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Dr Vasiliy V. Rosen
vasiliyr@savion.huji.ac.il
The Plasma Spectrochemistry Laboratory
The Scientific Service Core Facility
The Faculty of Agriculture
The Hebrew University of Jerusalem
July 4, 2023
Elemental analysis of plant material by
ICP-OES/MS
AGENDA
Introduction to ICP-OES/MS:
Principles and Instrumentation
Essential
Toxic
Major Micronutrients
Carbon (C)
Oxygen (O)
Hydrogen (H)
Nitrogen (N)
Phosphorus (P)
Potassium (K)
Sodium (Na)
Silica (Si)
Calcium (Ca)
Magnesium (Mg)
Sulfur (S)
Boron (B),
Chlorine (Cl)
Copper (Cu)
Iron (Fe)
Manganese (Mn)
Molybdenum (Mo)
Zinc (Zn)
Nickel (Ni)
Cobalt (Co)
Chromium (Cr)
Selenium (Se)
Vanadium (V)
Silver (Ag)
Aluminum (Al)
Arsenic (As)
Barium (Ba)
Beryllium (Be)
Cadmium (Cd)
Mercury (Hg)
Lead (Pb)
Lithium (Li)
And all
micronutrients at
critical
concentration
The role of chemical elements in plants
(adopted from Munson R., 1997, and Macnicol R., 1984)
EA
ICP
The levels of major elements and micronutrients in mature
leaf tissue (after Munson R., 1997)
4
Sampling
Decontamination
Drying
Grinding
Non-representative sample
Remained surface pollutants
Loss of volatile compounds
and weight
Contamination by mill blades
(Fe, Ni, Cr, Mo…)
Procedure Danger
Storage
Contamination by wrong
package (Sn from can; B, Si,
etc – from Pyrex Glass)
Sample
naming
Complicated names lead to
human mistake
Dry Ashing
Equipment
Porcelain or platinum crucible
Electric or microwave muffle
Open Wet Digestion
Hot-block open
digestion
Digestion on aluminum block in glass tubes with HNO3
Oven cavity
Rotor
Teflon vessel
Closed Microwave-assisted Wet Digestion
Closed digestion
Open digestion
Max. temperature 200 – 300 °C
Max. temperature limited by
solution boiling point
Low acid consumption
High acid consumption
Lower sample weight
Higher sample weight
No loss of volatile elements
Loss of volatile elements (Hg, I)
Risk of contamination from
digestion vessels
Risk of contamination from air and
cross-contamination
Digestion duration: 15-60 min
Digestion duration: 2 – 15 h (or
even more)
Matrices and Matrix Effect
HNO3 HCl
HF
https://www.inorganicventures.com/
Cd, 1 mg/L, in weak acid
Cd, 1 mg/L, in base
Analyte concentrations are equal, but intensities are different
Matrices and Matrix Effect
Dry Ashing
The sample (0.5 g dry
wt.) digested at 500 °
C 4-6 h, then
dissolved in acid(s)
Advantages: cheap
method; sample
weight may be
increased.
Disadvantages: loss of
volatile elements (Cl, As,
Se, Mo, Hg); (cross)-
contamination; formation
of non-soluble silicates.
Wet Digestion
(Hot-Block)
The sample (0.5 g dry
wt.) digested with
acid(s) in glass or
Teflon tube on the
Hotplate or Digestion
block
Advantages: less loss
and contamination than
in Dry Ashing Method;
high throughput.
Disadvantages: some
acids are extremely
dangerous (HClO4, HF);
method is time-consuming.
Wet Digestion with
Microwave Oven
The sample (0.5 g dry wt.)
digested with acid(s) in closed
Teflon vessel in microwave
oven.
Advantages: No volatile
compounds lost (closed
digestion); contamination is
minimized; digestion conditions
are strong (temperature, acid
and pressure); digestion is quick
(about 30 min).
Disadvantages: expensive
equipment; the throughput is
usually low.
