11

Determination of the Levels of Fungicides on Citrus Fruits

By Miseung Borgers

Chem 4101 Fall 2008

22

Introduction

Fungicides applied on the surface of post-harvest citrus fruits are possible health threats since they can be easily absorbed into the body through peeling and eating with hands Controlling and monitoring the appropriate levels of fungicides would reduce the possible harmful effects by the ingestion

3

Maximum residue limits for five fungicides and their structures

Benomyl 10 ppm

Carbendazim 05 ppm

Imazalil 10 ppm

Thiabendazole 10 ppm

Thiophanate methyl

05 ppm

(Ref 1 3)

44

Method

bull HPLCMS

- Since the fungicides have different polarities they are retained to the stationary phase by different degrees This allows separation by HPLC

- High sensitivity proved by the limit of detection (lt 003 ppm)

- No derivatization is needed - Fastness (No additional clean-up for sample preparation) - Highly automated (reduced manual labor and human

intervention)

(Ref 2)

5

Procedure

Homogenization

Extraction

LCMSD system

wwwthesuncouk wwwbuchicom

ic3dbouedu

wwwhhscacuk

6

Preparation of standards controls and samples

(1) Homogenization

- Blend chopped oranges with ethyl acetate anhydrous sodium sulphate

- For controls use oranges without post harvest fungicide treatment

(2) Extraction

- Use sodium sulphate for filtering

- Add acetonitrile Dry - Dissolve in acetonitrile 2

mM ammonium formate (5050 vv) - For standards controls

mixed with fungicide solutions

- Standard Concentrations 005 05 5 ppm

(Ref 3)

7

Conditions for HPLC

bull Injection volume 5 microL

bull Solvents used for mobile phase methanol 2 mM ammonium formate in water

bull Gradient elution with flow rate of 1 mLmin

Methanol 50

Methanol 80

Methanol 30

(Ref 3)

8

Conditions for interface and MS

bull APCI Interface

- Vaporizing temp 325 ordmC- Nebulizer gas ndash Nitrogen

at 40 bar - Drying gas ndash Nitrogen at 4

Lmin at 350 ordmC- Capillary voltage ndash4000 V- Corona current ndash 4 microA- LOD 001 microgmL except for

benomyl at 01 microgmL

bull MS

- Use SIM mode fragmentor voltage 70 V

(Ref 3)

Compound mz

Carbendazim benomyl

192

Thiabendazole 202

Imazalil 297

Thiophanate methyl

343

9

Chromatogram

A) Standard with concentration of 05 ppm B) Control C) SamplePeak identification (1) Carbendazim (2) thiabendazole (3) thiophanate methyl (4) benomyl and (5) imazalil (Ref 3)

10

Results and Conclusions

bull HPLCMS provides a reliable sensitive analytical tool in determining the levels of fungicides on citrus fruits

bull By monitoring the levels of fungicides the

possible health threats can be minimized (Ref3)

1111

Instrument Specificationsbull HP-1100 Series LCMSD

- quadrupole based LCMSD system

- HPLC column- Zorbax Rapid Resolution SB-C18 (30 x 46 mm ID 35 microm) column with a guard column of LiChrosorb RP-8 (10 x 46 mm 5 microm)

- standard API source that can be configured as APCI or ES (Ref 3) - mass range 2-3000 mz - mass accuracy plusmn013 u within the calibrated mass range in scan mode - mass axis stability plusmn013 u over 8 hours - scan speed 2500 us standard mode 5250 us fast scan mode - SIM sensitivity ESI at 400 microLmin APCI at 1000 microLmin - multiple signal acquisition ability to cycle through four different acquisition modes on a scan-by-scan basis

within a single run (Ref 4)

1212

References 1) httppostharvestifasufleduMRLsFL20Citrus20 MRLs20-209208pdf accessed December 2nd 20082) httpcvuasxn--untersuchungsmter-bwnzbdepdf poster_israel1999_orgzinnpdf accessed December 3rd 20083) Fernaacutendez M Rodriacuteguez R Picoacute Y Mantildees J J Chromatogr A 2001 912 301-3104) httpwwwgentechscientificcom10156htm accessed December 3rd 2008

