The Tool Box â€“ Getting More Productivity from Your GC and GC/MSD

Productivity Tool Box for GC & GC/MSOverview 5975C MSDAgilent RestrictedPage 2

The Tool Box – Getting More Productivity from Your GC and GC/MSD

M RameshApplication Engg – GC/GCMSAgilent Technologies India Pvt. Ltd.

Productivity Tool Box for GC & GC/MS

Page 4

Defining Productivity

• More analyses per unit time . . . – Faster and more efficient separations

– Shorter analysis cycle time (time from injection to injection)

– Reduced requirement for sample preparation

• with consistently better results . . . – Quantitative: accuracy, precision, detection limits, linear range

– Qualitative: response ratios, confirmatory retention times, library searches (eliminate false negatives and false positives)

• while using less operator time for each analysis.– Less time performing maintenance

– Extensive computer control and data processing

– Simplified operator review/approval


Page 5

Productivity: the Sum of Many Technologies

• Low Thermal Mass GC

• Capillary Flow Technology - Backflush

• Hydrogen carrier gas for GC and GC/MS

• Retention Time Locking (RTL) Editor (& Methods)– With more RTL Databases

• Trace Ion Detection

• Fast Electronic - Synchronous SIM/Scan

• Method Translator

• High signal-to-noise Triple-Axis Detector– Gain Normalization of the EM

– EM Saver

• 2nd Generation Deconvolution Reporting Software (DRS)

Return home(this slide)

Nextslide

Return tosection menu


Page 6

Productivity: Low Thermal Mass GC

• More analyses per unit time . . . – Faster and better separations– Shorter analysis cycle time (time from injection to injection)– Reduced requirement for sample preparation







Page 7

“LTM” (L ow Thermal M ass) Technology

•Direct heating of fused silica GC columns


Page 8

LTM Cooldown Times (Standard Size)

0

50

100

150

200

250

300

350

400

0 50 100 150 200 250

Time (sec)

Tem

p (o

C) 2m LTM

5m LTM

10m LTM

7890 GC

Additional Cooldown times available on website for LTM column modules up to 30m

Rapid Cooldown


Page 9

Low Thermal Mass Technology for 7890/6890 GC

Satisfies the need for high through-put, especially for low margin samples, where analytical cycle times are a critical element in making a profit

Independent and simultaneous temperature programming of:1-2 column modules (fastest cooldown), or 1-4 column modules (small format)

Note: Front thermal shield removed to show LTM column modulesEntire thermal shield should always be in place during operation

LTM Column Modules(standard width shown)

LTM Control System w/ Keypad User Interface… Agilent LTM Control SW now available


Page 10

LTM Also Greatly Reduces Cool-Down Times,� 9x Faster Heating/Cooling Cycle Times

40 min + 5.4 min

20 min + 5.4 min

3 min + 2 min

GC Run Time Cool Down

LTM/78909x Faster Cycle Time(200oC/min)


Page 11

Obtaining Faster GC, GC/MS Analytical Cycle Times – A Variety of Approaches to Fit Your Lab and Your N eeds

Post-RunBake-Out

ChromatographicRun

Post-RunCool-Down

Pre-RunALS Set-Up

Included “ALS Sample Overlap” 7890 GCin in ChemStation SW Faster Cool-down

HW/SW

Faster GC Column Selection,Analyses Method Translation

GC Add-Ons Capillary Flow TechnologySignficantly Faster GC (Backflush)Analytical Cycle Times

LTM Technology LTM Technology(Rapid Heating) (Rapid Cooling )


Page 12

6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00Time-->

3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00Time-->

Reference method – Standard Oven30 m x 0.25 mm ID x 0.25 µm DB-5MS

Fast Analysis - LTM20 m x 0.18 mm ID x 0.18 µm DB-5MS

PAH by LTM/GC/MS


Page 13

Column contamination from just one prior run

5.00 5.20 5.40 5.60 5.80 6.00 6.20 6.40 6.60 6.80Time (min)

Contamination free when using backflush

PAH by LTM/GC/MS


Page 14

LTM Productivity

• High efficiency temperature programming– Fast cooling

– High throughput

– Fast method development

• High performance chromatography– Utilize commercially available columns (typically < 30 m)

• Easily integrates with conventional GC instruments– Utilize existing injectors, detectors, software

• Multicolumn temperature control

– Independent optimization

– Multidimensional GC

– High throughput parallel systems


Page 15

Hydrocarbon Processing Industry (HPI)•HPIShorter analytical cycle times get data back to operations faster, or allow more samples/shift (eg. Simulated Distillation) •Key Sales Tools5990-3174EN Fast Hydrocarbon & Sulfur Simulated Distillation Using the

Agilent Low Thermal Mass (LTM) System on the 7890A GCand 355 Sulfur Chemiluminescence Detector(Wang,Firor,Tripp - Fast Simdist (< 3 mins), 6x faster than conventional Simdist- Dual channel, simultaneous SimDist for hydrocarb’s & sulfur

•Article Multidimensional Gas Chromatography with Capillary Flow(Fall’08) Technology and LTM-GC

J Luong (et. al) Dow Chemical, R Mustacich (RVM)Published:Journal of Separation Science, ISSN 0021-9665,Volume 31, pp. 3385-3394 http://www3.interscience.wiley.com/journal/121406899/abstract- HPI examples 2D LTM/CFT/GC using Deans Switch andLTM’s simultaneous/independent temperature programmingof multiple LTM column modules

•


Page 16

Environmental/Food Safety/Forensics

•Need for high through-put, especially for low margin samples, where analytical cycle times are a critical element in making a profit

•Key Sales Tools5990-3201EN Ultra-Fast Total Petroleum Hydrocarbons (TPH) Analysis with

