Determination of Nutritional Elements in Plant Leaves using an Inductively Coupled Plasma Optical Emission Spectrometer with a Dual Plasma View Elena Chernetsova, Sanja Asendorf, Matthew Cassap, Thermo Fisher Scientific, Bremen, Germany

2 Determination of Nutritional Elements in Plant Leaves using an Inductively Coupled Plasma Optical Emission Spectrometer with a Dual Plasma View

Conclusion The iCAP 7000 Series ICP-OES with dual view provides the unprecedented flexibility of analysis for determining the macro- and microelements in plant matrices. Driven by the Qtegra ISDS Software, it is easily adjusted to the peculiarities of sample matrix and at the same time allows to increase the analysis productivity using multiple tools, integrated into such systems. The complete set of data for ionomics studies can be easily collected by ICP-OES combined with workflow-driven Software.

This brings your analysis to the new level of robustness and confidence.

References 1. Husted S et al (2011) J Anal Atom Spectrom 26:52-79 2. Salt D et al (2008) Annu Rev Plant Biol 59:709-733 3. Hansen T et al (2013) Methods Mol Biol 953:121-141 4. Technical Note 43225 “Increasing productivity in ICP-OES with

Qtegra ISDS Software” 5. Technical Note 43173 “Thermo Scientific iCAP 7000 Series ICP-

OES Instrument Detection Limits”

In Normal mode the analysis is carried out per replicate, while the Speed mode allows a faster analysis by minimizing the number of transitions between different plasma views and slits, like shown on the scheme below as an example (where R is Replicate and represents the transition between plasma views):

Normal: R1 (Vis) R1 (UV) R2 (Vis) R2 (UV) R3 (Vis) R3 (UV) Speed: R1 (Vis) R2 (Vis) R3 (Vis) R1 (UV) R2 (UV) R3 (UV)

The Sprint mode operates similarly to Speed mode, this mode is typically used for trend analysis.

In the Speed mode one analysis per sample took a minimum of at least 20 seconds less than in Normal mode. Analyzing a sequence of 50 multielement samples in the Speed mode provided an increased sample throughput with the recovery at the level of 100% over the whole analysis duration (Fig. 4). Due to this, analysis of leaf digests was carried out in the Speed mode.

Overview Purpose: To demonstrate the advantages of an Inductively Coupled Plasma – Optical Emission Spectrometer (ICP-OES) with a Dual Plasma view for the determination of nutritional elements in plant leaves.

Methods: ICP-OES, microwave digestion.

Results: Concentrations of elements were determined in tobacco and apple leaves, the found concentrations were in the good agreement with the values, given for the Standard Reference Materials (SRMs).

Introduction The commercial value of a plant as an edible product or as a raw material for preparing medicines depends on the balance of different chemical constituents in the plant, including the elemental composition. Based on the element concentrations in the different plant parts, e.g., in plant leaves, the quality of plants can be evaluated for scientific and commercial purposes. Typically the major elements (macronutrients, including calcium, potassium, phosphorous, nitrogen, magnesium, sulfur) as well as the trace elements (micronutrients, including boron, manganese, iron, nickel, cupper, and zinc) are assessed. This allows the identification of nutrient deficiency, which can lead to the reduced quality of flowers, fruits or vegetables, produced by the plant. Besides, the information on the elemental composition of the plant may explain its medical or toxic properties.

In recent years with the development of OMICS-techniques, the study of plant ionomes has become important. Ionomics involves the quantitative and simultaneous measurement of plant elemental composition and changes in this composition in response to physiological effects and genetic modifications [1-3]. Due to this, development of new advanced techniques for the quantitation of plant ionoms or their parts is of high importance.

ICP-OES has the high potential for being employed within such studies. In this study the advantages of Thermo ScientificTM iCAPTM 7000 Series ICP-OES dual view for the determination of nutritional elements in plant matrices are discussed, using plant leaf analysis as an example.

Methods

Sample Preparation

SRMs of dried apple leaves (SRM 1515, NIST, USA) and dried tobacco (Institute of Nuclear Chemistry and Technology, Poland) were used. Both materials were analyzed for moisture before each analysis.

All samples were digested in a microwave oven using 0.5 g of each sample in 7 mL concentrated HNO3, 2 mL concentrated H2O2 and 3 mL deionized water.

The digestion included 4 steps: 1. 3 minutes at 700 W 2. 9 minutes at 500 W 3. 4 minutes at 1000 W 4. 14 minutes at 1000 W.

After digestion an aliquot of the sample was diluted 10-fold with 6% (v/v) HNO3 directly before analysis, the remaining sample was analyzed directly. As a blank sample, 6% (v/v) HNO3 was used for all measurements. Duplicate method blanks were analyzed for each microwave digestion batch.

Single element stock standards (Fisher Scientific) were used to prepare multielement standard solutions with the necessary element concentrations. The standard solutions were used for productivity studies and for calibration curve generation. Al, Ca, Fe, K, Mg, Na, P and S were present at 0.5 to 75 mg L-1; Ba, B, Cr, Cu, Li, Mn, Pb, V, and Zn were present at 50 mg L-1 to 10 mg L-1 in the standard solutions.

Instrumentation

The Thermo Scientific™ iCAP™ 7600 ICP-OES Duo (Fig.01) has been used in all experiments described in this paper. It was coupled to a Teledyne CETAC ASX-520 Random Access Intelligent Autosampler. Method parameters, listed in Table 1, were applied for all analyses (unless stated).

Tygon is a registered trade mark of Saint-Gobain Corporation. CETAC is a registered trademark of Teledyne CETAC Technologies. All other trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manners that might infringe the intellectual property rights of others.

Presented at the eWCPS, Münster, Germany, 2/2015.

FIGURE 4. Recoveries at P 177.495 nm (axial) and at Mg 279.553 nm (radial) for 50 samples, analyzed in Speed mode.

TABLE 2. Concentrations of elements in leaves and DLs.

The determined concentrations of elements in apple and tobacco leaves were in the good agreement with the specified values for the used SRMs (see Table 2). The deviations from the specified values were less than 10% in all cases. The detection limits (DL) for the majority of the elements were much lower than 1 mg L-1 [5].

Determination of Nutritional Elements in Plant Leaves using an Inductively Coupled Plasma Optical Emission Spectrometer with a Dual Plasma View Elena Chernetsova, Sanja Asendorf, Matthew Cassap Thermo Fisher Scientific, Bremen, Germany

Results Maximizing the Analysis Productivity

Using autosamplers for ICP-OES analysis reduces user intervention and is the first step towards increasing sample throughput. Autosamplers allow implementing unattended analysis with higher reproducibility of measurements, with precise uptake and wash cycles applied to each sample.

Sample uptake was preformed via a Teledyne CETAC ASX-520 autosampler. All parameters relating to the autosampler can be set directly in the Thermo Scientific™ QtegraTM ISDSTM Software (Fig. 2). Timings and layout can be modified directly in the respective Labbooks, with flexible rack configurations available.

With the Qtegra ISDS Software it is possible to apply three different modes of analysis (Fig. 3): Normal, Speed, and Sprint. These modes relate to the sequencing of the plasma views.

TABLE 1. ICP-OES method parameters.

Parameter Setting

Pump Tubing Sample TygonTM white/white/white

Drain TygonTM yellow/blue/yellow

Pump Speed 50 rpm

Spray Chamber Glass cyclonic

Nebulizer Glass concentric

Center Tube 2 mm

Nebulizer Gas Flow 0.5 L min-1

Auxiliary Gas Flow 0.5 L min-1

Coolant Gas Flow 12 L min-1

RF Power 1150 W

Number of replicates 3

Exposure Time UV 10 s and Vis 5 s for both Axial and Radial

modes Wavelengths Measured Axially

Al 167.079 nm, B 249.773 nm, Ba 455.403 nm, Co 228.616 nm, Cr 283.563 nm, Ni 221.647 nm, P 177.495 nm, Pb 220.353 nm, Zn 202.548 nm

Wavelengths Measured Radially

Ca 393.366 nm, Cu 324.754 nm, Fe 259.940 nm, K 766.490 nm, Mg 279.553 nm, Mn 257.610 nm, Na 589.592 nm, S 180.731 nm, V 292.402 nm

FIGURE 2. ASX-520 Autosampler view on the Dashboard of Qtegra ISDS Software.

