Agilent MassHunter Quantitative Data Analysis - Batch Table, Compound Information Setup, Calibration Curve and Globals Settings
Souhrn
Význam tématu
MassHunter Quantitative Analysis je klíčovým nástrojem pro kvantitativní analýzu hmotnostních spekter v chemii, farmaceutickém výzkumu, ochraně životního prostředí i v průmyslové kontrolní kvalitě. Umožňuje komplexní nastavení metodiky, řízení dávkování standardů, detailní zpracování dat a zajišťuje reprodukovatelné a validovatelné výsledky.
Cíle a přehled studie / článku
Cílem prezentace je představit hlavní moduly a pracovní postupy v softwaru MassHunter Quantitative Analysis, včetně orientace v Batch Table, nastavení informací o sloučeninách, tvorby kalibračních křivek, práce ve “Compounds at a Glance” a správy globálních parametrů metody.
Použitá metodika a instrumentace
Software podporuje zpracování dat z různých typů hmotnostních analyzátorů:
- Triple quadrupole (QQQ) pro MS/MS kvantifikaci
- Time-of-Flight (TOF) a QTOF pro přesnou měrnou hmotnost
- Nastavení okna extrakce iontů (Mass Extraction Setup)
- Definici přechodů (precursor → produkt ion)
- Retenční časy, polarity iontů a kvalifikační kritéria
- Konfiguraci standardního přídavku, interního standardu a multiplikátorů dávkování
Hlavní výsledky a diskuse
Software nabízí tři klíčová zobrazení:
- Batch-at-a-Glance pro rychlý přehled výsledků
- Method Editor pro editaci metod a globálních parametrů
- Compounds-at-a-Glance pro vysokopropustné hodnocení jednotlivých sloučenin napříč vzorky
Přínosy a praktické využití metody
MassHunter Quantitative Analysis nabízí:
- Zvýšenou produktivitu díky přehlednému uživatelskému rozhraní
- Flexibilitu v definici vzorků a standardů (skupiny vzorků, multiplikátory)
- Automatizované a manuální nástroje pro integraci píků
- Podporu validace metod podle regulativních požadavků
Budoucí trendy a možnosti využití
Vývoj softwaru bude směřovat k ještě větší automatizaci AI-podporovaných funkcí, adaptivnímu řízení kvality dat a rozšíření integrace s LIMS a cloudovými platformami. Možnosti využití se posunují k real-time analýze a prediktivní údržbě přístrojů založené na statistické analýze výkonu.
Závěr
Agilent MassHunter Quantitative Analysis představuje robustní a flexibilní prostředí pro kvantitativní zpracování dat z hmotnostních spektrometrů. Díky širokému spektru nastavení, dynamickým nástrojům pro integraci a pokročilé podpoře kalibrací je ideálním řešením pro moderní laboratoře vyžadující vysokou přesnost a spolehlivost.
Reference
- Agilent Technologies: MassHunter Quantitative Analysis Webinar Series, 1/17/2018
Agilent MassHunter
Quantitative Data
Analysis
Presenters: Howard Sanford
Stephen Harnos
MassHunter Quantitation:
Batch Table, Compound
Information Setup,
Calibration Curve and
Globals Settings
1/17/2018
MassHunter Webinar Series
1
MassHunter Quantitative Software
Review and Quant Method Optimization
Topics
• Brief Review
• Batch Table Navigation
• Compound Information
- Data review and manual integration
• Calibration Curve
• Working in Compounds At a Glance
• Method Editor Globals Setting
1/17/2018
MassHunter Webinar Series
2
Review
Three major views in MassHunter Quantitative Analysis.
• Batch At a Glance
• Method Editor
• Compounds At a Glance
Handling MS/MS data QQQ and QTOF.
Handling accurate mass data TOF and QTOF.
1/17/2018
MassHunter Webinar Series
3
MassHunter Quantitative Software
Batch-at-a-Glance View
1/17/2018
MassHunter Webinar Series
4
Batch Table
Calibration Curve
Compound Information
Method
Tasks
MassHunter Quantitative Software
Method Editor View
Method > Edit menu or F10 key
1/17/2018
MassHunter Webinar Series
5
Method Table
Sample
Information
Compound Information
MassHunter Quantitative Software
Compounds at a Glance View
View > Compounds-at-a-Glance
1/17/2018
MassHunter Webinar Series
6
Editing a Quantitation Method for TOF
Mass Extraction Setup
Method > Edit > Advanced Tasks > Mass Extraction Setup
Allows for a mass range for the extraction of the accurate mass (MZ).