Agilent Low Thermal Mass (LTM) GC and Simultaneous Dual-Tower Injection (Wei & Szelewski)- 9x faster TPH throughput using LTM- 18x faster TPH using LTM and dual simultaneous injection

•Pub No. TBA Polyaromatic Hydrocarbons (PAH) by LTM/GC/MS(~end Jan’09) - 2x faster PAH runs using LTM, even faster with Backflush

- Enviro and Food Safety samples can be dirty. LTM columns cannot be trimmed (not Agilent, not Thermo…)

- LTM uses guard columns…and now Backflush/CFTbackflush delivers multiple benefits for LTM analyses including. reduced potential for carryover, longer column life, shorter analysistime


Page 17

Food/Flavor/Fragrance and Pharmaceutical

•Chiral Separations• Chiral GC is most effective at low temperatures, yet complex samples usually require a broad temperature program to separate target compounds from background and to elute highly retained sample components.• LTM/GC/MS uses a traditional column in the GC oven to separate target compounds, then heart-cut of target compounds to a chiral column in LTM module at a lower temp. � optimal chiral separations are achieved while decreasing analysis cycle times and increasing chiral column lifetime.

Pub No. TBA Chiral Separations by LTM/GC/MS(~end Jan’09) - 2D LTM/GC/MS, simultaneous/independent temperature

programming of GC and LTM column module(s)

Article Multidimensional Gas Chromatography with Capillary Flow(Fall’08) Technology and LTM-GC, J Luong (et. al) Dow Chemical,R Mustacich

Published:Journal of Separation Science, ISSN 0021-9665,Volume 31, pp. 3385-3394253-261- Examples 2D LTM/CFT/GC using Deans Switch andLTM’s simultaneous/independent temperature programmingof multiple LTM column modules


Page 18

Productivity: CFT Backflush


– Shorter analysis cycle time (time from injection to injection) – Reduced requirement for sample preparation

• with consistently better results . . . – Quantitative: accuracy, precision, detection limits , linear range

– Qualitative: response ratios, confirmatory retention times, libr ary searches (eliminate false negatives and false posit ives)

• while using less operator time for each analysis.– Less time performing maintenance– Extensive computer control and data processing



Page 19

Backflush – Complete Column

1 psi

45 psi

During GC Run

After GC Run

S/S Inlet

QuickSwap or 3-way SplitterColumn

25 psi

4 psi

MSD

Aux EPCSplit Vent Trap

S/S Inlet

QuickSwap or 3-way SplitterColumn

MSD

Aux EPCSplit Vent Trap

performance turbo recommended


Page 20

Backflush – Mid-Column (Analysis Mode)

Vent

Pressure / Flow

Controller

15-m HP-5ms (0.25mm id x0.25um)

15-m HP-5ms (0.25mm id x0.25um)

Capillary Flow Technology Device

5975C MSDEI mode

7890A GC

Split/Splitless Injection Port

Y ml/min

Y+Z ml/min

Z ml/min

Z mL/min very low flow; set against column backpressure, not short restrictor as with QuickSwap

Column 1 and 2 could be different phases and different dimensionsPCT available Oct’08


Page 21

Backflush – Mid-Column (Backflush Mode)

Vent

Pressure / Flow

Controller

15-m HP-5ms (0.25mm id x0.25um)

15-m HP-5ms (0.25mm id x0.25um)

5975C MSDEI mode

7890A GC


Decrease inlet pressure during backflush

Pressure Controlled Tee


Page 22

Backflush – Mid-Column (Backflush Mode)

Vent

Pressure / Flow

Controller

15-m HP-5ms (0.25mm id x0.25um)

15-m HP-5ms (0.25mm id x0.25um)

5975C MSDEI mode

7890A GC


Pressure Controlled Tee

Decrease inlet pressure during backflush

Purged UltimateUnion From 1st

column

To 2nd

Column & MSD

From P control


Page 23

9.00 9.50 10.00 10.50 11.00 11.50 12.00

9.00 9.50 10.00 10.50 11.00 11.50 12.00

9.00 9.50 10.00 10.50 11.00 11.50 12.00

a b

Backflush – Mid-Column (Timing Considerations)

a = begin backflush (last peak exits column 1)

b = increase flow 4 mL/min

Sample - no backflush

Sample - with backflush

Solvent blank - no backflush


Page 24

Comparison of Backflush Configurations

T=300°C; head pressure =0.5psi

4x faster & lower flow!

* smallest restrictor => best QS case

Backflush (BF) Pressure (psi)

BF flow (mL/min)

1 column vol (min)

MDS flow (mL/min)

BF flow (mL/min)

1 column vol (min)

MSD flow (mL/min)

10.0 0.3 3.82 1.5 0.6 0.94 0.9 4.120.0 0.7 1.89 2.9 1.5 0.47 1.8 4.030.0 1.3 1.28 4.8 2.6 0.32 3.0 4.040.0 2.1 0.98 7.2 4.1 0.25 4.5 3.9

PCT time savings

(N x)

QuickSwap (QS) Pressure Controlled Tee (PCT)30 m column; restrictor (0.092mm*) Two 15 m columns


Page 25

Backflush Impact on Data Quality

– Late eluters are more than just a timing problem • Bake-out may push the limits of phase stability

• Matrix may not bake out well even at the upper limit of the column

– For ‘high boiling’ matrices, backflush offers:• Improved retention time precision

• Improved spectra quality

• Consistent detector response

• Increased column lifetime


Page 26

GC/MS Industry Standard: Low Bleed Stationary Phase sLow bleed means less background (and background ion s)