FIGURE 3. Analysis mode drop-down menu.

Element

l (nm)

Apple leaves Tobacco leaves DL, mg L-1

Cref., mg kg-1 Cexp., mg kg-1 Cref., mg kg-1 Cexp., mg/kg-1

Al 167.079 290±30 280 200±30 190 0.3

B 249.773 27±3 25 34±3 33 5.1

Ba 455.403 49±5 49 67±7 64 0.03

Ca 393.366 15200±150

0 14800 40000±4000 40400 1.6

Co 228.616 - - 1.0±0.1 1.0 0.4

Cr 283.563 - - 6.3±0.6 5.7 0.4

Cu 324.754 5.6±0.6 5.7 10.1±1.0 10.0 2.4

Fe 259.940 83±8 82 - - 0.8

K 766.490 16100±200

0 17800 23000±8000 29000 5.1

Mg 279.553 2700±300 2670 8500±900 8400 0.04

Mn 257.610 54±5 54 180±20 180 0.2

Na 589.592 24±2 24 - - 1.8

Ni 221.647 0.9±0.1 0.9 8.5±0.9 8.7 0.3

P 177.495 1600±200 1700 1700±200 1800 5.7

Pb 220.353 0.47±0.05 0.48 2.0±0.3 1.7 1.1

S 180.731 - - 4600±900 5100 2.2

V 292.402 - - 4.1±0.6 4.6 4.9

Zn 202.548 12.5±0.3 12.2 52±5 51 1.8

Quantitation of Elements in Tobacco and Apple Leaves

Using the iCAP 7000 Series ICP-OES with a Dual Plasma View, it was possible to determine the elements of interest in the wavelength range from 166 to 847 nm in one run using radial and axial views. This allows selecting the most beneficial wavelengths and plasma view for each analyte and minimizing matrix effects. Resulting calibration curves are exemplarily shown (Fig. 5)

FIGURE 1. iCAP 7600 Duo ICP-OES and its optical system.

More information of increasing the analysis productivity using the iCAP 7000 Series ICP-OES with Qtegra ISDS Software can be found in our Technical Note 43225 [4].

FIGURE 5. Calibration curves obtained for 4 different types of signals (axial vs. radial + low slit vs. high slit).

0

20

40

60

80

100

120

0 20 40 60 80 100 120 140 160 180

Rec

over

y, %

Time, min

P 177.495 nm axial

Mg 279.553 nm radial

3Thermo Scienti� c Poster Note • eWPC • PN43236-EN 0315S

Conclusion The iCAP 7000 Series ICP-OES with dual view provides the unprecedented flexibility of analysis for determining the macro- and microelements in plant matrices. Driven by the Qtegra ISDS Software, it is easily adjusted to the peculiarities of sample matrix and at the same time allows to increase the analysis productivity using multiple tools, integrated into such systems. The complete set of data for ionomics studies can be easily collected by ICP-OES combined with workflow-driven Software.

This brings your analysis to the new level of robustness and confidence.

References 1. Husted S et al (2011) J Anal Atom Spectrom 26:52-79 2. Salt D et al (2008) Annu Rev Plant Biol 59:709-733 3. Hansen T et al (2013) Methods Mol Biol 953:121-141 4. Technical Note 43225 “Increasing productivity in ICP-OES with

Qtegra ISDS Software” 5. Technical Note 43173 “Thermo Scientific iCAP 7000 Series ICP-

OES Instrument Detection Limits”

In Normal mode the analysis is carried out per replicate, while the Speed mode allows a faster analysis by minimizing the number of transitions between different plasma views and slits, like shown on the scheme below as an example (where R is Replicate and represents the transition between plasma views):

Normal: R1 (Vis) R1 (UV) R2 (Vis) R2 (UV) R3 (Vis) R3 (UV) Speed: R1 (Vis) R2 (Vis) R3 (Vis) R1 (UV) R2 (UV) R3 (UV)

The Sprint mode operates similarly to Speed mode, this mode is typically used for trend analysis.

In the Speed mode one analysis per sample took a minimum of at least 20 seconds less than in Normal mode. Analyzing a sequence of 50 multielement samples in the Speed mode provided an increased sample throughput with the recovery at the level of 100% over the whole analysis duration (Fig. 4). Due to this, analysis of leaf digests was carried out in the Speed mode.

Overview Purpose: To demonstrate the advantages of an Inductively Coupled Plasma – Optical Emission Spectrometer (ICP-OES) with a Dual Plasma view for the determination of nutritional elements in plant leaves.

Methods: ICP-OES, microwave digestion.

Results: Concentrations of elements were determined in tobacco and apple leaves, the found concentrations were in the good agreement with the values, given for the Standard Reference Materials (SRMs).

Introduction The commercial value of a plant as an edible product or as a raw material for preparing medicines depends on the balance of different chemical constituents in the plant, including the elemental composition. Based on the element concentrations in the different plant parts, e.g., in plant leaves, the quality of plants can be evaluated for scientific and commercial purposes. Typically the major elements (macronutrients, including calcium, potassium, phosphorous, nitrogen, magnesium, sulfur) as well as the trace elements (micronutrients, including boron, manganese, iron, nickel, cupper, and zinc) are assessed. This allows the identification of nutrient deficiency, which can lead to the reduced quality of flowers, fruits or vegetables, produced by the plant. Besides, the information on the elemental composition of the plant may explain its medical or toxic properties.

In recent years with the development of OMICS-techniques, the study of plant ionomes has become important. Ionomics involves the quantitative and simultaneous measurement of plant elemental composition and changes in this composition in response to physiological effects and genetic modifications [1-3]. Due to this, development of new advanced techniques for the quantitation of plant ionoms or their parts is of high importance.

ICP-OES has the high potential for being employed within such studies. In this study the advantages of Thermo ScientificTM iCAPTM 7000 Series ICP-OES dual view for the determination of nutritional elements in plant matrices are discussed, using plant leaf analysis as an example.

Methods

Sample Preparation

SRMs of dried apple leaves (SRM 1515, NIST, USA) and dried tobacco (Institute of Nuclear Chemistry and Technology, Poland) were used. Both materials were analyzed for moisture before each analysis.

All samples were digested in a microwave oven using 0.5 g of each sample in 7 mL concentrated HNO3, 2 mL concentrated H2O2 and 3 mL deionized water.

The digestion included 4 steps: 1. 3 minutes at 700 W 2. 9 minutes at 500 W 3. 4 minutes at 1000 W 4. 14 minutes at 1000 W.

After digestion an aliquot of the sample was diluted 10-fold with 6% (v/v) HNO3 directly before analysis, the remaining sample was analyzed directly. As a blank sample, 6% (v/v) HNO3 was used for all measurements. Duplicate method blanks were analyzed for each microwave digestion batch.

Single element stock standards (Fisher Scientific) were used to prepare multielement standard solutions with the necessary element concentrations. The standard solutions were used for productivity studies and for calibration curve generation. Al, Ca, Fe, K, Mg, Na, P and S were present at 0.5 to 75 mg L-1; Ba, B, Cr, Cu, Li, Mn, Pb, V, and Zn were present at 50 mg L-1 to 10 mg L-1 in the standard solutions.

Instrumentation

The Thermo Scientific™ iCAP™ 7600 ICP-OES Duo (Fig.01) has been used in all experiments described in this paper. It was coupled to a Teledyne CETAC ASX-520 Random Access Intelligent Autosampler. Method parameters, listed in Table 1, were applied for all analyses (unless stated).

