SBSE in a routine

environment, does it really

work?

A testimonial

2

TDU set-up

Cooled Injection System CIS

Thermal Desorption Unit TDU

CIS liner

Twister

TDU Liner

Coolant

Twister extraction in routine analysis:

Batch sample enrichment

Twister tray is fixed and followed-up

50 analyses with one Twister

“Transferline”

EQC in The Coca-Cola Company

The Coca-Cola Company

Concentrate plants Coca-Cola Technical

R&D QSE

GQAS Labs

APTC

Shanghai, China

EQC

Brussels, Belgium

Packaging & sales equipment lab

Trade sample lab

Analytical services lab

ATC

Johannesburg, SA

LAAS,

Mexico City, Mexico

EAS

Pune, IndiaNAAS,

Atlanta, USA

Coca-Cola International

North America

Analytical

Services Atlanta, GA

Eurasia Analytical

Services Pune, India

(June 2009

Europe : Assets Worth Euro 4.9 MM

Latin America

Analytical Services

Mexico City, MX

(July 2009)

Africa Technical

Center

Johannesburg, SA

(March 2008)

European Quality Center

Brussels, Belgium

(2001)

Asia Pacific

Technical Center

Shanghai, China

(March 2009)

Global Analytical Services Worldwide

Classified -

Unclassified

EQC provides laboratory testing services and expertise:

✓ To enhance and sustain quality performance and food safety

✓ To support innovations of beverage, packages, equipment and technologies

✓ To asses and address risks and support corrective actions

✓ To quickly resolve quality incidents

EUROPEAN QUALITY CENTER (EQC) – why and what?

EQC has strong expertise in:

- Analytical beverage testing

- Ingredients (CO2, sugar, water) testing

- Microbiology

- Packaging

- Immediate Consumption Equipment

- Sensory

- Beverage and Package Quality parameters

- Label claims and legislation

- Troubleshooting

EUROPEAN QUALITY CENTER (EQC) – How?

INGREDIENTS MONITORING PROGRAMS

Due diligence monitoring by in-house designed and organized programs:

• To ensure ongoing legal and TCCC compliance• To call for corrective actions in case of non-compliance

Following monitoring programs are in place:

• (Raw) and Treated water• Bottled water• Nutritional sweeteners• CO2

WEBU

CSE

R&D

EU

RUB

TCCAR

MENA

CEWA

EAG

Corporate quality

SRA

CPS

McDo Div

Other

Ingredient suppliers

Bottlers

McDo

….

External

EUROPEAN QUALITY CENTER (EQC) – customers?

8

The way to Q&A analysis – Compounds to be analyzedCoca – Cola is following WHO and European guidelines

To be analyzed:

- Pesticides

- PAH’s

- Phtalates

- phenols

History:

→ 3 different methods

- PAH”s: SBSE (1 cm x 0.5 µm Df, 24 µl PDMS)

- Phenols: derivatization (acylation/SBSE)

- Pesticides: SBSE (2 cm x 1 µm Df, 126 µl PDMS)

→ Simplification

- PAH’s together with pesticides

- Phenols

→ Current methodology – “all in one”

- SBSE for PAH’s, pesticides, phtalates and phenols

9

The way to Q&A analysis – Origin of the samples

“raw” water: depends on origin (ground water / tap water (storage))

→ desinfected (chlorinated) / purification

process water:

→ After purification / de-chlorinated

commercial water (mineral water) = “package water”

Actions to be made:

Acidification (phenolic compounds) / de-chlorination (Na-thiosulfate)

Analytical procedure → simplified and optimized

Water sample, 500 mL + 0.3 mL 10 % Na2S2O3

→ 60 mL sample + IS + “biggest” Twister (2 cm x 1 µm Df) GC MS

→ 20 mL sample + IS + “smallest” Twister (1 cm x 0.5 µm Df) GC QQQ

Already present

→ Complete validation (GCMS) – Currently ca. 80 samples / month

(2 samples can be analyzed together / risk of duplicate analysis afterwards)

→ Everything done in a Quality Assured environment

- ISO 17025 guidelines

→ Method conversion towards higher selectivity (GC QQQ)

- Sensitivity increased (pesticides, phenols)

- More difficult matrices (beverages)

- Miniaturization (robustness improved): Sample / Twister

60 mL to 20 mL → 2 cm x 1 µm Df to 1 cm x 0.5 µm Df

Future

→ SBSE GC QQQ in routine

→ Applying SBSE and GC QQQ to beverages

→ Package water – wall effect study:

Addition of organic modifier in SBSE enrichment

→ More compounds to be followed

→ Accreditation (Belac) – Audits

→ ...