DB-35ms

8.00 10.00 12.00 14.00 16.00 18.00 20.00

320°C

DB-35

8.00 10.00 12.00 14.00 16.00 18.00 20.00

300°C

CLP Pesticides Analysis

S/N =10

DB-35ms, 320°C

S/N =3

DB-35, 300°C

51

78 96119

144

207

197

214

250

253

286

315

346

356 377

405

428

455470

498

m/z-->50 100 150 200 250 300 350 400 450 500

0

Primary IonDB-35ms

51

78 96119

144

207

197

214

250

253

286

315

346

356 377

405

428

455470

498

m/z-->50 100 150 200 250 300 350 400 450 500

m/z-->50 100 150 200 250 300 350 400 450 500

0

Primary IonDB-35ms

52

78

96 119

135156

197

207

249

253

281

315331

356

377

405

428

451

470

498

m/z-->50 100 150 200 250 300 350 400 450 500

0

Primary Ion

DB-35

52

78

96 119

135156

197

207

249

253

281

315331

356

377

405

428

451

470

498

m/z-->50 100 150 200 250 300 350 400 450 500

m/z-->50 100 150 200 250 300 350 400 450 500

0

Primary Ion

DB-35

Low bleed column

Standard column

Better detection limits

Better spectra

Column bleed = chemical noise


Page 27

Typical GC/MS Separation Sequence

• Injection• Filament ON after the solvent peak• Separation and detection of the analytes• Filament OFF after the last analyte peak and before bake-out

– In an effort to ‘save’ the electron multiplier (EM), the operator is ‘blind’ to the success of bakeout

– Bake-out may NOT be adequate for all samples

This means the typical method will always have a potential for matrix carryover that

the user does not expect.


Page 28

Without Backflush: A Serious Problem

Abundance

4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.000

2000000400000060000008000000

1e+071.2e+071.4e+071.6e+071.8e+07

2e+072.2e+072.4e+072.6e+072.8e+07

3e+073.2e+073.4e+073.6e+073.8e+07

4e+074.2e+074.4e+074.6e+07

Time

A: TIC: lettuce_blank.D\data.ms

B: TIC: lettuce_blank3.D\data.ms

B

A

Overlay of two chromatograms of a blank extract inj ected BEFORE (A) and AFTER (B) three injections wit hout backflush

Data provided by MSD user in Almeria, Spain

After only 3 samples , the background is significantly higher (increase chemical noise is every spectrum)


Page 29

Without Backflush: Changes in Retention Time


Abundance

4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.000

2000000400000060000008000000

1e+071.2e+071.4e+071.6e+071.8e+07

2e+072.2e+072.4e+072.6e+072.8e+07

3e+073.2e+073.4e+073.6e+073.8e+07

4e+074.2e+074.4e+074.6e+07

Time



B

A


Highly retained matrix is altering the column selectivity and changing the retention time

After only 3 samples


Page 30

Without Backflush: Increased Background

Abundance

4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.000

2000000400000060000008000000

1e+071.2e+071.4e+071.6e+071.8e+07

2e+072.2e+072.4e+072.6e+072.8e+07

3e+073.2e+073.4e+073.6e+073.8e+07

4e+074.2e+074.4e+074.6e+07

Time



B

A



Increased background due to carryover of concentrated matrix

After only 3 samples

Sample matrix = chemical noise


Page 31

With Backflush: Consistent Retention Times and Baselines (No Increase in Spectral Noise)

Stable retention times and baseline . . . less chemical noise

4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.000

2000000400000060000008000000

1e+071.2e+071.4e+071.6e+071.8e+07

2e+072.2e+072.4e+072.6e+072.8e+07

3e+073.2e+073.4e+073.6e+073.8e+07

4e+074.2e+074.4e+074.6e+07

Time

Abundance

TIC: lettuce_10_ppb.D\data.ms



Overlay of three chromatograms of lettuce extract r un with 2 min of back flush

Data provided by user in Almeria, Spain


Page 32

What Real Value Does High Performance EPC Have If Matrix Carryover Destroys Rt Precision?

m in5 .3 5 .4 5 .5 5 .6 5 .7 5 .8 5 .9 6 6 . 1

N or m .

0

2 0 0 0

4 0 0 0

6 0 0 0

8 0 0 0

1 0 0 0 0

1 2 0 0 0

1 4 0 0 0

1 6 0 0 0

m in5 .3 5 .4 5 .5 5 . 6 5 .7 5 .8 5 .9 6 6 .1 6 .2

N or m .

2 0 0 0

4 0 0 0

6 0 0 0

8 0 0 0

1 0 0 0 0

1 2 0 0 0

1 4 0 0 0

10 Runs with Backflushing

10 Runs without BackflushingRetention times shift ~4-5 secduring 10 runs

Fish oil analysis


Page 33

Backflush with Purged Ultimate Union: Lipid Peroxidation Products in Blood

App Note 5989-9359EN

4-hydroxy-2,3-nonenal (HNE), an indicator of oxidative stress, and its metabolite, 1,4-dihydroxynonene (DHN)

PCI with NH 3

Late eluting peaks were quickly contaminating the source


Page 34

Backflush with Purged Ultimate Union: Loss of Response without Backflush

without backflush

with backflush

These were the first backflush results; the system is getting clearer

App Note 5989-9359EN


Page 35

Productivity Benefits of Backflush

• More samples per day per instrument

• Longer column life

• Lower operating costs

• Less frequent and faster GC & MSD maintenance

• Less chemical background– More consistent retention times

– More consistent baselines

– Higher quality spectra (no increase in noise during analysis sequence)

– Higher quality quantitation (no increase in interfering ions during analysis sequence)