Tygon is a registered trade mark of Saint-Gobain Corporation. CETAC is a registered trademark of Teledyne CETAC Technologies. All other trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manners that might infringe the intellectual property rights of others.

Presented at the eWCPS, Münster, Germany, 2/2015.

FIGURE 4. Recoveries at P 177.495 nm (axial) and at Mg 279.553 nm (radial) for 50 samples, analyzed in Speed mode.

TABLE 2. Concentrations of elements in leaves and DLs.

The determined concentrations of elements in apple and tobacco leaves were in the good agreement with the specified values for the used SRMs (see Table 2). The deviations from the specified values were less than 10% in all cases. The detection limits (DL) for the majority of the elements were much lower than 1 mg L-1 [5].

Determination of Nutritional Elements in Plant Leaves using an Inductively Coupled Plasma Optical Emission Spectrometer with a Dual Plasma View Elena Chernetsova, Sanja Asendorf, Matthew Cassap Thermo Fisher Scientific, Bremen, Germany

Results Maximizing the Analysis Productivity

Using autosamplers for ICP-OES analysis reduces user intervention and is the first step towards increasing sample throughput. Autosamplers allow implementing unattended analysis with higher reproducibility of measurements, with precise uptake and wash cycles applied to each sample.

Sample uptake was preformed via a Teledyne CETAC ASX-520 autosampler. All parameters relating to the autosampler can be set directly in the Thermo Scientific™ QtegraTM ISDSTM Software (Fig. 2). Timings and layout can be modified directly in the respective Labbooks, with flexible rack configurations available.

With the Qtegra ISDS Software it is possible to apply three different modes of analysis (Fig. 3): Normal, Speed, and Sprint. These modes relate to the sequencing of the plasma views.

TABLE 1. ICP-OES method parameters.

Parameter Setting

Pump Tubing Sample TygonTM white/white/white

Drain TygonTM yellow/blue/yellow

Pump Speed 50 rpm

Spray Chamber Glass cyclonic

Nebulizer Glass concentric

Center Tube 2 mm

Nebulizer Gas Flow 0.5 L min-1

Auxiliary Gas Flow 0.5 L min-1

Coolant Gas Flow 12 L min-1

RF Power 1150 W

Number of replicates 3

Exposure Time UV 10 s and Vis 5 s for both Axial and Radial

modes Wavelengths Measured Axially

Al 167.079 nm, B 249.773 nm, Ba 455.403 nm, Co 228.616 nm, Cr 283.563 nm, Ni 221.647 nm, P 177.495 nm, Pb 220.353 nm, Zn 202.548 nm

Wavelengths Measured Radially

Ca 393.366 nm, Cu 324.754 nm, Fe 259.940 nm, K 766.490 nm, Mg 279.553 nm, Mn 257.610 nm, Na 589.592 nm, S 180.731 nm, V 292.402 nm

FIGURE 2. ASX-520 Autosampler view on the Dashboard of Qtegra ISDS Software.

FIGURE 3. Analysis mode drop-down menu.

Element

l (nm)

Apple leaves Tobacco leaves DL, mg L-1

Cref., mg kg-1 Cexp., mg kg-1 Cref., mg kg-1 Cexp., mg/kg-1

Al 167.079 290±30 280 200±30 190 0.3

B 249.773 27±3 25 34±3 33 5.1

Ba 455.403 49±5 49 67±7 64 0.03

Ca 393.366 15200±150

0 14800 40000±4000 40400 1.6

Co 228.616 - - 1.0±0.1 1.0 0.4

Cr 283.563 - - 6.3±0.6 5.7 0.4

Cu 324.754 5.6±0.6 5.7 10.1±1.0 10.0 2.4

Fe 259.940 83±8 82 - - 0.8

K 766.490 16100±200

0 17800 23000±8000 29000 5.1

Mg 279.553 2700±300 2670 8500±900 8400 0.04

Mn 257.610 54±5 54 180±20 180 0.2

Na 589.592 24±2 24 - - 1.8

Ni 221.647 0.9±0.1 0.9 8.5±0.9 8.7 0.3

P 177.495 1600±200 1700 1700±200 1800 5.7

Pb 220.353 0.47±0.05 0.48 2.0±0.3 1.7 1.1

S 180.731 - - 4600±900 5100 2.2

V 292.402 - - 4.1±0.6 4.6 4.9

Zn 202.548 12.5±0.3 12.2 52±5 51 1.8

Quantitation of Elements in Tobacco and Apple Leaves

Using the iCAP 7000 Series ICP-OES with a Dual Plasma View, it was possible to determine the elements of interest in the wavelength range from 166 to 847 nm in one run using radial and axial views. This allows selecting the most beneficial wavelengths and plasma view for each analyte and minimizing matrix effects. Resulting calibration curves are exemplarily shown (Fig. 5)

FIGURE 1. iCAP 7600 Duo ICP-OES and its optical system.

More information of increasing the analysis productivity using the iCAP 7000 Series ICP-OES with Qtegra ISDS Software can be found in our Technical Note 43225 [4].

FIGURE 5. Calibration curves obtained for 4 different types of signals (axial vs. radial + low slit vs. high slit).

0

20

40

60

80

100

120

0 20 40 60 80 100 120 140 160 180

Rec

over

y, %

Time, min

P 177.495 nm axial

Mg 279.553 nm radial

4 Determination of Nutritional Elements in Plant Leaves using an Inductively Coupled Plasma Optical Emission Spectrometer with a Dual Plasma View

Conclusion The iCAP 7000 Series ICP-OES with dual view provides the unprecedented flexibility of analysis for determining the macro- and microelements in plant matrices. Driven by the Qtegra ISDS Software, it is easily adjusted to the peculiarities of sample matrix and at the same time allows to increase the analysis productivity using multiple tools, integrated into such systems. The complete set of data for ionomics studies can be easily collected by ICP-OES combined with workflow-driven Software.

This brings your analysis to the new level of robustness and confidence.

References 1. Husted S et al (2011) J Anal Atom Spectrom 26:52-79 2. Salt D et al (2008) Annu Rev Plant Biol 59:709-733 3. Hansen T et al (2013) Methods Mol Biol 953:121-141 4. Technical Note 43225 “Increasing productivity in ICP-OES with

Qtegra ISDS Software” 5. Technical Note 43173 “Thermo Scientific iCAP 7000 Series ICP-

OES Instrument Detection Limits”

In Normal mode the analysis is carried out per replicate, while the Speed mode allows a faster analysis by minimizing the number of transitions between different plasma views and slits, like shown on the scheme below as an example (where R is Replicate and represents the transition between plasma views):

Normal: R1 (Vis) R1 (UV) R2 (Vis) R2 (UV) R3 (Vis) R3 (UV) Speed: R1 (Vis) R2 (Vis) R3 (Vis) R1 (UV) R2 (UV) R3 (UV)

The Sprint mode operates similarly to Speed mode, this mode is typically used for trend analysis.

In the Speed mode one analysis per sample took a minimum of at least 20 seconds less than in Normal mode. Analyzing a sequence of 50 multielement samples in the Speed mode provided an increased sample throughput with the recovery at the level of 100% over the whole analysis duration (Fig. 4). Due to this, analysis of leaf digests was carried out in the Speed mode.

Overview Purpose: To demonstrate the advantages of an Inductively Coupled Plasma – Optical Emission Spectrometer (ICP-OES) with a Dual Plasma view for the determination of nutritional elements in plant leaves.

Methods: ICP-OES, microwave digestion.

Results: Concentrations of elements were determined in tobacco and apple leaves, the found concentrations were in the good agreement with the values, given for the Standard Reference Materials (SRMs).

