selective extraction and analysis of chemical migrants ... · pdf filechemical migrants from...

©2015 Waters Corporation 1

Selective Extraction And Analysis of Chemical Migrants from Packaging Material

using a Supercritical Fluids (SFE)

Guy WilsonE&L 2015


Regulatory aspects

The pharmaceutical drug companies have to prove that nothing harmful gets transferred from the packaging into drug product. Or that whatever does leach into sample is below the thresholds established by the guidelines.

For food industry only certain chemicals must be checked/monitored e.g. bisphenols A, B and E, phthalates. But other have to be determined as well.

Under the Federal Food, Drug, and Cosmetic Act, cosmetic products and ingredients do not require FDA approval before they go on the market. The exception is color additives .

FDA periodically buys cosmetics and analyzes them, especially if aware of a potential problem. The information obtained can be used to alert consumers, support regulatory actions, or issue guidance for industry. FDA does not have the resources to sample and analyze all cosmetics on the market.


Typical extractables & leachables

Chemical additives, plasticizers, antioxidants and contaminants present in individual polymers

Monomers and oligomers from incomplete polymerization reactions

Volatile compounds from the secondary packaging such as inks and adhesives

Residual compounds from the surfaces of the molding equipment, antistatics etc


Sample PreparationMajor Source of Laboratory Costs

Sample preparation is the most often cited area of improvement to save time and operating costs

Most sample preparation involves being in an organic phase – Liquid/Liquid, PPT, Soxhlet, Distillation, Evaporation and

Reconstitution

Many matrices will respond best to organic phases (gels, blisters, ointments, synthesis solvents, etc.)

image from dyapharma.com image from sefetec.net image from tasnee.com


Comparison study of 4 different extractions techniques

Compare 4 different extraction techniques of 4 common packaging materials :

o Liquid extraction• Water

o Microwave• Hexane, Isopropanol

o Soxhlet• Hexane , Isopropanol

o Supercritical Fluid Extraction (SFE)• Isopropanol

Compare extraction profiles of the same packaging materials by using UPC² (SFC)


Samples

High Density Polyethylene bottle (HDPE)

Low Density Polyethylene container (LDPE)

Ethylene Vinyl Acetate plasma bag (EVA)

Polyvinyl Chloride blister pack (PVC)

Analytes :

o Irgafos 168, 5-chloro-2-hydroxy-4-methylbenzophenone (5-Cl-2-OH-4-

methyl BP), 4-hydroxy-2-octyloxybenzophenone (4-OH-2-octyloxy BP),

Irganox 245, Lowinox 44B25, Naugard 445, Diphenyl phthalate, Tinuvin

328, Uvitex OB


Extractions conditions

2g sample1 cm² piecesTeflon Vessel

10 mL Hexane + stirrer and

heating element

10 mL IPA+ stirrer

2g sample1 cm² pieces

20 mL Headspace Vial

20 mL Water

3 hours 50°C

3 hours 50°C

72 hours 50°C

Microwave extraction Water extraction

3 g of PVC 5 g of HDPE, LDPE, and EVA

1 cm² piecesWhatman 33 mm x 94mm cellulose

extraction thimble

175 mL Hexane

8 hours

175 mL IPA

8 hours

Dry 15mL Hexane

Dry 15mL IPA

Soxhlet


Supercritical Fluid Extraction (SFE)


What Is Supercritical Fluid Extraction

Supercritical Fluid Extraction is the process of separating one or multiple components (the extractant) from another (the matrix) using supercritical fluids as the extracting solvent.

Extraction is usually from a solid matrix

SFE can be used as :– a sample preparation step for analytical purposes– or on a larger scale to either strip unwanted material from a product (e.g.

decaffeination) or collect a desired product (e.g. essential oils).

Carbon dioxide (CO2) is the most used supercritical fluid, sometimes modified by co-solvents such as ethanol or methanol. – > 31°C and 74 bar (1073 psi)

It is based on the principle that solubility in a supercritical fluid increase dramatically with increasing density, and different solutes have different solubility at the same condition.


Why A Supercritical Fluid?

Why do Supercritical fluids make good mobile phases for chromatography?

Diffusivity describes the rate at which one substance can move through another

Viscosity is resistance to flow

High diffusivity, and low viscosity combine in SFC to give fast, efficient chromatography


Supercritical Fluid Applications

SupercriticalCO2

Extraction (SFE) Bioactive compounds Nutraceuticals Spices and aromatic Decaffeination Decontamination or cleaning Valuable molecules from waste

Reaction Hydrogenation Hydroformylation Carboxylation

SF Particle Design(RESS & SAS) Polymerization Cristallization Impregnation

Chromatography (SFC) Analysis & Purification Chiral Separation Normal Phase


Why A Supercritical Fluid?


