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June 14, 2017 ASMS 2017 1

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Validation of New GC and GC/MS Technology with Real World Matrix

Rebecca Veeneman, PhD Validation of New GC and GC/MS Technology with Real World Matrix June 7, 2017

June 14, 2017 ASMS 2017 2

• Introducing Intuvo

• A perfect partner for Mass Spectrometry

• Validation through matrix applications

• 8270 in soil and sediment

• Pesticides in food

• Additional benefits from Intuvo

• Conclusion

June 14, 2017 ASMS 2017 3

Outline

4

Shifting Demographics

— Seasoned operators retiring; highly

skilled replacements harder to find

— GC troubleshooting skills not

necessarily located at point of use

— Operators assuming multiple

responsibilities

Challenging Economics

— Budgets squeezed, expectations

increased

— Capital purchase decisions made on

business NOT technical basis

June 14, 2017 ASMS 2017

Challenges Facing The GC Lab Time for a change

June 14, 2017 5

Round 1 Round 2

ASMS 2017

Scouring The Globe Listening to customers

June 14, 2017 6

Improve the User Experience

• Install and setup

• Use and maintain

• Make GC more practical for today’s busy lab enterprise

Prepare for Next Generation of Users

Its not only about better analytical performance – its about better lab outcomes

ASMS 2017

The GC Community Voice We listened and responded

The GC Usability Gap Usability lags features and performance

• Over the years, GC features and performance have matured and largely serve today’s needs

• Usability has lagged and does not meet today’s expectations, especially as expertise more scarce

June 14, 2017 7

Features and performance Usability

Time

ASMS 2017

June 14, 2017 8

Make GC Easier

ASMS 2017

Operator Frustration

“I’m spending too much downtime trimming columns, recalibrating, and maintaining GC systems.

That’s far too many billable samples we’re not running”

June 14, 2017 9

The GC Community Voice Too much user intervention

ASMS 2017

Managing Challenge

“Sometimes we can’t be sure what’s in a sample.

Our GC systems need to handle whatever comes through the lab, nasty or nice”

June 14, 2017 10

The GC Community Voice Welcome to the real world

ASMS 2017

June 14, 2017 11

Accelerating Success

• Increased revenue

• Improved on-time delivery to customers

• Reduced opportunity loss

• Reduced training requirements

• Reduced business uncertainty

The GC Community Voice Drive better business outcome

ASMS 2017

Better Business Outcome

• Less maintenance and downtime, more samples run • Streamlined operation with lower facilities impact

June 14, 2017 12

Simplicity

• Simpler operation, less training, fewer mistakes • Transformed user interface requiring less training

Innovation

• GC re-imagined: the future of how we GC • Instrument, consumables, software, services

The GC Community Voice Driving Agilent strategy

ASMS 2017

Introducing Intuvo 9000 GC System

• A completely re-imagined integrated system of GC consumables, MS services and software

