Latest Update for Food and Environmental from Waters · 2019. 12. 16. · Refreshed Xevo TQ-S micro provides –Update to the detector to improve performance for small negative ionizing

©2019 Waters Corporation 1COMPANY CONFIDENTIAL

Latest Update for Food and Environmental from Waters

2019 Food and Water Summit, 11-12th September 2019

Dr. Peter Hancock


Leading Scientific OrganizationTechnology Provider

Understand market needs

Monitor & assess market trends

Meet regulatory requirements

Address analytical challenges

Introduce innovative technology

Waters’ Role in Food and Environmental Testing

Shared responsibility

Capacity building

Educational programs

Network creation

Partnerships


Regulatory

Requirements

Future Directions

and Trends

Emerging Risks

and scares

Current Analytical

Challenges

• Sample diversity and complexity

• Sensitivity, selectivity and speed of analysis

• Direct or point of use analysis

• Contaminant scares

• Consumer/Trade Protection

• Toxicological assessments

• Researching links between food, nutrition and health

• Technological developments for analyte detection

• Improving data management/use of digital information

Understanding Needs

• Permitted limits; banned contaminants

• Validation and Analytical QC

• Reference methods


Regulatory

Requirements

Future Directions

and Trends

Emerging Risks

and scares

Current Analytical

Challenges










Understanding Needs





TQ-S cronos

New

Accessible reliable performance

TQ-S micro

Refreshed

Sensitive, flexible and compact

What is the new TQ family for 2019?

Ultimate sensitivity and flexibility

TQ-XS


Xevo TQD, Xevo TQ-S cronos, Xevo TQ-S micro

Sensitivity comparison for some pos ion pesticides

5x

15x

4x

15x

3x

12x

Xevo TQ-S micro peak area

Xevo TQ-S cronos peak area

Xevo TQD peak area

1x

1x

1x

Aztrazine-desethyl

Dicrotophos

Methamidophos


Xevo TQD, Xevo TQ-S cronos, Xevo TQ-S micro

Sensitivity comparison for some neg ion pesticides

11x

55x

13x

47x

8x

34x

Xevo TQ-S micro peak area

Xevo TQ-S cronos peak area

Xevo TQD peak area

1x

1x

1x

Fluazinam

Fludioxynil

Lufenuron


Why is glyphosate (and other polar pesticides) such a

challenge to analyse?

Method Step Challenges

Sample Preparation

Glyphosate is very polar, water soluble and insoluble in organic

solvents, and will bind to metal ions, therefore making the extraction

possibilities limited. Derivatization can be very time consuming

SeparationInsufficient retention on column using reverse phase, requires

derivatization, HILIC or IEX stationary phase

Analysis

Within the extract, other water soluble matrix compounds can also be

found (proteins, sugars, amino acids, salts, etc.) that interfere with the

determination of glyphosate. The use of derivatization is not compound

specific and can lead to selectivity issues

Glyphosate used as a desiccant on cereal crops to aid harvest-results in increased

frequency of residues in products such as bread and breakfast

cereals and beer


AMPAGlufosinateGlyphosate

Extended scope of analytes related to glyphosate

Parents:

Glyphosate

Glufosinate

Ethephon

Fosetyl aluminum

Metabolites:

N-acetylglyphosate, aminomethylphosphonic acid (AMPA), n-acetylAMPA

N-acetylglufosinate, 3-methylphosphinicopropionic acid (MPP)

Regulated residue definitions:

mg/kg in

wheatGlyphosate Glufosinate* Fosetyl al**

EU 10 0.03 2

US 30 - -

China 5 - -

* Glufosinate residue definition must report the sum of glufosinate, its salts

MPP and NAG, expressed as glufosinate equivalents.

** Fosetyl aluminum must report the sum of fosetyl, phosphonic acid and their

salts, expressed as fosetyl.

Anionic polar pesticide compounds

https://ec.europa.eu/food/plant/pesticides/eu-pesticides-database/public/?event=pesticide.residue.selection&language=EN

https://bcglobal.bryantchristie.com/db#pesticides/query

https://gain.fas.usda.gov/Pages/Default.aspx


Our journey to the NEW Anionic Polar Pesticide Column

➢ Mixed mode

Retention

Separation of critical pairs

➢ HILIC type: DEA

Retention

Excellent separation of critical pairs

Retention time stable

Perchlorate compatible

➢ HILIC type (polyvinyl alcohol)

Retention

Acceptable separation of critical pairs

Retention time unstable

Perchlorate incompatible (5 min wide peak)

Retention

RSDs > 20% in different commodities

Lemon Wheat flour

AMPA

Fos Al

Retention time stability

Glyphosate

13.94 min 12.70 min

Separation


Waters Anionic Polar Pesticide Column

– 5 µm BEH DEA Particle Technology

– 2.1 x 100mm UPLC Hardware

Enhancements have enabled:

– Extended Peak Shape Integrity

– Maintain Key Analyte Separations

– Retention Without Derivatization

– Reliable Low-Level Detection

– No Sodium EDTA wash

– No need for high levels of buffers

What it is the APP column and why?

