phthalates - Food Review Indonesia

PHTHALATES, DIOXINS/FURANS & PAH
TESTING METHOD ON BEVERAGES
Melissa Dita, M.Sc
AGENDA
•
•
•
•
PHTHALATES :
INTRODUCTION
SAMPLE STORAGE,
HOMOGENISATION
and EXTRACTION
•
DIOXINS/FURANS &
PAH :
MEASUREMENT
TECHNIQUE
•
•
REGULATION
•
ANALYSIS
SOURCE OF
CONTAMINATION,
CARCINOGENIC &
EXPOSURE
PHTHALATES
INTRODUCTION
• 1,2-Benzendicarboxylic acid esters, commonly
denoted as phthalates, form a group of
compounds that is mainly used as plasticisers
for polymers such as polyvinilchloride (PVC)
• Others application are adhesives, paints, films,
glues, cosmetics and so forth.
• The derivative of phthalate is infinite. Despite
only a few phthalate are produced in the
industrial scale, the annual production of
phthalates was estimated by the WHO to
approach 8 million tons.
• The most important congeners are in that
respect DEHP (50% of the world production of
phthalates), DIDP and DINP.
Phthalate
PTHALATE CHARACTERISTIC
•
•
•
•
Phthalates are stable in solution
Able to resist high temperature
Degrade under exposure to sunlight
Metabolised under aerobic microbial activity
• Humans are exposed to phthalates via food, air, water, and other
sources such as cosmetics or pharmaceutical products.
• Fatty and oily foods are primarily contaminated with phthalates due
to their lipophilic character.
PTHALATE DERIVATIVE
DEHP – Di(2-ethylhexyl) phthlate
BBP – Benzyl butyl phthalate
DIDP – Di-isodecyl phthalate
DBP – Dibutyl phthalate
DINP – Di-isononyl phthalate
DNOP – Di-n-octyl phthalate
PHTHALATE INVESTIGATED
FOOD & BEVERAGE MATRICES INVESTIGATED
SAMPLE STORAGE, HOMOGENISATION AND
EXTRACTION
Sampel Storage
•
•
A potential source of contamination are sampling containeres, which for
reasons of convenience (thight, unbrakeable, resistant to low temperature
etc) are frequently made of plastics.
Original food packaging should be used for sample storage if
possible, also those intended for phthalate analysis are stored
cooled.
Sample Homogenisation
•
•
•
Sample have to be homogenised prior to extraction.
This can be achieve by shaking, stirring or mixing.
In case of solid samples, mixers are used to pulpify them, which is
frequently enhanced by the addition of distilled water or polar organic
solvents.
FOOD & BEVERAGES TESTING
Sample Preparation
Weight Homogenized Sample in a Centrifuge Tube
Add Solvent, MgSO4, NaCL and International Standard
Vortex
Centrifuge
Pipette to GC Vial
Direct inject GCMS
Continue….
FOOD TYPE
EXTRACTION
Non-fatty liquid :
water, soft drink,
alcoholic beverages
Liquid-liquid (L/L) extraction, extracted with non-polar organic
solvent. The solvents employed are chloroform, n-hexane, nheptane, or isooctane.
Non- fatty solid foods:
Extracted with acetonitril, or mixtures of acetonitril and water,
to lower viscosity of the sample.
Fatty solid food:
Total diet samples.
Phthalates are extracted from the matrix either together with
the fat by application of solvents such as dichloromethane,
mixtures of dichloromethane with cyclohexane, n-hexane and
mixture of n-hexane with acetone, or acetonitril. The extraction
is mostly accomplished by simply shaking.
SAMPLE CLEAN UP
Two technique are applied for the clean up of the food extract,
liquid-liquid partioning and gel permeation chromatography (GPC).
GPC employed if the extract contained large amounts of fat.
FOOD MATRICE TYPE
CLEAN UP
Non-fatty liquid :
water, soft drink,
alcoholic beverages
Not required
Non- fatty solid foods:
Co-extracted interference were removed by them from the
acetonitril solution by partioning inti n-hexane.