10

The way to Q&A analysis – Present and Future plans

11

Method Configuration

CCD 2

+ TIC Scan pest 180216002.D

Blank Twister + ISTD

12

Twister in QC environmentPosition # of uses Position # of uses Position # of uses Position # of uses Position # of uses Position # of uses Position # of uses

1 15 10 29 10 43 9 57 9 71 9 85 9

2 10 16 10 30 10 44 9 58 9 72 9 86 9

3 10 17 10 31 10 45 9 59 9 73 9 87 9

4 10 18 10 32 10 46 9 60 9 74 9 88 9

5 10 19 10 33 10 47 9 61 9 75 9 89 9

6 10 20 10 34 10 48 9 62 9 76 9 90 9

7 10 21 10 35 10 49 9 63 9 77 9 91 9

8 10 22 10 36 9 50 9 64 9 78 9 92 9

9 10 23 10 37 9 51 9 65 9 79 9 93 9

10 10 24 10 38 9 52 9 66 9 80 9 94 9

11 10 25 10 39 9 53 9 67 9 81 9 95 9

12 10 26 10 40 9 54 9 68 9 82 9 96 9

13 10 27 10 41 9 55 9 69 9 83 9 97 9

14 10 28 10 42 9 56 9 70 9 84 9 98 9

# times used

(Max set at 50)

position in tray

Batch:

Lot:

Date # samples twister untill twister

10/15/2015 32 1

10/16/2015 26 1

10/20/2015 5 1

10/22/2015 33 1

11/4/2015 26 1

11/9/2015 37 1

11/18/2015 15 1

11/19/2015 48 1

11/24/2015 33 1

12/1/2015 31 1

12/4/2015 36 1

12/8/2015 30 1

12/14/2015 37 1

12/17/2015 5 1

12/18/2015 15 1

12/23/2015 37 1

1/4/2016 56 1

1/13/2016 40 1

1/22/2016 42 1

2/3/2016 37 1

2/18/2016 28 1

3/1/2016 24 1

3/10/2016 5 1

3/17/2016 37 1

3/29/2016 5 1

3/30/2016 34 1

4/8/2016 9 1

4/11/2016 29 1

4/19/2016 23 1

4/27/2016 14 1

4/29/2016 18 1

5/3/2016 28 1

5/18/2016 22 4 2 25

ISO 17025

Tracebility !!

Date – # of samples – tray number (start / end)

Staying aware of Twister usage → method robustness

13

Twister in QC environmentFollow-up internal standard response

Outlier !

ISO 17025

Tracebility !! Outcome:

- If 2x times out of spec → Redrawn from Twister batch

- Various causes: broken Twister, “bad” bleeding Twister, sample enrichment phenomena, wrong ISTD spike, etc.

Automatically “recheck” analysis for this Twister

0

2000

4000

6000

8000

10000

12000

14000

16000

18000

20000

0

100000

200000

300000

400000

500000

600000

700000

800000

900000

1000000

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26

01/03/2016 18:43:00

Acenaphtene D10 Phenanthrene D10 Chrysene D12 Benzyl butyl phthalate d4 Perylene D12

Staying aware of Twister condition → method robustness

# Samples in sequence

Validation elementMajor

Componen

t Assay

Trace

Analysis/

Quantificati

on

Presence/

Absence

Method

Transfer

Reference

Method

Physical

Test /

Characteristi

cs

Qualitative

Test/

Identificatio

n

Selectivity - Specificity Required Required Required No No No Required

Bias Required Required No Required Required Required No

Recovery Required Required No Required Required Required No

Precision - Repeatability Required Required Required Required Required Required No

Precision - Reproducibility Required Required Required Required Required Required No

Limit of Detection (LOD) No No Required No No No Required

Limit of Quantitation (LOQ) No Required No No No No No

Working (Linear) Range Required Required No No No Required No

Robustness Required (*)Required (*) Required (*) Required (*) Required (*) Required (*) Required (*)

Uncertainty Required Required Required No No No No

Method validation form

Creation Date: 18-Jun-2015 EQC, Brussels Form

Effective Date: Form DOC-06103

14

Twister in QC environment – EQC documentationWhat needs to be validated ?