5 µm

1.7 µm

8.5 X

5 µm

1.7 µm4.5 X

AMPA

Glyphosate


Retention: Analyte retention, without derivatisationRetaining analytes to maximise performance

Time1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 10.50 11.00 11.50 12.00 12.50

%

0

100

t0 = 0.48min

Minimum acceptable RT for the analyte(s) should be at minimum twice the RT

corresponding to the void volume of the column (SANTE/11813/2017)


Separation: Critical separations to avoid false detectionsIsobaric interferences from targeted analytes

AMPA

fosetyl-al

phosphonic acid

Separation of fosetyl isobaric

interference in AMPA and

phosphonic acid MRM transitions.

110 < 63

109 < 63

81 < 63


min5.050 5.100 5.150 5.200 5.250 5.300 5.350 5.400 5.450 5.500 5.550 5.600 5.650 5.700 5.750

%

0

100

HPEU_021_220819_120

QuPPe Flour 2.5 ng/mLGlyphosate

5.35

1378

min

%

0

100

HPEU_021_220819_120

QuPPe Flour 2.5 ng/mLGlyphosate

5.35

1580

Spike equivalent to 0.010 mg/kg

min2.850 2.900 2.950 3.000 3.050 3.100 3.150 3.200 3.250 3.300 3.350

%

0

100

HPEU_021_220819_120

QuPPe Flour 2.5 ng/mLGlufosinate

3.05

233

2.85

min

%

0

100

HPEU_021_220819_120

QuPPe Flour 2.5 ng/mLGlufosinate

3.05

307

Detection: Detecting, Identifying and QuantifyingAccurately, well below current MRLs

0.010 mg/kg – 0.400 mg/kg wheat flour matrix matched

Glu

fos

ina

teG

lyp

ho

sa

te

Compound name: Glyphosate

Correlation coefficient: r = 0.999119, r^2 = 0.998238

Calibration curve: 530.208 * x + 72.8129

Response type: External Std, Area

Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None

Standard Addition Concentration : 0.137329

Conc-0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100

Res

pons

e

-0

20000

40000

Conc

Res

idua

l

-5.0

0.0

5.0

10.0

Compound name: Glufosinate


Calibration curve: 129.083 * x + -39.8463



Conc-0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100

Res

pons

e

-0

2500

5000

7500

10000

12500

Conc

Res

idua

l

-5.0

-2.5

0.0

2.5

5.0


Refreshed Xevo TQ-S micro provides

– Update to the detector to improve performance for small

negative ionizing daughter ions (<m/z 150)

– Enhancements to MassLynx quan workflow

Time3.40 3.60 3.80 4.00 4.20 4.40 4.60 4.80 5.00 5.20 5.40 5.60 5.80 6.00

%

0

100 Glyphosate m/z: 168 > 63

3.5 fold increase in peak area

Xevo TQ-S micro

Refreshed Xevo TQ-S micro

Detection: Achieving default MRL on Xevo TQ-S microEnhanced Xevo TQ-S micro and column showing small can be powerful

Gly1: Gly


Reference retention time (TRef) TRef± 0.1

Tomato | Cucumber | Wheat flour

Reliability: Retention time stabilityRetention times you can rely on

Reliable retention times, even between matrices, without significant column conditioning or maintenance.


