Cameron George Technical Support Engineer 26 April 2001 An Optimized Dual Column System for the...

Cameron GeorgeTechnical Support Engineer26 April 2001

An Optimized Dual Column System for the Analysis of Chlorinated Pesticides, Herbicides and PCBs by GC-ECD

Cameron GeorgeApplications ChemistApril 26, 2001

11:00 a.m. ESTTelephone Number: 904-779-4715Chair Person: Lisa Lloyd

Starts in Five Minutes

Starts in One Minute

CLP Pesticides B-0447March 8, 2001

AREAS OF FOCUS

• Injector

• Detector

• Column (including guard column)

TRACE ANALYSIS INJECTION TECHNIQUES5-2000 pg On-Column

• Megabore direct

• Splitless

• PTV

• On-Column (cold & hot)

• Large volume

SAMPLE INJECTIONGoals

• Introduce sample into the column

• Reproducible

• No efficiency losses

• Representative of sample

ShortConcentrated

LongDiffuse

Solute Bands

Same column, same chromatographic conditions

INFLUENCE OF INJECTION EFFICIENCY

FIRST, SOME BASIC DEFINITIONS:

Regarding Inlets

• Backflash

• Discrimination

A FEW WORDS ABOUT BACKFLASH

Definition:

In a vaporization injection, a phenomenon wherein a portion of the sample expands beyond the boundary of the injection port liner towards the septum face and incoming gas line

BACKFLASHEffects

• Ghost peaks

• Erratic quantitation

• Poor accuracy

BACKFLASHCauses

• Highly volatile solvent

• Excessive inlet temperature

• Excessive injection volume

• Small liner volume

BACKFLASHPreventative Measures

• Lower inlet temperature

• Less volatile solvent

• 1-2 µL injection volume

• Chambered liner

• Using pulsed split or pulsed splitless

Yes NO!!!

INJECTION PORT LINERSSplitless

INLET DISCRIMINATION

• Injected sample Sample into the column

• Due to compound volatility differences

• Higher volatility = More into the column

INLET DISCRIMINATIONIn Pesticide Analyses

Generally inlet discrimination is not a concern for

pesticide analyses when utilizing appropriate GC

inlets such as Splitless, Pulsed Splitless and PTV

OTHER INLET CONSIDERATIONS

• Silylation

• Using glass wool

• Cleaning and reusing liners

A FEW WORDS ABOUT RESIDUES

• Semivolatile vs nonvolatile

• Worst offenders

• Sample prep

GUARD COLUMNSUse 0.53mm Or 0.32mm ID Deactivated Tubing

• Fit all inlets

• Enhanced deposition of residues

• Easy to work with

• Minimum 1 meter

Usually 1-5 meters long and same diameter as the column

Detector

DeactivatedFused SilicaTubing

UnionColumn

Injector

GUARD COLUMNTraps non-volatile sample residues

COLUMN CONNECTORSThermal Mass, Inertness, Seal Integrity

• Stainless steel

• VU-Tight

• Press-fits

• Integral guard columns (Duraguard)

Let’s Be SensitiveANALYTE DETECTION

The minimum amount of an analyte that can still be confidently identified as a peak (S/N > 4)

SENSITIVITYAnalytical Definition

• Detector Sensitivity: No sample influence (standard)

• Method Sensitivity: Matrix influence (sample)

"Normal" response

1. High noise, detector, background

2. Absorption, breakdown of analyte

3. Sample prep losses

"Improved" response

1. High quality circuit components and reagents

2. Inertness

3. Method optimization

RESPONSE TO A CONSTANT ANALYTE AMOUNT2 Cases

HP 6890 Series Micro-ECD Design

Comparison of HP 6890 ECDs

Standard

< 0.040

> 10^4

no spec.

5 - 80

Micro-ECD

< 0.008

> 5 x 10^5

> 5 x 10^4

1 - 500

pg/sec lindane

lindane

pg (ppb)

Dynamic range

Linear range

Linear Range, CLP pesticides

Maximum data rate

Break Number 1

For Questions and Answers

Press *1 on Your Phone to

Ask a Question

LETS GET TO THE CHROMATOGRAPHY

WHAT EXACTLY ARE WE TRYING TO ACHIEVE

• The best resolution possible

• Minimize analysis time

• Get MDLs as low as possible

RESOLUTION VS SEPARATION

• Separation: Time between the 2 peaks

• Resolution: Describes how well 2 peaks are separated with regard to their widths

10 11 12 13 14 15

11.24 12.72

(tm = 95.5)

11.6111.14

WHICH PAIR OF SOLUTES HAVE BETTER SEPARATION?