Introduction The commercial value of a plant as an edible product or as a raw material for preparing medicines depends on the balance of different chemical constituents in the plant, including the elemental composition. Based on the element concentrations in the different plant parts, e.g., in plant leaves, the quality of plants can be evaluated for scientific and commercial purposes. Typically the major elements (macronutrients, including calcium, potassium, phosphorous, nitrogen, magnesium, sulfur) as well as the trace elements (micronutrients, including boron, manganese, iron, nickel, cupper, and zinc) are assessed. This allows the identification of nutrient deficiency, which can lead to the reduced quality of flowers, fruits or vegetables, produced by the plant. Besides, the information on the elemental composition of the plant may explain its medical or toxic properties.

In recent years with the development of OMICS-techniques, the study of plant ionomes has become important. Ionomics involves the quantitative and simultaneous measurement of plant elemental composition and changes in this composition in response to physiological effects and genetic modifications [1-3]. Due to this, development of new advanced techniques for the quantitation of plant ionoms or their parts is of high importance.

ICP-OES has the high potential for being employed within such studies. In this study the advantages of Thermo ScientificTM iCAPTM 7000 Series ICP-OES dual view for the determination of nutritional elements in plant matrices are discussed, using plant leaf analysis as an example.

Methods

Sample Preparation

SRMs of dried apple leaves (SRM 1515, NIST, USA) and dried tobacco (Institute of Nuclear Chemistry and Technology, Poland) were used. Both materials were analyzed for moisture before each analysis.

All samples were digested in a microwave oven using 0.5 g of each sample in 7 mL concentrated HNO3, 2 mL concentrated H2O2 and 3 mL deionized water.

The digestion included 4 steps: 1. 3 minutes at 700 W 2. 9 minutes at 500 W 3. 4 minutes at 1000 W 4. 14 minutes at 1000 W.

After digestion an aliquot of the sample was diluted 10-fold with 6% (v/v) HNO3 directly before analysis, the remaining sample was analyzed directly. As a blank sample, 6% (v/v) HNO3 was used for all measurements. Duplicate method blanks were analyzed for each microwave digestion batch.

Single element stock standards (Fisher Scientific) were used to prepare multielement standard solutions with the necessary element concentrations. The standard solutions were used for productivity studies and for calibration curve generation. Al, Ca, Fe, K, Mg, Na, P and S were present at 0.5 to 75 mg L-1; Ba, B, Cr, Cu, Li, Mn, Pb, V, and Zn were present at 50 mg L-1 to 10 mg L-1 in the standard solutions.

Instrumentation

The Thermo Scientific™ iCAP™ 7600 ICP-OES Duo (Fig.01) has been used in all experiments described in this paper. It was coupled to a Teledyne CETAC ASX-520 Random Access Intelligent Autosampler. Method parameters, listed in Table 1, were applied for all analyses (unless stated).

Tygon is a registered trade mark of Saint-Gobain Corporation. CETAC is a registered trademark of Teledyne CETAC Technologies. All other trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manners that might infringe the intellectual property rights of others.

Presented at the eWCPS, Münster, Germany, 2/2015.

FIGURE 4. Recoveries at P 177.495 nm (axial) and at Mg 279.553 nm (radial) for 50 samples, analyzed in Speed mode.

TABLE 2. Concentrations of elements in leaves and DLs.

The determined concentrations of elements in apple and tobacco leaves were in the good agreement with the specified values for the used SRMs (see Table 2). The deviations from the specified values were less than 10% in all cases. The detection limits (DL) for the majority of the elements were much lower than 1 mg L-1 [5].

Determination of Nutritional Elements in Plant Leaves using an Inductively Coupled Plasma Optical Emission Spectrometer with a Dual Plasma View Elena Chernetsova, Sanja Asendorf, Matthew Cassap Thermo Fisher Scientific, Bremen, Germany

Results Maximizing the Analysis Productivity

Using autosamplers for ICP-OES analysis reduces user intervention and is the first step towards increasing sample throughput. Autosamplers allow implementing unattended analysis with higher reproducibility of measurements, with precise uptake and wash cycles applied to each sample.

Sample uptake was preformed via a Teledyne CETAC ASX-520 autosampler. All parameters relating to the autosampler can be set directly in the Thermo Scientific™ QtegraTM ISDSTM Software (Fig. 2). Timings and layout can be modified directly in the respective Labbooks, with flexible rack configurations available.

With the Qtegra ISDS Software it is possible to apply three different modes of analysis (Fig. 3): Normal, Speed, and Sprint. These modes relate to the sequencing of the plasma views.

TABLE 1. ICP-OES method parameters.

Parameter Setting

Pump Tubing Sample TygonTM white/white/white

Drain TygonTM yellow/blue/yellow

Pump Speed 50 rpm

Spray Chamber Glass cyclonic

Nebulizer Glass concentric

Center Tube 2 mm

Nebulizer Gas Flow 0.5 L min-1

Auxiliary Gas Flow 0.5 L min-1

Coolant Gas Flow 12 L min-1

RF Power 1150 W

Number of replicates 3

Exposure Time UV 10 s and Vis 5 s for both Axial and Radial

modes Wavelengths Measured Axially

Al 167.079 nm, B 249.773 nm, Ba 455.403 nm, Co 228.616 nm, Cr 283.563 nm, Ni 221.647 nm, P 177.495 nm, Pb 220.353 nm, Zn 202.548 nm

Wavelengths Measured Radially

Ca 393.366 nm, Cu 324.754 nm, Fe 259.940 nm, K 766.490 nm, Mg 279.553 nm, Mn 257.610 nm, Na 589.592 nm, S 180.731 nm, V 292.402 nm

FIGURE 2. ASX-520 Autosampler view on the Dashboard of Qtegra ISDS Software.

FIGURE 3. Analysis mode drop-down menu.

Element

l (nm)

Apple leaves Tobacco leaves DL, mg L-1

Cref., mg kg-1 Cexp., mg kg-1 Cref., mg kg-1 Cexp., mg/kg-1

Al 167.079 290±30 280 200±30 190 0.3

B 249.773 27±3 25 34±3 33 5.1

Ba 455.403 49±5 49 67±7 64 0.03

Ca 393.366 15200±150

0 14800 40000±4000 40400 1.6

Co 228.616 - - 1.0±0.1 1.0 0.4

Cr 283.563 - - 6.3±0.6 5.7 0.4

Cu 324.754 5.6±0.6 5.7 10.1±1.0 10.0 2.4

Fe 259.940 83±8 82 - - 0.8

K 766.490 16100±200

0 17800 23000±8000 29000 5.1

Mg 279.553 2700±300 2670 8500±900 8400 0.04

Mn 257.610 54±5 54 180±20 180 0.2

Na 589.592 24±2 24 - - 1.8

Ni 221.647 0.9±0.1 0.9 8.5±0.9 8.7 0.3

P 177.495 1600±200 1700 1700±200 1800 5.7

Pb 220.353 0.47±0.05 0.48 2.0±0.3 1.7 1.1

S 180.731 - - 4600±900 5100 2.2

V 292.402 - - 4.1±0.6 4.6 4.9

Zn 202.548 12.5±0.3 12.2 52±5 51 1.8

Quantitation of Elements in Tobacco and Apple Leaves

Using the iCAP 7000 Series ICP-OES with a Dual Plasma View, it was possible to determine the elements of interest in the wavelength range from 166 to 847 nm in one run using radial and axial views. This allows selecting the most beneficial wavelengths and plasma view for each analyte and minimizing matrix effects. Resulting calibration curves are exemplarily shown (Fig. 5)

FIGURE 1. iCAP 7600 Duo ICP-OES and its optical system.

More information of increasing the analysis productivity using the iCAP 7000 Series ICP-OES with Qtegra ISDS Software can be found in our Technical Note 43225 [4].

FIGURE 5. Calibration curves obtained for 4 different types of signals (axial vs. radial + low slit vs. high slit).

0

20

40

60

80

100

120

0 20 40 60 80 100 120 140 160 180

Rec

over

y, %

Time, min

P 177.495 nm axial

Mg 279.553 nm radial

5Thermo Scienti� c Poster Note • eWPC • PN43236-EN 0315S

Conclusion The iCAP 7000 Series ICP-OES with dual view provides the unprecedented flexibility of analysis for determining the macro- and microelements in plant matrices. Driven by the Qtegra ISDS Software, it is easily adjusted to the peculiarities of sample matrix and at the same time allows to increase the analysis productivity using multiple tools, integrated into such systems. The complete set of data for ionomics studies can be easily collected by ICP-OES combined with workflow-driven Software.