Available MZ Extraction Window Units
1/17/2018
MassHunter Webinar Series
7
Editing a Quantitation Method for QTOF
Compound Setup & Mass Extraction Setup
QTOF is a combination of MS/MS and accurate mass data
Name
– Compound name
TS
– time segments may be multiple
Transition
– Precursor ion Product ion scan
Scan
– Product Ion
Type
– Target, ISTD, Surrogate or Matrix Spike
Precursor Ion
– mass of the ion
Product Ion
– mass of the ion for the target ion to monitor
RT
– retention time of compound of interest
Ion Polarity
– usually positive
Criteria
– Close RT, Close RT with Qualifiers, Greatest Response or
Greatest Q-Value
1/17/2018
MassHunter Webinar Series
8
Batch Table
Navigation
Click in a sample row to display data on that sample
Click on Next and Previous icons to move through a batch or use
Hotkeys:
Next Sample
= Alt + Down
Next Compound
= Alt + Right
Previous Sample= Alt + Up
Previous Compound
= Alt + Left
Compound list are frequently long, try the Compound drop down list to go
directly to the desired compound.
1/17/2018
MassHunter Webinar Series
9
Batch Table
Alphabetizing the Compound List
From within the Batch Table
right click, select Arrange
Compounds By > Name
1/17/2018
MassHunter Webinar Series
10
Batch Table
Messages and Outliers
Messages
Outliers
Red Outlier
– High
(above upper limit)
Select
Outliers
for Display
Blue Outlier
– Low
(below lower limit)
Hover cursor over the outlier or message to display details
1/17/2018
MassHunter Webinar Series
11
Batch Table Layout Modification
Add/Remove/Move Columns
Use the Add/Remove Columns function to
customize the Batch table.
Note that the columns of data in the Batch
Table are organized into sections.
Columns can be moved only within a section.
1/17/2018
MassHunter Webinar Series
12
Batch Table Layout Modification
Single and Multiple Compound Modes
Single Compound Mode
Multiple Compound Mode
1/17/2018
MassHunter Webinar Series
13
Flat Table compounds
across the top.
Batch Table Layout Modification
Compound Table Modes
Compound Table lists by compound rather than sample.
Select Flat Table to see list by sample.
1/17/2018
MassHunter Webinar Series
14
Batch Table
Compounds Groups
It may be useful to group compounds to organize Batch Table.
Look at parent compound and metabolites.
Group hydrocarbons to Group 1, aromatics to Group 2 and so forth.
Compound groups are generated in the Method Editor.
1/17/2018
MassHunter Webinar Series
15
Batch Table
Compounds Groups
Yields an Additional Toolbar.
Activated with a right click
to the right of the ‘flags’ or View
> Toolbars > Filtering.
1/17/2018
MassHunter Webinar Series
16
Batch Table
Samples Groups
Sample Groups are specified in the Batch Table through Add/Remove
Columns.
Each Sample is
assigned to a
group, then only
samples specific
to a group are
displayed and
evaluated.
Samples groups are distinctively different from compound groups.
1/17/2018
MassHunter Webinar Series
17
Batch Table Layout Modification
Format Columns
How to change the
number of decimal
places.
Alter number formats
1) Exponential
(scientific notation)
2) Fixed point
3) General
4) Significant Figures
Alter Date formats
1/17/2018
MassHunter Webinar Series
18
Batch Table Layout Modification
Context Menu Properties
Change the Font size
Change the Color scheme
1/17/2018
MassHunter Webinar Series
19
Export Batch Table
Easy to layout columns and format data, then export into Excel.
1/17/2018
MassHunter Webinar Series
20
Let’s take a moment
for questions on
Batch Table
Navigation.
1/17/2018
21
MassHunter Webinar Series
Time for a demo on
Batch Table
Navigation
Next up:
Compound Information
1/17/2018
22
MassHunter Webinar Series
Compound Information
Target
Quantifier
ISTD
Quantifier
Target
Qualifiers
ISTD Qualifiers
Target
Spectrum
ISTD
Spectrum
Display and access one compound in one sample at a time.
