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RESIDUE ANALYSIS

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PESTICIDE RESIDUE
ANALYSIS
Reliable data on residues are essential in
evaluating pesticide safety to man and the
environment.
Analytical methods are required for
Identification of trace levels of pesticides and their
degradation products in crops, soils, water and
biological tissues
Determination and enforcement of tolerances
Fate of chemical studies

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PESTICIDE RESIDUE
METHODS
Methods must be
Specific each case is a particular problem
Sensitive 10 ppm acceptable in 1930s, presently
down to ppt for dioxins
Analytical steps involved under careful
scrutiny and quality assurance (QA)
Sampling
Extraction and cleanup
Measurement
Validation, calculation and interpretation
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SAMPLING
Results only as valid as representative
character of sample
Sample integrity must be preserved
Sampling variables include:
source, size and heterogeneity, storage stability,
chemical characteristics, intended use of the crop,
behavior of chemical on sample
Untreated crops or "blanks" important for
interfering materials

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EXTRACTION AND
CLEANUP
Extraction
Separate residue from interfering materials without loss
Extraction of solids or liquids with organic solvents
Cleanup
The separated portion may be used as is or after chemical
modification (derivatization) to improve sensitivity, specificity or to
enhance removal of extraneous materials
Liquidliquid partition concentrates the residues or eliminates
interfering materials
Chromatography:
Liquid-solid column most often used (e.g., SepPak)
Solidsolvent combination that selectively retains or passes the residue sought
Liquidliquid and gasliquid partition chromatography also important
Partition: hexane / acetonitrile or ether / ethanol / (NH 4)2SO4
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BIOASSAY METHODS FOR
RESIDUE ANALYSIS
Immunoassays
Antibodies based on protein conjugates of the
pesticides in a highly specific manner
Allows a rapid screen to be followed by more
specific chemical analysis
Enzyme or receptor assay
Utilizes isolated biological systems
Toxicity tests with living organisms
Daphnia, etc. normally not specific

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SPECTROPHOTOMETRIC
METHODS FOR RESIDUE
ANALYSIS
Based on light absorption, transmission or
emission characteristics
Ultraviolet and infrared
Requires appropriate chromophore and thorough clean
up, low sensitivity
Fluorescence
Requires conversion to fluorescent derivative, usually
after chemical transformation of the pesticide
(hydrolysis, oxidation)
Very sensitive (ng/ml or ppb) and specific.
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THIN-LAYER
CHROMATOGRAPHY (TLC)
Sample is absorbed on
silica gel surface
Separation is achieved
by differential mobility
Used primarily for
qualitative analysis or
cleanup
Can be fairly specific
when used in
combination with spray
reagents
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SEPARATION AND
IDENTIFICATION
Processes with on-line combinations and
sensitivities

Sensitivity: MS or GC or UV > IR or NMR 9


HIGH PERFORMANCE LIQUID
CHROMATOGRAPHY (HPLC)
Sample is partitioned
between the carrier
solvent and a liquid-
phase coating a solid
support
Pressure is applied to the
column to obtain greater
speed and separation
Mass spectrometry, UV
absorbance and
refractive index are used
as detection methods
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GAS-LIQUID
CHROMATOGRAPHY (GLC)
Very sensitive and good for
qualitative and quantitative
work
Sample is vaporized into a
constant stream of inert gas
A capillary column coated
with a liquid
Components move through
the column at varying rates
The position (time) of a
response identifies the
component and the height
or area measures it
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TEMPERATURE PROGRAM

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CAPILLARY COLUMN

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QUANTITATION MILK
SAMPLE

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NUCLEAR MAGNETIC
RESONANCE (NMR)
Chemicals are identified on
the basis of the frequency
at which a resonance
signal is observed as a
function of their electronic
environment
Nuclei which can be
observed routinely include
1H, 13C, 31P and 19F

Not normally useful for


residue analysis because
of lack of sensitivity

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MASS SPECTROMETRY
Provides greatest specificity and good sensitivity
Sample is ionized on emergence from GLC column
by electron impact (M+) or chemical ionization [M+1]+
or from HPLC (LC) column by electrospray
Ions are accelerated through a magnetic/electrostatic
analyzer or quadruple filter and detected after
enhancement with an electron multiplier
Results are presented as total ion current (TIC)
chromatograms for which a full mass spectrum or a
selected ion (SIM) correspond to each acquisition
point

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GC total ion current reference DDE
shows everything sample

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MS SIM shows DDE from milk
only m/e full MS spectrum

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GC-MS INTERNAL
STANDARD
2H or 13C label
m/e increased by 1.00 versus contaminant to
be analyzed

C12 residue C13 standard C13 standard


must be known amount 1 ppm
1 ppm 1 ppm

C12 residue
0.25 ppm

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VALIDATION, CALCULATION
AND INTERPRETATION
Analyst must evaluate such critical factors as
sensitivity, specificity, capability or need for
determining toxic degradation products, and
productivity (efficiency)
Analyst must interpret the analytical data
obtained.
An ideal method does not give a response for
non-residue components and gives a sensitive
response only for the component(s) sought.

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