This brings your analysis to the new level of robustness and confidence.

References 1. Husted S et al (2011) J Anal Atom Spectrom 26:52-79 2. Salt D et al (2008) Annu Rev Plant Biol 59:709-733 3. Hansen T et al (2013) Methods Mol Biol 953:121-141 4. Technical Note 43225 “Increasing productivity in ICP-OES with

Qtegra ISDS Software” 5. Technical Note 43173 “Thermo Scientific iCAP 7000 Series ICP-

OES Instrument Detection Limits”

In Normal mode the analysis is carried out per replicate, while the Speed mode allows a faster analysis by minimizing the number of transitions between different plasma views and slits, like shown on the scheme below as an example (where R is Replicate and represents the transition between plasma views):

Normal: R1 (Vis) R1 (UV) R2 (Vis) R2 (UV) R3 (Vis) R3 (UV) Speed: R1 (Vis) R2 (Vis) R3 (Vis) R1 (UV) R2 (UV) R3 (UV)

The Sprint mode operates similarly to Speed mode, this mode is typically used for trend analysis.

In the Speed mode one analysis per sample took a minimum of at least 20 seconds less than in Normal mode. Analyzing a sequence of 50 multielement samples in the Speed mode provided an increased sample throughput with the recovery at the level of 100% over the whole analysis duration (Fig. 4). Due to this, analysis of leaf digests was carried out in the Speed mode.

Overview Purpose: To demonstrate the advantages of an Inductively Coupled Plasma – Optical Emission Spectrometer (ICP-OES) with a Dual Plasma view for the determination of nutritional elements in plant leaves.

Methods: ICP-OES, microwave digestion.

Results: Concentrations of elements were determined in tobacco and apple leaves, the found concentrations were in the good agreement with the values, given for the Standard Reference Materials (SRMs).

Introduction The commercial value of a plant as an edible product or as a raw material for preparing medicines depends on the balance of different chemical constituents in the plant, including the elemental composition. Based on the element concentrations in the different plant parts, e.g., in plant leaves, the quality of plants can be evaluated for scientific and commercial purposes. Typically the major elements (macronutrients, including calcium, potassium, phosphorous, nitrogen, magnesium, sulfur) as well as the trace elements (micronutrients, including boron, manganese, iron, nickel, cupper, and zinc) are assessed. This allows the identification of nutrient deficiency, which can lead to the reduced quality of flowers, fruits or vegetables, produced by the plant. Besides, the information on the elemental composition of the plant may explain its medical or toxic properties.

In recent years with the development of OMICS-techniques, the study of plant ionomes has become important. Ionomics involves the quantitative and simultaneous measurement of plant elemental composition and changes in this composition in response to physiological effects and genetic modifications [1-3]. Due to this, development of new advanced techniques for the quantitation of plant ionoms or their parts is of high importance.

ICP-OES has the high potential for being employed within such studies. In this study the advantages of Thermo ScientificTM iCAPTM 7000 Series ICP-OES dual view for the determination of nutritional elements in plant matrices are discussed, using plant leaf analysis as an example.

Methods

Sample Preparation

SRMs of dried apple leaves (SRM 1515, NIST, USA) and dried tobacco (Institute of Nuclear Chemistry and Technology, Poland) were used. Both materials were analyzed for moisture before each analysis.

All samples were digested in a microwave oven using 0.5 g of each sample in 7 mL concentrated HNO3, 2 mL concentrated H2O2 and 3 mL deionized water.

The digestion included 4 steps: 1. 3 minutes at 700 W 2. 9 minutes at 500 W 3. 4 minutes at 1000 W 4. 14 minutes at 1000 W.

After digestion an aliquot of the sample was diluted 10-fold with 6% (v/v) HNO3 directly before analysis, the remaining sample was analyzed directly. As a blank sample, 6% (v/v) HNO3 was used for all measurements. Duplicate method blanks were analyzed for each microwave digestion batch.

Single element stock standards (Fisher Scientific) were used to prepare multielement standard solutions with the necessary element concentrations. The standard solutions were used for productivity studies and for calibration curve generation. Al, Ca, Fe, K, Mg, Na, P and S were present at 0.5 to 75 mg L-1; Ba, B, Cr, Cu, Li, Mn, Pb, V, and Zn were present at 50 mg L-1 to 10 mg L-1 in the standard solutions.

Instrumentation

The Thermo Scientific™ iCAP™ 7600 ICP-OES Duo (Fig.01) has been used in all experiments described in this paper. It was coupled to a Teledyne CETAC ASX-520 Random Access Intelligent Autosampler. Method parameters, listed in Table 1, were applied for all analyses (unless stated).

Tygon is a registered trade mark of Saint-Gobain Corporation. CETAC is a registered trademark of Teledyne CETAC Technologies. All other trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manners that might infringe the intellectual property rights of others.

Presented at the eWCPS, Münster, Germany, 2/2015.

FIGURE 4. Recoveries at P 177.495 nm (axial) and at Mg 279.553 nm (radial) for 50 samples, analyzed in Speed mode.

TABLE 2. Concentrations of elements in leaves and DLs.

The determined concentrations of elements in apple and tobacco leaves were in the good agreement with the specified values for the used SRMs (see Table 2). The deviations from the specified values were less than 10% in all cases. The detection limits (DL) for the majority of the elements were much lower than 1 mg L-1 [5].

Determination of Nutritional Elements in Plant Leaves using an Inductively Coupled Plasma Optical Emission Spectrometer with a Dual Plasma View Elena Chernetsova, Sanja Asendorf, Matthew Cassap Thermo Fisher Scientific, Bremen, Germany

Results Maximizing the Analysis Productivity

Using autosamplers for ICP-OES analysis reduces user intervention and is the first step towards increasing sample throughput. Autosamplers allow implementing unattended analysis with higher reproducibility of measurements, with precise uptake and wash cycles applied to each sample.

Sample uptake was preformed via a Teledyne CETAC ASX-520 autosampler. All parameters relating to the autosampler can be set directly in the Thermo Scientific™ QtegraTM ISDSTM Software (Fig. 2). Timings and layout can be modified directly in the respective Labbooks, with flexible rack configurations available.

With the Qtegra ISDS Software it is possible to apply three different modes of analysis (Fig. 3): Normal, Speed, and Sprint. These modes relate to the sequencing of the plasma views.

TABLE 1. ICP-OES method parameters.

Parameter Setting

Pump Tubing Sample TygonTM white/white/white

Drain TygonTM yellow/blue/yellow

Pump Speed 50 rpm

Spray Chamber Glass cyclonic

Nebulizer Glass concentric

Center Tube 2 mm

Nebulizer Gas Flow 0.5 L min-1

Auxiliary Gas Flow 0.5 L min-1

Coolant Gas Flow 12 L min-1

RF Power 1150 W

Number of replicates 3

Exposure Time UV 10 s and Vis 5 s for both Axial and Radial

modes Wavelengths Measured Axially

Al 167.079 nm, B 249.773 nm, Ba 455.403 nm, Co 228.616 nm, Cr 283.563 nm, Ni 221.647 nm, P 177.495 nm, Pb 220.353 nm, Zn 202.548 nm

Wavelengths Measured Radially

Ca 393.366 nm, Cu 324.754 nm, Fe 259.940 nm, K 766.490 nm, Mg 279.553 nm, Mn 257.610 nm, Na 589.592 nm, S 180.731 nm, V 292.402 nm

FIGURE 2. ASX-520 Autosampler view on the Dashboard of Qtegra ISDS Software.

FIGURE 3. Analysis mode drop-down menu.