• New paradigm streamlining the way GC is done

• Better Business Outcome

June 14, 2017 ASMS 2017 13

• Guard chip protection of column and MS ion source

• Retention time stability after changing guard chip

• Easy and reliable backflush

• Leak-free click-and-run connections

• Fast direct heating and cooling

June 14, 2017 ASMS 2017 14

An Ideal Partner for Mass Spectrometry

15

Direct heating

A whole new way to GC

Innovating a New Path to GC Productivity

June 14, 2017 ASMS 2017

16

No-trim column

Direct heating



June 14, 2017 ASMS 2017

17

No-trim column

Direct heating

Ferrule-free click-and-run connections



June 14, 2017 ASMS 2017

18

Disposable Guard chip

No-trim column

Direct heating




June 14, 2017 ASMS 2017

19

Disposable Guard chip

No-trim column

Direct heating

Modular Intuvo flow chips




June 14, 2017 ASMS 2017

• SSL, MMI, GSV, LSV inlets

• FID, TCD, ECD, NPD, FPD, NCD, SCD detectors

• SQ and TQ mass spectrometers

• Headspace, thermal desorption, purge and trap samplers, CTC PAL 3

• 16-, 50-, 150-position auto-injectors and trays

June 14, 2017 ASMS 2017 20

Configurable to any application

Flexible Compatible Design

June 14, 2017 ASMS 2017

Click-and-run direct

connections

To detector

To detector

Conventional flow path

Intuvo flow path

Inlet Inlet

Guard chip

Intuvo planar column

Gold seal

Nuts and ferrules

Classic capillary column

Flow chip

21

Innovating The GC Flow Path

22

• Inert

• Modular, configurable

• Click-and-run connections

Intuvo Flow Chip cross section

Thermally conductive proprietary alloy

High purity silica channel

Intuvo ultra inert treatment

Carrier gas

High purity inert silica flow path

Modular Intuvo Flow Chips

June 14, 2017 ASMS 2017

• No more ferrules

• Direct face seal connections

• Audible and tactile click lets you know connection is made

• Easier to train

• Less unplanned downtime

• Fewer batch reruns and precious samples lost

June 14, 2017 ASMS 2017 23

Eliminating connection uncertainty

Click-and-Run Direct Connections

Microfluidic-enabled 6th generation EPC modules allow hands-free leak checking to confirm and document leak-free operation autonomously.

June 14, 2017 ASMS 2017 24

Avoiding unplanned downtime

Autonomous Leak Checking

• Fast, efficient heating

• Ballistic cooling

• Smaller footprint

June 14, 2017 ASMS 2017 25

Every single minute counts

Fast Direct Heating And Cooling

26 June 14, 2017 ASMS 2017

Summary Of Intuvo Applications

Environmental • EPA Method 8270 – Semivolatile Organic Compounds (SVOC) • Ultra-Fast Total Petroleum Hydrocarbons (TPH)

Energy and Chemical • Ultra-Fast SimDis using ASTM D7798 • Monocyclic Aromatic Purity using ASTM D7504 • Sulfur in Light Petroleum Liquids

Food • Multiresidue Pesticide Analysis in Food

Pharma • Extractables and Leachables

Consumer Products • Longevity of Phthalates • Fragrance Impurities using Intuvo with OpenLab Match/Compare

27 June 14, 2017 ASMS 2017







Resilience to Matrix and

Difficult Compounds

28 June 14, 2017 ASMS 2017








Difficult Compounds

June 14, 2017 ASMS 2017

8270 Calibration Standard – 77 Acids/Bases/Neutrals (PAHs), 6 ISTDs

Compound

Number

Compound

1 N-Nitrosodimethylamine

2 Pyridine

3 2-Fluorophenol (surrogate)

4 Phenol-d5 (surrogate)

5 Phenol

6 Aniline

7 Bis(2-chloroethyl) ether

8 2-Chlorophenol

9 1,3-Dichlorobenzene


11 Benzyl alcohol


13 2-Methylphenol

14 Bis(2-chloroisopropyl) ether

15 4-Methylphenol

16 N-Nitrosodi-n-propylamine

17 Hexachloroethane

18 Nitrobenzene-d5 (surrogate)

19 Nitrobenzene

20 Isophorone

21 2-Nitrophenol

22 2,4-Dimethylphenol

23 Benzoic acid

24 Bis(2-chloroethoxy)methane

25 2,4-Dichlorophenol

26 1,2,4-Trichlorobenzene

27 Naphthalene

28 4-Chloroaniline

29 Hexachlorobutadiene

30 4-Chloro-3-methylphenol

31 2-Methylnaphthalene

32 Hexachlorocyclopentadiene

33 2,4,6-Trichlorophenol

34 2,4,5-Trichlorophenol

35 2-Fluorobiphenyl (surrogate)

Compound

Number

Compound

36 2-Chloronaphthalene

37 2-Nitroaniline

38 Dimethyl phthalate

39 2,6-Dinitrotoluene

40 Acenaphthylene

41 3-Nitroaniline

42 Acenaphthene

43 2,4-Dinitrophenol

44 4-Nitrophenol

45 2,4-Dinitrotoluene

46 Dibenzofuran

47 Diethyl phthalate

48 4-Chlorophenyl-phenyl ether

49 Fluorene

50 4-Nitroaniline

51 4,6-Dinitro-2-methylphenol

52 N-Nitrosodiphenylamine

53 Azobenzene

54 2,4,6-Tribromophenol (surrogate)