22

Introduction

Fungicides applied on the surface of post-harvest citrus fruits are possible health threats since they can be easily absorbed into the body through peeling and eating with hands Controlling and monitoring the appropriate levels of fungicides would reduce the possible harmful effects by the ingestion

3

Maximum residue limits for five fungicides and their structures

Benomyl 10 ppm

Carbendazim 05 ppm

Imazalil 10 ppm

Thiabendazole 10 ppm

Thiophanate methyl

05 ppm

(Ref 1 3)

44

Method

bull HPLCMS

- Since the fungicides have different polarities they are retained to the stationary phase by different degrees This allows separation by HPLC

- High sensitivity proved by the limit of detection (lt 003 ppm)

- No derivatization is needed - Fastness (No additional clean-up for sample preparation) - Highly automated (reduced manual labor and human

intervention)

(Ref 2)

5

Procedure

Homogenization

Extraction

LCMSD system

wwwthesuncouk wwwbuchicom

ic3dbouedu

wwwhhscacuk

6

Preparation of standards controls and samples

(1) Homogenization

- Blend chopped oranges with ethyl acetate anhydrous sodium sulphate

- For controls use oranges without post harvest fungicide treatment

(2) Extraction

- Use sodium sulphate for filtering

- Add acetonitrile Dry - Dissolve in acetonitrile 2

mM ammonium formate (5050 vv) - For standards controls

mixed with fungicide solutions

- Standard Concentrations 005 05 5 ppm

(Ref 3)

7

Conditions for HPLC

bull Injection volume 5 microL

bull Solvents used for mobile phase methanol 2 mM ammonium formate in water

bull Gradient elution with flow rate of 1 mLmin

Methanol 50

Methanol 80

Methanol 30

(Ref 3)

8

Conditions for interface and MS

bull APCI Interface

- Vaporizing temp 325 ordmC- Nebulizer gas ndash Nitrogen

at 40 bar - Drying gas ndash Nitrogen at 4

Lmin at 350 ordmC- Capillary voltage ndash4000 V- Corona current ndash 4 microA- LOD 001 microgmL except for

benomyl at 01 microgmL

bull MS

- Use SIM mode fragmentor voltage 70 V

(Ref 3)

Compound mz

Carbendazim benomyl

192

Thiabendazole 202

Imazalil 297

Thiophanate methyl

343

9

Chromatogram

A) Standard with concentration of 05 ppm B) Control C) SamplePeak identification (1) Carbendazim (2) thiabendazole (3) thiophanate methyl (4) benomyl and (5) imazalil (Ref 3)

10

Results and Conclusions

bull HPLCMS provides a reliable sensitive analytical tool in determining the levels of fungicides on citrus fruits

bull By monitoring the levels of fungicides the

possible health threats can be minimized (Ref3)

1111

Instrument Specificationsbull HP-1100 Series LCMSD

- quadrupole based LCMSD system

- HPLC column- Zorbax Rapid Resolution SB-C18 (30 x 46 mm ID 35 microm) column with a guard column of LiChrosorb RP-8 (10 x 46 mm 5 microm)

- standard API source that can be configured as APCI or ES (Ref 3) - mass range 2-3000 mz - mass accuracy plusmn013 u within the calibrated mass range in scan mode - mass axis stability plusmn013 u over 8 hours - scan speed 2500 us standard mode 5250 us fast scan mode - SIM sensitivity ESI at 400 microLmin APCI at 1000 microLmin - multiple signal acquisition ability to cycle through four different acquisition modes on a scan-by-scan basis

within a single run (Ref 4)

