Solid phase extraction is the very popular technique currently available for rapid and selective sample preparation. The versatility of SPE allows use of this technique for many purposes, such as purification, trace enrichment, desalting, and class fractionation and etc.

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In the name of God

2. Solid phase extraction & cold vapor
atomic absorption spectrometry
December 2014
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3. 3
Introduction & background
Methods
Application
Outline
Conclusion

4. Introduction
Solid phase extraction is the very popular technique
currently available for rapid and selective sample
preparation. The versatility of SPE allows use of this
technique for many purposes, such as purification,
trace enrichment, desalting, and class fractionation
and etc.
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5. SPE procedure
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6. The formats in SPE
•Free disks
•Cartridge
•96-well plates
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7. Cartridges
The SPE cartridge is a small plastic or glass open-
ended container filled with adsorptive particles of
various types and adsorption characteristics.
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8. Discs
Discs consist of a 0.5 mm thick membrane
where the adsorbent is immobilized in a web of
microfibrils.The sorbent (on polymer or silica) is
embedded in a web of PTFE or glass fibre.
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9. 96-well plates
The new format of SPE is 96-Well SPE Plates.
Parallel sample processing allows 96 samples to be
extracted in approximately one hour or less.
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10. Advantage for SPE
Solid phase extraction has been widely used
in the separation and preconcentration of
various species. There are many advantages
to this technique, such as simplicity, reliability,
reduction in analysis time, reduction or
elimination of the use of organic solvents and
a high potential for automation.
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11. Atomic absorption spectrometry
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12. CV-AAS
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13. Paper1: A new method for preconcentration and
determination of mercury in fish, shellfish and
saliva by CV-AAS
1. separation and preconcentration of mercury
2. measurement of mercury was performed in
CV-AAS
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14. Sample preparations for the
determination of mercury in human saliva
1. Samples were collected from healthy volunteers aged 20-25 years.
2. 2 ml of sample was transferred to teflon cup of acid digestion bomb
3. HNO3(65% w/v, 4 ml) and H2O2 (30% v/v, 1ml) were added.
4. The digestion bomb flask was placed in an oven at 75ᵒc for 6 h.
5. Cooling and adjusting the pH with a NaOH solution
6. The digest was transferred to a 100 ml volumetric flask containing
20 ml of buffer solution
7. Finally, the flask was filled to volume with ultrapure water.
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15. Sample preparations for the determination of
mercury in fish, shellfish:
1. 0.1 g of sample was weighed
2. Transferred to teflon cups in order to microwave the acid digestion
bombs.
3. 2.0 ml of 65% (w/v) HNO3 and 1.0 ml of ultrapure water were added.
4. The system subjected to microwave radiation for 60 s at a power of
100 w.
5. Cooling and adjusting the pH with a NaOH solution
6. The digest was transferred to a 100 ml flask containing 20 ml of buffer
solution.
7. Finally, the flask was filled to volume with ultrapure water.
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16. Optimization of conditions for the
preconcentration
A 1.0 µg L−1 Hg (II) solution was used in the optimization of
variables. The extraction of the element by XAD-BTAC was
calculated using the following equation: E = (CV/C∘V∘) x
100, where C is the concentration of mercury in the eluent,
V is the volume of the eluent,C∘ is the initial concentration of
mercury in the solution passed through the minicolumn, and
V∘ is the volume of the mercury solution that was passed
through the minicolumn.
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17. Effect of pH
pH plays an important role in the ligand–metal
interaction
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18. Effect of concentration of eluent
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1.75 mol L−1concentration of
HCl was chosen

19. Effect of flow rate of eluent
• The effect of the flow of eluent was examined in
the range from 1.0 to 8.1 mL min−1
• Employing flow values between 5.5 and 8.1 mL
min−1, no significant variation on response was
observed.
the value of 6.6 ml min−1
for the flow of the
eluent was chosen.
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20. Sample flow rate
●The sorption of Hg (II) on functionalized resin
was studied at flow rates varying from 1.0 to 10.1
ml min−1
● The best results were obtained when the flow
rates were from 1.0 to 6.6 mL min−1
.
● A flow of 5.5 ml min−1
was established to
provide a high sampling rate without compromising
retention of hg (II).
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21. Results for determination of mercury in
certified reference material and saliva
samples.
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Results for determination of mercury in seafood
samples using the proposed procedure.