Fatty solid food:
Total diet samples.
Dichloromethane applied for the isolation of lipid fraction from
acetone extract. The lipid fraction was evaporated, recontituted
in cyclohexane/ethylacetate(1/1) and cleaned up by GPC.
MEASUREMENT
Diethylphthalate – SIM mode
Summary of phthalates analyzed
Phthalates type
Abb.
Retention
Time (min)
SIM ions detail
Dimethylphthalate
DMP
7.474
163.05, 77.05, 50.05
Diethylphthalate
DEP
8.171
149.00, 150.00, 65.00,
75.95
Di-n-butylphthalate
DIBP
9.796
149.00, 150.00, 205.05
Benzyl butyl phthalate
BBP
11.293
149.00, 91.05, 206.05
Bis-2-ethylhexyl
phthalate
DEHP
11.858
148.95, 167.00, 57.05,
71.05
Di-n-octyl phthalate
DNOP
12.475
149.00, 279.90, 150.70
DETECTION LIMIT
•
•
•
•
•
•
•
•
DMP
DEP
DIBP
BBP
DEHP
DNOP
DINP
DIDP
LOR set at -1ppm under scan mode
- 0.5ppm under sim mode
QA/QC
• BLANK VALUES
Hardly constant, need to be well
controlled
• Quality control charts
• Replicate sample
• Avoiding the use of plastic material such as PVC during
sample preparation
• Checking chemicals before use, distillation, heating of
glassware in a furnace 2h; 400oC; store it until use in
dessicator, rinsing glass ware with solvents, exchanging
frequently wash solvents, performing runs without
injection to clean the instrument
DIOXINS/FURANS
and
Polycyclic Aromatic Hydrocarbon (PAH)
Regulation
Chemical Contamination
Dioxins
PAH
It is necessary to accurately determine the level priority
PAHs and Dioxins in food to respond to Badan
Pengawasan Obat dan Makanan (BPOM) legislation and
ensure food safety.
Poly Aromatic Hydrocarbon (PAH)
INTRODUCTION
• Known as poly-aromatic hydrocarbons or polynuclear
aromatic hydrocarbons
• Potent atmospheric pollutants that consist of fused aromatic
rings and do not contain heteroatoms or carry substituents
• Exist in oil, coal, tar deposits, and as byproducts of fuel
burning (whether fossil fuel or biomass)
• Identified as carcinogenic and mutagenic
• PAHs are also found in cooked foods, studies have shown
that high levels of PAHs are found in meat cooked at high
temperatures such as grilling or barbecuing, and in smoked
fish
Benzo (a) pyrene
•
•
•
•
PAHs compound used as indicator as safety level
caused by identified as the most high level of
carcinogenicity.
Mutagenic and highly carcinogenic, listed as a
Group 1 carcinogen by the IARC.
Benzopyrenes, formed by a benzene ring fused
to pyrene.
The result of incomplete combustion at
temperatures between 300 °C (572 °F) and 600
°C (1,112 °F).
Carcinogenic of PAHs
• Banyak senyawa-senyawa aromatik, termasuk PAHs, yang bersifat
karsinogenik.
• Hidrofobik dan tidak memiliki gugus metil atau gugus reaktif.
• Sangat sulit diekskresi dari dalam tubuh dan biasanya terakumulasi
pada jaringan hati, ginjal, maupun adiposa atau lemak tubuh.
• Struktur molekul yang menyerupai basa nukleat (adenosin, timin,
guanin, dan sitosin), molekul PAH dapat dengan mudah menyisipkan
diri pada untaian DNA.
• Akibatnya fungsi DNA akan terganggu dan apabila kerusakan ini tidak
dapat diperbaiki dalam sel, maka akan menimbulkan penyakit kanker.
Analysis PAH
Food &
Beverage
Sample
Preparation
Identification
METHOD
• Preparation method 3510, 3520 (water)
• Analysis 8270 C (GCMS)
Food Sample Preparation
DIOXINS/FURANS
Description
•
•
•
The polychlorinated dibenzo-para-dioxins (PCDD) and polychlorinated
dibenzofurans (PCDF) are chlorinated, planar, aromatic compounds
containing two benzene rings.