Page 36

References for Backflush

• 5989-9359EN Capillary Flow Technology for GC/MS: Efficacy of the Simple Tee Configuration for Robust Analysis Using Rapid Backflushing for Matrix Elimination (Prest, Foucault, and Aubut, Aug 08)

• 5989-9341EN Screening for 430 Pesticide Residues in Traditional Chinese Medicine Using GC/MS: From Sample Preparation to Report Generation in One Hour (Luan, Xu, Sept08)

• 5989-8664EN Capillary Flow Technology for GC/MS: a Simple Tee Configuration for Analysis at trace Concentrations with Rapid Backflushing for Matrix Elimination (June 08, Prest)

• 5989-8588EN The Use of Automated Backflush on the 7890A/5975C GC-MS System• 5989-8582EN Improved Forensic Toxicology Screening Using A GC/MS/NPD System with a 725-Compound DRS Database (Quimby, May08)• 5989-7670EN, Replacing Multiple 50-Minute FPD/ELCD/SIM Analyses with One 15-Minute Full-Scan Analysis for 10x Productivity Gain (Meng/Szelewski, Nov07)


Page 37

References for Backflush

• 5989-6460EN Analysis of Suspected Flavor and Fragrance Allergens in Cosmetics Using the 7890A GC and Capillary Column Backflush (March 2007)• 5989-6095EN Direct Injection of Fish Oil for the GC-ECD Analysis of PCBs: Results Using a Dean Switch with Backflushing (Jan 2007)• 5989-6066EN Rapid Forensic Toxicology Screening Using an Agilent 7890A/NPD/5975C/DRS GC/MSD System (Jan 2007)• 5989-6018EN Improving Productivity and Extending Column Life with Backflush (Dec 2006)• 5989-5111EN Simplified Backflush Using Agilent 6890 GC Post Run Command (June 2006)• 5989-1716EN New Tools for Rapid Pesticide Analysis in High Matrix Samples (October 2004)


Page 38

Productivity: Retention Time Locking





– Qualitative: response ratios, confirmatory retention times , library searches (eliminate false negatives and false positives)




Page 39

What is Retention Time Locking (RTL)?The ability to precisely match chromatographic retention times in any systems to those in another chromatographic system with the same nominal method and column.

How is Retention Time Locking Done?By adjusting column head pressure via EPC using interactive ChemStation software.

Retention Time Locking


Page 40

Pesticides on Two Different GCs Before RTL

23 23.5 24 24.5 25 25.5

23 23.5 24 24.5 25 25.5


Page 41

Pesticides on Two Different GCs After RTL

23 23.5 24 24.5 25 25.5

23 23.5 24 24.5 25 25.5


Page 42

RTL Calibration - Done Once for a Given Method

� Make 5 runs at 5 different inlet pressures:� target -20%

� target -10%

� target

� target +10%

� target +20%

� Determine RT of your target compound in each chromatogram� Enter values into the RTL Software


Page 43

Acquire RTLock Calibration Data..


Page 44

RTL Calibration Runs Are Collected Automatically

5 10 15 20 25 30

34 psi

31 psi

28 psi

25 psi

22 psi

Dichlorvos Methyl chlorpyrifos

Mirex

15.192 min

15.771 min

16.433 min

17.201 min

18.121 min

Target Compound for RTL


Page 45

RTL Calibration Runs Are Collected Automatically

And times can be edit manually


Page 46


Page 47

RTL Re-Locking Procedure for Trimmed Column

0

50

100

Original RTL Head pressure 16.00 psi

16.50 min.

16.34 min.Trim front of column

New RTL Head pressure 15.71 psi

0

50

100

0

50

100

16.50 min.

Re-lock method


Page 48

Benefits of Locked Methods

• Faster method transfer & validation

• Faster startup after column maintenance

• Reduces / eliminates need to update RTs in calibration tables

• Easy set up of windows for SIM groups and MS/MS transitions

• Equivalent chromatography for all labs sharing a common method

• More efficient troubleshooting

• Easier review of data

• Faster identification of unknowns


Page 49

Productivity: Trace Ion Detection




• with consistently better results . . . – Quantitative: accuracy, precision, detection limits , linear range






Page 50

Trace Ion Detection Technology

• Proprietary algorithm– Digital filtering

• Reduced noise level • Improved peak shape

– Smoothes ‘spikes’ on trace peaks

– Especially under-sampled peaks

• Improved library matchS/N = 92

WithoutTrace Ion Detection

S/N = 53

WithTrace Ion Detection

… better detection at trace levels


Page 51

Improve Library Match QualityFenthion in strawberry extract

Match Quality30

Trace Ion Detection off

Trace Ion Detection on Match Quality92


Page 52

Trace Ion Detection Reduces False Negatives


Page 53

Literature for Trace Ion Detection

• Technical Overview 5989-6425EN , The 5975C Series MSD: Method Optimization and Trace Ion Detection (Roushall/Prest, Mar 07)

• App Note 5989-7670EN , Replacing Multiple 50-Minute FPD/ELCD/SIM Analyses with One 15-Minute Full-Scan Analysis for 10x Productivity Gain (Meng/Szelewski, Nov07)


Page 54

Productivity: Fast Electronic - SIM/Scan Mode

• More analyses per unit time . . . – Faster and more efficient separations– Shorter analysis cycle time (time from injection to injection)


• with consistently better results . . . – Quantitative: accuracy, precision, detection limits, linear range– Qualitative: response ratios , confirmatory retention times, library

searches (eliminate false negatives and false posit ives)





Page 55

High Performance, Fast Electronics

Higher percentage of time monitoring ions = more signal

Higher data rate for narrow “fast” peaks = better area precision


Page 56

High Performance Electronics for High Performance Synchronous SIM/Scan

5989-3108EN: Improving Productivity with Synchronous SIM/Scan


Page 57

Fast Electronics: Synchronous SIM/Scan

• The best of both modes in a single run– SIM = maximum sensitivity for target compounds– Scan = best identification of non-targets (unknowns )