10 11 12 13 14 15

11.24 12.72

11.6111.14

Better Separation

= 1.17 Rs = 0.6

Better Resolution

= 1.05 Rs = 2.7

K = 7.00K = 6.07

K = 6.00 K = 6.30

RESOLUTION VS SEPARATION

RESOLUTION AND ANALYSIS TIME

• Improving resolution often results in the opportunity to shorten analysis times

• Many variables can affect resolution

N = (L, rc)

k = (T, df, rc)

= (T, phase)

RN kks =

RESOLUTION

The column must provide sufficient retention of the early eluting compounds without excessive retention of the late eluting compounds

IMPROVING RESOLUTION

Retention

IMPROVING RESOLUTIONFilm Thickness

Decreasing Film Thickness Results In:

• Increased efficiency

• Elution of analytes at lower temperatures

• Decreased analysis time

• Decreased bleed interference

• Increased column activity

• Decreased capacity

IMPROVING RESOLUTIONEfficiency

• High column efficiency is necessary to resolve large numbers of compounds

• Improperly operated injectors and/or improperly optimized carrier gas can result in efficiency losses

IMPROVING RESOLUTIONColumn Length

Increasing Column Length Results In:

• Increased analysis time

• Increased bleed

• Big increase in cost

IMPROVING RESOLUTION Column Inner Diameter

Decreasing Column Inner Diameter Results In:

• Increased head pressure

• Decreased capacity

• Decreased carrier gas flow rates

IMPROVING RESOLUTION Stationary Phase

• Stationary phase selectivity has the largest impact on separation, thus resolution

• Optimization of stationary phase selectivity should be approached cautiously

For Years Environmental Laboratories Have Suffered With Pesticide Analyses Using Non-

Optimal Stationary Phases

TRADITIONAL STATIONARY PHASES

•5% Phenyl-methylpolysiloxane

•35-50% Phenyl-methylpolysiloxane

•Trifluoropropyl-methylpolysiloxane

•14% Cyanopropylphenyl-methylpolysiloxane

PROBLEMS WITH TRADITIONAL PHASES

•Long analysis times (Over 30 minutes!)

•High bleed resulting in decreased sensitivity

•Poor resolution and confirmation capabilities

•Poor inertness of some phases

TRADITIONAL PESTICIDE COLUMNS

High Bleed at 280°C

0 10 20 30 40Time (min.)

608 Type Phase

1701 Type Phase

Coelution of peaks 10 & 11

40 min

Poor peak shape for Endrin aldehyde

EFFORTS TO IMPROVE PESTICIDE ANALYSES

Application Specific Phases

Stationary phases designed with a primary focus placed upon maximizing separation () for a specific group of target analytes

APPLICATION SPECIFIC PHASES

C2H4CF

CH3m n

Si SiO O

Phase 1Trifluoropropyl-dimethylpolysiloxane

Si SiO O

C2H4CF

Common CLP Pesticide Phases

Phase 2Trifluoropropyl-diphenyl-dimethylpolysiloxane

Dimethyl functionality of Phase 2 not shown

EFFECT OF PHASE POLARITY

Polarity

Thermal Stability

DRAWBACKS OF APPLICATION SPECIFIC PHASES

•Limited thermal stability resulting in longer analysis times and hampered sensitivity

•Excessive column conditioning times leading to increased column activity

•Decreased column lifetimes

PHASES DESIGNED WITH OPTIMUM PERFORMANCE IN MINDArylene Phase Technology

OBiphenyl

CH3 CH

Arylene

LOW BLEED STATIONARY PHASESArylene Structure

DB-35ms

8.00 10.00 12.00 14.00 16.00 18.00 20.00

320°C

300°C

CLP Pesticides Analysis

LOW BLEED STATIONARY PHASESDB-35ms vs. DB-35

STATIONARY PHASE AFFECT ON SENSITIVITY

Both 30 m x 0.25 mm I.D., x 0.25 µm5 ng Decachlorobiphenyl

S/N =10 S/N =3

DB-35ms, 320°C DB-35, 300°C

LOW BLEED STATIONARY PHASESSpectral Purity

m/z-->

78 96 119 144

356 377

455470

50 100 150 200 250 300 350 400 450 500

Primary IonDB-35ms

m/z-->

96 119

135156

315 331

50 100 150 200 250 300 350 400 450 500

Primary Ion

BENEFITS OF LOW BLEED STATIONARY PHASES• Improved sensitivity

• Improved spectral purity

• Reduced analysis times

• Potential increase in column lifetimes

• Rigorous inertness testing

• Less Detector maintenance

Break Number 1

For Questions and Answers

Press *1 on Your Phone to

Ask a Question

DEMANDING PESTICIDE APPLICATIONS BENEFIT FROM ADVANCED PHASE TECHNOLOGY

DB-35ms and DB-XLB capillary columns contain second generation arylene stationary phases giving them enhanced thermal stability