The terms PCDD/F and dioxins carry the same meaning and are used
interchangeably
A dioxin molecule is bonded by two oxygen atoms, furan molecule by a
single oxygen atom and a direct bond.
Introduction
• By products of various industrial processes
• Have been in the environmental spotlight for
over two decades
• Regarded as highly toxic compound
• Environmental pollutants
• Persistent Organic Pollutants (POPs)
• Extensive global research has concluded to
these compounds can result biochemical and
biological effects in animal and human
Dioxins and Furans come from:
• Not made for any specific purpose but formed when
products are burned
•
Byproduct from :
** pulp and paper industry from chlorine bleaching the wood pulp
** burn household waste
** burn wood
** municipal solid waste incinerator
** copper smelter
** production of iron and steel
** cement kiln
** electrical power generation
** fuel burning
** coal fired power plant
Cement Kiln
Copper Smelter
Coal fired power plant
Electrical power generation
How Can Human Exposed to Dioxins and Furans
*Dioxins and Furans are enter your body :
• Breathing contaminated air
• Drinking contaminated water
• Eating contaminated food
*90 % of exposure to dioxins and furans is from
eating contaminated food.
*Build up in fatty tissue of the animals
*Contaminated beef, pork, poultry, fish and dairy
product can be a source of exposure
• Dioxins and Furans have been found in air, soil and food
• Mainly distributed by air
• Fat Soluble, it bio accumulates, climbing up the food
chain
• Hydrophobic and Lipophilic
Dioxins and Furans analysis
Analytical method Water/Solid/Tissue by:
• USEPA 1613 A Tetra- through Octa-Chlorinated Dioxins
and Furans by Isotope Dilution HRGC/HRMS
• SW846 8290 Polychlorinated Dibenzodioxins (PCDDs)
and Polychlorinated Dibenzofurans (PCDFs) by HighResolution Gas Chromatograph/High-Resolution Mass
Spectrometry (HRGC/HRMS)
Practical sampling guide
Food/Feed & Beverages
Foodstuffs/Feedingstuffs
Amount
Food & Feed: 100 grams
Beverages: 1 L
Container
Aluminium foil / Amber glass with Teflon lined cap
Preservation
None required
Storage
conditions
4°C or frozen in darkness
Holding times
Frozen, up to 1 year
Sampling
Sample
Extraction standard
Extraction
Cleanup
Fractionation
Final extract
HRGC-HRMS analysis
Syringe standard
Determination of PCDD/F
HRGC-HRMS instruments
2 HRMS instruments
HRMS Finnigan MAT 95XP high
resolution (R > 10 000) with two capillary
gas chromatographs Agilent 6890N
(HRGC) equipped with auto samplers
CTC GCPAL
HRMS ThermoElectron DFS– GC Trace
Ultra equiped with a TriPlus autosampler
ALS QC scheme for dioxin food/feed &
beverages samples is following:
•
1. The extraction standard recoveries are determined for each
sample, the recovery have to meet our criteria
•
•
•
•
2. The method blank is measured for each batch of 20 samples
3. The duplicate is measured for each batch of 20 samples
4. Certified reference material is measured every 3 months
5. Take part in interlaboratory comparisons with minimal
frequency 1 / 2 years, but usually 1-2 times/year
Excellence in HRMS analyses
ALS offers the most
comprehensive range of
services including:
•
Dioxins (PCDD/Fs) and co-planar
PCBs – All enviro matrices (incl.
Emissions/Air) and bio matrices (incl.
Food/Feed/Tissue)
•
Polybrominated diphenyl ethers
(PBDE) – All enviro matrices and bio
matrices (incl. Food/Feed/ Tissue)
•
Polyaromatic hydrocarbons (PAH)-All
enviro matrices (incl. Emissions/ Air)
and bio matrices (incl.
Food/Feed/Tissue)
Thank You
any question?
Contact us : [email protected]
Phone : 0251-8253813