Increasing PolarityNon-polars

Alkanes

EthersEsters

AlcoholsAmides

AcidsAmines Highly polar organics Inorganic

ions

Neat CO2 SFECO2 + modifier

CO2 + modifier + ternary additives

CO2 + modifier + ternary additives + water

Liquid – based extraction methods

Small molecules Peptides Large proteins

Increasing Molecular Weight

Extractability Based on Polarity

One of the largest advantages of SFE: Selectivity


Advantages of SFE:

Increased selectivity and specificity• Fine tune the extraction with changes in co-solvents (Method Dev)

Decreased cost per sample• Minimal procurement or disposal cost of CO2 in comparison to organic solvents• Improves extraction efficiency and reduces extraction time vs. other sample

preparation techniques

Minimize exposure to organic solvents• Lack of residual organic solvents• Is environmentally friendly

Accelerate the extraction process • Extract analytes faster than comparative techniques• Eliminate cumbersome traditional solid/liquid extraction (ie. Sohxlet or solvent soak)

Ability to handle thermally labile compounds• Operates at lower temperatures than PSE, MAE and soxhlet


An extraction technique complementary/alternative to Soxhlet or liquid/liquid extraction – CO2 in combination with an organic solvent, most commonly alcohols, is used as

the extraction solvent

Instrumentation


Extraction Modes

Extractions are done in dynamic, static, or combination modes.

In a dynamic extraction the supercritical fluid continuously flows through the sample in the extraction vessel and out the restrictor to the trapping vessel.

In static mode the supercritical fluid is held in the extraction vessel for some period of time before being released through the restrictor to the trapping vessel.

In combination mode, a static extraction is performed for some period of time, followed by a dynamic extraction.


SFE conditions

2g sample

10 mL Hexane

10 mL IPA

2g sample

20 mL Water

3 hours 50°C

3 hours 50°C

72 hours 50°C

Microwave extraction

Water extraction

3 g of PVC 5 g of HDPE,

LDPE, and EVA

175 mL Hexane

8 hours

175 mL IPA

8 hours

Dry 15mL

Hexane

Dry 15mL IPA

Soxhlet

2 g of PVC 3 g of HDPE, LDPE, and EVA

1 cm² pieces

CO2:IPA 98:24.9 mL/min CO20.1 mL/min IPA

CO2:IPA 80:204 mL/min CO2

1.0 mL/min IPA

50°C & 300 Bars30 min Dynamic 20 min Static 10 min Dynamic

Supercritical Fluid Extraction

2 X

Dry10 mL IPA for PVC,

9 mL IPA for HDPE, LDPE, and EVA


What About The Analysis?


Supercritical Fluid Chromatography


Broad Applicability

Courtesy of A. Grand-Guillaume Perrenoud, D. Guillarme, Pr J-L. Veuthey, University of Geneva


The ACQUITY UPC2

Inject valve

AuxiliaryInject valve

Column Manager

PDA detector

Back Pressure Regulator(Dynamic and Static)

Waste Modifier CO2Supply CO2

PumpModifier

Pump

mixerThermo-electric heat exchanger

Make-upPump

Mass Spec

Splitter


UPC2: Compatibility with all MS Technologies

For ultimate CC-MS performance, ACQUITY UPC2 System coupled with:

ACQUITY QDa- Single quadrupole detector for robust and routine

performance

Xevo TQ-S - Ultimate sensitivity

Xevo G2-S Qtof and Synapt G2-S- Qualitative and quantitative results from a single