EQC documentation

Outcome:

Everything put in macro containing Excel files

15

Twister in QC environment – EQC documentationRecovery mg/l

g/l

Method:

Parameter:

Matrix: Units

Period validation µg/L

People involved

Relevant matrices are measured non-fortified and fortified

Matrix description BBN Fortification Matrix Blank Matrix Fortified % Recovery

Milli Q 0.0008 0.0000 0.0009 110.508

TW (mix S15090062-79-86-95) 0.0017 0.0000 0.0012 73.416

RW (mix S15090054-55-56-57) 0.0025 0.0000 0.0027 107.34

PW NC (S15100139) 0.0033 0.0000 0.0033 98.925

PW C (CHF red 254 CH1 06:48-30 MAR16) 0.0042 0.0000 0.0044 104.7744

Average Recovery % = 99.0

Acceptable recoveries (%)

% w/w Acceptable Recovery %

1.00E+02 1000 g/l 98 - 101

1.00E+01 100 g/l 95 - 102

1.00E+00 10 g/l 92 - 105

1.00E-01 1 g/l 90 -108

1.00E-02 100 mg/l 85 - 110

1.00E-03 10 mg/l 80 -115

1.00E-04 1 mg/l 75 -120

1.00E-06 10 µg/l 70 -125

Concentration

0

Benzo(ghi) perylene

0

#

# 72compounds

Parameter LOD LOQ Repeat. (%) Reprod. (%) Rec. (%)

