Iva
Juranovic
a,
Patrick
Breinhoelder
b and
Ilse
Steffan
*b
aDepartment of Analytical Chemistry, Faculty of Science, University of Zagreb, Strossmayerov trg 14, HR-10000, Zagreb, Croatia
bInstitute of Analytical Chemistry, University of Vienna, Waehringerstrasse 38, A-1090, Vienna, Austria
First published on 11th December 2002
Inductively coupled plasma atomic emission spectrometry (ICP-AES) was used for the determination of the elements present in pumpkin seed oil and pumpkin seeds. After extraction of the oil from pumpkin seeds a residue was obtained. For the determination of elements three types of samples were used: oil, seeds and the residue (after oil extraction). Three different digestion types were applied: open vessel and closed vessel digestion in a steel bomb as well as microwave digestion in a closed system. For most elements of interest the measured concentration differs depending on the digestion method used. After microwave digestion a good reproducibility for ICP-AES measurements for all elements was found. Determination of Zn by ICP-AES is possible both after open and closed vessel digestion (steel bomb or microwave digestion). No loss of volatile compounds in pumpkin oil during preparation or microwave digestion prior to ICP-AES analysis could be observed. In general, the recoveries for all elements in pumpkin seed oils and seeds were >95%, for S only they were <50%. Differences in the element concentration were found for seed oil obtained from the same seeds but prepared by two different procedures, extracted by Soxhlet and commercially produced after roasting of the seeds. The differences of the measured element concentrations after application of different types of dissolution procedures are discussed. The closed vessel dissolution was found to be the best procedure prior to the ICP-AES determination of metals. The method is evaluated by application of the standard addition method and by recovery experiments. Addition of salt during the oil production procedure causes approx. 10 times the Ca, K, Mg, and Na amounts compared to the Soxhlet extracted oil produced in the laboratory. Higher amounts of Na could be registered in the residues as well. The LODs were <0.1 µg g−1 for Ca, Cd, Mg, Mn, Ti, and Zn, in the range of 0.1 to 0.8 µg g−1 for Co, Cu, Fe, K, Mo, Na, Ni, Pb, and V, and >0.8 µg g−1 for Al, Cr and P in pumpkin oil samples.
Hydrogenation of edible seed oils and fats has been performed using nickel catalysts. The presence of copper and iron can be caused by the processing equipment as well. Because of the metabolic functions of the metals, a development of fast and accurate analytical methods for trace element determination in oils is important from the point of view of both quality control of production and food analysis. The authenticity of products is important from the standpoints of both commercial value and health aspects. Over the years, a high degree of sophistication has evolved for chromatographic methods4–6 for the analysis of components of oils and fats. At the same time spectroscopic methods7–10 are emerging as potential tools for a rapid screening of samples for the detection of their adulteration. Inductively coupled plasma atomic emission spectrometry (ICP-AES) and atomic absorption spectrometry (AAS) are the most commonly used techniques for the determination of metals in different samples.11,12 Fats and oils are particularly difficult to analyze for their trace metal contents. Different digestion methods were applied for oil digestion prior to spectrometric measurements. Many of the used wet or dry digestion methods are not recommended for use in high fat material because of the associated safety hazards.13 A direct determination method like dilution with an appropriate organic solvent followed by direct aspiration into an atomic absorption flame or an ICP-excitation unit is sometimes not sensitive enough. In most cases the analytical method exhibits changes in the excitation power for organic media compared to aqueous solutions.14 Also calibration is more difficult in organic media.15,16 Optional methods of sample introduction into ICP-AES are the emulsions-formation techniques. Their application does not require destruction of the organic matter or the use of large amounts of organic solvent. Their disadvantage compared to aqueous systems is the difficulty occurring in calibration since aqueous standard solution cannot be used in this case. The stability of emulsions, selection of the surfactant, the proportions of the phases, the preparation procedure, and the nature of the analyte compounds present in the matrix are usually limitations for such a method.8,17,18
Procedures based on trace element extraction by different extractants are often time consuming and prone to losses and contamination.19 An alternative methodology is microwave-enhanced dissolution chemistry. It enables fast, efficient and reproducible sample preparation. Sample solutions are heated so effectively that the reaction time can be reduced drastically, often from days to minutes. Furthermore the level of the reaction and process control offered by microwave heating is better than by any other heating method.8,20
The aim of this work was the determination of elements in pumpkin seed oil and seeds by ICP-AES. Furthermore differences in element concentrations between oil extracted by Soxhlet (self extracted) and commercially pressed pumpkin seed oil from roasted seeds should be registered. Self extracted oil was gained by Soxhlet extraction without salt addition during the procedure. The resulting product was used for comparison to commercially pressed pumpkin oil.