1212

References 1) httppostharvestifasufleduMRLsFL20Citrus20 MRLs20-209208pdf accessed December 2nd 20082) httpcvuasxn--untersuchungsmter-bwnzbdepdf poster_israel1999_orgzinnpdf accessed December 3rd 20083) Fernaacutendez M Rodriacuteguez R Picoacute Y Mantildees J J Chromatogr A 2001 912 301-3104) httpwwwgentechscientificcom10156htm accessed December 3rd 2008

3

Maximum residue limits for five fungicides and their structures

Benomyl 10 ppm

Carbendazim 05 ppm

Imazalil 10 ppm

Thiabendazole 10 ppm

Thiophanate methyl

05 ppm

(Ref 1 3)

44

Method

bull HPLCMS

- Since the fungicides have different polarities they are retained to the stationary phase by different degrees This allows separation by HPLC

- High sensitivity proved by the limit of detection (lt 003 ppm)

- No derivatization is needed - Fastness (No additional clean-up for sample preparation) - Highly automated (reduced manual labor and human

intervention)

(Ref 2)

5

Procedure

Homogenization

Extraction

LCMSD system

wwwthesuncouk wwwbuchicom

ic3dbouedu

wwwhhscacuk

6

Preparation of standards controls and samples

(1) Homogenization

- Blend chopped oranges with ethyl acetate anhydrous sodium sulphate

- For controls use oranges without post harvest fungicide treatment

(2) Extraction

- Use sodium sulphate for filtering

- Add acetonitrile Dry - Dissolve in acetonitrile 2

mM ammonium formate (5050 vv) - For standards controls

mixed with fungicide solutions

- Standard Concentrations 005 05 5 ppm

(Ref 3)

7

Conditions for HPLC

bull Injection volume 5 microL

bull Solvents used for mobile phase methanol 2 mM ammonium formate in water

bull Gradient elution with flow rate of 1 mLmin

Methanol 50

Methanol 80

Methanol 30

(Ref 3)

8

Conditions for interface and MS

bull APCI Interface

- Vaporizing temp 325 ordmC- Nebulizer gas ndash Nitrogen

at 40 bar - Drying gas ndash Nitrogen at 4

Lmin at 350 ordmC- Capillary voltage ndash4000 V- Corona current ndash 4 microA- LOD 001 microgmL except for

benomyl at 01 microgmL

bull MS

- Use SIM mode fragmentor voltage 70 V

(Ref 3)

Compound mz

Carbendazim benomyl

192

Thiabendazole 202

Imazalil 297

Thiophanate methyl

343

9

Chromatogram

A) Standard with concentration of 05 ppm B) Control C) SamplePeak identification (1) Carbendazim (2) thiabendazole (3) thiophanate methyl (4) benomyl and (5) imazalil (Ref 3)

10

Results and Conclusions

bull HPLCMS provides a reliable sensitive analytical tool in determining the levels of fungicides on citrus fruits

bull By monitoring the levels of fungicides the

possible health threats can be minimized (Ref3)

1111

Instrument Specificationsbull HP-1100 Series LCMSD

- quadrupole based LCMSD system

- HPLC column- Zorbax Rapid Resolution SB-C18 (30 x 46 mm ID 35 microm) column with a guard column of LiChrosorb RP-8 (10 x 46 mm 5 microm)

- standard API source that can be configured as APCI or ES (Ref 3) - mass range 2-3000 mz - mass accuracy plusmn013 u within the calibrated mass range in scan mode - mass axis stability plusmn013 u over 8 hours - scan speed 2500 us standard mode 5250 us fast scan mode - SIM sensitivity ESI at 400 microLmin APCI at 1000 microLmin - multiple signal acquisition ability to cycle through four different acquisition modes on a scan-by-scan basis

within a single run (Ref 4)

1212

References 1) httppostharvestifasufleduMRLsFL20Citrus20 MRLs20-209208pdf accessed December 2nd 20082) httpcvuasxn--untersuchungsmter-bwnzbdepdf poster_israel1999_orgzinnpdf accessed December 3rd 20083) Fernaacutendez M Rodriacuteguez R Picoacute Y Mantildees J J Chromatogr A 2001 912 301-3104) httpwwwgentechscientificcom10156htm accessed December 3rd 2008