Regulatory

Requirements

Future Directions

and Trends

Emerging Risks

and scares

Current Analytical

Challenges










Understanding Needs





Mass Spectrometric Profiling

Direct MS

• Minimal sample preparation

• No chromatography, Ambient ionisation

• Spatially well defined

• Few hundred species in 1-5 sec

Traditional

• Sample preparation

• GC-MS, GC-MS/MS

• LC-MS, LC-MS/MS

• Few thousand species in 5-30 min


Direct analysis of;

– Solids & powders (black tea, spices, etc)

– Liquids (edible oils, whiskey, milk, etc)

– Tablets

– Vapour / aroma

Automation using 12 position

Quick strip cards, Dip-it and SPE-it sampling

DART QDa sample introduction techniques

https://www.google.co.uk/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&uact=8&ved=2ahUKEwjB297BysPaAhUIaRQKHXMMDYUQjRx6BAgAEAU&url=https://www.livestrong.com/article/68702-pros-cons-powdered-milk/&psig=AOvVaw2v2TjwN_7nN3ylTwemtGag&ust=1524132590373377

http://www.google.co.uk/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&uact=8&ved=2ahUKEwjdlqL-zMPaAhUCxxQKHX6eAGcQjRx6BAgAEAU&url=http://jamestownarts.com/2018/01/creative-resolutions/red-white-wine-120111/&psig=AOvVaw0cFd_TXEdBhkm54yhNZZ9Y&ust=1524133250750822

http://www.google.co.uk/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&ved=2ahUKEwirqIaUvsjZAhWthOAKHW39DIwQjRx6BAgAEAY&url=http://www.dailymail.co.uk/health/article-4486130/Cinnamon-reverse-high-fat-diet.html&psig=AOvVaw1yHZGfuXRb1JCnwZ_K9E3q&ust=1519903004371100


Sample preparation

0.2 g honey

Add 1 mL of ultrapure water

Warm (45oC), vortex mix 30 s

Add 9 mL of 50:50 MeCN: MeOH

Vortex mix 30 s

Roller shake ~ 5mins

Centrifuge @ 6000 rpm for 5 mins

Remove 1 mL supernatant

Dispense 3 µL onto Quickstrip and

allow to dry

Honey samples

1. Light brown / set

2. Yellow / set

3. Yellow / set

4. Dark brown / runny

5. Honey supplement

(light brown / runny)

DART/QDa, Honey authenticity example


Honey sample 4 and solvent blank overlay

Showing unique feature regions

Monosaccharides[M+H]+ , [M+nH2O]+

Disaccharides[M+H]+ , dehydration products

Oligosaccharides[M+H]+ ,dehydration productsFlavonoids

Helium

DART+

Desorption temp: 350oC

Rail speed: 0.8 mm/min

Cone voltage: 10 V

Mass range: m/z 100 – 600

Acquisition: Continuum

Sampling frequency: 2 Hz

Gain: 1


Honey profile

DART QDa positive ionisation mode He gas @ 350oC

Adulterated honey sample

Authentic honey sample 1





LiveID 1D Loadings plot, PCA/LDA 3D Scores Plots

PCA/LDA model

PCA model


Extracted ion chromatograms for ion at 390.1

Sample 1 (authentic honey)

Sample 5 (adulterated honey)

Full scan spectral overlay

380 – 397 m/z

Solvent blank – red

Authentic honey – green

Adulterated honey - purple


Regulatory

Requirements

Future Directions

and Trends

Emerging Risks

and scares

Current Analytical

Challenges










Understanding Needs





Perfluoroalkylated Substances (PFAS)

PFAS = PFC = AFFF

First created in the 1930s

Widespread applications

– Non stick coatings, surfactants, packaging, firefighting foams

– Polymerization aid for polytetrafluoroethylene (PTFE) and

other fluoropolymers

Stable and persistent in the environment (POP)

– Bio-accumulative

Identified in environmental samples worldwide

– Found in arctic polar bears

– Most humans have a range of PFAS in their blood

PFOS and PFOA most studied PFAS compounds but 1000’s have

been created

Perfluoro Carboxylic Acid

(PFOA)

Perfluoro Sulfonic Acid

(PFOS)


Worldwide Interest in PFAS


EPA Method

537*

ISO

25101

ASTM

7979

Sample Prep SPE SPE Dilute, Filter

Injection Volume 1 - 10 µL 1 - 10 µL 30 µL

Instrument sensitivity

requiredXevo TQ-S micro Xevo TQ-S micro Xevo TQ-XS

Number of

compounds18 2 24

Comparison of analysis methods

*EPA 537 is for drinking water analysis only


463>369

463>369

Time2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50

%

0

100

2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50

%

0

100

100 pptPFOA standard

Blank injection

BackgroundPFOA

PFOA

PFAS’s Interferences/Contamination


Solvent

Flow

PEEK

Tubing

Stainless

Steel Coil

Isolator

Column

Waters PFC Kit


Time2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00

%

0

100

2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00

%

0

100

Nopeaks

Background PFOA contaminant

Background PFOA contaminant

PFOA

Blank Injection

300 pptPFOA standard

0.5min

MRM Chromatograms of PFOA

With PFC Kit Installed


ASTM 7979-17

Sample Prep:

5 mL sample + 5 mL

methanol

2 g sample + 10 mL

1:1 water : methanol

Adjust pH to ~ 9-10

Shake 1 hour

Syringe filter entire

sample

Acidify and transfer to

polypropylene vial

WA

TE

R

AS

TM

7979

SO

IL

AS

TM

7968

Sample Pre-

Treatment

WaterSurface

Ground

Influent

Effluent

SoilsSand

Silt

Lean Clay

Fat Clay


Compounds Included in Methods

Carboxylates

C4 – C14, C16, C18

Sulfonates

C4 – C10

Others

4:2/6:2/8:2 FTS FHEA/FOEA/FDEA

FHUEA FHpPA

FOUEA

PFecHS

diPAP

Emerging

GenX ADONA

11Cl-PF3OUdS 9Cl-PF3ONS

PFEESA

PFMBA

NFHDA

FOSA

NMeFOSAA/NEtFOSAA

NMeFOSA/NEtFOSA


Source Parameters

Instrument: Xevo TQ-XS

Ion Mode: ESI-

Capillary Voltage: 1.0 kV

Desolvation Temperature: 500°C

Desolvation Flow: 1100 L/hr

Cone Flow: 150 L/hr

LC Method

Instrument: Acquity I Class with PFC Kit

Column: CSH Phenyl Hexyl 2.1mm x 100 mm, 1.7 µm

Mobile Phase A: 95:5 H2O:MeOH + 2 mMammonium acetate

Mobile Phase B: MeOH + 2 mM ammonium acetate

Injection Volume: 30 uL

Gradient:

Instrument Methods

Time

(min)

Flow

(mL/min)

%A %B

0 0.3 100 0

1 0.3 80 20

6 0.3 55 45

13 0.3 20 80

14 0.4 5 95

17 0.4 5 95

18 0.3 100 0

22 0.3 100 0

MS Method

• Developed using QuanOptimize

• MRMs, CV, CE

• Divert flow to waste from 15 – 21 mins


Cal Curve pt 8

Time3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00 14.00

%

0

100

PFAS_1Feb18_010 68: MRM of 1 Channel ES- TIC (M4_82diPAP)

9.93e6

Overall Method Summary

ERA certified QC sample injected as control throughout

analysis

Reporting Limit Check

Blank sample + targets &

surrogates. Taken through sample

prep

Detectable recovery

35-150%

Reagent Blanks

50:50 water : methanol + 0.1% acetic

acid

Concentration must be < half reporting limit

Method Blank

Blank sample + surrogates.

Taken through sample prep

Concentration must be < half reporting limit

Lab Control

Blank sample + targets &

surrogates. Taken through sample

prep

Target recovery

70-150%


Method Detection Limits (MDL)

Well below the necessary reporting limits for the compounds defined in

ASTM 7979

Reporting Range is the required range the method must be able to cover as defined in ASTM 7979 method


Compound name: PFOA





Conc-0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200

Re

sp

on

se

-0

25000

50000

75000

100000

125000

150000

175000

200000

225000

250000

275000

Curve 3

Time10.90 11.00 11.10 11.20

%

0

100

10.90 11.00 11.10 11.20

%

0

100

PFAS_30Jan19_09 Sm (Mn, 2x2) 34: MRM of 2 Channels ES- 498.9 > 98.9 (PFOS)

9.56e3Area

11.01;566


5.71e3Area

11.01

454

498.9 >

98.9

Linearity and Sensitivity

2.5 ng/L injection

R2 = 0.9992

Curve 3

Time9.75 9.80 9.85 9.90 9.95

%

0

100

9.75 9.80 9.85 9.90 9.95

%

0

100

PFAS_30Jan19_09 Sm (Mn, 2x2) 20: MRM of 2 Channels ES- 412.9 > 368.9 (PFOA)