DB-35MS PRIMARY COLUMNCLP Pesticides

•Should provide adequate resolution of all 22 CLP Pesticides and surrogates

•Short analysis time ideal

•Easily realized analysis conditions preferable

•Should give minimal column bleed

1. Tetrachloro-m-xylene (SS)2. -BHC3. -BHC4. -BHC5. Heptachl

or6. -BHC7. Aldri

n8. Heptachlor

epoxide9. -

Chlordane

10. -Chlordane

11. Endosulfan I12. 4,4’-DDE13. Dieldrin14. Endrin15. 4,4’-DDD16. Endosulfan II17

. 4,4’-DDT18

. Endrin

aldehyde19. Endosulfan

sulfate20. Methoxychlor21. Endrin

ketone 22.

Decachlorobiphenyl (SS)

CLP PESTICIDESColumn: DB-35ms

30 m x 0.32 mm I.D., 0.25 µmP/N: 123-3832

Carrier: Helium at 45 cm/sec (EPC in constant flow mode)Oven: 110°C for 0.5 min

110-320°C at 15°C/min320°C for 2 min

Injector: Splitless, 250°C30 sec purge activation time50 pg per component

Detector: µECD, 350°CNitrogen makeup gas (column + makeup flow = 30 mL/min constant flow)

min2 4 6 8 10 12 14 16

6 10 14 168 12

Short Analysis Time

Low Bleed At 320ºC

DB-XLB CONFIRMATION COLUMNWith Regard To The Primary Column

•Should resolve any co-elutions

•Needs to provide adequate resolving power when run under the same analytical conditions

•Should possess at least equivalent thermal stability

•Should provide a different relative elution pattern

•Ideally, elution order changes should be realized

CLP PESTICIDESColumn: DB-XLB

30 m x 0.32 mm I.D., 0.50 µmP/N: 123-1236

110-320°C at 15°C/min320°C for 2 min

min2 4 6 8 10 12 14 166 10 14 168 12

100% Confirmation In Under 16 Minutes

1. Tetrachloro-m-xylene (SS)2. -BHC3. -BHC4. -BHC5. Heptachl

or6. -BHC7. Aldri

n8. Heptachlor

epoxide9. -

Chlordane

10. -Chlordane

11. Endosulfan I12. 4,4’-DDE13. Dieldrin14. Endrin15. 4,4’-DDD16. Endosulfan II17

. 4,4’-DDT18

. Endrin

aldehyde19. Endosulfan

sulfate20. Methoxychlor21. Endrin

ketone 22.

Decachlorobiphenyl (SS)

OPTIMIZING FOR SPEED WITHOUT SACRIFICING ANALYTE RESOLUTION

•Use Hydrogen as the carrier gas

•Set linear velocity at 65 cm/sec (3.4 mL/min)

•Increase oven ramp rate from 15°/min to 25°/min

1. Tetrachloro-m-xylene (SS)2. -BHC3. -BHC4. -BHC5. Heptachlor6. -BHC7. Aldrin8. Heptachlor epoxide9. -Chlordane

10. -Chlordane11. Endosulfan I12. 4,4’-DDE13. Dieldrin14. Endrin15. 4,4’-DDD16. Endosulfan II17. 4,4’-DDT18. Endrin aldehyde19. Endosulfan sulfate20. Methoxychlor21. Endrin ketone

22. Decachlorobiphenyl

CLP PESTICIDES USING HYDROGEN CARRIER GASColumn: DB-35ms

30 m x 0.32 mm I.D., 0.25 µmP/N: 123-3832

3 4 5 6 7 8 9 1010

4 85 6 7 9

Baseline Resolution In Under 9 Minutes!