55 4-Bromophenyl phenyl ether

56 Hexachlorobenzene

57 Pentachlorophenol

58 Phenanthrene

59 Anthracene

60 Carbazole

61 Di-n-butylphthalate

62 Fluoranthene

63 Benzidine

64 Pyrene

65 p-Terphenyl-d14

66 Butylbenzylphthalate

67 3,3'-Dichlorobenzidine

68 Benzo[a]anthracene

69 Bis(2-ethylhexyl)phthalate

70 Chrysene

Compound

Number

Compound

71 Di-n-octyl phthalate

72 Benzo[b]fluoranthene

73 Benzo[k]fluoranthene

74 Benzo[a]pyrene

75 Indeno[1,2,3-cd]pyrene

76 Dibenzo[a,h]anthracene

77 Benzo[g,h,i]perylene

78 1,4-Dichlorobenzene-d4 (internal standard)

79 Naphthalene-d8 (internal standard)

80 Acenaphthalene-d10 (internal standard)

81 Phenanthrene-d10 (internal standard)

82 Chrysene-d12 (internal standard)

83 Perylene-d12 (internal standard)

29

Semivolatiles Selection Of Analytes

Injection volume 1 µL

Inlet Split/Splitless 300 C

Pulsed splitless 60 psi until 0.5 min

Purge 50 mL/min at 0.5 min

Septum purge switched flow mode 3mL/min

Liner UI Splitless liner with a single taper and glass wool at the bottom

Guard chip (Intuvo only) 60 C for 2 min, 20 C/min to 260 C, 6 C/min to 330 C hold 1.333

min (standards) or 10.333 min (soil extract)

Column Agilent DB-5ms UI 30 m x 0.25 mm x 0.5 µm

Flow 2 mL/min constant flow

Column temperature 40 C for 2 min, 20 C/min to 260 C, 6 C/min to 330 C hold 1.333

min (standards) or 10.333 min (soil extract)

Transferline temperature 330 C

Drawout plate 6 mm

Ion source temperature 330 C

Quadrupole temperature 200 C

Scan 35 to 550 m/z

Gain factor 1

Threshold 50

A/D samples 2

GC (7890 & Intuvo)

MSD (5977)

30 June 14, 2017 ASMS 2017

Semivolatiles GC Method

8270 Calibration Standard – 77 Acids/Bases/Neutrals (PAHs), 6 ISTDs

31

Intuvo

7890

June 14, 2017 ASMS 2017

Chromatographic Comparison: Which Is Intuvo?

0.5

0.6

0.7

0.8

0.9

1

1.1

1.2

0 10 20 30 40 50 60 70 80

Re

lati

ve R

ete

nti

on

Tim

e

Compound Number

7890

Relative Retention Time = RTtarget / RTISTD

0.5

0.6

0.7

0.8

0.9

1

1.1

1.2

0 10 20 30 40 50 60 70 80

Re

lati

ve R

ete

nti

on

Tim

e

Compound Number

7890 Intuvo

Average relative retention time difference = 0.0006

32 June 14, 2017 ASMS 2017

Retention Time Comparison

Response Comparison

Response Factor = (Areatarget * ConcentrationISTD)/(AreaISTD * Concentrationtarget)

Average response factor difference = 4.6%

0.00

0.50

1.00

1.50

2.00

2.50

0 10 20 30 40 50 60 70 80

Re

sp

on

se

Fa

cto

r

Compound Number

7890 Intuvo

33 June 14, 2017 ASMS 2017

Test system acid/base activity, inertness and MSD tuning ratios (25 µg/mL)

Pentachlorophenol

Criteria: TF < 2 @ 10% height

DFTPP

Criteria: IE fragments within specified ion ratios

Benzidine

Criteria: TF < 2 @ 10% height

DDT

Criteria: Breakdown to DDD and DDE < 20%

0

10

20

30

40

50

60

70

80

90

100

11.50 12.00 12.50 13.00 13.50 14.00 14.50 15.00 15.50

No

rma

ilze

d R

es

po

ns

e

Time (min)

34 June 14, 2017 ASMS 2017

Semivolatiles Suitability Standard

Test system acid/base activity, inertness and MSD tuning ratios (25 µg/mL)

Pentachlorophenol

Pass TF = 1.0

DFTPP

Pass Ion ratios in spec

Benzidine

Pass TF = 0.8

DDT

Pass Breakdown = 1.4%

0

10

20

30

40

50

60

70

80

90

100

11.50 12.00 12.50 13.00 13.50 14.00 14.50 15.00 15.50

No

rma

ilze

d R

es

po

ns

e

Time (min)

35 June 14, 2017 ASMS 2017

Semivolatiles Suitability Standard

Decision Flow Diagram

Initial QC and

Calibration

Inject Matrix

Set (N=20)

Check

QC / CCV /

ISTD

Pass Check?