Introduction to ICP-OES/MS: Principles and Instrumentation
This is how AI thinks the ICP instrument looks like
ICP-OES
720/725, Agilent
USA
Arcos, Spectro,
Germany
Analytik Jena,
Germany
iCAP 7000 Plus,
Thermo Scientific
USA
Optima,
Perkin-Elmer, USA
ICPE 9800,
Shimadzu, Japan
Introduction to ICP-OES/MS: Principles and Instrumentation
This is what the ICP-OES really looks
The Plasma Spectrochemistry (ICP) Lab
1989
1994
2007
2007
2018
Radial, Sequential Axial, Simultaneous
Axial, Simultaneous
Axial, Simultaneous
Radial, Simultaneous
Dual-view,
Sequential
Mass-Spectrometer (ICP-MS)
Introduction to ICP-OES/MS: Principles and Instrumentation
Ernest
Rutherford
(1871 – 1937)
Niels Bohr
(1885 – 1962)
Introduction to ICP-OES/MS: Principles and Instrumentation
Introduction to ICP-OES/MS: Principles and Instrumentation
This is how AI perceives the
appearance of the argon
plasma
This is what the
argon plasma really
looks like
Energy absorption
by ground-state
atoms
Atomic Absorption
(AA, GFAA)
Singly charged
ion formation
ICP-MS
Energy emission by
excited atoms and
ions
Adapted from Boss and Freeden, 1997
Flame photometry,
ICP-OES
Introduction to ICP-OES/MS: Principles and Instrumentation
sampling
disposal
argon
argon
argon
T, °C of the excitation source
Instrument LOQ
Low High
High Low
Introduction to ICP-OES/MS: Principles and Instrumentation
Introduction to ICP-OES/MS: Principles and Instrumentation
Introduction to ICP-OES/MS: Principles and Instrumentation
ICP-OES vs MS :
Adjustment
Ion Optics
Scan settings
Element selection
ICP-OES
ICP-OES: Cd 224 nm
LOQ 0.2 µg/L (ppb)
Introduction to ICP-OES/MS: Principles and Instrumentation
RSD = 3.91 %
RSD = 0.76 %
LOQ 0.0054
µg/L (ppb)
Fresh animal liver,
digested in HNO3
(0.5 g in 25 mL)
Introduction to ICP-OES/MS: Principles and Instrumentation
Method Advantages Method Limitations
❑ Multi-element ❑ Adjustment of measurement
conditions; Spectral
Interference
❑ High sensitivity = Low
LOQ
❑ Sample preparation
❑ Wide linear dynamic
range
❑ Matrix effect and Spectral
Interference
Introduction to ICP-OES/MS: Principles and Instrumentation
❖ Objects: soil, plants, waters,
tissues, food, and drinks
❖ Nutrient analysis
❖ Trace elements analysis (including
toxic elements)
❖ Food authentication (geographical
origin through mineral profiling
and/or isotope ratio analysis)
❖ Speciation (HPLC+ICP-MS)
Introduction to ICP-OES/MS: Principles and Instrumentation
Brown seaweeds (Kombu and Wakame) display a greater capacity to accumulate total As and iodine than the red
seaweed (Nori)
Rosen, Shimshoni, et al, 2023 (unpublished)
Introduction to ICP-OES/MS: Principles and Instrumentation
Quality Control in ICP-OES/MS analysis
(after Munson R., 1997)
Ycps = a*Xmg/L+b
The slope is a measure of sensitivity:
how much the signal changes for a
change in concentration.
Ycps = a*Xmg/L+b
a
Ca K Cu Pb Cr
Quality Control in ICP-OES/MS analysis
Concentration Units
Accuracy is how close a
measured value is to the actual
(true) value.
Precision is how close the measured
values are to each other.
Quality Control in ICP-OES/MS analysis
LOQ is the lowest concentration at
which a measurement is quantitatively
meaningful (Mitra, 2003)
LOD is the concentration at
which we can decide whether an
element is present or not
(Thomsen, 2003)
Quality Control in ICP-OES/MS analysis
The combined uncertainty Uc in analytical
measurement is a single numerical value that
represents the total estimation of uncertainty,
taking into account various sources of error and
variability associated with the measurement process.
Uncertainty
Uexp = Uc*k, k is coverage factor
(appr. 2 for CI=95%)
Take home message
• Choose your spectrometry
technique/instrument based
on your research goals.