platform


Workflow Benefit of ACQUITY UPC2 for the Analysis of Polymer Extracts

Non-Polar Solvent Extraction

Polar Solvent Extraction

Inject direct on GC

Evaporate and reconstitute in a more polar solvent for

LC injection

Inject direct on LC

Back-extract with a non-polar solvent for GC injection

Polar or Non-Polar Extraction

Inject directOn UPC2


4 min separation by UPC2 vs. 9.5 min by UPLC

UPC2

BH

T

5-Cl-2-

OH

-4-m

ethy

l BP

Tinu

vin

P

Tinu

vin

328

Irga

fos

168

2-O

H-4

-oct

ylox

y BP

Irga

nox

1076

Dip

heny

lpht

hala

te

Uvi

tex

OB

Nau

gard

445

Irga

nox

1330

Irga

nox

1010

Irga

nox

245

Low

inox

44B

25

AU

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50

Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00

5-Cl-2-

OH

-4-m

ethy

l BP

UPLC

Tinu

vin

P

Dip

heny

l pht

hala

te

BH

T

Irga

nox

245

Low

inox

44B

25

4-O

H-2

-oct

ylox

y BP

Uvi

tex

OB

Nau

gard

445

Tinu

vin

328

Irga

nox

1076

Irga

nox

1330

Irga

nox

1010

Irga

fos

168A

U

-0.15

-0.10

-0.05

0.00

0.05

0.10

0.15

0.20

0.25

0.30

0.35

0.40

0.45

0.50

0.55

0.60

Minutes0.00 0.50 1.00 1.50 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

Chromatographic separations


UPC2 results - Microwave and Water extraction profiles for LDPE

Column Name: 2-EP SampleName: LDPE hex mw Date Acquired: 9/6/2012 3:47:25 PM EDT Instrument Method Id: 1953 Injection Id: 1992

AU

0.000

0.010

0.020

0.030

Column Name: 2-EP SampleName: LDPE IPA mw Date Acquired: 9/6/2012 5:50:59 PM EDT Instrument Method Id: 1953 Injection Id: 2107

AU

-0.005

0.000

0.005

0.010

Column Name: 2-EP SampleName: LDPE water Date Acquired: 9/6/2012 7:42:54 PM EDT Instrument Method Id: 1953 Injection Id: 2211

AU

-0.006

-0.004

-0.002

0.000

0.002

0.004

Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60 4.80 5.00

Hexane

IPA

Water


Soxhlet and Microwave IPA extractions

Column Name: 2-EP SampleName: PET IPA mw Date Acquired: 9/6/2012 6:14:33 PM EDT Instrument Method Id: 1953 Injection Id: 2129

AU

-0.005

0.000

Column Name: 2-EP SampleName: EVA IPA mw Date Acquired: 9/6/2012 6:02:46 PM EDT Instrument Method Id: 1953 Injection Id: 2118

AU

-0.005

0.000

0.005


AU0.000

0.010

Column Name: 2-EP SampleName: HDPE IPA mw Date Acquired: 9/6/2012 5:39:12 PM EDT Instrument Method Id: 1953 Injection Id: 2096

AU

-0.005

0.000

Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60 4.80 5.00

PVC

EVA

LDPE

HDPE

Column Name: 2-EP SampleName: PET IPA sox Date Acquired: 9/6/2012 7:01:40 PM EDT Instrument Method Id: 1953 Injection Id: 2173

AU

-0.005

0.000

0.005

Column Name: 2-EP SampleName: EVA IPA sox Date Acquired: 9/6/2012 6:49:54 PM EDT Instrument Method Id: 1953 Injection Id: 2162

AU

-0.005

0.000

0.005

Column Name: 2-EP SampleName: LDPE IPA sox Date Acquired: 9/6/2012 6:38:07 PM EDT Instrument Method Id: 1953 Injection Id: 2151

AU

0.000

0.010

0.020

Column Name: 2-EP SampleName: HDPE IPA sox Date Acquired: 9/6/2012 6:26:20 PM EDT Instrument Method Id: 1953 Injection Id: 2140

AU

-0.005

0.000

Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60 4.80 5.00

PVC

EVA

LDPE

HDPE

Soxhlet Microwave


SFE extracts

Column Name: 2-EP SampleName: PET low IPA SFE Date Acquired: 9/6/2012 9:05:18 PM EDT Instrument Method Id: 1953 Injection Id: 2288

AU

-0.006

-0.004

-0.002

0.000

0.002

0.004

Column Name: 2-EP SampleName: EVA low IPA SFE Date Acquired: 9/6/2012 8:53:32 PM EDT Instrument Method Id: 1953 Injection Id: 2277

AU

-0.006

-0.004

-0.002

0.000

0.002

0.004

Column Name: 2-EP SampleName: LDPE low IPA SFE Date Acquired: 9/6/2012 8:41:45 PM EDT Instrument Method Id: 1953 Injection Id: 2266

AU

-0.005

0.000

0.005

Column Name: 2-EP SampleName: HDPE low IPA SFE Date Acquired: 9/6/2012 8:29:57 PM EDT Instrument Method Id: 1953 Injection Id: 2255