44

Method

bull HPLCMS

- Since the fungicides have different polarities they are retained to the stationary phase by different degrees This allows separation by HPLC

- High sensitivity proved by the limit of detection (lt 003 ppm)

- No derivatization is needed - Fastness (No additional clean-up for sample preparation) - Highly automated (reduced manual labor and human

intervention)

(Ref 2)

5

Procedure

Homogenization

Extraction

LCMSD system

wwwthesuncouk wwwbuchicom

ic3dbouedu

wwwhhscacuk

6

Preparation of standards controls and samples

(1) Homogenization

- Blend chopped oranges with ethyl acetate anhydrous sodium sulphate

- For controls use oranges without post harvest fungicide treatment

(2) Extraction

- Use sodium sulphate for filtering

- Add acetonitrile Dry - Dissolve in acetonitrile 2

mM ammonium formate (5050 vv) - For standards controls

mixed with fungicide solutions

- Standard Concentrations 005 05 5 ppm

(Ref 3)

7

Conditions for HPLC

bull Injection volume 5 microL

bull Solvents used for mobile phase methanol 2 mM ammonium formate in water

bull Gradient elution with flow rate of 1 mLmin

Methanol 50

Methanol 80

Methanol 30

(Ref 3)

8

Conditions for interface and MS

bull APCI Interface

- Vaporizing temp 325 ordmC- Nebulizer gas ndash Nitrogen

at 40 bar - Drying gas ndash Nitrogen at 4

Lmin at 350 ordmC- Capillary voltage ndash4000 V- Corona current ndash 4 microA- LOD 001 microgmL except for

benomyl at 01 microgmL

bull MS

- Use SIM mode fragmentor voltage 70 V

(Ref 3)

Compound mz

Carbendazim benomyl

192

Thiabendazole 202

Imazalil 297

Thiophanate methyl

343

9

Chromatogram

A) Standard with concentration of 05 ppm B) Control C) SamplePeak identification (1) Carbendazim (2) thiabendazole (3) thiophanate methyl (4) benomyl and (5) imazalil (Ref 3)

10

Results and Conclusions

bull HPLCMS provides a reliable sensitive analytical tool in determining the levels of fungicides on citrus fruits

bull By monitoring the levels of fungicides the

possible health threats can be minimized (Ref3)

1111

Instrument Specificationsbull HP-1100 Series LCMSD

- quadrupole based LCMSD system

- HPLC column- Zorbax Rapid Resolution SB-C18 (30 x 46 mm ID 35 microm) column with a guard column of LiChrosorb RP-8 (10 x 46 mm 5 microm)

- standard API source that can be configured as APCI or ES (Ref 3) - mass range 2-3000 mz - mass accuracy plusmn013 u within the calibrated mass range in scan mode - mass axis stability plusmn013 u over 8 hours - scan speed 2500 us standard mode 5250 us fast scan mode - SIM sensitivity ESI at 400 microLmin APCI at 1000 microLmin - multiple signal acquisition ability to cycle through four different acquisition modes on a scan-by-scan basis

within a single run (Ref 4)

1212

References 1) httppostharvestifasufleduMRLsFL20Citrus20 MRLs20-209208pdf accessed December 2nd 20082) httpcvuasxn--untersuchungsmter-bwnzbdepdf poster_israel1999_orgzinnpdf accessed December 3rd 20083) Fernaacutendez M Rodriacuteguez R Picoacute Y Mantildees J J Chromatogr A 2001 912 301-3104) httpwwwgentechscientificcom10156htm accessed December 3rd 2008