• High speed electronics is an Agilent strength

… maximum information from a single run

Scan data points

SIM data points

Chromatographic peak time

Scan time SIM time

Tech Overview 5989-3108EN


Page 58

Under-Sampled Peaks Acceptable for MS

• Acceptable precision can be achieved with fewer poi nt per peak

– 6 samples/peak width for area; 8 samples/peak width for height

• Compromises to evaluate

– Affect of a slower data rate on quant precision

– Affect of shorter dwell time decrease SIM S/N

– Affect of faster scan rates decrease scan S/N

For many applications . . . thedecrease in precision and S/N are small compared to the benefits of SIM sensitivity andscan library searches


Page 59

Literature for Synchronous SIM/Scan (slide 1 of 2 )

• Tech Note 5989-3108EN, Improving Productivity with Synchronous SIM/Scan (Meng, May’05 )• Tech Note 5989-5669EN, Strategies for Developing Optimal Synchronous SIM-Scan Acquisition Methods—AutoSIM/Sca n Setup and Rapid SIM ( Sept’06 )• App Note 5989-6066EN , Rapid Forensic Toxicology Screening Using an Agilent 7890A/NPD/5975C/DRS GC/MSD System (Quim by, Jan’07)• App Note 5989-4184EN , Synchronous SIM/Scan Low-Level PAH Analysis Using the Agilent Technologies 6890/5975 i nert GC/MSD (Szelewski, Nov’05 )• App Note 5989-3571EN, Determination of Trace Levels of Aromatic and Chlorinated Hydrocarbons in Water Using the G18 88 Headspace Sample/6890N and 5975 inert MSD ( Firor, Oct’05 )


Page 60

Literature for Synchronous SIM/Scan (slide 2 of 2 )

• App Note 5989-3196EN, The Determination of Residual Solvents in Pharmaceuticals Using the Agilent G1888 Headspace/6 890N GC/5975 inert MSD system ( Firor/Gudat, June’05 )• App Note 5989-5494EN, The Determination of Extractables and Leachables in Pharmaceutical Packaging Material Usi ng Headspace GC/MS (Gudat/Firor, Aug’06 )•App Note 5989-4834EN , Screening for Hazardous Chemicals in Homeland Security and Environmental Samples Using a GC/MS/ECD/FPD with a 731 Compound DRS Database (Quimby/Szelewski, Feb’06 )• App Note 5989-3299EN, Identifying Pesticides with Full Scan, SIM, uECD, and FPD from a Single Injection ( Meng/Quimby, July’05 )• Tech Note 5988-4188EN (AutoSIM), New Approaches to the Development of GC/MS Selected Ion Monitoring Acquis ition and Quantitation Methods ( Prest/Peterson, Nov’01 )


Page 61

Productivity – Method Translator

• More analyses per unit time . . . – Faster and more efficient separations– Shorter analysis cycle time (time from injection to injection) – Reduced requirement for sample preparation

• with consistently better results . . .– Quantitative: accuracy, precision, MDL, linearity, consistent sensitivity






Page 62

What is Method Translation?

MTL is a freeware program that determines new method conditions such that relative elution order is constant, and relative run time is known.

MTL works:

g when changing column outlet pressure (e.g., from FID to MSD)

g when changing carrier gas type (e.g., from He to H2) or flow rate

g when changing column dimensions (same phase, different diameter, length, or phase ratio)


Page 63

Screen capture showing the Method Translation (MTL) Software data entry used in a 4X speed gain translation.

Method Translator


Page 64

250 µm(X1)30250

0.25018.0

01.970

2.0025.01500.003.02000.008.0280

10.00

100 µm(X2)10100

0.10036.55

00.470

1.0050.01500.006.02000.0016.02805.00

100 µm(X3)10100

0.10063.17

00.870

0.6775.01500.009.02000.0024.02803.30

100 µm(X4)10100

0.10090.00

01.5700.51001500.0012.02000.0032.02802.50

Length (m)I.D. (µm)D.F. (µm)Inlet P (psig)Outlet P (psia)Flow (mL/min)Init Temp (°C) Init Hold (min)Ramp 1 (°C/min)Final Temp1 (°C)Final Time 1 (min)Ramp 2 (°C/min)Final Temp 2 (°C)Final Time 2 (min)Ramp 3 (°C/min)Final Temp 3 (°C)Final Time 3 (min)

Speed GainColumn id.

Method Translations to 100 um Column


Page 65

Method Translations to 15 m Column

30 m(X1)

19091S-4332500.2518.0

01.970

2.0025.01500.003.02000.008.0280

10.00

Part NumberI.D. (µm)D.F. (µm)Inlet P (psig)Outlet P (psia)Flow (mL/min)Init Temp (°C) Init Hold (min)Ramp 1 (°C/min)Final Temp1 (°C)Final Time 1 (min)Ramp 2 (°C/min)Final Temp 2 (°C)Final Time 2 (min)Ramp 3 (°C/min)Final Temp 3 (°C)Final Time 3 (min)

Speed GainColumn Length (m) 15 m

(X2.5)

2500.255.74

01.4970

0.8062.51500.007.52000.00202804.00

19091S-431

15 m(X4)

2500.2518.0

03.870

0.501001500.00122000.00322802.5

19091S-431


Page 66

Productivity – Triple Axis Detector with Gain Normalization and EM Saver• More analyses per unit time . . .

