolerance   0.015
Retention time factor   0.5
MS1 factor   0.5
Peak count filter   0
N% detected in at least one group   0
Remove feature based on peak height fold-change   False
Sample max / blank average   5
Sample average / blank average   5
Keep identified and annotated metabolites   True
Keep removable features and assign the tag for checking   True
Gap filling by compulsion   True

#Tracking of isotope labels
Tracking of isotopic labels   FALSE

#Ion mobility
Ion mobility data   FALSE

7
MS-DIAL / How to analyze Agilent 6560 ion mobility AIF data?
Last post by kljw001 -
Hi,

I have a question about using MSDial to analyze the Agilent 6560 ion mobility AIF data. For example, in attached picture 1, when I select chromatography as the separation type, "SWATH-MS" and "All ion with multiple CES" option are available in the MS Method type.

However, in attached picture 2, when I select the "Ion mobility" in the separation type, there is no option about "all ion with multiple CES"

Thus, I am wondering how to use the MSDial to analyze the data which run under ion mobility and AIF using Agilent 6560.

Thanks a lot

 
9
MS-DIAL / Re: Not all my internal standards are detected
Last post by BBonnefille -
Hi Hiroshi,

Here is a link to download two of my .raw data and an excel file with the information you requested. I have the same problem in ESI negative, but with more IS not picked up by MS DIAL (7/24 not found), so I attached a .raw file in both modes:
grosfi.ch/adm4hAq6bpD

Thank you for your help,
Bénilde