1/17/2018
MassHunter Webinar Series
23
Compound Information - Toolbar
Sample
and
Compound
Navigation
Next /
Pervious
Un-zooms,
Peak scaling,
Auto Scales x
and y axes
Manual Integration
Functions
Display/Hide
Target,
Qualifiers,
Spectra,
ISTD
1/17/2018
MassHunter Webinar Series
24
Manual Integrations and MI Toolbar
Click on the Manual Integration icon to place Compound Information in
Manual Integration mode.
Each signal is placed in its own window (no overlay of qualifiers).
1/17/2018
MassHunter Webinar Series
25
Manual Integration Toolbar
Enable Manual Integration
– this
tool activates the manual integration
toolbar
.
Clear Manual Integration
– this tool
becomes active once a manual
integration is present.
Zero Peak
– this tool sets start and
stop to the same value.
Merge Right/Left Peak
– this tool
merges the peak to the right/left of
the selected peak.
Split Peak and Pick Right/Left
–
this tool splits the peak and then
selects the right or left peak.
Snap Baseline
– this tool places
start/stop integration points on the
baseline.
Drop Baseline
– this tool finds the
lowest end of the peak, then drops
the baseline on the other side of the
peak from it, thereby creating a flat
baseline that avoids negative area.
1/17/2018
MassHunter Webinar Series
26
Compound Information
Peak Annotation
Activated from Tools > Add-
Ins…
NI
– The peak was not integrated at all by the
computer software.
LT
– The peak in question was inappropriately
integrated to an area less than what it should be
(e.g., Peak area was cut).
GT - The peak in question was inappropriately
integrated to an area greater than what it should be
(e.g., Peak Tailing).
BA
– The baseline had to be adjusted correctly by the analyst.
CO
–The analyst had to split to co-eluting peaks apart that were not (or could
not be) separated by the computer system.
RT
– The retention time for the peak in question has shifted from the expected
retention time.
INT
– There was electronic interference (e.g., Noise).
1/17/2018
MassHunter Webinar Series
27
Compound Information
Peak Annotation
The editable file is located in C:\Program
Files\Agilent\MassHunter\Workstation\Quant\bin\AddIns\PeakAnnotations.xml.
1/17/2018
MassHunter Webinar Series
28
Compound Information
Peak Annotation
The Peak Annotation can also be displayed in the Batch Table.
When the batch is saved, the values are retained.
The MI flag is checked in the Compound
Results.
1/17/2018
MassHunter Webinar Series
29
Compound Information
Context Menu Integration Parameters
Right click to expose the
context menu.
Most of the features are toolbar
icons.
Integration Parameters can be
used to change the integration
parameters.
Applies ONLY to this compound
in this sample.
Restore Integration Parameters
reverts to the method
integration values.
1/17/2018
MassHunter Webinar Series
30
Compound Information
Context Menu Properties
Properties allows for
customization of the display.
Can change Fill colors.
Can change Peak labels.
Can change Titles.
1/17/2018
MassHunter Webinar Series
31
Compound Information
Context Menu Properties
Qualifier Properties can be
changed.
Spectrum Properties can be
altered.
Manual integration
parameters can be varied.
1/17/2018
MassHunter Webinar Series
32
Signal to Noise
Five algorithms are available
Consult online help for more
information on the algorithms.
Noise regions can be
automatically determined or
individually specified.
The Noise regions can be
displayed in the Compound
Information window under
Properties > Compound
Information (1) > Baseline
Calculation Points.
In this example, 2 noise regions
were determined
—one before
the peak and the other after.
1/17/2018
MassHunter Webinar Series
33
Let’s take a moment
for questions on
Compound
Information
1/17/2018
34
MassHunter Webinar Series
Let’s take a moment
for a demo on the
features of Compound
Information
Next up:
Calibration Curve and
Curve Fit Assistant
1/17/2018
35
MassHunter Webinar Series
Calibration Curve
View ISTD Responses
Change Curve Fit.
R2
Fit to
Levels
(Scaling)
Concentration can be set as relative (to ISTD) or actual.
QC Samples
CC Levels
1/17/2018
MassHunter Webinar Series
36
Calibration Curve
Disable calibration points
Click on calibration point once to disable and remove from curve.
1/17/2018
MassHunter Webinar Series
37
Curve Fit Assistant
1/17/2018
MassHunter Webinar Series
38
Curve Fit Assistant
Sorting Columns
Curve Fit Assistant calculates the mathematical curves.
Best curve fit may be ranked using by
R2, Standard Error, or Max % Residual.
1/17/2018
MassHunter Webinar Series
39