AU

-0.006

-0.004

-0.002

0.000

0.002

0.004

Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60 4.80 5.00

PVC

EVA

LDPE

HDPE

Column Name: 2-EP SampleName: PET high IPA SFE Date Acquired: 9/6/2012 9:52:24 PM EDT Instrument Method Id: 1953 Injection Id: 2332

AU

-0.005

0.000

0.005

Column Name: 2-EP SampleName: EVA high IPA SFE Date Acquired: 9/6/2012 9:40:39 PM EDT Instrument Method Id: 1953 Injection Id: 2321

AU

-0.005

0.000

0.005

0.010

Column Name: 2-EP SampleName: LDPE high IPA SFE Date Acquired: 9/6/2012 9:28:52 PM EDT Instrument Method Id: 1953 Injection Id: 2310

AU-0.005

0.000

0.005

0.010

0.015

Column Name: 2-EP SampleName: HDPE high IPA SFE Date Acquired: 9/6/2012 9:17:05 PM EDT Instrument Method Id: 1953 Injection Id: 2299 AU

-0.006

-0.004

-0.002

0.000

0.002

0.004

Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60 4.80 5.00

PVC

EVA

LDPE

HDPE

Low IPA High IPA


LDPE, all IPA extracts

Low IPA SFE

High IPA SFE

Soxhlet

Microwave

Column Name: 2-EP SampleName: LDPE high IPA SFE Date Acquired: 9/6/2012 9:28:52 PM EDT Instrument Method Id: 1953 Injection Id: 2310

AU

-0.005

0.000

0.005

0.010

0.015

Column Name: 2-EP SampleName: LDPE low IPA SFE Date Acquired: 9/6/2012 8:41:45 PM EDT Instrument Method Id: 1953 Injection Id: 2266

AU

-0.006

-0.004

-0.002

0.000

0.002

0.004

0.006

Column Name: 2-EP SampleName: LDPE IPA sox Date Acquired: 9/6/2012 6:38:07 PM EDT Instrument Method Id: 1953 Injection Id: 2151

AU

-0.005

0.000

0.005

0.010

0.015

0.020


AU

-0.005

0.000

0.005

0.010

Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60 4.80 5.00


Irgaf

os 1

68

Irgan

ox 1

076 Irg

anox

101

0

AU

-0.008

-0.006

-0.004

-0.002

0.000

0.002

0.004

0.006

0.008

0.010

0.012

0.014

0.016

0.018

0.020

0.022

0.024

0.026

0.028

0.030

Minutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60 4.80 5.00

UV chromatogram of LDPE SFE extract analysed by UPC2


Inte

nsity

0.0

5.0x105

1.0x106

1.5x106

2.0x106

2.5x106

3.0x106

3.5x106

Minutes0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00

Combined - SQ 1: MS Scan 1: 200.00-1200.00 ES+, Centroid, CV=Tune

475.5

476.5 531.6

548.6

549.6

553.6

554.6 569.5

Inte

nsity

0.0

2.0x106

4.0x106

6.0x106

8.0x106

1.0x107

1.2x107

1.4x107

1.6x107

1.8x107

2.0x107

m/z450.00 460.00 470.00 480.00 490.00 500.00 510.00 520.00 530.00 540.00 550.00 560.00 570.00 580.00 590.00 600.00

Confirmation of identity using MSIn

tens

ity

0.0

2.0x106

4.0x106

6.0x106

8.0x106

1.0x107

Minutes0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00

Combined - SQ 1: MS Scan 1: 200.00-1200.00 ES+, Centroid, CV=Tune

475.5

476.5 531.6

548.6

549.6

553.6

554.6 572.6

Inte

nsity

0.0

2.0x106

4.0x106

6.0x106

8.0x106

1.0x107

1.2x107

1.4x107

1.6x107

1.8x107

2.0x107

2.2x107

m/z450.00 460.00 470.00 480.00 490.00 500.00 510.00 520.00 530.00 540.00 550.00 560.00 570.00 580.00 590.00 600.00

Peak in LDPE extract

Peak from Irganox 1076 std


Conclusion

All extraction techniques provided similar extractablesprofiles, but…

SFE consumes much less solvent and is quicker than Soxhlet extraction.

The MV-10 SFE System has automated method development and extractions on 10 samples to simplify the process.

UPC2 gives a fast, high resolution separation and has wide sample diluent compatibility.


Thank You!

selective extraction and analysis of chemical migrants ... · pdf filechemical migrants from...

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