5

Procedure

Homogenization

Extraction

LCMSD system

wwwthesuncouk wwwbuchicom

ic3dbouedu

wwwhhscacuk

6

Preparation of standards controls and samples

(1) Homogenization

- Blend chopped oranges with ethyl acetate anhydrous sodium sulphate

- For controls use oranges without post harvest fungicide treatment

(2) Extraction

- Use sodium sulphate for filtering

- Add acetonitrile Dry - Dissolve in acetonitrile 2

mM ammonium formate (5050 vv) - For standards controls

mixed with fungicide solutions

- Standard Concentrations 005 05 5 ppm

(Ref 3)

7

Conditions for HPLC

bull Injection volume 5 microL

bull Solvents used for mobile phase methanol 2 mM ammonium formate in water

bull Gradient elution with flow rate of 1 mLmin

Methanol 50

Methanol 80

Methanol 30

(Ref 3)

8

Conditions for interface and MS

bull APCI Interface

- Vaporizing temp 325 ordmC- Nebulizer gas ndash Nitrogen

at 40 bar - Drying gas ndash Nitrogen at 4

Lmin at 350 ordmC- Capillary voltage ndash4000 V- Corona current ndash 4 microA- LOD 001 microgmL except for

benomyl at 01 microgmL

bull MS

- Use SIM mode fragmentor voltage 70 V

(Ref 3)

Compound mz

Carbendazim benomyl

192

Thiabendazole 202

Imazalil 297

Thiophanate methyl

343

9

Chromatogram

A) Standard with concentration of 05 ppm B) Control C) SamplePeak identification (1) Carbendazim (2) thiabendazole (3) thiophanate methyl (4) benomyl and (5) imazalil (Ref 3)

10

Results and Conclusions

bull HPLCMS provides a reliable sensitive analytical tool in determining the levels of fungicides on citrus fruits

bull By monitoring the levels of fungicides the

possible health threats can be minimized (Ref3)

1111

Instrument Specificationsbull HP-1100 Series LCMSD

- quadrupole based LCMSD system

- HPLC column- Zorbax Rapid Resolution SB-C18 (30 x 46 mm ID 35 microm) column with a guard column of LiChrosorb RP-8 (10 x 46 mm 5 microm)

- standard API source that can be configured as APCI or ES (Ref 3) - mass range 2-3000 mz - mass accuracy plusmn013 u within the calibrated mass range in scan mode - mass axis stability plusmn013 u over 8 hours - scan speed 2500 us standard mode 5250 us fast scan mode - SIM sensitivity ESI at 400 microLmin APCI at 1000 microLmin - multiple signal acquisition ability to cycle through four different acquisition modes on a scan-by-scan basis

within a single run (Ref 4)

1212

References 1) httppostharvestifasufleduMRLsFL20Citrus20 MRLs20-209208pdf accessed December 2nd 20082) httpcvuasxn--untersuchungsmter-bwnzbdepdf poster_israel1999_orgzinnpdf accessed December 3rd 20083) Fernaacutendez M Rodriacuteguez R Picoacute Y Mantildees J J Chromatogr A 2001 912 301-3104) httpwwwgentechscientificcom10156htm accessed December 3rd 2008

6

Preparation of standards controls and samples

(1) Homogenization

- Blend chopped oranges with ethyl acetate anhydrous sodium sulphate

- For controls use oranges without post harvest fungicide treatment

(2) Extraction

- Use sodium sulphate for filtering

- Add acetonitrile Dry - Dissolve in acetonitrile 2

mM ammonium formate (5050 vv) - For standards controls

mixed with fungicide solutions

- Standard Concentrations 005 05 5 ppm

(Ref 3)

7

Conditions for HPLC

bull Injection volume 5 microL

bull Solvents used for mobile phase methanol 2 mM ammonium formate in water

bull Gradient elution with flow rate of 1 mLmin

Methanol 50

Methanol 80

Methanol 30

(Ref 3)