EPTC 2.07 6.91 2.8 3.1 107.5

Dichlobenil 3.19 10.62 5.7 4.7 85.7

Biphenyl 1.37 4.56 2.6 4.6 96.0

Etridiazole 1.85 6.16 3.8 3.9 113.8

Molinate 2.62 8.73 2.5 5 98.0

Tecnazene 1.65 5.49 2.5 4.1 95.9

Trifluralin 1.18 3.93 2.3 6.7 96.0

Alpha HCH 2.17 7.24 1.6 2.5 101.7

Hexachlorobenzene 0.86 2.87 1.5 3.2 103.0

terbufos 1.54 5.12 4.4 5.8 105.9

Trietazine 3.05 10.15 3.5 4.5 94.7

Terbuthylazine 3.77 12.58 6.1 9.2 97.6

Pentachlorophenol 2.89 9.64 10.6 12.3 93.7

Lindane 2.36 7.87 1.5 3.1 96.2

disulfoton 1.72 5.75 4.5 12.3 83.0

Beta HCH 2.59 8.63 4.1 6.1 90.7

Chlorpyrifos Methyl 1.82 6.08 1.6 3.8 99.0

2.4.4-trichlorobifenyl 1.13 3.76 1.1 2.6 99.7

Delta HCH 2.24 7.45 2.4 5.2 87.7

Heptachlor 0.95 3.18 2.2 6.8 110.4

Parathion methyl 2.09 6.96 3.8 10.5 93.7

ethofumesate 3.05 10.16 4.6 6.3 84.3

Malathion 2.33 7.76 5.3 7.1 84.9

Fenitrothion 2.13 7.09 4.1 9.3 88.0

2.2.5.5-tetrachlorobifenyl 1.20 4.00 1.5 3.2 99.1

Aldrin 1.06 3.53 1.9 3.6 98.8

Chlorpyrifos 1.67 5.55 1.4 4 97.1

Chlorthal dimethyl 1.96 6.53 1.4 3.6 96.9

Parathion ethyl 2.12 7.06 3.3 7.3 101.9

isodrin 1.06 3.54 1.8 4.4 94.4

chlozolinate 2.47 8.22 2.1 4.3 90.7

Pendimethalin 1.75 5.83 3.6 6.1 91.2

phenthoate 1.62 5.40 3.3 7.6 98.8

Heptachlor epoxide B 1.57 5.23 1.6 3.5 97.1

penconazole 2.85 9.49 3.7 6.5 107.6

Heptachlor epoxide A 1.64 5.47 1.7 3.5 91.7

2,4 DDE 1.40 4.66 1.6 4.1 96.1

Fluoranthene 0.14 0.45 3.4 5.6 99.8

2.2.4.5.5-pentachlorobifenyl 1.07 3.58 1.5 3.9 98.8

Trans chlordan 1.26 4.19 1.9 3.9 97.2

Cis chlordan 1.26 4.20 1.8 3.8 96.1

endosulfan I 1.57 5.23 2.3 5.1 92.1

prothiofos 1.52 5.06 1.6 4.1 95.2

4,4 DDE 1.19 3.96 1.4 3.8 96.7

Dieldrin 1.49 4.98 2.0 8.6 85.1

2,4 DDD 1.56 5.21 1.7 4.5 92.6

Endrin 2.00 6.65 5.8 5.7 99.8

ethion 1.47 4.89 2.8 7.1 93.7

2,4 DDT 1.00 3.34 3.3 6.5 104.9

4,4 DDD 1.87 6.23 1.2 6.4 92.9

2.2.4.4.5.5-hexachlorobifenyl 1.05 3.51 1.7 6.3 100.2

endosufan II 3.42 11.41 4.5 7.6 89.2

carbofenothion 1.92 6.39 4.0 9.2 94.7

Bis(2-ethylhexyl)adipate 0.03 0.10 2.1 11.2 103.1

4,4 DDT 1.10 3.65 3.6 8.6 103.1

2.2.3.4.4.5-hexachlorobifenyl 1.07 3.57 2.1 7.3 100.2

endosulfan sulfate 1.69 5.64 3.3 13.5 79.2

fenpropathrin 1.48 4.94 3.2 12.2 89.3

Methoxychlor 1.43 4.75 3.8 12.5 95.4

Bis(2-ethylhexyl)phthalate 0.05 0.18 1.2 11.2 97.5

Endrin Ketone 2.46 8.21 5.4 6.3 88.1

2,2,3,4,4,5,5-heptachlorobiphenyl 1.47 4.92 4.0 7.1 102.9

Pyriproxyfen 2.45 8.16 1.7 6.1 94.8

fenarimol 2.87 9.56 6.0 15.1 89.6

cis permethrin 2.07 6.90 2.6 9.6 89.6

trans permethrin 2.06 6.88 3.0 11.3 88.1

cyfluthrin 2.49 8.29 4.4 21.9 80.6

cypermethrin 1.57 5.23 4.3 22.6 79.0

Benzo(b,k)fluoranthene 0.32 1.08 1.6 9.7 110.7

Benzo(a)pyrene 0.09 0.31 1.5 4.3 101.8

Indeno(1,2,3-cd) pyrene 0.08 0.27 3.1 14.4 94.2

Benzo(ghi) perylene 0.08 0.27 4.5 13.8 99.0

16

Twister in QC environment – EQC documentation

Acenapthene-d10

Phenanthrene-d10

Chrysene-d12

Perylene-d12

Bis(2-ethylhexyl) phthalate-d4

17

All-in-one method transferred to GC-QQQ analysis

0

10

20

30

40

50

60

70

80

90

100

1 10 100 1000 10000 100000 K(o/w)

Re

co

ve

ry (

%)

SBSEParathion ethyl

20 mL, 1cm x 0.5mm

dichlobenil

20 mL, 1cm x 0.5mm

Parathion ethyl

60 mL, 2cm x 1mm

dichlobenil

60 mL, 2cm x 1mm

Miniaturization (sample amount / Twister size)

QQQ

Blank Twister (small)

Blank Twister (large)

1.1x104

6.0x105

GCMS

GCQQQ

50 ppt S/N @

5 ppt

GC QQQ

Rel. Conc.0.2 0.6 1 1.4 1.8

0.2

0.4

0.6

R2 = 0.9997

Re

l. R

esp

.

GC MS

0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8

-1x10

0.4

0.8

1.2

R2 = 0.9992

Re

l. R

esp

.

Rel. Conc.

Malathion (5 – 100 ppt)

8x10

LLE dirty aquoues sample – LVI inj.

0

+ TIC SIM 0416-0145.D

Counts vs. Acquisition Time (min)7 7.5 8 8.5 9 9.5 10 10.5 11 11.5 12 12.5 13 13.5 14 14.5 15 15.5 16 16.5 17 17.5 18 18.5 19 19.5 20 20.5 21 21.5 22 22.5 23 23.5 24 24.5 25

Scan

SIM

All-in-one method transferred to GC-QQQ analysisKeeping in mind future plans to analyze also beverages

7x10

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

+EI TIC MRM (** -> **) Coca_cola-twister-1.D

1 2 2 3 3 4 4 5 5 6 6 7 7 8 8 9 9 10

Counts vs. Acquisition Time (min)

5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36

TIC MRM

24 twisters over 24 hours

24 twisters over 24 hours

2.5 ppt

PAH

276 → 274

50 ppt

Cyfluthrin

163 → 91

241#

Response Benzo(g,h,i)perylene + ISTD

New features and improvements

Quick and easy

- 3 guiding poles for easier installation

- “slide in” carrier gas entrance line

→ flexibility towards TDU – Liquid injection

- PTV liner check tool

Hardware improvements

- Cooling ventilator is put outside the TDU heater body

- Thicker internal diameter for cooling lines

- UPC plus Peltier with standard installed filters for cooling

lines

- Better design – easier access inside of TDU

- More robust internal design of TDU

21

TDU, a product with already a long history, and even further improved by Gerstel !

22

ATEX

Thermal

Extraction in

µ-vials

Dynamic Headspace DHS

Twister Desorption

Thermal

Desorption of

Adsorbent tubes

Thermal

Extraction

of solids

SPME, Headspace- and

Liquid injection in dual step

PTV System HIT Headspace

Pyrolysis of solid and

liquid samples PYRO

Thanks to TDU 2 ...

Twicester®

mSBSE