– Faster and more efficient separations



• with consistently better results . . . – Quantitative: accuracy, precision, MDL, linearity, consistent sensitivity– Qualitative: response ratios, confirmatory retention times, library

searches (eliminate false negatives and false positives)




Page 67

Conventional Off-Axis Detector

hyperbolic quartztransmission

quadrupoleanalyzer

ion beam

energetic neutrals

Although not directly striking the HED-EM, excited helium neutrals can ionize other molecules and create secondary particles from collisions with surfaces (sputtered ions from surface contamination, ions from the surface, photons and electrons)

high energy dynode (high voltage pulls the ions away from the neutrals)

electron multiplier


Page 68

Triple-Axis Detector

ion beam X

Y

OffsetZ

With a combination of three axes and appropriate shielding, noise from metastable helium, photons, etc. can be reduced to a minimum

This aperture is larger than the off-axis detector which means more ions and energetic neutrals pass to the detector shield blocking

secondary ions formed by metastable helium after the analyzer


Page 69

Triple-Axis Detector: Higher Signal – Lower MDL

Lower detection limits !


Increased slope = higher sensitivity

Dashed line show 95% confidence limits

fg


Page 70

Triple Channel EM

•Triple channels improve signal amplification

– Lower initial EM voltage required

•Triple channels increase life– Lower cost of operation

Exit of the triple channels

Collector removed to show exit passages

Collector


Page 71

Tuning – Optimization of Multiple Components

• Tuning ion transmission characteristics– Optimize potentials on lens and analyzer

– Effects the relative abundance of ions WITHIN a spectrum

• For example, the intensity ratio of m/z 100 to 200 or 100 to 300

– Effects the absolute number of ions entering the EM

– Improves spectra comparisons between instruments

– US EPA standards made this a critical issue in many areas of the world

• Gain Normalization of ion amplification (EM only)– Effect the ion ‘count’ or absolute abundance of each ion

– Does NOT affect the number of ions entering the EM

– Changes the amplification of charge passing through the EM


Page 72

Gain Normalized EM Adjustment for Sensitivity

I

EMV: 1kV to 3kV

High Energy Dynode (10kV)QuadrupoleIon source

EMVElectron Multiplier (EM)

Ii

GAIN

Amplification of signalnow, user-selectable!!!

i

Tuning EM voltage changes sensitivity for all ions -- NOT the relative abundances



Page 73

S igna l vs E MV

0

50000

100000

150000

200000

250000

300000

1000 1200 1400 1600 1800 2000 2200 2400EM V

Sig

nal

Sensitivity Limitation of ATUNE + nV•During aging, the same EMV setting will not give the same signal!

ATUNE (new) ATUNE (aged)

ATUNE+400VATUNE + 400V

“New” detector “Aged” detector

Decreasingsignal with age

+ 600V


As signal decreases, it appears the MS has lost sensitivity In reality, the

MS was OK, but the EMV was too low


Page 74

Sensitivity Advantage of Gain Normalization

Sign a l v s G AIN

0

5000

10000

15000

20000

25000

30000

35000

40000

0 2 4 6 8 10 12 14 16 18 20

G ain Facto r as GA IN x 10^ 5

Sig

nal

“New”

“Aged”

•During aging, the same GAIN setting will give the same signal!


With Gain Normalization, the operation does not have to think about aging, sensitivity is the same for a new or aged EM at any Gain setting – simple!


Page 75

“ATune + nV” versus Gain Normalized Methods

9.80 9.90 10.00 10.10 10.20 10.30

1000

9.80 9.90 10.00 10.10 10.20 10.30

1000

“New” “Aged”

Consistent sensitivity over the life of the electron multiplier!Consistent sensitivity from MSD to MSD and lab to lab!


GAIN Tune GAIN TuneATUNE+ n V

ATUNE+ n V

New triple channel EM ages more slowly


Page 76

Benefits of Gain Normalization (EM Optimization)

New Option: GAIN Normalized methods

Gain Factors are set & saved as a method parameter !

Finer Gain Adjustment for better optimization

Greater reproducibility over time

Greater consistency -for any instrument, or -between a group of instruments

No linearity errors with new EM

Easier tuning & system troubleshooting

Six excellent reasons to use Gain Factors!!


Page 77

Literature for Gain Normalization

• Technical Overview 5989-7654EN , Enhancements to Gain Normalized Instrument Tuning: Understanding the Benefits and Features• Technical Overview 5989-7655EN , The Triple-Axis Detector: Attributes and Operating Advice• Technical Overview 5989-6050EN , The 5975C Series MSD: Normalized Instrument Tuning


Page 78

EM Saver

• When too many ions are striking the EM, the response saturates and the overload can damage the EM• EM Saver and fast electronics quickly alters the analyzer voltages to decrease ion transmission

• A simple concept with real benefits to EM life– Highly concentrated peaks cannot damage the Agilent EM


Page 79

Productivity – Hydrogen Carrier Gas

• More analyses per unit time . . . – Faster and more efficient separations– Shorter analysis cycle time (time from injection to injection) – Reduced requirement for sample preparation







Page 80

Helium Shortage Means Higher Operating Costs

www.cen-online.orgPg 31October 8, 2007


Page 81

H2 Requires Different Setup and Extra Care

• Lower column head pressure experienced with H2– H2 is upwards of 100% faster than with helium; the column internal diameter may

need to decrease to have optimal pressure control – There is a downloadable Method Translation Software to guide the customer

• H2 is reactive towards some compounds. – This can be an advantage or a disadvantage depending on the application – Sometimes very reactive compounds can be measured as their hydrogenated

counterparts • Diffusion is much faster in H2

– The van Deemter curve will shift and the HETP (height equivalent of a theoretical plate; column efficiency) will be different compared to helium

