Introduction to Chromatography software - Mohsin …...2017/03/04 · Introduction to...

©2007 Waters Corporation

Introduction to Chromatography software


Agenda

• Chromatography Theory

• Basic Chromatography Sequence

• Data Management

• Data Processing

• Reporting

• Software Option

• Instrument Control


Chromatography Theory

• Everything is relative– From Area ( Response) Concentration (Amount)– Or Amount = B x (Area)

• But what affects B?– Absorption capabilities of the molecule’s chemistry– UV lamp or other detector qualities (age, efficiency, energy, cell size)– Sensitivity of the recording device– Mobile phase (composition and flow rate)

• Therefore the pharmacopoeia specifies that you should run standards of known concentrations to determine B.– On the same LC– With the same conditions– On the same day– With the same integration algorithms

∝


Amount (sam) = Amount (std)

Response (sam) Response (std)

Amount (sam) = Amount (std) X Response (sam)

Response (std)

Quantitation Theory

First Principals

This is B:- the slope of a plot


Quantitation Theory

• Plot amount against response through origin

• For any measured Sample RESPONSE

• We can read off the appropriate Sample AMOUNT

Amount Standard

Res

pons

e


Identifying StandardsStandard Replicates

• How many standards identified as replicates in the Sample Set Method– No. of vials

Multiple vials of the same standard concentrationMultiple vials with different standard concentrations

– No. of injections

Sample Set Method


Quantitation Theory

• How do we plot the Std measurements?

• One strength solution multiple injections

• Similar strength single injection

• Multiple strengths

Amount Standard

Res

pons

e


Position of Standards

• Traditionally placed at the beginning of a run

• Response can change during a long sequence– Particularly with early detectors– With the long run times needed with previous generation

separation columns– Depends on solution stability

• Habit of placing standards through the run was established– Even when up to date columns and equipment removes the

need– Either “every 5 samples” or “every 45 minutes”


Performing CalibrationBracketing Standards

• No Bracket– Runs all standards first and

– Create Calibration curve

– Run all samples

– Quantitate Samples



• Overall Bracket– Standards are injected at the start

and end of the sample set.

– Entire sequence collected

– Standards Calibrated

– Samples Quantitated



• Modified Overall Bracket– Standards run AROUND GROUPS of n

samples

– Entire sequence collected

– Standards Calibrated

– Samples Quantitated



• Sliding, No Overlap– Standards run AROUND GROUPS of

n samples– One an entire group is collected

Calibrate the Stds of the groupQuantitate the Samples of the group

– Standards are used only once during calibration.



• Sliding, with Overlap– Standards run AROUND GROUPS

of n samples– One an entire group is collected

Calibrate the Stds of the groupQuantitate the Samples of the group

– Standards are used multiple times during calibration.


Performing CalibrationSliding Bracket-No Overlap

Bracket 1

Bracket 2

Bracket 3


Pre Sequence Checks or System Suitability

• Blank Injection

• Replicate Injections

• Test Quantitation – control samples


Basic Chromatographic Workflow Process


1. Login.2. Instrument Method.3. Sequence/Run Samples.4. Process Data.5. Review/Preview6. Print

1.

5.

Basic Sequence

2. 3.

4.

6.


Single Injection


Create Sample Set


Log In

• Log into Empower using your user name– demo user– sdms60

• Be sure you are in the QuickStart Interface


Methods

• Method Set– Instrument Method– Processing Method– Report Method– Export Method


Instrument Method



Collect Chromatographic Data


Processing Method



Process the Data to Create Results


Report Method



1. Login.2. Instrument Method.3. Sequence/Run Samples.4. Process Data.5. Review/Preview6. Print

1.

5.

Basic Sequence

2. 3.

4.

6.


Data Management


Empower 2 Data ViewsQuick Sort - Click on any column heading to sort ascending or descending


Empower 2 Data ViewsRaw Data = Sample Sets, Injections and Channels


Empower 2 Data Views

Sample Sets

Injections

Channels

AU

0.00

0.02

0.04

0.06

Minutes0.50 1.00 1.50 2.00 2.50 3.00

AU

0.00

0.02

0.04

0.06

Minutes0.50 1.00 1.50 2.00 2.50 3.00

AU

0.00

0.02

0.04

0.06

Minutes0.50 1.00 1.50 2.00 2.50 3.00

AU

0.00

0.02

0.04

0.06

Minutes0.50 1.00 1.50 2.00 2.50 3.00

AU

0.00

0.02

0.04

0.06

Minutes0.50 1.00 1.50 2.00 2.50 3.00

AU

0.00

0.02

0.04

0.06

Minutes0.50 1.00 1.50 2.00 2.50 3.00

A Sample Set contains multiple injectionsEach injection may contain multiple channels


Empower 2 Data ViewsSample Sets -> Injections -> Channels


Empower 2 Data ViewsMethods = Instrument, Processing, Report, Method Set and Sample Set



• Instrument Method - Specifies instrument control and data acquisition parameters, such as flow rate and wavelength

• Processing Method - Includes a set of instructions defining how Empower processes raw data

• Report Method - Serves as a template to organize the data, results, calibration curves, and method contents in a generated report

• Export Method - Specifies the data file format you want to use to export data.