Element

l (nm)

Apple leaves Tobacco leaves DL, mg L-1

Cref., mg kg-1 Cexp., mg kg-1 Cref., mg kg-1 Cexp., mg/kg-1

Al 167.079 290±30 280 200±30 190 0.3

B 249.773 27±3 25 34±3 33 5.1

Ba 455.403 49±5 49 67±7 64 0.03

Ca 393.366 15200±150

0 14800 40000±4000 40400 1.6

Co 228.616 - - 1.0±0.1 1.0 0.4

Cr 283.563 - - 6.3±0.6 5.7 0.4

Cu 324.754 5.6±0.6 5.7 10.1±1.0 10.0 2.4

Fe 259.940 83±8 82 - - 0.8

K 766.490 16100±200

0 17800 23000±8000 29000 5.1

Mg 279.553 2700±300 2670 8500±900 8400 0.04

Mn 257.610 54±5 54 180±20 180 0.2

Na 589.592 24±2 24 - - 1.8

Ni 221.647 0.9±0.1 0.9 8.5±0.9 8.7 0.3

P 177.495 1600±200 1700 1700±200 1800 5.7

Pb 220.353 0.47±0.05 0.48 2.0±0.3 1.7 1.1

S 180.731 - - 4600±900 5100 2.2

V 292.402 - - 4.1±0.6 4.6 4.9

Zn 202.548 12.5±0.3 12.2 52±5 51 1.8

Quantitation of Elements in Tobacco and Apple Leaves

Using the iCAP 7000 Series ICP-OES with a Dual Plasma View, it was possible to determine the elements of interest in the wavelength range from 166 to 847 nm in one run using radial and axial views. This allows selecting the most beneficial wavelengths and plasma view for each analyte and minimizing matrix effects. Resulting calibration curves are exemplarily shown (Fig. 5)

FIGURE 1. iCAP 7600 Duo ICP-OES and its optical system.

More information of increasing the analysis productivity using the iCAP 7000 Series ICP-OES with Qtegra ISDS Software can be found in our Technical Note 43225 [4].

FIGURE 5. Calibration curves obtained for 4 different types of signals (axial vs. radial + low slit vs. high slit).

0

20

40

60

80

100

120

0 20 40 60 80 100 120 140 160 180

Rec

over

y, %

Time, min

P 177.495 nm axial

Mg 279.553 nm radial

6 Determination of Nutritional Elements in Plant Leaves using an Inductively Coupled Plasma Optical Emission Spectrometer with a Dual Plasma View

Conclusion The iCAP 7000 Series ICP-OES with dual view provides the unprecedented flexibility of analysis for determining the macro- and microelements in plant matrices. Driven by the Qtegra ISDS Software, it is easily adjusted to the peculiarities of sample matrix and at the same time allows to increase the analysis productivity using multiple tools, integrated into such systems. The complete set of data for ionomics studies can be easily collected by ICP-OES combined with workflow-driven Software.

This brings your analysis to the new level of robustness and confidence.

References 1. Husted S et al (2011) J Anal Atom Spectrom 26:52-79 2. Salt D et al (2008) Annu Rev Plant Biol 59:709-733 3. Hansen T et al (2013) Methods Mol Biol 953:121-141 4. Technical Note 43225 “Increasing productivity in ICP-OES with

Qtegra ISDS Software” 5. Technical Note 43173 “Thermo Scientific iCAP 7000 Series ICP-

OES Instrument Detection Limits”

In Normal mode the analysis is carried out per replicate, while the Speed mode allows a faster analysis by minimizing the number of transitions between different plasma views and slits, like shown on the scheme below as an example (where R is Replicate and represents the transition between plasma views):

Normal: R1 (Vis) R1 (UV) R2 (Vis) R2 (UV) R3 (Vis) R3 (UV) Speed: R1 (Vis) R2 (Vis) R3 (Vis) R1 (UV) R2 (UV) R3 (UV)

The Sprint mode operates similarly to Speed mode, this mode is typically used for trend analysis.

In the Speed mode one analysis per sample took a minimum of at least 20 seconds less than in Normal mode. Analyzing a sequence of 50 multielement samples in the Speed mode provided an increased sample throughput with the recovery at the level of 100% over the whole analysis duration (Fig. 4). Due to this, analysis of leaf digests was carried out in the Speed mode.

Overview Purpose: To demonstrate the advantages of an Inductively Coupled Plasma – Optical Emission Spectrometer (ICP-OES) with a Dual Plasma view for the determination of nutritional elements in plant leaves.

Methods: ICP-OES, microwave digestion.

Results: Concentrations of elements were determined in tobacco and apple leaves, the found concentrations were in the good agreement with the values, given for the Standard Reference Materials (SRMs).

Introduction The commercial value of a plant as an edible product or as a raw material for preparing medicines depends on the balance of different chemical constituents in the plant, including the elemental composition. Based on the element concentrations in the different plant parts, e.g., in plant leaves, the quality of plants can be evaluated for scientific and commercial purposes. Typically the major elements (macronutrients, including calcium, potassium, phosphorous, nitrogen, magnesium, sulfur) as well as the trace elements (micronutrients, including boron, manganese, iron, nickel, cupper, and zinc) are assessed. This allows the identification of nutrient deficiency, which can lead to the reduced quality of flowers, fruits or vegetables, produced by the plant. Besides, the information on the elemental composition of the plant may explain its medical or toxic properties.

In recent years with the development of OMICS-techniques, the study of plant ionomes has become important. Ionomics involves the quantitative and simultaneous measurement of plant elemental composition and changes in this composition in response to physiological effects and genetic modifications [1-3]. Due to this, development of new advanced techniques for the quantitation of plant ionoms or their parts is of high importance.

ICP-OES has the high potential for being employed within such studies. In this study the advantages of Thermo ScientificTM iCAPTM 7000 Series ICP-OES dual view for the determination of nutritional elements in plant matrices are discussed, using plant leaf analysis as an example.

Methods

Sample Preparation

SRMs of dried apple leaves (SRM 1515, NIST, USA) and dried tobacco (Institute of Nuclear Chemistry and Technology, Poland) were used. Both materials were analyzed for moisture before each analysis.

All samples were digested in a microwave oven using 0.5 g of each sample in 7 mL concentrated HNO3, 2 mL concentrated H2O2 and 3 mL deionized water.

The digestion included 4 steps: 1. 3 minutes at 700 W 2. 9 minutes at 500 W 3. 4 minutes at 1000 W 4. 14 minutes at 1000 W.

After digestion an aliquot of the sample was diluted 10-fold with 6% (v/v) HNO3 directly before analysis, the remaining sample was analyzed directly. As a blank sample, 6% (v/v) HNO3 was used for all measurements. Duplicate method blanks were analyzed for each microwave digestion batch.

Single element stock standards (Fisher Scientific) were used to prepare multielement standard solutions with the necessary element concentrations. The standard solutions were used for productivity studies and for calibration curve generation. Al, Ca, Fe, K, Mg, Na, P and S were present at 0.5 to 75 mg L-1; Ba, B, Cr, Cu, Li, Mn, Pb, V, and Zn were present at 50 mg L-1 to 10 mg L-1 in the standard solutions.

Instrumentation

The Thermo Scientific™ iCAP™ 7600 ICP-OES Duo (Fig.01) has been used in all experiments described in this paper. It was coupled to a Teledyne CETAC ASX-520 Random Access Intelligent Autosampler. Method parameters, listed in Table 1, were applied for all analyses (unless stated).

Tygon is a registered trade mark of Saint-Gobain Corporation. CETAC is a registered trademark of Teledyne CETAC Technologies. All other trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manners that might infringe the intellectual property rights of others.

Presented at the eWCPS, Münster, Germany, 2/2015.

FIGURE 4. Recoveries at P 177.495 nm (axial) and at Mg 279.553 nm (radial) for 50 samples, analyzed in Speed mode.

TABLE 2. Concentrations of elements in leaves and DLs.