– GC resolution can be improved under optimal conditions• H2 is explosive

– Auto-ignition temperature is 400ºC and the LEL/UEL are very wide– Special cautions may be required for safety– Hydrogen generators can greatly reduce safety concerns


Page 82

Agilent Tests for Hydrogen Safety


Page 83

Hydrogen Generators Make the Cost Very Low and Ensure High Quality Gas


Page 84

Literature for Hydrogen Carrier Gas

• Manual G3170-90010 , 5975C MSD Hydrogen Safety (Oct ’05)• Technical Overview 5988-4971EN , A Complete Solution for Chlorinated Pesticides and Herbicides Using DB-35ms and DB-XLB Columns (hydrogen and helium comparison) (Dec ’01)


Page 85

Productivity – Deconvolution Reporting Software




• with consistently better results . . . – Quantitative: accuracy , precision, detection limits , linear range

– Qualitative: response ratios, confirmatory retention times, library searches (eliminate false negatives and false posit ives)


– Extensive computer control and data processing– Simplified operator review/approval


Page 86

What is Deconvolution Reporting Software (DRS)?

• A second opportunity for accurate result without another analysis

– A post-run calculation based on the NIST algorithm

– A solution to the unpredictable matrix interferences

• DRS makes optimal use of both GC and MS information– Combines peak profiles and spectra

– Extract ‘pure component’ information from the coeluting peaks

– Uses RTL database spectra to filter analytes of interest from vast number of matrix peaks

– Confirms against complete NIST library spectra

– ChemStation QEdit summary of standard and deconvoluted data


Page 87

Component 2

Component 3

Deconvolution

Deconvoluted peaks and spectra

matrix

target

interference

TIC & Spectrum

TIC

Peak detected with Trace Ion Detection

How Does AMDIS Work?

Some m/z values “ pure ”Some m/z values mixed

m/z

Mathematical “Separation”

Component 1

Uses Difference in Spectra, Retention Time and Peak Shape


Page 88

Pulling a Useable Spectrum Out of a Mess

Analyte ions are buried in the matrix ions!!

‘Cleaned’ analyte spectrum is easy to match!


Page 89

NISTConfirm+

ChemStationfor Quant

+AMDIS 32Deconvolute and ID

What is the Agilent DRS?

CONSOLIDATED QUANTITATIVEand

QUALITATIVE HTML REPORT


Page 90

Reduced Errors and Less Faster Results

32 minutes~ 8 hoursProcessing Time

01False Positives

Same 37+ 99 additional

37Targets Found

Agilent DRS*CDFA

*CDFA is the California Department of Food and AgricultureData files courtesy of Dr. Mark Lee and Steve Siegel

Save about 7.5 hours to do other jobs

Summary of Pesticide Analysis in 17 Surface Water S amples


Page 91

QEdit Summary of Qual and Quant (DRS A.04)

Overlay of target(s) and Deconvolutedion plots

Spectral review: Before AMDISAfter AMDISAMDIS Library

Areas & amounts from target ion andDeconvoluted ion

Deconvoluted ion plot

Target ion plot

Hits X = MSDA = AMDIS


Page 92

Summary Report with MSD and AMDIS Results

partial report


Page 93

Graphics Report

MSD and AMDIS 5-ion overlay

Raw, deconvoluted and library spectra

MSD and AMDIS areas and amounts


Page 94

Supporting Literature for DRS (slide 1 of 3)

• App Note 5989-8582EN , Improved Forensic Toxicology Screening Using a GC/MS/NPD System with a 725-Compound DRS Database (Quimby, May08)– Capillary Flow Technology for ‘solvent vent’ and NPD– Backflush to shorten analysis time and increase column lifetime– Synchronous SIM/scan (max sensitivity and max information)– Trace Ion Detection to reduce noise before DRS– Next generation of DRS with AMDIS results in QEdit and Agilent database– Real blood extracts from NMS Lab, Willow Grove, PA, USA

• App Note 5989-7670EN , Replacing Multiple 50-Minute FPD/ELCD/SIM Analyses with One 15-Minute Full-Scan Analysis for 10x Productivity Gain (Meng/Szelewski, Nov07)– Capillary Flow Technology for multiple detectors (3-way splitter)– Backflush to shorten analysis time and increase column lifetime– Trace Ion Detection to reduce noise before DRS– Next generation of DRS with AMDIS results in QEdit and Agilent database– Real results from US Food and Drug Administration/Center for Food Safety and

Applied Nutrition (FDA/CFSAN)


Page 95


• App Note 5989-7436EN , Screening for Pesticides in Food Using the Japanese Positive List Pesticide Method: Benefits of Using GC/MS with Deconvolution Reporting Software and a Retention Time Locked Mass Spectral Database (Wylie, Sept07)

• App Note 5989-6677EN , Reducing Analysis Time Using GC/MSD and Deconvolution Reporting Software (May07)

• App Note 5989-6066EN , Rapid Forensic Toxicology Screening Using an Agilent 7890A/NPD/5975/DRS GC/MSD System (Quimby, Jan07)

• App Note 5989-5435EN , Screening for 171 Volatile Organic Air Pollutants Using GC/MS with Deconvolution Reporting Software and a New Indoor Air Toxics Library (Wylie, Aug06)

• App Note 5989-5076EN , Screening for 926 Pesticides and Endocrine Disruptors by GC/Ms with Deconvolution Reporting Software and a New Pesticide Library (Wylie, Apr06)


Page 96


• App Note 5989-4834EN , Screening for Hazardous Chemicals in Homeland Security and Environmental Samples Using a GC/MS/ECD/FPD with a 731 Compound DRS Database (Quimby, Szelewski, Feb06)