Change

septum, liner

Corrective

Maintenance

Attempt 1?

Corrective

Maintenance

Attempt 2

Replace Guard

Chip

Yes

No No

Yes YesNo

New Column

Check

QC / CCV /

ISTD

Pass Check?

MSD

Maintenance

Check

QC / CCV /

ISTD

Yes

No

36 June 14, 2017 ASMS 2017

Matrix Study For Semivolatiles

Composite soil extracts in methylene chloride from ESC Lab Sciences (Mt. Juliet, TN)

20 sequential injections on a wool packed liner

Typical of their “Worst Matrix”

37 June 14, 2017 ASMS 2017

Semivolatiles Matrix Extracts

June 14, 2017 ASMS 2017 38

Breakdown Limit

0

10

20

30

40

50

60

70

80

90

100

0

20

40

60

80

10

0

12

0

14

0

16

0

18

0

20

0

22

0

24

0

24

0

26

0

28

0

30

0

32

0

34

0

36

0

38

0

40

0

42

0

44

0

46

0

48

0

50

0

52

0

54

0

54

0

56

0

58

0

60

0

62

0

64

0

66

0

68

0

4,4

'-DD

T %

Bre

akd

ow

n

Number of Matrix Injections

Contaminated liner

New liner

Suitability Test – Tuning Standard DDT breakdown

Guard Chip Replacement

Tailing Factor Limit

0

0.5

1

1.5

2

2.5

3

0

20

40

60

80

10

0

12

0

14

0

16

0

18

0

20

0

22

0

24

0

26

0

28

0

30

0

32

0

34

0

36

0

38

0

40

0

42

0

44

0

46

0

48

0

50

0

52

0

54

0

56

0

58

0

60

0

62

0

64

0

66

0

68

0

70

0

Ta

ilin

g F

ac

tor

at

10

% P

ea

k H

eig

ht


Pentachlorophenol Benzidine

June 14, 2017 ASMS 2017 39

Semivolatiles Calibration Verification – Matrix Study Pentachlorophenol and benzidine tailing factor – after liner change

-40

-30

-20

-10

0

10

20

30

40

0 10 20 30 40 50 60 70 80

Ca

libra

tio

n C

he

ck

% E

rro

r

Compound Number

After Calibration

40 June 14, 2017 ASMS 2017

Semivolatiles Calibration Verification – Matrix Study Continuing calibration verification

Hexachlorocyclopentadiene

4-Nitrophenol

Pentachlorophenol

-40

-30

-20

-10

0

10

20

30

40

0 10 20 30 40 50 60 70 80

Ca

libra

tio

n C

he

ck

% E

rro

r

Compound Number

After Calibration

After 680 Matrix Injections

41 June 14, 2017 ASMS 2017


Benzidine

4-Nitrophenol

Pentachlorophenol

Hexachlorocyclopentadiene -40

-30

-20

-10

0

10

20

30

40

0 10 20 30 40 50 60 70 80

Ca

libra

tio

n C

he

ck

% E

rro

r

Compound Number

After Calibration

After 680 Matrix Injections

After Column Replacement

42 June 14, 2017 ASMS 2017


0.00

0.20

0.40

0.60

0.80

1.00

1.20

1.40

1.60

0 100 200 300 400 500 600 700

No

rma

lize

d I

nte

rna

l Sta

nd

ard

Are

a


1,4-Dichlorobenzene-d4 Naphthalene-d8

Acenaphthene-d10 Phenanthrene-d10

Chrysene-d12 Perylene-d12

43 June 14, 2017 ASMS 2017

Semivolatiles ISTDs– Matrix Study Internal standard response

44 June 14, 2017 ASMS 2017








Difficult Compounds

Injection volume Sandwich injection

1 µL standard + 0.5 µL analyte protectants + 1 µL matrix

Inlet Split/Splitless 280 C

Pulsed splitless 30 psi until 0.5 min

Purge 15 mL/min at 0.5 min

Septum purge flow mode 3mL/min

Guard chip (Intuvo only) 60 C for 1.5 min, 50 C/min to 310 C

Column Agilent HP-5ms UI 15 m x 0.25 mm x 0.25 µm

Flow 1.3 mL/min constant flow

Column temperature 60 C for 1.5 min, 50 C/min to 160 C, 8 C/min to 240 C, 50 C/min

to 280 C, 100 C/min to 290 C hold 1.1 min

Transferline temperature 310 C

Ion source temperature 280 C

Quadrupole temperature 150 C

MRM P&EP MRM database

GC (7890 & Intuvo)