The determined concentrations of elements in apple and tobacco leaves were in the good agreement with the specified values for the used SRMs (see Table 2). The deviations from the specified values were less than 10% in all cases. The detection limits (DL) for the majority of the elements were much lower than 1 mg L-1 [5].

Determination of Nutritional Elements in Plant Leaves using an Inductively Coupled Plasma Optical Emission Spectrometer with a Dual Plasma View Elena Chernetsova, Sanja Asendorf, Matthew Cassap Thermo Fisher Scientific, Bremen, Germany

Results Maximizing the Analysis Productivity

Using autosamplers for ICP-OES analysis reduces user intervention and is the first step towards increasing sample throughput. Autosamplers allow implementing unattended analysis with higher reproducibility of measurements, with precise uptake and wash cycles applied to each sample.

Sample uptake was preformed via a Teledyne CETAC ASX-520 autosampler. All parameters relating to the autosampler can be set directly in the Thermo Scientific™ QtegraTM ISDSTM Software (Fig. 2). Timings and layout can be modified directly in the respective Labbooks, with flexible rack configurations available.

With the Qtegra ISDS Software it is possible to apply three different modes of analysis (Fig. 3): Normal, Speed, and Sprint. These modes relate to the sequencing of the plasma views.

TABLE 1. ICP-OES method parameters.

Parameter Setting

Pump Tubing Sample TygonTM white/white/white

Drain TygonTM yellow/blue/yellow

Pump Speed 50 rpm

Spray Chamber Glass cyclonic

Nebulizer Glass concentric

Center Tube 2 mm

Nebulizer Gas Flow 0.5 L min-1

Auxiliary Gas Flow 0.5 L min-1

Coolant Gas Flow 12 L min-1

RF Power 1150 W

Number of replicates 3

Exposure Time UV 10 s and Vis 5 s for both Axial and Radial

modes Wavelengths Measured Axially

Al 167.079 nm, B 249.773 nm, Ba 455.403 nm, Co 228.616 nm, Cr 283.563 nm, Ni 221.647 nm, P 177.495 nm, Pb 220.353 nm, Zn 202.548 nm

Wavelengths Measured Radially

Ca 393.366 nm, Cu 324.754 nm, Fe 259.940 nm, K 766.490 nm, Mg 279.553 nm, Mn 257.610 nm, Na 589.592 nm, S 180.731 nm, V 292.402 nm

FIGURE 2. ASX-520 Autosampler view on the Dashboard of Qtegra ISDS Software.

FIGURE 3. Analysis mode drop-down menu.

Element

l (nm)

Apple leaves Tobacco leaves DL, mg L-1

Cref., mg kg-1 Cexp., mg kg-1 Cref., mg kg-1 Cexp., mg/kg-1

Al 167.079 290±30 280 200±30 190 0.3

B 249.773 27±3 25 34±3 33 5.1

Ba 455.403 49±5 49 67±7 64 0.03

Ca 393.366 15200±150

0 14800 40000±4000 40400 1.6

Co 228.616 - - 1.0±0.1 1.0 0.4

Cr 283.563 - - 6.3±0.6 5.7 0.4

Cu 324.754 5.6±0.6 5.7 10.1±1.0 10.0 2.4

Fe 259.940 83±8 82 - - 0.8

K 766.490 16100±200

0 17800 23000±8000 29000 5.1

Mg 279.553 2700±300 2670 8500±900 8400 0.04

Mn 257.610 54±5 54 180±20 180 0.2

Na 589.592 24±2 24 - - 1.8

Ni 221.647 0.9±0.1 0.9 8.5±0.9 8.7 0.3

P 177.495 1600±200 1700 1700±200 1800 5.7

Pb 220.353 0.47±0.05 0.48 2.0±0.3 1.7 1.1

S 180.731 - - 4600±900 5100 2.2

V 292.402 - - 4.1±0.6 4.6 4.9

Zn 202.548 12.5±0.3 12.2 52±5 51 1.8

Quantitation of Elements in Tobacco and Apple Leaves

Using the iCAP 7000 Series ICP-OES with a Dual Plasma View, it was possible to determine the elements of interest in the wavelength range from 166 to 847 nm in one run using radial and axial views. This allows selecting the most beneficial wavelengths and plasma view for each analyte and minimizing matrix effects. Resulting calibration curves are exemplarily shown (Fig. 5)

FIGURE 1. iCAP 7600 Duo ICP-OES and its optical system.

More information of increasing the analysis productivity using the iCAP 7000 Series ICP-OES with Qtegra ISDS Software can be found in our Technical Note 43225 [4].

FIGURE 5. Calibration curves obtained for 4 different types of signals (axial vs. radial + low slit vs. high slit).

0

20

40

60

80

100

120

0 20 40 60 80 100 120 140 160 180

Rec

over

y, %

Time, min

P 177.495 nm axial

Mg 279.553 nm radial

PN43236-EN 0315S

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www.thermoscientific.com©2015 Thermo Fisher Scienti� c Inc. All rights reserved. ISO is a trademark of the International Standards Organization. Tygon is a registered trade mark of Saint-Gobain Corporation. CETAC is a registered trademark of Teledyne CETAC Technologies. All other trademarks are the property of Thermo Fisher Scienti� c and its subsidiaries. This information is presented as an example of the capabilities of Thermo Fisher Scienti� c products. It is not intended to encourage use of these products in any manners that might infringe the intellectual property rights of others. Speci� cations, terms and pricing are subject to change. Not all products are available in all countries. Please consult your local sales representative for details.

Thermo Fisher Scienti� c (Bremen) GmbHManagement System Registered to ISO 9001:2008

Conclusion The iCAP 7000 Series ICP-OES with dual view provides the unprecedented flexibility of analysis for determining the macro- and microelements in plant matrices. Driven by the Qtegra ISDS Software, it is easily adjusted to the peculiarities of sample matrix and at the same time allows to increase the analysis productivity using multiple tools, integrated into such systems. The complete set of data for ionomics studies can be easily collected by ICP-OES combined with workflow-driven Software.

This brings your analysis to the new level of robustness and confidence.

References 1. Husted S et al (2011) J Anal Atom Spectrom 26:52-79 2. Salt D et al (2008) Annu Rev Plant Biol 59:709-733 3. Hansen T et al (2013) Methods Mol Biol 953:121-141 4. Technical Note 43225 “Increasing productivity in ICP-OES with

Qtegra ISDS Software” 5. Technical Note 43173 “Thermo Scientific iCAP 7000 Series ICP-

OES Instrument Detection Limits”

In Normal mode the analysis is carried out per replicate, while the Speed mode allows a faster analysis by minimizing the number of transitions between different plasma views and slits, like shown on the scheme below as an example (where R is Replicate and represents the transition between plasma views):

Normal: R1 (Vis) R1 (UV) R2 (Vis) R2 (UV) R3 (Vis) R3 (UV) Speed: R1 (Vis) R2 (Vis) R3 (Vis) R1 (UV) R2 (UV) R3 (UV)

The Sprint mode operates similarly to Speed mode, this mode is typically used for trend analysis.

In the Speed mode one analysis per sample took a minimum of at least 20 seconds less than in Normal mode. Analyzing a sequence of 50 multielement samples in the Speed mode provided an increased sample throughput with the recovery at the level of 100% over the whole analysis duration (Fig. 4). Due to this, analysis of leaf digests was carried out in the Speed mode.

Overview Purpose: To demonstrate the advantages of an Inductively Coupled Plasma – Optical Emission Spectrometer (ICP-OES) with a Dual Plasma view for the determination of nutritional elements in plant leaves.

Methods: ICP-OES, microwave digestion.

Results: Concentrations of elements were determined in tobacco and apple leaves, the found concentrations were in the good agreement with the values, given for the Standard Reference Materials (SRMs).