8

Conditions for interface and MS

bull APCI Interface

- Vaporizing temp 325 ordmC- Nebulizer gas ndash Nitrogen

at 40 bar - Drying gas ndash Nitrogen at 4

Lmin at 350 ordmC- Capillary voltage ndash4000 V- Corona current ndash 4 microA- LOD 001 microgmL except for

benomyl at 01 microgmL

bull MS

- Use SIM mode fragmentor voltage 70 V

(Ref 3)

Compound mz

Carbendazim benomyl

192

Thiabendazole 202

Imazalil 297

Thiophanate methyl

343

9

Chromatogram

A) Standard with concentration of 05 ppm B) Control C) SamplePeak identification (1) Carbendazim (2) thiabendazole (3) thiophanate methyl (4) benomyl and (5) imazalil (Ref 3)

10

Results and Conclusions

bull HPLCMS provides a reliable sensitive analytical tool in determining the levels of fungicides on citrus fruits

bull By monitoring the levels of fungicides the

possible health threats can be minimized (Ref3)

1111

Instrument Specificationsbull HP-1100 Series LCMSD

- quadrupole based LCMSD system

- HPLC column- Zorbax Rapid Resolution SB-C18 (30 x 46 mm ID 35 microm) column with a guard column of LiChrosorb RP-8 (10 x 46 mm 5 microm)

- standard API source that can be configured as APCI or ES (Ref 3) - mass range 2-3000 mz - mass accuracy plusmn013 u within the calibrated mass range in scan mode - mass axis stability plusmn013 u over 8 hours - scan speed 2500 us standard mode 5250 us fast scan mode - SIM sensitivity ESI at 400 microLmin APCI at 1000 microLmin - multiple signal acquisition ability to cycle through four different acquisition modes on a scan-by-scan basis

within a single run (Ref 4)

1212

References 1) httppostharvestifasufleduMRLsFL20Citrus20 MRLs20-209208pdf accessed December 2nd 20082) httpcvuasxn--untersuchungsmter-bwnzbdepdf poster_israel1999_orgzinnpdf accessed December 3rd 20083) Fernaacutendez M Rodriacuteguez R Picoacute Y Mantildees J J Chromatogr A 2001 912 301-3104) httpwwwgentechscientificcom10156htm accessed December 3rd 2008

7

Conditions for HPLC

bull Injection volume 5 microL

bull Solvents used for mobile phase methanol 2 mM ammonium formate in water

bull Gradient elution with flow rate of 1 mLmin

Methanol 50

Methanol 80

Methanol 30

(Ref 3)

8

Conditions for interface and MS

bull APCI Interface

- Vaporizing temp 325 ordmC- Nebulizer gas ndash Nitrogen

at 40 bar - Drying gas ndash Nitrogen at 4

Lmin at 350 ordmC- Capillary voltage ndash4000 V- Corona current ndash 4 microA- LOD 001 microgmL except for

benomyl at 01 microgmL

bull MS

- Use SIM mode fragmentor voltage 70 V

(Ref 3)

Compound mz

Carbendazim benomyl

192

Thiabendazole 202

Imazalil 297

Thiophanate methyl

343

9

Chromatogram

A) Standard with concentration of 05 ppm B) Control C) SamplePeak identification (1) Carbendazim (2) thiabendazole (3) thiophanate methyl (4) benomyl and (5) imazalil (Ref 3)

10

Results and Conclusions

bull HPLCMS provides a reliable sensitive analytical tool in determining the levels of fungicides on citrus fruits

bull By monitoring the levels of fungicides the

possible health threats can be minimized (Ref3)

1111

Instrument Specificationsbull HP-1100 Series LCMSD

- quadrupole based LCMSD system

- HPLC column- Zorbax Rapid Resolution SB-C18 (30 x 46 mm ID 35 microm) column with a guard column of LiChrosorb RP-8 (10 x 46 mm 5 microm)

- standard API source that can be configured as APCI or ES (Ref 3) - mass range 2-3000 mz - mass accuracy plusmn013 u within the calibrated mass range in scan mode - mass axis stability plusmn013 u over 8 hours - scan speed 2500 us standard mode 5250 us fast scan mode - SIM sensitivity ESI at 400 microLmin APCI at 1000 microLmin - multiple signal acquisition ability to cycle through four different acquisition modes on a scan-by-scan basis

within a single run (Ref 4)