MSD (7000 Series)

45 June 14, 2017 ASMS 2017

Pesticides in Food GC Method

June 14, 2017 ASMS 2017

Calibration

Cal Check

60 matrix injections

Maintenance

Cal Check

8 level calibration curve (1-1000ppb)

Calibration curve accuracy check with

50ppb standard

Honey, rice, orange, black tea, olive oil,

onion, or cucumber

Replace septum, liner, guard chip

Calibration curve accuracy check with

50ppb standard

46

Preemptive Maintenance Protocols For Pesticides In Food

0

20

40

60

80

100

120

140

160

Rec

ove

ry

Honey

Rice

Orange

Black Tea

Olive Oil

Onion

Cucumber

June 14, 2017 ASMS 2017

• Average cal check accuracy for 60 injections with the seven extracts are shown.

• Average accuracy for the 50ppb calibration standard was 82%

• RSD was 6.3%

• 70% and 120% accuracy thresholds are noted

Ac

cu

rac

y

47

Average Calibration Accuracy For 60 Matrix Injections

0

20

40

60

80

100

120

140

Rec

ove

ry

Honey

0

20

40

60

80

100

120

Rec

ove

ry

Black Tea

June 14, 2017 ASMS 2017

0

20

40

60

80

100

120

140

Rec

ove

ry

Olive Oil

0

20

40

60

80

100

120

Rec

ove

ry

Cucumber

Ac

cu

rac

y

Ac

cu

rac

y A

cc

ura

cy

Ac

cu

rac

y

Honey Black Tea

Olive Oil Cucumber

48

Despite Matrix Difficulty Calibration Accuracy Is Maintained

Chromatograms from the 50ppb cal check, after 60 extract injections, and after maintenance are overlaid. Retention times and peak shapes do not change.

June 14, 2017 ASMS 2017

21

3

4

5

6

7

8

9

10

1112

13 14

15

16

17

1819

20

21

22,23

24

25

26

27

Honey extract

21

3

45

6

7

8

9

10

11 12

1314

15

16

17

18 19

20

2122,23

24

25

26

27

Cucumber extract

Over 500 injections were made on a single 15m column without column or source maintenance Symmetric peaks and consistent recoveries were observed

Honey Rice Orange Tea Olive Oil Onion Cucumber

49

Chromatograms Are Consistent Beginning To End

June 14, 2017 ASMS 2017

Inject matrix (N=30)

Initial calibration

Check calibration accuracy

Pass? Maintenance #1

Maintenance #2

yes

no

yes

Replace liner

yes

Replace guard chip

no

New column

Check calibration accuracy

Pass? yes

Clean source

50

Reactive Maintenance Protocol With Spinach

June 14, 2017 ASMS 2017

Injection 1 Injection 2 Injection 3

Injection 174 Injection 175 Injection 176

51

Intuvo Longevity Of Deltamethrin

52

0.5 min

Shifts beyond retention time

window

Pesticide mix injected before and after 300 mm column trim

0.5 min

Maintaining retention times

Pesticide mix injected before and after guard chip change

Conventional Column Intuvo Column

June 14, 2017 ASMS 2017

Intuvo Guard Chip And No-Trim Columns No retention time shifts after maintenance

June 14, 2017 Intovo Applications 53

Intuvo – Guard Chip Replacement

7890 – Column Trim

Intuvo Guard Chip – A New Way To Maintain The System Column trimming compared to guard chip replacement

54 June 14, 2017 ASMS 2017

Conclusions

• The Intuvo GC presents a straight-forward system well-suited to address the requirements of matrix methods

• Highly consistent chromatography was observed even with heavy matrix load

• With regular maintenance (guard chip and liner changes), the column can withstand over 500 injections without loss of chromatographic integrity

– Peak shape was symmetric and consistent

– Retention times were unchanged

– Response was maintained

– Suitability standards continue to pass

– Source remained clean

• Modular flow path simplifies method development

• Small footprint allows more instruments/bench

Thank You!

Questions?

June 14, 2017 ASMS 2017 55

validation of new gc and gcms technology with real world ... · validation of new gc and gc/ms...

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