23. Characteristics of methods using solid
phase extraction for preconcentration of
mercury
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LOD: limit of detection; EF: enrichment factor; PDCA: 2,6-pyridine dicarboxylic acid; AATP: 2-aminoacetylthiophenol; 1,2-BAE: 1,2-
bis(o-minophenylthio)ethane; PSDVB: polystyrene–divinylbenzene; ACPG: aminopropyl-controlled pore glass; PSTH: 1,5-bis(2
pyridyl)-3-sulphophenyl methylene thiocarbonohydrazyde; DPTH: 1,5-bis(di-2-pyridyl)methylene thiocarbohydrazide; TS:
methylthiosalicylate; BG: Brilliant Green; MB: 2-mercaptobenzimidazole; TC: thiosemicarbazide

24. Amberlite XAD-4 (polystyrene–
divinylbenzene) resin
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25. BTAC: 2-(2-benzothiazolylazo)-p-
cresol
Molecular formula: C14H11N3OS
Systematic name: 2-[(E)-1,3-Benzothiazol-2-yldiazenyl]-
4-methylphenol
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26. Conclusion
• The proposed method for the determination of mercury proved to be
simple, efficient and easy to perform in the matrices analysed.
• The Amberlite XAD-4 functionalized with BTAC reagent, which was
efficient for the complexation of Hg (II) allowing
extraction/preconcentration with a high enrichment factor.
• The proposed procedure also presented analytical features (e.g., limit
of detection, precision and accuracy) suitable for the determination of
Hg(II) in both saliva samples and in fish and shellfish.
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27. SPE of ultra traces mercury (II) using octadecyl
silica membrane disks modified by 1,3-bis(2-
ethoxyphenyl)triazene (EPT) ligand and
determination by CV-AAS
• pH=3.5
• Amount of ligand=7.5 mg
• Eluent= use 1.5 M perchloric acid solution
• Flow rate= 3 mL min−1
for the eluent and 50 mL
min−1
for the sample
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28. Chemical structure of 1,3-di(2-ethoxyphenyl)
triazene (EPT).
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29. Determination of Hg(ǁ) ions in 500 ml of
real samples after applying
recommended procedure.
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30. Comparing the performance of this method
with other methods for determination of Hg2+
ions.
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31. SPE of Cd on 2-mercaptobenzothiazole loaded on sulfur
powder in the medium of ionic liquid 1-butyl-3-
methylimidazolium hexafluorophosphate and CVG-AAS
determination
• pH=8.5 using either 0.1 mol L−1 HNO3 & NaOH solutions
• parameters using either 2.5 ml of 2 mol L−1
of HCl
• flow rate a flow rate of 8 mL min−1 was selected for the
eluent and 12 mL min−1
for the sample
• sample volume the maximum sample volume is 200mL
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32. Chemical structure of ligands:
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1-butyl-3-methylimidazolium hexafluorophosphate
2-mercaptobenzothiazole

33. Determination of cadmium in water
samples
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34. Determination of cadmium in fish
samples
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35. Comparison of papers
sample element sample
flow rate
adsorben
t
pH
paper1
Water &
fish
mercury 6 ml
min−1
Agar
modified
2.5
paper2 Water &
fish
mercury 4 ml
min−1
Dowex
Optipore
V-493
2
paper3 Water &
fish
mercury 16 ml
min−1
sulfur 8.5
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36. Comparison of papers
ligand
eluent
solution DL
Linear
rang
paper1
2-
mercaptoben
zimidazole
HCl 0.2
ng ml−1
0.04-2.4
ng ml−1
paper2
S. aureus HCl __ __
paper3 N-(2-chloro
benzoyl)-N-
phenylthioure
a
HCl __ __
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37. Structure of ligands
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N-(2-chlorobenzoyl)-N-phenylthiourea 2-mercaptobenzimidazole

38. Refrences:
• V. A. Lemos, L. O. dos Santos, 2014, Food Chemistry.,
149, 203–207.
• M. K. Rofouei, A. Sabouri, A. Ahmadalinezhad, H.
Ferdowsi, 2011, Journal of Hazardous Materials., 192,
1358–1363.
• N. Pourreza, K. Ghanemi, 2010, Journal of Hazardous
Materials., 178, 566–571.
• N. Pourreza, H. Parham, A. R. Kiasat, K. Ghanemi, N.
Abdollahi, 2009, Talanta., 78, 1293–1297.
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39. • N. Pourreza, K.Ghanemi, 2009, Journal of
Hazardous Materials., 161, 982–987.
• M. Tuzen, I. Karaman, D. Citak, M. Soylak, 2009,
Food and Chemical Toxicology., 47, 1648–1652.
• A. Żwir-Ferenc, M. Biziuk, 2006, Polish J. of
Environ. Stud., 15, 677-690.
• www.mercury-instrumentsusa.com
• www.sigmaaldrich.com
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