1212

References 1) httppostharvestifasufleduMRLsFL20Citrus20 MRLs20-209208pdf accessed December 2nd 20082) httpcvuasxn--untersuchungsmter-bwnzbdepdf poster_israel1999_orgzinnpdf accessed December 3rd 20083) Fernaacutendez M Rodriacuteguez R Picoacute Y Mantildees J J Chromatogr A 2001 912 301-3104) httpwwwgentechscientificcom10156htm accessed December 3rd 2008

8

Conditions for interface and MS

bull APCI Interface

- Vaporizing temp 325 ordmC- Nebulizer gas ndash Nitrogen

at 40 bar - Drying gas ndash Nitrogen at 4

Lmin at 350 ordmC- Capillary voltage ndash4000 V- Corona current ndash 4 microA- LOD 001 microgmL except for

benomyl at 01 microgmL

bull MS

- Use SIM mode fragmentor voltage 70 V

(Ref 3)

Compound mz

Carbendazim benomyl

192

Thiabendazole 202

Imazalil 297

Thiophanate methyl

343

9

Chromatogram

A) Standard with concentration of 05 ppm B) Control C) SamplePeak identification (1) Carbendazim (2) thiabendazole (3) thiophanate methyl (4) benomyl and (5) imazalil (Ref 3)

10

Results and Conclusions

bull HPLCMS provides a reliable sensitive analytical tool in determining the levels of fungicides on citrus fruits

bull By monitoring the levels of fungicides the

possible health threats can be minimized (Ref3)

1111

Instrument Specificationsbull HP-1100 Series LCMSD

- quadrupole based LCMSD system

- HPLC column- Zorbax Rapid Resolution SB-C18 (30 x 46 mm ID 35 microm) column with a guard column of LiChrosorb RP-8 (10 x 46 mm 5 microm)

- standard API source that can be configured as APCI or ES (Ref 3) - mass range 2-3000 mz - mass accuracy plusmn013 u within the calibrated mass range in scan mode - mass axis stability plusmn013 u over 8 hours - scan speed 2500 us standard mode 5250 us fast scan mode - SIM sensitivity ESI at 400 microLmin APCI at 1000 microLmin - multiple signal acquisition ability to cycle through four different acquisition modes on a scan-by-scan basis

within a single run (Ref 4)

1212

References 1) httppostharvestifasufleduMRLsFL20Citrus20 MRLs20-209208pdf accessed December 2nd 20082) httpcvuasxn--untersuchungsmter-bwnzbdepdf poster_israel1999_orgzinnpdf accessed December 3rd 20083) Fernaacutendez M Rodriacuteguez R Picoacute Y Mantildees J J Chromatogr A 2001 912 301-3104) httpwwwgentechscientificcom10156htm accessed December 3rd 2008

9

Chromatogram

A) Standard with concentration of 05 ppm B) Control C) SamplePeak identification (1) Carbendazim (2) thiabendazole (3) thiophanate methyl (4) benomyl and (5) imazalil (Ref 3)

10

Results and Conclusions

bull HPLCMS provides a reliable sensitive analytical tool in determining the levels of fungicides on citrus fruits

bull By monitoring the levels of fungicides the

possible health threats can be minimized (Ref3)

1111

Instrument Specificationsbull HP-1100 Series LCMSD

- quadrupole based LCMSD system

- HPLC column- Zorbax Rapid Resolution SB-C18 (30 x 46 mm ID 35 microm) column with a guard column of LiChrosorb RP-8 (10 x 46 mm 5 microm)