Carrier: Hydrogen at 65 cm/sec (EPC in constant flow mode)Oven: 110°C for 0.5 min

110-320°C at 25°C/min320°C for 2 min

1. Tetrachloro-m-xylene (SS)2. -BHC3. -BHC4. -BHC5. Heptachlor6. -BHC7. Aldrin8. Heptachlor epoxide9. -Chlordane

10. -Chlordane11. Endosulfan I12. 4,4’-DDE13. Dieldrin14. Endrin15. 4,4’-DDD16. Endosulfan II17. 4,4’-DDT18. Endrin aldehyde19. Endosulfan sulfate20. Methoxychlor21. Endrin ketone

22. Decachlorobiphenyl

Carrier: Hydrogen at 65 cm/sec (EPC in constant flow mode)Oven: 110°C for 0.5 min

110-320°C at 25°C/min320°C for 2 min

3 4 5 6 7 8 9 10

105 7 96 8

CLP PESTICIDES USING HYDROGEN CARRIER GASColumn: DB-XLB

30 m x 0.32 mm I.D., 0.5 µmP/N: 123-1236

Excellent Resolution And Confirmation In Under 10 Minutes

OTHER COMMON ECD APPLICATIONS THAT THIS DUAL COLUMN SHOULD BE ABLE TO PERFORM

•EPA 8151A (Phenoxy acid herbicides)

•EPA 508.1 (Pesticides)

•EPA 552.2 (Haloacetic acids)

•EPA 8082 (PCB congeners and Aroclors)

2 4 6 8 10 12 14 16Time (min)

EPA 8151A PHENOXY ACID HERBICIDES

DB-XLB

2 4 6 8 10 12 14 16Time (min)

DB-35ms

2 4 6 8 10 12 14 16

12 131415

min2 4 6 8 10 12 14 16

1 23 4

121314 15

1819 20

EPA 8151A PHENOXY ACID HERBICIDESDB-35ms

30m x 0.32 mm I.D., 0.25 µmP/N: 123-3832

DB-XLB30m x 0.32 mm I.D., 0.50 µmP/N: 123-1236

50-100°C at 25°C/min100-320°C at 12°C/min320°C for 2 min

Injector: Splitless, 250°C

30 sec purge activation time50 pg per component

1. Dalapon2. 3,5-Dichlorobenzoic acid3. 4-Nitrophenol4. Methyl-2,4-dichlorophenylacetate (SS)5. Dicamba6. MCPP7. MCPA8. 4,4´, Dibromooctafluorobiphenyl (IS)9. Dichloroprop

10. 2,4-D11. Pentachlorophenol12. 2,4,5-T,P13. 2,4,5-T14. Chloramben15. Dinoseb16. 2,4-DB17. Bentazone18. DCPA19. Picloram20. Acifluorofen

EPA 508.1 PESTICIDES

DB-XLB

DB-35ms

8 10 12 14 16 18 20 22 24Time (min)

min8 10 12 14 16 18 20 22 24

min8 10 12 14 16 18 20 22 24 8 10 12 14 16 18 20 22 24

Time (min)

303132

262728

EPA 508.1 PESTICIDES

DB-35ms30m x 0.32 mm I.D., 0.25 µmP/N: 123-3832

75-300°C at 10°C/min300°C for 2 min

1. Hexachloropentadiene2. Etridiazole3. Chloroneb4. Trifluralin5. Propachlor6. Hexachlorobenzene7. a-BHC8. Atrazine9. Pentachloronitrobenzene (IS)

10. Simazine11. g-BHC12. b-BHC13. Hepatachlor14. Alachlor15. d-BHC16. Chlorothalonil17. Aldrin18. Metribuzin19. Metolachlor20. DCPA

21. 4,4´-Dibromobiphenyl (SS)22. Heptachlor epoxide23. Cyanazine24. g-Chlordane25. a-Chlordane26. Endosulfan I27. 4,4´-DDE28. Dieldrin29. Chlorobenzilate30. Endrin31. 4,4´-DDD32. Endosulfan II33. 4,4´-DDT34. Endrin Aldehyde35. Endosulfan sulfate36. Methoxychlor37. cis-Permethrin38. Trans-Permethrin

min4 5 6 7 8

EPA 552.2 HALOACETIC ACIDS

DB-XLB

DB-35ms

4 5 6 7 8Time (min)

min4 5 6 7 8

4 5 6 7 8Time (min)

EPA 552.2 HALOACETIC ACIDS

1. Chloroacetic acid2. Bromoacetic acid3. Dichloroacetic acid4. Dalapon5. Trichloroacetic acid6. 1,2,3-Trichloropropane (IS)7. Bromochloroacetic acid8. Bromodichloroacetic acid9. Dibromoacetic acid

10. 2,3-Dibromopropionic acid (SS)11. Chlorodibromoacetic acid12. Tribromoacetic acid

DB-35ms30m x 0.32 mm I.D., 0.25 µmP/N: 123-3832

40-200°C at 15°C/min200°C for 2 min

EPA 8082 PCB CONGENERS

DB-XLB

DB-35ms

6 8 10 12 14 16Time (min)

min6 8 10 12 14 16

6 8 10 12 14 16Time (min)

