determination of fungicide residues in grape by gc/ms...determination of fungicide residues in grape...
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DETERMINATION OF FUNGICIDE RESIDUES IN GRAPE BY GC/MS
SanjaLazić, Vedrana Komlen, Dragana Šunjka, Nada Grahovac, Jadranka Pejčić, Alma Rahimić, Milenko Blesić
Faculty of Agriculture, Novi Sad, SERBIA Agromediterannian faculty, Mostar, BiH
Plant protection products (pesticides) are
used worldwide within agriculture to protect
crops and ensure the quantity and quality of
the harvest.
Fungicides represent one of the most relevant
groups of pesticides applied to vineyards.
These compounds are sprayed directly on
fruit and leaves to prevent the attack of fungi,
which reduce the yield of fruit.
If fungicides are not degraded, some of them will
penetrate plant tissues and appear in the pulp and juice.
Once present in the pulp, pesticides are difficult to
completely be removed.
Residues of fungicides are one of the most important
pollution sources in food production and might pose
potential threat to public health.
The concentration of pesticide residues may increase
during post processing, where in general pesticide
concentrations in processed juices are higher than that in
the natural fruit.
Having in mind the health risk of pesticide residues in fruit and wine, it is
of particular importance to provide precise, accurate and reliable test
result of residues as the scientific basis for ensuring food safety and fair
practice in international trade.
• single component
• multi-residue analysis method (different
components of different types of pesticides)
Fungicide residues
Analysis of fungicide residues
PRESENCE OF BENALAXYL, DIMETHOMORPH AND
TETRACONAZOLE RESIDUES IN GRAPE SAMPLES
Dimethomorph
Tetraconazole
Benalaxyl
Aim of the study
Field experiment
The vineyards choice was done to research on four different
locations in the vineyards of Mostar.
• Žilavka
• Blatina
Grape varieties
• famous Hercegovinian grape varieties
• Special clime conditions
• 2290 sunshine hours per year
Vineyards location
Area of vineyard 2 ha
Age of vineyard 9 years
Variety Žilavka
Blatina
Humčine – plantation vineyard
Area of vineyard 13 ha
Age of vineyard 5 years
Variety Žilavka
Mukoša – plantation vineyard
Area of vineyard 30 ha
Age of vineyard 7 years
Variety Blatina
Žitomislić – plantation vineyard
Area of vineyard 0,4 ha
Age of vineyard 6 years
Variety Žilavka
Blatina
Hodbina – small vineyard
Protection program
Treatments were carried out air assistance sprayers (atomizer) with
300 l with average interval between treatments was of 10 days.
The protection program in the vineyard Mukoša and Žitomislić
Date treatment Product Active substance Quantity
kg or l/ha
May 5th
Delan Dithianon 0.3
Sulfur Sulfur 1.5
May 15th Delan Dithianon 0.4
Thiovit jet Sulfur 2.0
May 25th Pergado F Mandipropamid+ folpet 2.0
Talendo Proquinazid 0.25
June 5th Fantic F Benalaxyl + folpet 2.5
Domark Tetraconazole 0.75
June 18th Forum star Dimethomorph + folpet 2.0
Collis Kresoxim-methyl + boskalid 0.4
June 30th Mikal premium Fosetyl + folpet+ iprovalicarb 3.0
Postalon Quinoxyfen + myclobutanil 0.125
July 12th Cabrio top Pyraclostrobin + methiram 2.0
August 9th Mythos Pirimethanil 2.5
The protection program in the vineyard Hodbina
Date treatment Product Active substance Quantity
kg or l/ha
April 20th Cuprablau Copper 3.0
Chromosul Sulfur 2.5
April 28th Chromosul Sulfur 2.5
May 7th Chromosul Sulfur 2.5
May 20th
Ridomil gold Metalaxyl + mankozeb 2.5
Chromosul Sulfur 2.5
Topas Penconazole 0.25
June 1st Ridomil gold Metalakxyl + mankozeb 2.5
Chromosul Sulfur 2.5
June 11th
Fantic F Benalaxyl + folpet 2.5
Domark Tetraconazole 0.75
Pirus Pirimetanil 2.5
June 24th Acrobat Dimethomorph + mankozeb 2.5
Systane 24 E Myclobutanil 1.0
July 4th Cuprablau Copper 3.0
Thiovit jet Sulfur 4.0
July 20th Nordoks 75W Copper 1.25
Thiovit jet Sulfur 4.0
August 5th Mythos Pirimetanil 2.5
The protection program in the vineyard Humčine
Date treatment Product Active substance Quantity
kg or l/ha
April 21st
Folpan Folpet 2.05
Kossan WG Sulfur 4.1
May 6 th Folpan Folpet 2.5
Kossan WG Sulfur 4.9
May 14 th Manfil 75 WG Mancozeb 3.3
Karathane Gold 350 Metyldinocap 0.9
May 23rd Manfil 75 WG Mancozeb 3.3
Karathane Gold 350 Meptyldinocap 0.9
May 27 th Fantic M Benalaxyl + mancozeb 4.9
Domark Tetraconazole 1.4
June 16 th Fantic F Benalaxyl + folpet 4.9
Crystal Quinoxyfen 0.4
June 29 th Fantikc F Benalaxyl + folpet 4.9
Crystal Quinoxyfen 0.4
July 15th Mythos Pirimetanile 2.5
The field sampling was conducted during the technological maturity of the
grape varieties of "Žilavka" and "Blatina“.