After the oil is extracted from pumpkin seeds a residue is obtained. The concentrations of the elements in oil, seeds and residue were determined, applying three different digestion types: open vessel and closed vessel digestion in a steel bomb as well as microwave digestion in a closed system.
Prior to the analysis of the elements by ICP-AES the oil was extracted from the ground seeds using a Soxhlet apparatus (petroleum ether, bp 60–90 °C, 24 h). After extraction of the oil from the seeds a residue was left. For the determination of the elements three types of samples were used: oil, seeds and residue.
For this purpose 1 g each of the oils, seeds and residues were weighed for the open vessel digestion. Open vessel digestions of the samples were performed in glass vessels after addition of 5 mL of HNO3 conc. and 15 drops of H2O2 at approx. 120 °C and heating of the samples for approximately 12 h. During the digestion procedure the same amount of acid and hydrogen peroxide was added to the samples again twice. After digestion all the samples were transferred into 10 or 20 mL volumetric flasks and filled to volume. For each series of digestions a reagent blank was prepared.
For the closed vessel digestion of the samples steel bombs equipped with a PTFE inlet were used. To 0.25 g oil, seeds or residue 5 mL of HNO3 conc. and 15 drops of H2O2 were added and the sample was heated to 100 °C for approx. 12 h. After digestion all the samples were transferred into 10 mL volumetric flasks and diluted to volume with 1 M HNO3 for measurement.
For the microwave digestion procedure oil (0.5 g), seeds (0.5 g) and residue (0.5 g) were weighed into the digestion vessels. The digestions were performed by adding 4 mL of HNO3 conc. and 2 mL H2O2 (30% v/v) to the sample. The microwave program applied is listed in Table 2 (see later). A rotating turntable was used to insure homogeneous distribution of the microwave radiation in the oven. Temperature control required a temperature sensor in one vessel during the entire decomposition. The oil, seeds and residues were digested according to the following optimized program (Power in W/time in min): 250/2, 0/1, 250/2, 600/1, 400/5, ventilation 3.0 min. The internal temperature was limited to 240 °C during the last step and ventilation. After cooling all the digests were transferred into 10 mL volumetric flasks and diluted to volume with HNO3 (1% v/v). Reagent blanks were prepared similarly to the samples.
The element content of commercially produced pumpkin seed oil (“roasted seeds”), self extracted oil (Soxhlet apparatus) and salad oil products available at local markets were used for comparison. In the laboratory the different oils were extracted by a Soxhlet apparatus from ground seeds without addition of salt.
Element | LOD/µg g−1 pumpkin oil | Wavelength/nm |
---|---|---|
Al | 0.920 | 396.1 |
Ca | 0.041 | 393.3 |
Cd | 0.044 | 226.5 |
Co | 0.356 | 239.8 |
Cr | 0.880 | 267.7 |
Cu | 0.293 | 327.3 |
Fe | 0.320 | 259.9 |
K | 0.248 | 766.5 |
Mg | 0.030 | 279.5 |
Mn | 0.032 | 257.6 |
Mo | 0.161 | 202.0 |
Na | 0.139 | 589.6 |
Ni | 0.164 | 341.4 |
P | 1.101 | 178.3 |
Pb | 0.166 | 220.3 |
S | — | 181.0 |
Ti | 0.042 | 337.2 |
V | 0.114 | 310.2 |
Zn | 0.051 | 206.2 |
The background correction position was selected as +0.14 nm. The integration time used was 5 s. For the determination of K, Na, P, and S sequential spectral lines were used, while for the other elements a simultaneous program was applied.
