New GC & GC/MS Technologies to Improve the Productivity of … · Ghost Peaks Eliminate Ghost Peaks...

New GC & GC/MS Technologies to Improve the Productivity of Your Laboratory

Agilent Science and Technology Symposium

May 2014

May 2014

1

Presentation Overview GC and GC/MS Productivity Tools

Challenges for the Current Analytical Laboratory

Capillary Flow Technology

Retention Time Locking

Deconvolution and Reporting Software

Self-Cleaning Ion Source

May 2014

Agilent Science and Technology Symposium 2014 2

Challenges faced by Today’s Customers Expecting More from Less

• Increasing cost of ownership

• Trace analysis in complex matrices Business

Challenges

• Less time for method development

• Limited technical experience Resource Allocation

• Demands for higher throughput • Quicker return on capital investment

Optimizing Efficiency

May 2014


Laboratory Challenges Analytical Requirements

Challenges Samples of varying nature

Identify closely related analytes:

isomers, analogs, etc.

Variable concentrations: column overload

to trace concentrations

Complex matrices

Cost of Operation

May 2014


Laboratory Challenges Analytical Requirements

Throughput Analyze complex mixtures

Efficient workflow

Speed of analysis

Quality data/results

Efficient data

processing

Simple sample

prep

May 2014


Technologies to Improve the Productivity of Your Laboratory

Increasing Throughput and Efficiency

May 2014

6

GC and MS Technologies Improving Sample Throughput

Thro

ughp

ut

CFT Backflush

Retention Time Locking

Self-Cleaning Ion Source

May 2014


Capillary Flow Technology Select Modes of Operation

Detector Splitting

Backflush

Solvent Venting

May 2014


CFT Backflush Eliminates less volatile matrix components from the GC column by reversing the column flow at a pressure junction point:

Inlet

Flow

MS

Flow

Inlet

Flow

MS

Flow

May 2014


C.-K. Meng, Agilent Application Brief 5989-6018EN

• Elimination of long “baked out” at a high temperature to remove less volatile, late eluting matrix components

• Reduced analysis time • Increased column life time • Prevention of the MS source contamination • Less frequent MS source maintenance

Increased sample

throughput

CFT Backflush - Benefits

May 2014


CFT Backflush - Benefits

Using a 1.0 minute backflush prevents heavy matrix compounds from carrying over to next run

TIC: Blnk_After_Mshrms_BF_5min.D\data.ms

TIC: Blnk_After_Mshrms_BF_1min.D\data.ms (*)

2 3 4 5 6 7 8 9

2 3 4 5 6 7 8 9

Blank following mushroom sample, 0.5 min backflush (too short)

Blank following mushroom sample, 1.0 min backflush (correct length)

Ghost Peaks

Eliminate Ghost Peaks

May 2014


Column Backflush - Benefits

M. Mezcua, M.A. Martinez-Uroz, P.L. Wylie, A.R. Fernandez-Alba, J. AOAC Int. 92 (2009) 1790-1806.

Improved Chromatography

May 2014


Overlays of GC-MS/MS chromatograms for selected analytes (at 50 ng/g ) obtained within a 2.5-day sequence of 125 dietary supplement sample injections

Deltamethrinm/z 253>174

GinsengRootPowder

Saw Palmetto Berry Powder

ScutellariaPowderedExtract

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Dichlorvosm/z 185>93

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Malathionm/z 173>99

Ethionm/z 231>129

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Deltamethrinm/z 253>174

GinsengRootPowder

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ScutellariaPowderedExtract

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Malathionm/z 173>99

Ethionm/z 231>129

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Column Backflushing - Benefits

K. Mastovska and P.L. Wylie, J. Chromatogr. A 1265 (2012) 155-164

Accurate DB Match

May 2014


Retention Time Locking How Does It Work? For each method: • A set of five runs of retention time vs inlet pressure for a

single locking compound is collected (only once per method) • The calibration is stored with the method • When locking a new instrument, the locking compound is run and its RT is

measured • RTL software calculates new inlet pressure that makes the RT of the

locking compound and all analytes precisely match that of the original method

• Updating RTs of individual cal compounds and timed events unnecessary • If a table of hundreds of RTs is collected under RTL conditions, anyone

anywhere can lock to the same conditions and get the same RTs. This is basis of Agilent screening databases for Tox compounds, pesticides, etc.

May 2014


Retention Time Locking RTL Example

Initial run 4.72 psi

Trim 1 meter 4.72 psi

Relock 4.42 psi

4.296 min.

4.064 min.

4.297 min.

Original locked method

Column trimmed, locking compound run at original pressure (RT is too short)

RTL calculates new inlet pressure to restore RTs

RTL keeps the retention times of all analytes typically within 0.030 min absolute. Locking is a simple procedure using 1 compound. After pressure adjustment, all compounds fall within their RT recognition window.