• Always remember the sample
pre-treatment and preparation
steps.
• Check the reliability of your
analysis

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Elemental Analysis of Plants using ICP-OES(2023)

  • 1. Dr Vasiliy V. Rosen vasiliyr@savion.huji.ac.il The Plasma Spectrochemistry Laboratory The Scientific Service Core Facility The Faculty of Agriculture The Hebrew University of Jerusalem July 4, 2023 Elemental analysis of plant material by ICP-OES/MS
  • 3. Essential Toxic Major Micronutrients Carbon (C) Oxygen (O) Hydrogen (H) Nitrogen (N) Phosphorus (P) Potassium (K) Sodium (Na) Silica (Si) Calcium (Ca) Magnesium (Mg) Sulfur (S) Boron (B), Chlorine (Cl) Copper (Cu) Iron (Fe) Manganese (Mn) Molybdenum (Mo) Zinc (Zn) Nickel (Ni) Cobalt (Co) Chromium (Cr) Selenium (Se) Vanadium (V) Silver (Ag) Aluminum (Al) Arsenic (As) Barium (Ba) Beryllium (Be) Cadmium (Cd) Mercury (Hg) Lead (Pb) Lithium (Li) And all micronutrients at critical concentration The role of chemical elements in plants (adopted from Munson R., 1997, and Macnicol R., 1984) EA ICP
  • 4. The levels of major elements and micronutrients in mature leaf tissue (after Munson R., 1997) 4
  • 5. Sampling Decontamination Drying Grinding Non-representative sample Remained surface pollutants Loss of volatile compounds and weight Contamination by mill blades (Fe, Ni, Cr, Mo…) Procedure Danger Storage Contamination by wrong package (Sn from can; B, Si, etc – from Pyrex Glass) Sample naming Complicated names lead to human mistake
  • 6. Dry Ashing Equipment Porcelain or platinum crucible Electric or microwave muffle
  • 7. Open Wet Digestion Hot-block open digestion Digestion on aluminum block in glass tubes with HNO3
  • 8. Oven cavity Rotor Teflon vessel Closed Microwave-assisted Wet Digestion
  • 9. Closed digestion Open digestion Max. temperature 200 – 300 °C Max. temperature limited by solution boiling point Low acid consumption High acid consumption Lower sample weight Higher sample weight No loss of volatile elements Loss of volatile elements (Hg, I) Risk of contamination from digestion vessels Risk of contamination from air and cross-contamination Digestion duration: 15-60 min Digestion duration: 2 – 15 h (or even more)
  • 10. Matrices and Matrix Effect HNO3 HCl HF https://www.inorganicventures.com/
  • 11. Cd, 1 mg/L, in weak acid Cd, 1 mg/L, in base Analyte concentrations are equal, but intensities are different Matrices and Matrix Effect
  • 12. Dry Ashing The sample (0.5 g dry wt.) digested at 500 ° C 4-6 h, then dissolved in acid(s) Advantages: cheap method; sample weight may be increased. Disadvantages: loss of volatile elements (Cl, As, Se, Mo, Hg); (cross)- contamination; formation of non-soluble silicates. Wet Digestion (Hot-Block) The sample (0.5 g dry wt.) digested with acid(s) in glass or Teflon tube on the Hotplate or Digestion block Advantages: less loss and contamination than in Dry Ashing Method; high throughput. Disadvantages: some acids are extremely dangerous (HClO4, HF); method is time-consuming. Wet Digestion with Microwave Oven The sample (0.5 g dry wt.) digested with acid(s) in closed Teflon vessel in microwave oven. Advantages: No volatile compounds lost (closed digestion); contamination is minimized; digestion conditions are strong (temperature, acid and pressure); digestion is quick (about 30 min). Disadvantages: expensive equipment; the throughput is usually low.