Pumpkin oil | Al | Fe | Zn | Pumpkin seeds | Al | Fe | Zn |
---|---|---|---|---|---|---|---|
Microwave digestion | 31.9 | 67.83 | 12.21 | Microwave digestion | 69.86 | 139.13 | 59.64 |
Steel bomb | 32.03 | 60.16 | 11.64 | Steel bomb | 72.66 | 151.08 | 53.15 |
Open vessel | 24.76 | 47.19 | 12.74 | Open vessel | 59.87 | 93.53 | 54.23 |
Ca | Cd | Cr | Cu | Fe | Mg | Mo | Na | Ti | V | Zn | Al, Co, Ni, Mn, P, K, Pb | ||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
a A—commercially produced pumpkin oil. B—commercially produced pumpkin oil was diluted with petroleum ether and the solvent was evaporated on a rotary evaporator at 45 °C. C—commercially produced pumpkin oil was filled in a round bottom flask, connected to a vacuum pump at room temperature for 30 min. Results are given in µg g−1. b The LOD were 0.1 µg g−1 for Co, K, Mn, Ni, Pb and >0.8 µg g−1 for Al and P. | |||||||||||||||||||||||||
A a | 1.46 | 1.74 | 6.63 | 9.76 | 15.45 | 46.04 | 0.84 | 34.41 | 3.87 | 24.65 | 3.22 | <DLb | |||||||||||||
A b | 6.13 | 1.72 | 4.93 | 9.91 | 14.4 | 41.12 | 0.81 | 33.35 | 3.84 | 22.26 | 2.88 | <DLb | |||||||||||||
B a | 7.85 | 1.76 | 8.6 | 14.77 | 17.75 | 48.62 | 0.91 | 37.82 | 3.91 | 26.51 | 3.55 | <DLb | |||||||||||||
B b | 5.43 | 1.71 | 5.05 | 10.59 | 15.88 | 40.46 | 0.79 | 36.42 | 3.83 | 24.86 | 3.25 | <DLb | |||||||||||||
C a | 7.28 | 1.76 | 8.46 | 14.59 | 17.7 | 48.86 | 0.9 | 36.61 | 3.91 | 26.59 | 3.49 | <DLb | |||||||||||||
C b | 6.4 | 1.72 | 7.44 | 12.83 | 15.56 | 42.97 | 0.82 | 32.2 | 3.84 | 23.38 | 3.07 | <DLb | |||||||||||||
Mean | 5.8 | 1.7 | 6.8 | 12.1 | 16.1 | 44.7 | 0.8 | 35.1 | 3.9 | 24.7 | 3.2 | ||||||||||||||
SD | 2.30 | 0.02 | 1.61 | 2.30 | 1.34 | 3.70 | 0.05 | 2.20 | 0.04 | 1.71 | 0.25 | ||||||||||||||
%RSD | 39.4 | 1.1 | 23.5 | 19.0 | 8.3 | 8.3 | 5.9 | 6.2 | 1.0 | 6.9 | 7.7 |
In Table 4 the results obtained by the standard addition method and the recovery experiments for pumpkin oil (sample 2) are shown. The expected concentration of the elements was 10 mg L−1 (200 µg g−1). In general, recoveries for all elements in pumpkin oils were >95%. For Na and S they were <50% or 70%, respectively.
Al | Ca | Cd | Co | Cr | Cu | Fe | K | Mg | Mn | Mo | Na | Ni | P | Pb | S | Ti | V | Zn | |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
p.oil 2a | 9.3 | 10.0 | 9.6 | 9.2 | 9.5 | 9.3 | 10.0 | 10.6 | 10.6 | 10.0 | 9,7 | 3.4 | 9.2 | 9.2 | 10.2 | 6.6 | 13.1 | 9.8 | 9.2 |
p.oil 2b | 11.5 | 11.7 | 11.4 | 12.0 | 11.2 | 12.8 | 11.5 | 12.4 | 12.8 | 11.7 | 11.1 | 4.8 | 10.4 | 9.8 | 10.0 | 8.7 | 16.4 | 11.6 | 12.6 |
p.oil 2c | 10.3 | 10.6 | 10.3 | 10.0 | 10.1 | 10.4 | 10.5 | 9.9 | 11.7 | 10.6 | 10.1 | 4.8 | 9.6 | 13.0 | 9.1 | 7.2 | 15.1 | 10.6 | 13.0 |
Mean | 10.4 | 10.8 | 10.4 | 10.4 | 10.2 | 10.8 | 10.7 | 11.0 | 11.7 | 10.8 | 10.3 | 4.4 | 9.7 | 10.7 | 9.8 | 7.5 | 14.9 | 10.7 | 11.6 |
SD | 1.1 | 0.9 | 0.9 | 1.4 | 0.9 | 1.8 | 0.8 | 1.3 | 1.1 | 0.9 | 0.7 | 0.8 | 0.6 | 2.0 | 0.6 | 1.1 | 1.7 | 0.9 | 2.1 |
Results for spike experiments in pumpkin oil (sample 1) and pumpkin seeds (sample 1) are given in Table 5. The expected concentration of elements was 10 mg L−1 (200 µg g−1). Recoveries for all elements in pumpkin oil and pumpkin seeds were approx. 10 mg L−1, or >95%, except for S they were <50%. The results clearly demonstrate that this type of digestions and ICP-measurements are suitable for all elements except S.