May 2014


DB-5MS: Comparison of Different Speeds on MSD

1 x

2 x

3 x

4 x

30 m, 10C/min and 1.7 mL/min: 120V oven and Standard Turbo

15 m, 20C/min and 1.2 mL/min: 120V oven and Standard Turbo

15 m, 30C/min and 2.7 mL/min: 240V oven and Perf Turbo

15 m, 40C/min and 5.9 mL/min: 240V oven and Perf Turbo

4 8 12 16 20 24 28

2 4 6 8 10 12 14

1 2 3 4 5 6 7 8 9

1 2 3 4 5 6 7

4X method RTs are precisely the same as 1X divided by 4

May 2014


• MRM Spectrum Unchanged

• Troubleshooting

• Repeatability - Run-to-run - Operator-to-operator - Instrument-to-instrument

Retention Time Locking (RTL) Improve Confidence with Retention Time Locking

Initial run 4.72 psi

Trim 1 meter 4.72 psi

Relock 4.42 psi

0

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100

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100

0

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100

4.296 min.

4.064 min.

4.297 min.

Consistent Retention

Times

May 2014


GC/MS System Maintenance Potential disruption to workflow and productivity

System venting and disassembly of MS source

Manual cleaning requires expertise

Reassembly and system re-equilibration

System could be offline for full day of operation

May 2014


Self Cleaning Ios Source Add Hydrogen to the Source

Hydrogen In

Use CI transfer line on EI MSD. Hydrogen flows concentrically around end of column into ion volume, the same path used for CI reagent addition

May 2014


Self Cleaning Ion Source Add Hydrogen to the Source (top view)

Use CI transfer line on EI MSD. Hydrogen flows concentrically around end of column into ion volume, the same path used for CI reagent addition

Hydrogen in via CI transfer line

Repeller

Drawout (or extractor) lens

May 2014


Post Run Cleaning • Reduce frequency of cleaning ion source to less than once every 6 months • No changes to the user’s analytical method

In Situ Source Cleaning • High throughput users can reduce the need to vent their system to once every three months by

using In Situ Source Cleaning in combination with performing Post Run Clean once every two weeks.

• User would need to review analytical method for possible changes

Online vs Offline

May 2014


Self-Cleaning Ion Source Using H2 to Clean to MS Source

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252.2289.1

326.2360.2402.3440.3476.3

Very Dirty Source: Background Spectrum

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78.145.1 206.9 281.1134.9 355.1 445.5241.6168.1 320.6 388.9 477.9

After Cleaning with H2

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78.1

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281.1134.9 355.1

445.5241.6168.1 320.6 388.9 477.9

Expanded scale

50

May 2014


Cleaning Restores Analyte (OFN) Detection Comparable to manual cleaning?

4.104.124.144.164.184.204.224.244.264.284.304.324.344.364.38140000

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Time-->

( ) p p

4.104.124.144.164.184.204.224.244.264.284.304.324.344.364.380

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S:N > 50 pk-pk > 200 rms

S:N ≈ 1 pk-pk > 2 rms

Very dirty source before cleaning

After cleaning with H2

May 2014


Self-Cleaning Ion Source Stable ISTD RF for PAHs

May 2014


Additional calibration sets, standards in solvent, after system left idle for 12 days, with system modifications

Self-Cleaning Ion Source Stable calibrations for PAHs

May 2014


Self-Cleaning Ion Source On-Board GC/MS System Maintenance

On-Line bleed prolongs time between cleaning

Eliminates MS venting and source disassembly

Eliminates reassembly & system re-equilibration

System offline for 2 hours versus full day

Improves Workflow and Data Quality

May 2014


Effi

cien

cy Deconvolution Reporting Software

Peal Explorer

MRM Databases

Accurate Mass Databases

GC and MS Technologies Efficient Data Analysis

May 2014


Improving Efficiency Deconvolution Reporting Software

Deconvolution of mass spectra removes/reduces interferences from chromatographically overlapped peaks: • Improve identification and confirmation of analytes in high matrix samples • Reduces data review time, especially for large screening methods • Reduces false positives and false negatives in dirty samples • Identification based on matching entire spectrum cleaned of interferences

against library. Much more reliable than target/qualifier ratio method.

May 2014


Deconvolution Reporting Software Deconvolution Pulls Out Individual Components and Related Spectra

Deconvoluted components and spectra Component 1

Component 2

Component 3

TIC

Component 1 Component 2 Component 3

Deconvolution

Components and Mixed Spectra

May 2014


Deconvolution Challenge Standard Deconvolution Reporting Software (DRS) Familiarization

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What is this peak? Use conventional approach of average spectrum and PBM search to identify it

5.2 5.3 5.4

May 2014


Probability Based Matching Identifies THC Conventional Approach

• First hit is THC with 96 PBM match. • Second hit is hydrocodone with only

35 match

40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360

41 59 70 77 91 96 107

115

121 128

141 147 155 165 174

185 193 199

214 228

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284

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Average Spectrum

55 67 81 91 107 115 128 147

174 193 201 217

231

243 258

271

285

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40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360

THC, 96 Match

165

5.2 5.3 5.4

May 2014


Peak Is THC, Right? How confident are you?

