SEMI-QUANTITATIVE TEST FOR PARAQUAT IN PLASMA AND URINE Syngenta Crop Protection Basel
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Transcript of SEMI-QUANTITATIVE TEST FOR PARAQUAT IN PLASMA AND URINE Syngenta Crop Protection Basel
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SEMI-QUANTITATIVE TEST FOR PARAQUAT IN PLASMA AND URINE
Syngenta Crop Protection
Basel
Switzerland
Test developed at Syngenta CTL United Kingdom by Bruce Woollen
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WHAT SAMPLES ARE AVAILABLE ?
• Blood plasma is the preferred sample
• If only blood serum is available this can also be tested with limitations for interpretation of results (see slide 19)
• Urine samples are useful to confirm that a poisoning case involves paraquat
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N+
N+
CH3CH3 C NN+
CH3CH3
DITHIONITE REDUCTION OF PARAQUAT
Sodium
dithionite
alkali
PARAQUAT PARAQUAT RADICAL ION
(BLUE)
Paraquat is converted to a blue colour by sodium dithionite
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SUMMARY OF THE TEST
PART 1
A pre-screen will detect high concentrations of paraquat in plasma and urine
If Part 1 is negative
PART 2
A more sensitive blue band test will detect paraquat down to 0.1 µg/mL
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PLASTIC
MICROPIPETTESYRINGE ADAPTOR
15 ML PLASTIC TUBE
CEREX SILICA CARTRIDGE
WATER
WHAT IS NEEDED FOR THE TEST ?
1M NaOH( FOR URINE)
5 ML OR 10 ML SYRINGE
GLASS TUBES 1.2 ML
SYNGENTA TEST KIT
1M
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10 mL WATER
2 MINS
ALLOW TO SETTLE 2 mins
MIX
Sac
het
B
1g
N
a2S
204
Sachet A
2g
NaH
C0
3
PREPARATION OF DITHIONITE REAGENT
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Video clip preparation of dithionite
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approx 0.9 mL
X 20 drops
PLASMA
or URINE
DITHIONITE REAGENT
MIX
PART 1 : PRE-SCREEN IN PLASMA OR URINE
COMPARE COLOUR
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The limit of detection for paraquat in plasma and urine is about 2-3 µg/mL
0 3 10 30 100 µg/mL
COLOUR CHART FOR PRE-SCREEN
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Video clip pre-screen
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RESULTS OF THE PRE-SCREEN
• If a blue colour is produced the concentration of paraquat is estimated from the chart and no further testing is done
• If a very faint blue colour or no blue colour is produced the blue band test is carried out
• Urine samples require an additional step before the blue band test
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IF TESTING PLASMA PROCEED
TO SLIDE NO 14
PREPARATION OF URINE FOR THE BLUE BAND TEST
URINE
5 mL
1M NaOH
0.5 mL
+
CENTRIFUGE MIX
URINE FOR TESTING
10 MINS
3000 RPM
URINE ONLY
DISCARD PELLET1M
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PART 2 : BLUE BAND TEST FOR PARAQUAT IN PLASMA OR URINE
~ 3 mLPLASMA
or URINE
DITHIONITE REAGENT
2 MINS
~ 1 mL
WATER
BLUE
BAND
approx 1 mL
COMPARE COLOUR
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0 0.1 0.2 0.5 1 2 µg/mL
COLOUR CHART FOR BLUE BAND INTENSITY
Chart is applicable to both plasma and urine
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Video clip of blue band test
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Proudfoot curve – predictive line
Examples :
a) 1 µg/mL at 8h after ingestion
survival unlikely
b) 0.1µg/mL at 12h after ingestion
survival likely
INTERPRETATION OF PLASMA PARAQUAT RESULTS
a
b
Proudfoot A T, Stewart M S, Levitt T and Widdop B. Paraquat Poisoning : significance of plasma paraquat concentrations. Lancet 1: 330 1979
a
b
0.3
0.5
0.7
1.0
0.05
0.07
0.1
3
5
2 4 6 8 10 12 14 16 18 2220 24
Pla
sma
para
quat
con
cent
ratio
n µ
g/m
L
Time after ingestion (hours)
survival unlikely
survival likely
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From Syngenta paraquat treatment booklet
Example :
1 µg/mL at 8h after ingestion
approximately 30% chance of survival
INTERPRETATION OF PLASMA PARAQUAT RESULTS
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WARNING ABOUT SERUM
Plasma versus serum
Paraquat concentrations in plasma and serum are NOT equivalent (serum concentrations are about 3 times lower than plasma concentrations)
Proudfoot and Syngenta survival curves are based on plasma concentrations of paraquat
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No blue Colour
Pale blue
Light blue
Navy blue
Dark blue
From Schermann data
INTERPRETATION OF URINE PARAQUAT RESULTS
Schermann,J.M, Houze P, Bismuth C and Bourdon R. Prognostic value of Plasma and urine paraquat concentration. Human Toxicology : 6 91-93 1987
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LIST OF EQUIPMENT AND SUPPLIERS
• Paraquat test kit containing sodium bicarbonate and sodium dithionite (Syngenta)
• Glass tubes 40 x 6mm 1.2mL (NLG Scientific)
• Cerex silica cartridges 3mL 20mg Cat no 620-0203 ( Crawford Scientific)
• Micropipettes Mini-Pastette LW4231 (Alpha Laboratories)
• Syringe adaptor 120-1101 (Argonaut Technologies)