Al | Ca | Cd | Co | Cr | Cu | Fe | K | Mg | Mn | Mo | Na | Ni | P | Pb | S | Ti | V | Zn | |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
p. oil 1 | 1.6 | 0.7 | 10.6 | 2.9 | 1.7 | 9.2 | 3.5 | 2.1 | 0.9 | 0.3 | 9.3 | 11.1 | 11.0 | 5.9 | 0.2 | 0.0 | 13.9 | 0.0 | 0.7 |
p. oil 2 | 10.9 | 10.9 | 0.5 | 13.9 | 1.6 | 0.0 | 9.5 | 1.3 | 0.8 | 0.2 | 0.1 | 1.0 | 0.0 | 13.7 | 0.1 | 0.0 | 0.2 | 0.0 | 9.4 |
p. oil 3 | 1.0 | 0.7 | 0.6 | 3.0 | 10.5 | 0.0 | 3.4 | 11.8 | 11.4 | 11.7 | 0.0 | 1.2 | 0.0 | 5.6 | 10.2 | 0.0 | 0.2 | 10.0 | 0.6 |
p.seed 4 | 3.1 | 27.3 | 10.1 | 3.2 | 0.3 | 10.3 | 5.9 | 467.6 | 84.8 | 2.9 | 10.1 | 9.3 | 9.1 | 493.3 | 0.0 | 0.0 | 15.8 | 1.3 | 3.3 |
p.seed 5 | 11.3 | 31.8 | 1.7 | 14.6 | 0.3 | 0.8 | 15.9 | 500.6 | 95.8 | 2.1 | 0.2 | 1.9 | 0.0 | 514.6 | 0.0 | 0.0 | 3.8 | 1.3 | 15.2 |
p.seed 6 | 3.0 | 28.0 | 1.7 | 3.1 | 11.2 | 0.8 | 5.1 | 497.6 | 100.1 | 13.3 | 0.4 | 1.9 | 0.0 | 482.5 | 9.8 | 5.6 | 3.8 | 9.1 | 2.9 |
The limits of detection obtained for the determination of the elements in pumpkin oil are given in Table 1. LODs were determined also in pumpkin seeds and pumpkin oil residue samples. The limits of detection for the elements in those samples were the same as for the elements in pumpkin oil (Table 1). Limits of detection <0.1 µg g−1 were obtained for Ca, Cd, Mg, Mn, Ti, and Zn. For Co, Cu, Fe, K, Mo, Na, Ni, Pb, and V they were in the range from 0.1 to 0.8 µg g−1, and for Al, Cr, and P >0.8 µg g−1.
Further improvements of the method's performance, especially for the S determination are limited by the sample amount and the reagent volumes, necessary for the digestion procedure. Smaller volumes of nitric acid and hydrogen peroxide did not lead to complete digestion. Higher volumes of reagents can even cause excessive foaming and sample loss by the vapour removal system.
The clear sample digests obtained by the method described can be used not only for ICP-AES but also for ICP-MS, graphite furnace or flame atomic absorption spectrometry. Compared to direct analysis of samples after dilution with an organic solvent, the digestion method applied for this study allows a simultaneous determination of metals and avoids limitations due to the introduction of a carbonaceous matrix or contamination caused by the organic solvents used. Figs. 1 and 2 show results for the determination of elements in oil, seeds and residue after oil extraction and subsequent digestion using the microwave unit.
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Fig. 1 Determination of Al, Co, Cr, Cu, Fe, Mn, Na, Zn in pumpkin oil, seeds and residue after oil extraction and digestion in the microwave unit. |
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Fig. 2 Determination of Ca, K, Mg, P in pumpkin oil, seeds and residue after oil extraction and digestion in the microwave unit. |
For all residues investigated the concentrations of the elements of interest are higher compared to the ones measured for seeds and oils.
In all samples investigated the concentration of the toxic elements (Cd, Pb) was rather low; approx. 10 µg L−1 or less for Cd and Pb and approx. 6 µg L−1 or less for Mo, Ni and Ti.
Salad pumpkin oil is commercially produced from roasted seeds (PK is the so called “Presskuchen” i.e. molding cake). This type of oil was used also for comparison to the self extracted oil (“Soxhlet” apparatus). Higher concentrations of Ca, K, Mg, Na, and P in PK-oil samples were determined caused by addition of salt (NaCl) to the roasted seeds before pressing of the oil. Salt is added to obtain a higher amount of oil from the seeds. Self extracted oil contained approx. 10 times less Ca, K, Mg, Na, and P than the commercially produced pumpkin oil from “roasted seeds”. The amounts of Ca, K, Mg, Na, and P depend on the procedure of the oil production. For roasted seeds: 70 to 90 µg Ca g−1, 110 to 300 µg K g−1, 150 to 250 µg Mg g−1, 190 to 440 µg Na g−1, and 450 to 950 µg P g−1 were measured. Furthermore the results of the element determinations in the different residue samples were compared to those obtained in oils. No significant differences of the element concentrations were found for the residues after oil extraction by a Soxhlet apparatus and the residues after pressing of the oil. With one exception: for Na a difference of 23.78 to 31.04 µg Na g−1 for residues of ground seeds to approx. 10000 µg Na g−1 for the residues after pressing of the oil was found. As expected adding salt to seeds before pressing of the oil causes an increase of the Na amount of the oil. Self extracted oil was gained from ground seeds using a Soxhlet apparatus (without adding salt). Adding salt in commercially produced pumpkin seed oil causes high amounts of Ca, K, Mg, Na, and especially P.
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