101112

EPA 8082 PCB CONGENERS1. IUPAC 12. Tetrachloro-m-xylene (IS/SS)3. IUPAC 54. IUPAC 185. IUPAC 316. IUPAC 527. IUPAC 448. IUPAC 669. IUPAC 101

10. IUPAC 8711. IUPAC 11012. IUPAC 15113. IUPAC 15314. IUPAC 14115. IUPAC 13716. IUPAC 18717. IUPAC 18318. IUPAC 18019. IUPAC 17020. IUPAC 20621. Decachlorobiphenyl (IS/SS)

DB-35ms30m x 0.32 mm I.D., 0.25 µmP/N: 123-3832

110-320°C at 15°C/min320°C for 5 min

WHAT DOES THIS MEAN FOR ENVIRONMENTAL LABORATORIES?

•Improved productivity

•Increased sample throughput

•Superior results

•Total solution for ECD systems

CONCLUSION

•Excellent selectivity

•Exceptional inertness

•Superb thermal stability

DB-35ms and DB-XLB provide excellent resolution and confirmation for CLP Pesticides and other critical GC/ECD methods providing total performance:

CONCLUSION

When optimized columns are combined with effective injector setup and detector design improved testing results are realized while sample throughput and laboratory efficiency are improved dramatically

AGILENT’S J&W TECHNICAL SUPPORT

It’s Free!(800) 552-0413 (phone)(916) 985-7888 (phone)(916) 608-1964 (FAX)

web site:www.agilent.com/chem

Wrap-up E-Seminar Questions

Thank you for attending Agilent e-Seminars.

Our e-Seminar schedule is expanding, please

check our website frequently at:

www.agilent.com/chem

Cameron George Technical Support Engineer 26 April 2001 An Optimized Dual Column System for the...

Documents

de PCBs PCBs web site : ...2 . 0 INK PCBs MILK CAUTION PCBs . Title: ثنائي الفينيل متعدد الكلور PCBs (مطوية) Author: التنمية البيئية - المواد

PCBs - Presentación

MONITORING PLAN Revision1 - TCEQ - · 1.1 PCB Background and Properties PCBs are a group of 209 chlorinated organic compounds composed of two connected phenyl rings with 1 to 10

PCB Polychlorinated biphenyl. What are PCBs? PCBs belong to a broad family of man-made organic chemicals known as chlorinated hydrocarbons. PCBs were

Chlorinated biphenyls effect on estrogen-related receptor ...Yoshida et al. 1998). In rats after in utero or lactational expo-sure, PCBs 126 and 169 inhibited conversion of round sper-matidsbetweenstagesVIIandVIII.Ontheotherhand,PCBs

res__0222_151211 PCBS

Vancouver Lake PCBs, Chlorinated Pesticides, and Dioxins ... · Vancouver Lake collected in 1993 and 2002 could have been due to factors such as size of fish, lipid content, analytical

Disseny PCBs

Herbicides toxicity

Les herbicides

A TMDL Evaluation for Chlorinated Pesticides and PCBs in

herbicides oldset

Method 615: The Determination of Chlorinated Herbicides in ... › sites › production › files › 2015-10 › ...Method 615 . primary column. Section 15 provides gas chromatograph

Appli Biochem Biotechnol 2010 Bio Degradation Chlorinated Non Chlorinated Vocs Parmaceutical Industries

OCCURRENCE OF PCBs IN THE CZECH REPUBLIC. BIODEGRADATION OF BIPHENYL AS A MODEL STUDY FOR BIODEGRADATION OF ITS CHLORINATED DERIVATIVES. Jan Tříska Global

CHLORINATED SOLVENTSolinchlorinatedorganics.com/.../11/Chlorinated-Solvents...Digital.pdf · 2 Chlorinated Solvents Product Stewardship Manual ... Physical and Chemical Properties

METHOD 508.1 DETERMINATION OF CHLORINATED PESTICIDES, HERBICIDES

INTRODUCTION TO VALENT BIOSCIENCES · Products SCC Insecticides/ Herbicides SCC Insecticides/ Herbicides SCC Insecticides/ Herbicides/ Fungicides Microbial Biorational Insecticides

Potholes Reservoir: Screening Survey for Dieldrin, Other ......Potholes Reservoir: Screening Survey for Dieldrin, Other Chlorinated Pesticides, and PCBs in Fish, Water, and Sediments

CHLORINATED PARFINS · 2018. 6. 28. · CHLORINATED PARS 57 Table 2. Chemical and physicalproperties of representative chlorinated paraffnsa Paraffn Average Chlorine Density Refractive