• Method Set - User-defined collection of methods and data channels– Can include an instrument, processing, report and export method

• Sample Set Method - Contains a set of functions and related information that specifies the parameters to acquire data from each vial (standard or unknown) in a set of vials


Instrument Method

Wavelengths/Masses

Processing Method

Report Method

Sample Name

Standard or Sample

Vial – Run Time

Method Set to Use

Amounts of Standard

Sequence or Set of Samples to be Run



Empower 2 Data ViewsProcessed Data = Results and Result Sets


AU

0.00

0.02

0.04

0.06

Minutes0.50 1.00 1.50 2.00 2.50 3.00

0.51

4

0.88

3

1.31

6

2.03

7

AU

0.00

0.02

0.04

0.06

Minutes0.50 1.00 1.50 2.00 2.50 3.00

0.51

4

0.88

3

1.31

6

2.03

7

AU

0.00

0.02

0.04

0.06

Minutes0.50 1.00 1.50 2.00 2.50 3.00

0.51

4

0.88

3

1.31

6

2.03

7

AU

0.00

0.02

0.04

0.06

Minutes0.50 1.00 1.50 2.00 2.50 3.00

0.51

4

0.88

3

1.31

6

2.03

7


Result Sets

ResultsAU

0 . 0 0

0 . 0 2

0 . 0 4

0 . 0 6

M i n u t e s0 . 5 0 1 . 0 0 1 . 5 0 2 . 0 0 2 . 5 0 3 . 0 0

0.51

4

0.88

3

1.31

6

2.03

7

A Result Set contains multiple Results


View Filters

• View Filter - limits or customizes the types of information available for viewing in the Project window. – You can use View Filters to define the columns of information that appear

and their sequence and sorting. Empower provides several default view filters for project data.


View Filters


Find My Results


Custom Fields

Custom Fields - A custom field is a user-defined field in a table as opposed to a predefined field in Empower.Custom fields can be descriptors and formulas


Custom Fields

Descriptor Custom Field


Custom Fields

Formula Custom Field


Data Processing


Empower 2 Data Processing

Select a Sample Set, Injection or Channel and Click on Review



Click Integrate to perform default peak integration



Modify Integration Parameters or Add Integration Events



Manually Integration Peaks



Add Peak Names


Reporting


Empower 2 Reporting

Select a Result or Result Set and Click on Preview


Empower 2 Reporting

Select a Report Method from the list


Empower 2 Reporting

Edit Report Method


Empower 2 Reporting

Modify Peak Table


Empower 2 Options


Traditional Software Options

• Built In:– Photo Diode Array– Mass Spectrometry– CE/CIA


Traditional Built-in Option

Check peak purity and see the finest spectral details.

Single wavelength data,multi-wavelength data or complete spectra.

Integrated advance detection to minimize software training.

Photo Diode Array


Traditional Built-in Option

Data acquisition and instrument control of Mass Detectors.

Greater specificity of analysis.

Easily use MS and MS/MS in QC and Development lab environments.

Mass Detection

1st Chromatography Software to offer Tandem control and data processing in a network or

regulated environment


Traditional Software Option (Key Disk)

System SuitabilitySystem suitability testing is a means of checking that a chromatographic system and methodology are working within acceptable limits.

System Suitability calculations and limits are selected in the processing method and limits can be set so that a result will be flagged if it is outside the defined limit.


Integrity of your HPLC dissolution testing methods and results, helping increase lab productivity.

Combined software and hardware solution.

Automated Dissolution



Gel Permeation Chromatography (GPC) for Polymer AnalysisGPC is the only proven technique for characterizing the complete molecular weight distribution of a polymer, which is required for predicting material properties and performance.

Polymer analysis includes techniques such as Gel Permeation Chromatography (GPC)

Viscometry (GPC/V) Light Scattering(GPC/LS)



Integrated Chemical Structures

Integrate chemical structures to improve data management and reporting.

Perform structure searches, assign structures to components and connect to corporate structure databases.



Waters UPLC

• Full control of Waters binary and quaternary pumps.

• Full control of Multiple wavelength, Photodiode array, Fluorescence, and RI detectors.

• Full Control of sample compartment and column oven temperatures

• Full control of autosamplers.

• Full control of Waters Mass Spec detectors.


Waters HPLC’s

• Full control of Waters binary and quaternary pumps.

• Full control of Multiple wavelength, Photodiode array, Fluorescence, and RI detectors.

• Full Control of sample compartment and column oven temperatures

• Full control of autosamplers.

• Full control of Waters single quad mass detectors.


Waters Approach

Empower Software puts the right controls in the right hands for efficient, compliant lab work.

Introduction to Chromatography software - Mohsin …...2017/03/04 · Introduction to...

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