• App Note 5989-1716EN , New Tools for Rapid Pesticide Analysis in High Matrix Samples, (Szelewski, Quimby, Oct04)

• App Note 5989-1654EN , A Blind Study of Pesticide Residues in Spiked and Unspiked Fruit Extracts Using Deconvolution Reporting Software

• App Note 5989-1157EN , Comprehensive Pesticide Screening by GC/MSD using Deconvolution Reporting Software (Wylie/Szelewski/Meng, May04)


Page 97

RTL is more flexible now…

GC/MSD user can manually edit these times


Page 98

Largest Selection of Industry Specific Retention Ti me Locked Databases

modify a library to meet your need… or create your o wn

Product No. RTL DBL Description CompoundsG1671AA Hazardous Chemical 730G1672AA Pesticide 926G1673AA Indoor Air Toxics 171G1674AA Forensic Toxicology 723G1675AA JPN Positive List Pesticide 431G1676AA Fiehn Metabolomics 665G1677AA Environmental Semi-voas 273

Free Volatile Organic Compounds 65Free PCB Congeners 209Free Forensic Toxicology 277Free Fatty Acid Methyl Ester 37Free Flavors 409Free Organotin Derivatives Methyl, Ethyl, Pentyl

Library include GC method details, Getting Started manual, application notes, and HELP files


Page 99

Supporting Literature for RTL (slide 1 of 4)

• App Note 5989-7875EN, Semivolatiles Retention Time Locked (RTL) Deconvolution Databases for Agilent GC/MSD Systems (Szelewski, Feb’08) G1677AA

• App Note 5989-7670EN , Replacing Multiple 50-Minute FPD/ELCD/SIM Analyses with One 15-Minute Full-Scan Analysis for 10x Productivity Gain (Meng/Szelewski, Nov07) G1672AA

• App Note 5989-7436EN , Screening for Pesticides in Food Using the Japanese Positive List Pesticide Method: Benefits of Using GC/MS with Deconvolution Reporting Software and a Retention Time Locked Mass Spectral Database (Wylie, Sept07) G1675AA

• App Note 5989-6677EN , Reducing Analysis Time Using GC/MSD and Deconvolution Reporting Software (May07)

• App Note 5989-6066EN , Rapid Forensic Toxicology Screening Using an Agilent 7890A/NPD/5975/DRS GC/MSD System (Quimby, Jan07) free DBL


Page 100


• App Note 5989-5435EN , Screening for 171 Volatile Organic Air Pollutants Using GC/MS with Deconvolution Reporting Software and a New Indoor Air Toxics Library (Wylie, Aug06) G1673AA

• App Note 5989-5076EN , Screening for 926 Pesticides and Endocrine Disruptors by GC/Ms with Deconvolution Reporting Software and a New Pesticide Library (Wylie, Apr06) G1672AA

• App Note 5989-4834EN , Screening for Hazardous Chemicals in Homeland Security and Environmental Samples Using a GC/MS/ECD/FPD with a 731 Compound DRS Database (Quimby/Szelewski, Feb06) G1671AA

• App Note 5989-3347EN , Developing an eMethod for the Analysis of Volatile Organic Compounds in Water Using Purge and Trap/GC with Agilent’s New 5973 inert Mass Spectrometer (Wylie, Jul’05)

• App Note 5989-2850EN , Determination of Polybrominated Diphenyl Ethers in Polymeric Materials Using the 6890 GC/5973N inert MSD with Electron Impact Ionization (Tu/Prest, Apr05)


Page 101


• App Note 5989-1716EN , New Tools for Rapid Pesticide Analysis in High Matrix Samples, (Szelewski/Quimby, Oct04)

• App Note 5989-1654EN , A Blind Study of Pesticide Residues in Spiked and Unspiked Fruit Extracts Using Deconvolution Reporting Software (Sandy, Oct04)

• Tech Overview 5989-0916EN , Building and Editing RTL Screener/Quant Databases and Libraries (Szelewski/Weiner/Meng, June04)

• App Note 5989-1157EN , Comprehensive Pesticide Screening by GC/MSD using Deconvolution Reporting Software (Wylie/Szelewski/Meng, May04)

• App Note 5968-5871EN , Improving the Analysis of Fatty Acid Methyl Esters Using Retention Time Locked Methods and Retention Time Databases (David/Sandra/Wylie, Sept’03)

• App Note 5988-9256EN , Improving the Analysis of Organtin Compounds Using Retention Time Locked Methods and Retention Time Databases (David/Sandra/Wylie, Apr03)


Page 102


• App Note 5988-7150EN , Solid-phase Extraction and Retention Time Locked GC/MS Analysis of Selected Polycyclic Aromatic Hydrocarbons (PAHs) (Prest, July’02)

• App Note 5988-3934EN , Retention Time Locked GC-MS Analysis of Phenols (Reese/Prest, Apr02)

• App Note 5988-4392EN , Identification and Quantitation of Pesticides in the Part-per-Trillion Range Using Retention Time Locking and GC/MS (Meng, Nov’01)

• Tech Note 5988-4188EN (AutoSIM), New Approaches to the Development of GC/MS Selected Ion Monitoring Acquisition and Quantitation Methods (Prest/Peterson, Nov’01)

• App Note 5988-2244EN , A New Approach to the Analysis of Phthalate Esters by GC/MS (George/Prest, Mar’01)

• App Note 5988-9455EN , Ambient Headspace GC and GC-MSD Analysis of Non-Polar Volatiles in Water (Szelewski/Quimby, Feb’00)

• App Note 5968-8657E , Retention Time Locking: Creating Custom Retention Time Locked Screener Libraries (Weiner/Prest, Dec’99 )

Documents

The Tool Box â€“ Getting More Productivity from Your GC and GC/MSD