- standard API source that can be configured as APCI or ES (Ref 3) - mass range 2-3000 mz - mass accuracy plusmn013 u within the calibrated mass range in scan mode - mass axis stability plusmn013 u over 8 hours - scan speed 2500 us standard mode 5250 us fast scan mode - SIM sensitivity ESI at 400 microLmin APCI at 1000 microLmin - multiple signal acquisition ability to cycle through four different acquisition modes on a scan-by-scan basis

within a single run (Ref 4)

1212

References 1) httppostharvestifasufleduMRLsFL20Citrus20 MRLs20-209208pdf accessed December 2nd 20082) httpcvuasxn--untersuchungsmter-bwnzbdepdf poster_israel1999_orgzinnpdf accessed December 3rd 20083) Fernaacutendez M Rodriacuteguez R Picoacute Y Mantildees J J Chromatogr A 2001 912 301-3104) httpwwwgentechscientificcom10156htm accessed December 3rd 2008

10

Results and Conclusions

bull HPLCMS provides a reliable sensitive analytical tool in determining the levels of fungicides on citrus fruits

bull By monitoring the levels of fungicides the

possible health threats can be minimized (Ref3)

1111

Instrument Specificationsbull HP-1100 Series LCMSD

- quadrupole based LCMSD system

- HPLC column- Zorbax Rapid Resolution SB-C18 (30 x 46 mm ID 35 microm) column with a guard column of LiChrosorb RP-8 (10 x 46 mm 5 microm)

- standard API source that can be configured as APCI or ES (Ref 3) - mass range 2-3000 mz - mass accuracy plusmn013 u within the calibrated mass range in scan mode - mass axis stability plusmn013 u over 8 hours - scan speed 2500 us standard mode 5250 us fast scan mode - SIM sensitivity ESI at 400 microLmin APCI at 1000 microLmin - multiple signal acquisition ability to cycle through four different acquisition modes on a scan-by-scan basis

within a single run (Ref 4)

1212

References 1) httppostharvestifasufleduMRLsFL20Citrus20 MRLs20-209208pdf accessed December 2nd 20082) httpcvuasxn--untersuchungsmter-bwnzbdepdf poster_israel1999_orgzinnpdf accessed December 3rd 20083) Fernaacutendez M Rodriacuteguez R Picoacute Y Mantildees J J Chromatogr A 2001 912 301-3104) httpwwwgentechscientificcom10156htm accessed December 3rd 2008

1111

Instrument Specificationsbull HP-1100 Series LCMSD

- quadrupole based LCMSD system

- HPLC column- Zorbax Rapid Resolution SB-C18 (30 x 46 mm ID 35 microm) column with a guard column of LiChrosorb RP-8 (10 x 46 mm 5 microm)

- standard API source that can be configured as APCI or ES (Ref 3) - mass range 2-3000 mz - mass accuracy plusmn013 u within the calibrated mass range in scan mode - mass axis stability plusmn013 u over 8 hours - scan speed 2500 us standard mode 5250 us fast scan mode - SIM sensitivity ESI at 400 microLmin APCI at 1000 microLmin - multiple signal acquisition ability to cycle through four different acquisition modes on a scan-by-scan basis

within a single run (Ref 4)

1212

References 1) httppostharvestifasufleduMRLsFL20Citrus20 MRLs20-209208pdf accessed December 2nd 20082) httpcvuasxn--untersuchungsmter-bwnzbdepdf poster_israel1999_orgzinnpdf accessed December 3rd 20083) Fernaacutendez M Rodriacuteguez R Picoacute Y Mantildees J J Chromatogr A 2001 912 301-3104) httpwwwgentechscientificcom10156htm accessed December 3rd 2008

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References 1) httppostharvestifasufleduMRLsFL20Citrus20 MRLs20-209208pdf accessed December 2nd 20082) httpcvuasxn--untersuchungsmter-bwnzbdepdf poster_israel1999_orgzinnpdf accessed December 3rd 20083) Fernaacutendez M Rodriacuteguez R Picoacute Y Mantildees J J Chromatogr A 2001 912 301-3104) httpwwwgentechscientificcom10156htm accessed December 3rd 2008