Sampling procedure
Sampling procedure
The average weight of each laboratory sample of grapes was
approximately 2 kg with at least five clusters. The samples were packed
in plastic bags and delivered to the laboratory refrigerator.
Location Varieties Number of
samples
Humčine Žilavka
Blatina
4
4
Mukoša
Žitomislić
Žilavka
Blatina
4
4
Hodbina Žilavka
Blatina
4
4
50 g sample + 100 ml ACN - homogenize 5 min
Extraction procedure
add 10 g NaCl - homogenize 5 min
Clean up - Octadecyl (C18) SPE tubes
Clean up - Carbon SPE tubes / Aminopropyl SPE tubes
Evaporation to drynes
Extract diluted in acetone
GC/MS analisys
Metod by Fillion et al., 2002.
GS/MS analysis
Determinations were performed on Thermo type
Focus DSQ II GC/MS
Operating mode Splitless
Column HP-5MS, 5% phenyl methyl siloxan
(30m x 0.25mm x 0.25µm)
Helium flow rate 0.9 ml/min
Injector temperature 250 C
Temperature detector 285 C
Initial column temperature 70 C
Initial time 2 min
Speed of temperature rise 25 C/min
Final temperature 280 C
Determination conditions
Results
LOD 0.001 mg/kg
LOQ 0.003 mg/kg
Recovery values for
benalaxyl, dimethomorph
and tetraconazole
> 90 %
GS-MS chromatogram of standard mixture
Results
GS-MS chromatogram of grape samples
tetraconazole benalaxyl
Results
Locality Investigated
compound
Determinted
value (mg/kg)
LOD
(mg/kg)
Humčine
Benalaxyl < LOD 0.001
Dimethomorph < LOD 0.001
Tetraconazole 0.003 0.001
Mukoša
Benalaxyl 0.002 0.001
Dimethomorph < LOD 0.001
Tetraconazole 0.003 0.001
Hodbina
Benalaxyl < LOD 0.001
Dimethomorph < LOD 0.001
Tetraconazole 0.002 0.001
Determined value of fungicides in grape variety Žilavka by GC/MS
Locality Investigated
compound
Determinted
value (mg/kg)
LOD
(mg/kg)
Humčine
Benalaxyl 0.014 0.001
Dimethomorph < LOD 0.001
Tetraconazole 0.017 0.001
Žitomislić
Benalaxyl < LOD 0.001
Dimethomorph < LOD 0.001
Tetraconazole 0.001 0.001
Hodbina
Benalaxyl < LOD 0.001
Dimethomorph < LOD 0.001
Tetraconazole < LOD 0.001
Determined value of fungicides in grape variety Blatina by GC/MS
In grape samples variety Zilavka tetraconazole residues
were found in samples from all localities, while
benalaxyl residues were found only in samples from
locality Mukosa.
Conclusions
In grape samples variety Blatina tetraconazole residues
were found in samples from locality Humcine and
Zitomislic, whereas benalaxyl residues were found in
grape samples from locality Humcine.
Residues of dimethomorph in analyzed samples of
grape, variety Žilavka and Blatina were not detected.
The concentrations of benalaxyl in the analyzed grape
samples were 0.002-0.014 mg/kg.
Conclusions
The concentrations of tetraconazole in the analyzed
grape samples were 0.001-0.017 mg/kg.
The highest value of the fungicide residues were found
in grape variety Blatina, locality Humcine. It is can be
explain with intensive application of fungicide on this
locality.
Conclusions
The concentrations of fungicides were below the
permissible levels (MRL) set by EU regulations (The
Annexes of Commission Regulation (EC) No
396/2005) and by Serbian regulations (Official
gazette, No 25/2010 i 28/2011) for tetraconazole (0.5
mg/kg) and benalaxyl (0.3 mg/kg) in grape.
The obtained positive results for tetraconazole and
benalaxyl could be attributed to the high sensitivity of
the developed GC/MS method.