Introduction The commercial value of a plant as an edible product or as a raw material for preparing medicines depends on the balance of different chemical constituents in the plant, including the elemental composition. Based on the element concentrations in the different plant parts, e.g., in plant leaves, the quality of plants can be evaluated for scientific and commercial purposes. Typically the major elements (macronutrients, including calcium, potassium, phosphorous, nitrogen, magnesium, sulfur) as well as the trace elements (micronutrients, including boron, manganese, iron, nickel, cupper, and zinc) are assessed. This allows the identification of nutrient deficiency, which can lead to the reduced quality of flowers, fruits or vegetables, produced by the plant. Besides, the information on the elemental composition of the plant may explain its medical or toxic properties.

In recent years with the development of OMICS-techniques, the study of plant ionomes has become important. Ionomics involves the quantitative and simultaneous measurement of plant elemental composition and changes in this composition in response to physiological effects and genetic modifications [1-3]. Due to this, development of new advanced techniques for the quantitation of plant ionoms or their parts is of high importance.

ICP-OES has the high potential for being employed within such studies. In this study the advantages of Thermo ScientificTM iCAPTM 7000 Series ICP-OES dual view for the determination of nutritional elements in plant matrices are discussed, using plant leaf analysis as an example.

Methods

Sample Preparation

SRMs of dried apple leaves (SRM 1515, NIST, USA) and dried tobacco (Institute of Nuclear Chemistry and Technology, Poland) were used. Both materials were analyzed for moisture before each analysis.

All samples were digested in a microwave oven using 0.5 g of each sample in 7 mL concentrated HNO3, 2 mL concentrated H2O2 and 3 mL deionized water.

The digestion included 4 steps: 1. 3 minutes at 700 W 2. 9 minutes at 500 W 3. 4 minutes at 1000 W 4. 14 minutes at 1000 W.

After digestion an aliquot of the sample was diluted 10-fold with 6% (v/v) HNO3 directly before analysis, the remaining sample was analyzed directly. As a blank sample, 6% (v/v) HNO3 was used for all measurements. Duplicate method blanks were analyzed for each microwave digestion batch.

Single element stock standards (Fisher Scientific) were used to prepare multielement standard solutions with the necessary element concentrations. The standard solutions were used for productivity studies and for calibration curve generation. Al, Ca, Fe, K, Mg, Na, P and S were present at 0.5 to 75 mg L-1; Ba, B, Cr, Cu, Li, Mn, Pb, V, and Zn were present at 50 mg L-1 to 10 mg L-1 in the standard solutions.

Instrumentation

The Thermo Scientific™ iCAP™ 7600 ICP-OES Duo (Fig.01) has been used in all experiments described in this paper. It was coupled to a Teledyne CETAC ASX-520 Random Access Intelligent Autosampler. Method parameters, listed in Table 1, were applied for all analyses (unless stated).

Tygon is a registered trade mark of Saint-Gobain Corporation. CETAC is a registered trademark of Teledyne CETAC Technologies. All other trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manners that might infringe the intellectual property rights of others.

Presented at the eWCPS, Münster, Germany, 2/2015.

FIGURE 4. Recoveries at P 177.495 nm (axial) and at Mg 279.553 nm (radial) for 50 samples, analyzed in Speed mode.

TABLE 2. Concentrations of elements in leaves and DLs.

The determined concentrations of elements in apple and tobacco leaves were in the good agreement with the specified values for the used SRMs (see Table 2). The deviations from the specified values were less than 10% in all cases. The detection limits (DL) for the majority of the elements were much lower than 1 mg L-1 [5].

Determination of Nutritional Elements in Plant Leaves using an Inductively Coupled Plasma Optical Emission Spectrometer with a Dual Plasma View Elena Chernetsova, Sanja Asendorf, Matthew Cassap Thermo Fisher Scientific, Bremen, Germany

Results Maximizing the Analysis Productivity

Using autosamplers for ICP-OES analysis reduces user intervention and is the first step towards increasing sample throughput. Autosamplers allow implementing unattended analysis with higher reproducibility of measurements, with precise uptake and wash cycles applied to each sample.

Sample uptake was preformed via a Teledyne CETAC ASX-520 autosampler. All parameters relating to the autosampler can be set directly in the Thermo Scientific™ QtegraTM ISDSTM Software (Fig. 2). Timings and layout can be modified directly in the respective Labbooks, with flexible rack configurations available.

With the Qtegra ISDS Software it is possible to apply three different modes of analysis (Fig. 3): Normal, Speed, and Sprint. These modes relate to the sequencing of the plasma views.

TABLE 1. ICP-OES method parameters.

Parameter Setting

Pump Tubing Sample TygonTM white/white/white

Drain TygonTM yellow/blue/yellow

Pump Speed 50 rpm

Spray Chamber Glass cyclonic

Nebulizer Glass concentric

Center Tube 2 mm

Nebulizer Gas Flow 0.5 L min-1

Auxiliary Gas Flow 0.5 L min-1

Coolant Gas Flow 12 L min-1

RF Power 1150 W

Number of replicates 3

Exposure Time UV 10 s and Vis 5 s for both Axial and Radial

modes Wavelengths Measured Axially

Al 167.079 nm, B 249.773 nm, Ba 455.403 nm, Co 228.616 nm, Cr 283.563 nm, Ni 221.647 nm, P 177.495 nm, Pb 220.353 nm, Zn 202.548 nm

Wavelengths Measured Radially

Ca 393.366 nm, Cu 324.754 nm, Fe 259.940 nm, K 766.490 nm, Mg 279.553 nm, Mn 257.610 nm, Na 589.592 nm, S 180.731 nm, V 292.402 nm

FIGURE 2. ASX-520 Autosampler view on the Dashboard of Qtegra ISDS Software.

FIGURE 3. Analysis mode drop-down menu.

Element

l (nm)

Apple leaves Tobacco leaves DL, mg L-1

Cref., mg kg-1 Cexp., mg kg-1 Cref., mg kg-1 Cexp., mg/kg-1

Al 167.079 290±30 280 200±30 190 0.3

B 249.773 27±3 25 34±3 33 5.1

Ba 455.403 49±5 49 67±7 64 0.03

Ca 393.366 15200±150

0 14800 40000±4000 40400 1.6

Co 228.616 - - 1.0±0.1 1.0 0.4

Cr 283.563 - - 6.3±0.6 5.7 0.4

Cu 324.754 5.6±0.6 5.7 10.1±1.0 10.0 2.4

Fe 259.940 83±8 82 - - 0.8

K 766.490 16100±200

0 17800 23000±8000 29000 5.1

Mg 279.553 2700±300 2670 8500±900 8400 0.04

Mn 257.610 54±5 54 180±20 180 0.2

Na 589.592 24±2 24 - - 1.8

Ni 221.647 0.9±0.1 0.9 8.5±0.9 8.7 0.3

P 177.495 1600±200 1700 1700±200 1800 5.7

Pb 220.353 0.47±0.05 0.48 2.0±0.3 1.7 1.1

S 180.731 - - 4600±900 5100 2.2

V 292.402 - - 4.1±0.6 4.6 4.9

Zn 202.548 12.5±0.3 12.2 52±5 51 1.8

Quantitation of Elements in Tobacco and Apple Leaves

Using the iCAP 7000 Series ICP-OES with a Dual Plasma View, it was possible to determine the elements of interest in the wavelength range from 166 to 847 nm in one run using radial and axial views. This allows selecting the most beneficial wavelengths and plasma view for each analyte and minimizing matrix effects. Resulting calibration curves are exemplarily shown (Fig. 5)

FIGURE 1. iCAP 7600 Duo ICP-OES and its optical system.

More information of increasing the analysis productivity using the iCAP 7000 Series ICP-OES with Qtegra ISDS Software can be found in our Technical Note 43225 [4].

FIGURE 5. Calibration curves obtained for 4 different types of signals (axial vs. radial + low slit vs. high slit).

0

20

40

60

80

100

120

0 20 40 60 80 100 120 140 160 180

Rec

over

y, %

Time, min

P 177.495 nm axial

Mg 279.553 nm radial