  • 13. Introduction to ICP-OES/MS: Principles and Instrumentation This is how AI thinks the ICP instrument looks like
  • 14. ICP-OES 720/725, Agilent USA Arcos, Spectro, Germany Analytik Jena, Germany iCAP 7000 Plus, Thermo Scientific USA Optima, Perkin-Elmer, USA ICPE 9800, Shimadzu, Japan Introduction to ICP-OES/MS: Principles and Instrumentation This is what the ICP-OES really looks
  • 15. The Plasma Spectrochemistry (ICP) Lab 1989 1994 2007 2007 2018 Radial, Sequential Axial, Simultaneous Axial, Simultaneous Axial, Simultaneous Radial, Simultaneous Dual-view, Sequential Mass-Spectrometer (ICP-MS) Introduction to ICP-OES/MS: Principles and Instrumentation
  • 16. Ernest Rutherford (1871 – 1937) Niels Bohr (1885 – 1962) Introduction to ICP-OES/MS: Principles and Instrumentation
  • 17. Introduction to ICP-OES/MS: Principles and Instrumentation This is how AI perceives the appearance of the argon plasma This is what the argon plasma really looks like
  • 18. Energy absorption by ground-state atoms Atomic Absorption (AA, GFAA) Singly charged ion formation ICP-MS Energy emission by excited atoms and ions Adapted from Boss and Freeden, 1997 Flame photometry, ICP-OES Introduction to ICP-OES/MS: Principles and Instrumentation sampling disposal argon argon argon
  • 19. T, °C of the excitation source Instrument LOQ Low High High Low Introduction to ICP-OES/MS: Principles and Instrumentation
  • 20. Introduction to ICP-OES/MS: Principles and Instrumentation
  • 21. Introduction to ICP-OES/MS: Principles and Instrumentation ICP-OES vs MS : Adjustment Ion Optics Scan settings Element selection ICP-OES
  • 22. ICP-OES: Cd 224 nm LOQ 0.2 µg/L (ppb) Introduction to ICP-OES/MS: Principles and Instrumentation RSD = 3.91 % RSD = 0.76 % LOQ 0.0054 µg/L (ppb)
  • 23. Fresh animal liver, digested in HNO3 (0.5 g in 25 mL) Introduction to ICP-OES/MS: Principles and Instrumentation
  • 24. Method Advantages Method Limitations ❑ Multi-element ❑ Adjustment of measurement conditions; Spectral Interference ❑ High sensitivity = Low LOQ ❑ Sample preparation ❑ Wide linear dynamic range ❑ Matrix effect and Spectral Interference Introduction to ICP-OES/MS: Principles and Instrumentation
  • 25. ❖ Objects: soil, plants, waters, tissues, food, and drinks ❖ Nutrient analysis ❖ Trace elements analysis (including toxic elements) ❖ Food authentication (geographical origin through mineral profiling and/or isotope ratio analysis) ❖ Speciation (HPLC+ICP-MS) Introduction to ICP-OES/MS: Principles and Instrumentation
  • 26. Brown seaweeds (Kombu and Wakame) display a greater capacity to accumulate total As and iodine than the red seaweed (Nori) Rosen, Shimshoni, et al, 2023 (unpublished) Introduction to ICP-OES/MS: Principles and Instrumentation
  • 27. Quality Control in ICP-OES/MS analysis (after Munson R., 1997) Ycps = a*Xmg/L+b The slope is a measure of sensitivity: how much the signal changes for a change in concentration. Ycps = a*Xmg/L+b a Ca K Cu Pb Cr
  • 28. Quality Control in ICP-OES/MS analysis Concentration Units
  • 29. Accuracy is how close a measured value is to the actual (true) value. Precision is how close the measured values are to each other. Quality Control in ICP-OES/MS analysis LOQ is the lowest concentration at which a measurement is quantitatively meaningful (Mitra, 2003) LOD is the concentration at which we can decide whether an element is present or not (Thomsen, 2003)
  • 30. Quality Control in ICP-OES/MS analysis The combined uncertainty Uc in analytical measurement is a single numerical value that represents the total estimation of uncertainty, taking into account various sources of error and variability associated with the measurement process. Uncertainty Uexp = Uc*k, k is coverage factor (appr. 2 for CI=95%)
  • 31. Take home message • Choose your spectrometry technique/instrument based on your research goals. • Always remember the sample pre-treatment and preparation steps. • Check the reliability of your analysis