55 67 81 91 107 115 128 147 165

174 193 201 217

231

243 258

271

285

299

314

40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360

Hit 1:THC, 96 Match PBM

42 44 51 55

59 68 70 77 82

84 91

96

103

115

121

128

141 155 162 171

185

188 199

214 228

242

256 270 284

299

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Hit 2: Hydrocodone, 35 Match PBM

40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360

41 59 70 77 91 96 107

115

121 128

141 147 155 165 174

185

193 199 214 228

231

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258

271

284

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Average Spectrum

May 2014


Overlap of THC and Hydrocodone As viewed in AMDIS

THC 5.281 min Hydrocodone 5.283 min

THC 314 THC 271

Hydrocodone 185 Hydrocodone 96

• Deconvolution finds 2 compounds closely merged together. • Peaks are separated by only 0.002 min. (Less than 1 scan)

May 2014


DRS Report for Challenge Standard Automated Deconvolution and Reporting

MSD Deconvolution Report Sample Name: THC/HC Data File: D:\MassHunter\GCMS\1\data\HCOD_THC.D Date/Time: 17:18 Wednesday, Oct 2 2013

Adjacent Peak Subtraction = 1 Resolution = High Sensitivity = Medium Shape Requirements = Medium

The NIST library was searched for the components that were found in the AMDIS target library.

Retention Time (R.T.) Cas # Compound Name Chemstation AMDIS Match R.T.

DiffReverse Match

Hit Number

3.2702 51799327 N-Propylamphetamine 99 -1.3 91 1

4.6881 3158858 10,11-Dihydrodibenz(b,f)(1,4)oxazepin-11-one 97 -0.7 90 1

5.2479 521379 Cannabidiol 68 5.6 64 3

5.281 1972083 Delta-9-tetrahydrocannabinol (THC) 95 0.5 85 1

5.283 125291 Hydrocodone 95 0.2 88 1

5.3145 466999 Hydromorphone 78 -0.1 67 2

5.3637 521357 Cannabinol 67 0.8 55 4

Amount (ng) AMDIS NIST

Automated report generation took about ~ 60 sec

May 2014


Agilent Peak Explorer Another View on Data

May 2014


October 13, 2014 Confidentiality Label

36


Rapidly Review Multiple Data Sets

October 13, 2014 Confidentiality Label

37


Efficient Data Review

MRM Databases for GC/MS/MS G9250AA – Pesticides & Environmental Pollutants

Average and exact Molecular Weight

Each compound classified into two categories

Database has RTs (and RIs) to be used with three GC methods (CF-40min, CP-40min, and CF-20min)

May 2014


Importance of MRM Transitions Malathion Identification

Confident Identification

May 2014


Why are MRM Transitions Important? Endosulfan Sulfate (?) in Tea Extract

3.3 ppb, Equiv : 0.016 mg/Kg in Tea Tea EU MRL : Sum Endo a+b+sulfate = 30 mg/kg

Std 5 ppb Reagent Blank Tea Extract A

• Single Qualifier Transition • Ion Ratio outside 80-120%

confidence band • Unconfirmed

Agilent Restricted

40

• Rerun of data • Ion Ratio within 80-120%

confidence band • CONFIRMED!!

• Demonstrates the value of multiple optimized transitions in the MRM Database.

• Not just to avoid matrix interferences but also for additional confirmation!

Tea Extract A

Accurate Confirmation

May 2014


Mass Profiler Professional - Totarol Treatment When you must identify that unknown

The structure is pulled out from ChemSpider or .mol file

Structure of corresponding

fragment

Precursor formula, based on accurate mass information

Compatibility score

: Using Molecular Structure Correlator to Predict the Structure of Fragments

May 2014


Mass Profiler Professional with 7200 Data Facilitating Data Review - Filtering, PCA, ANOVA, Volcano Plot

Mass Profiler Professional (MPP) was used for statistical evaluation of the data including construction of class prediction model to correctly predict

whether the sample would pass or fail the sensory test

Accurate Identification of

Unknowns

May 2014


Technologies for GC and GC/MS Improving Laboratory Efficiency

Efficient Workflow

Reduced Operating Expense

Improved Sample

Throughput

Robust Analytical

Performance

Efficient Data

Analysis

Confident Target and Non-target

ID

Consistent Accurate Results

May 2014


Questions… Thank you for your attention

May 2014


New GC & GC/MS Technologies to Improve the Productivity of … · Ghost Peaks Eliminate Ghost Peaks...

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Transcript of New GC & GC/MS Technologies to Improve the Productivity of … · Ghost Peaks Eliminate Ghost Peaks...