9.64e4Area

9.82;4407


2.42e4Area

9.82;1327

412.9 >

368.9

Curve 3

Time9.75 9.80 9.85 9.90 9.95

%

0

100

9.75 9.80 9.85 9.90 9.95

%

0

100


9.64e4Area

9.82;4407


2.42e4Area

9.82;1327

412.9 >

168.9

PFOA

Compound name: PFOS


Calibration curve: 196.264 * x + -39.187



Conc-0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200

Re

sp

on

se

-0

5000

10000

15000

20000

25000

30000

35000

R2 = 0.9980

PFOS

Curve 3

Time10.90 11.00 11.10 11.20

%

0

100

10.90 11.00 11.10 11.20

%

0

100


9.56e3Area

11.01;566


5.71e3Area

11.01

454

498.9 >

79.9

2.5 ng/L injection

Cal 2

Time6.80 6.90 7.00 7.10

%

0

100

6.80 6.90 7.00 7.10

%

0

1006.95;417

6.95;602

Cal 2

Time6.80 6.90 7.00 7.10

%

0

100

6.80 6.90 7.00 7.10

%

0

1006.95;417

6.95;602

285 >

185

285 >

119

Compound name: GenX





ppt-0 20 40 60 80 100 120 140 160 180 200

Re

sp

on

se

-0

1000

2000

3000

4000

5000

6000

10 ng/L injection

R2 = 0.9988

GenX


Regulatory

Requirements

Future Directions

and Trends

Emerging Risks

and scares

Current Analytical

Challenges










Understanding Needs





SELECT SERIES Cyclic IMS is a unique ion mobility

mass spectrometer combining next generation cyclic

ion mobility with a new high performance TOF

Key features of Cyclic IMS

– Higher, variable IMS resolution

– IMS selection of ions

– IMSn multi stage IMS selection and fragmentation

Provides unlimited experimental opportunities

enabling leading researchers to solve the most

challenging analytical problems

Select Series Cyclic IMS


Universal Ion SourceESI & nESI

NEW Cyclic IMSHigh/ variable resolution Novel IMSn funcionality NEW Offset Tof

Extended flight path

Dual Gain ADCExtended

dynamic range

XS Collision CellEnhanced transmission

Stepwave XSEnhanced transmission

characteristics

NEW Customised MassLynxWith interactive instrument

control software


1 pass

(65 /)

2 passes

(100 /)

4 passes

(140 /)

5 passes

(158 /)

3 passes

(124 /)

IMS Performance Summary

Single and Multi Pass IMS

Trisaccharide mix - all 3 components have the same m/z [527.1588] and can’t be separated by mass spectrometry

Raffinose MaltotrioseMelezitose

*SYNAPT G2-Si, Resolution ~40


Delivering new levels of ion mobility performance

50 passes R(CCS/CCS) ~ 500

5 passes R(CCS/CCS) ~ 165

Time (ms)

38 39 40 41 42 43 44 45 46 47 48

360 370 380 390 400 410 420 430 440 450 460

Raffinose

Maltotriose

Melezitose

Cyclic IMS provides extraordinary ion mobility resolution

allowing highly detailed structural information to be obtained


Product ion mass spectra of mobility separated (Synapt)

danofloxacin protomers (I) and (II).

Observed mass [m/z]

Inte

nsity [

counts]

CH3

N N

N

F

O+

O

OH H

-H2O

-CH3NH2

N N

N+

F

O

O

O

CH3

H

H

H

-CO2

I

II

-HF

0

5000

10000

15000

20000

25000

30000

35000

40000

45000

1 2 3

Re

spo

nse

Replicate

Enrofloxacin Protomer Response Vs Replicate Spiked Porcine Extract

Enrofloxacin

Enrofloxacin_1

0

5000

10000

15000

20000

25000

1 2 3

Re

spo

nse

Replicate

Ciprofloxacin Protomer Response Vs Replicate Spiked Porcine Extract

Ciprofloxacin

Ciprofloxacin_1

https://doi.org/10.1002/rcm.8371



Danofloxacin IM protomer separation using Synapt

(N2 and CO2 IM gas) and cIM (N2 IM gas, 5 passes)

Arrival Time [ms]

4.54

5.56 82.89

92.42

95.96

Synapt N2cIM N2

II

I

II

III

I

II

III

Synapt CO2

5.9

3.84

4.20

I




Protein structureOligosaccharidesTechnology

Breakthrough innovation, driven by science

Multiple peer reviewed publications

Antibiotic residues


Waters has commitments as a technology provider and leading scientific

organisation for food and water safety, quality testing and research

Our goal is to understand your food and environmental testing needs and

provide technologies, knowledge and methods to address those needs

– Regulatory requirements

– Analytical challenges

– Emerging risks and food scares

– Future directions and trends

Summary

Documents

Latest Update for Food and Environmental from Waters · 2019. 12. 16. · Refreshed Xevo TQ-S micro provides –Update to the detector to improve performance for small negative ionizing