Journal of Analytical Methods in Chemistry

Journal of Analytical Methods in Chemistry / 2018 / Article

Research Article | Open Access

Volume 2018 |Article ID 2158407 |

Mateusz Kowalcze, Jan Wyrwa, Małgorzata Dziubaniuk, Małgorzata Jakubowska, "Voltammetric Determination of Anethole on La2O3/CPE and BDDE", Journal of Analytical Methods in Chemistry, vol. 2018, Article ID 2158407, 14 pages, 2018.

Voltammetric Determination of Anethole on La2O3/CPE and BDDE

Academic Editor: Pablo Richter
Received28 Aug 2017
Revised03 Nov 2017
Accepted09 Nov 2017
Published20 Feb 2018


In this work, DPV determination of anethole was presented using various carbon, two-diameter (1.5 and 3 mm) electrodes, that is, BDD, GC, CP, and CP doped by La2O3 and CeO2 nanoparticles. La2O3/CPE to our best knowledge was proposed first time. Cyclic voltammograms confirmed totally irreversible electrode electrooxidation process, controlled by diffusion, in which two electrons take part. The most satisfactory sensitivity 0.885 ± 0.016 µA/mg L−1 in 0.1 mol L−1 acetate buffer was obtained for La2O3/CPE with the correlation coefficient r of 0.9993, while for BDDE it was 0.135 ± 0.003 µA/mg L−1 with r of 0.9990. The lowest detection limit of 0.004 mg L−1 was reached on La2O3/CPE (3 mm), what may be compared with the most sensitive conjugate methods, but in the proposed approach, no sample preparation and analyte separation was needed. Anethole was successfully determined in specially prepared ethanol extracts of herbal mixtures of various compositions, which imitated real products. The proposed procedure was verified in analysis of commercial products, that is, anise essential oil, which contains a large concentration of anethole, and in alcohol drinks like Metaxa, Ouzo, and Rakija, in which the considered analyte occurs on trace levels. Structure and properties of the considered nanopowders and graphite pastes were investigated by EDX, SEM, and EIS.

1. Introduction

1.1. Anethole: Properties, Application, and Methods of Determination

Anethole—anise camphor, para-methoxyphenylpropene, CAS number: 104-46-1—is a colorless, crystalline aromatic terpenoid analogue with a characteristic sweet taste and pleasant aroma, occurring in many essential oils obtained from plants belonging to the family Apiaceae, such as anise (Pimpinella anisum), star anise (Illicium verum), fennel (Foeniculum vulgare), liquorice (Glycyrrhiza glabra), and caraway (Carum carvi) [14] (Table 1). It occurs naturally in the form of two isomers: trans- (CAS number: 4180-23-8) and cis- (CAS number: 25679-28-1), where naturally the trans-isomer is much more common [47].

Type of plantsCountry of originPart of plantsMethod of obtaining oil and analysisContent of anethole in the oil (%)Reference

FennelBrazilSeedSupercritical fluid extraction (SFE), GC-MS11.0–47.4-trans[1]
Star aniseIndiaFruitDistillation in Clevenger-type apparatus, GC-MS1.07-cis[7]
AniseGermanySeedDistillation in Clevenger-type apparatus, GC-MS0.14-cis[8]
CarawayDifferent European countriesFruitSteam-distillation, GC-FID0–2.2-trans[9]
Bitter orangeAlgeriaPeelDistillation in Clevenger-type apparatus, GC-MS2.3-trans[10]

Due to its organoleptic properties, pleasant aroma and sweet taste, essential oils containing anethole have been used for centuries in the perfume, pharmaceutical, and spirit industries [16, 11, 12]. In pharmaceutical applications, anethole properties such as oestrogenic action, depressive action to the central nervous system, psycholeptic, insecticide, bactericidal, anticarcinogenic, anti-inflammatory, and anesthetics activity [16, 11, 12] are very important. The bactericidal properties of anethole are, due to lack of free phenolic group, weaker than their natural analogue—eugenol (Figure 1) [6]. In the spirit industry, anethole is present in various types of alcoholic beverages based on anise, fennel, or licorice—mainly Absinthe, Pastise, Ouzo, Rakija, and Metaxa [6, 12]. In addition, some alcohols must contain exactly the specified amount of anethole; for example, Pastise contains 1.5–2.0 g L−1 of this compound [12]. So, accurate determination of anethole content is one of the important stages of drinks production.

Among the quantitative methods of anethole assays in various matrices (Table 2), chromatographic techniques are dominant. Voltammetric techniques are used to evaluate the antioxidant properties of the compounds containing anethole [13] and may be useful in the classification of alcoholic beverages [14]; therefore, developing the method of anethole determination seems to be justified and interesting.

Type of matrixAnalytical techniqueLinearity range (mg L−1)LOD (mg L−1)LOQ (mg L−1)Trans-anethole conc. (mg L−1)Reference

Fennel essential oilHPLC10–1000.953.00.2–30.5[15]
Fennel essential oilGC-MS10–5500.0020.0060.16–40[15]
Aniseed drinksHPLC2–160.00230.0077125–4040[16]
Human blood serumHS-SPME-GC-MS0.002–0.20.00360.00530.0054–0.0176[17]

In this work, we present the possibility of trans-anethole determination by differential pulse voltammetry (DPV) technique. Various carbon electrodes, that is, glassy carbon electrode (GCE), boron-doped diamond electrode (BDDE), carbon paste electrode (CPE), carbon paste electrode doped by cerium(IV) oxide (CeO2/CPE), and carbon paste electrode doped by lanthanum(III) oxide (La2O3/CPE), were used and tested. After designation of the analytical parameters of the method and optimization, quantitative and qualitative assays of anethole were applied in four specially prepared herbal matrices similar to anethole-containing beverages and in various commercially available products of natural origin.

The obtained results are very promising and can be used in determination of anethole in a variety of matrices without analyte separation or sample preparation.

2. Experimental

2.1. Measuring Apparatus and Software

A multipurpose Electrochemical Analyzer M161 with the electrode stand M164 (both MTM-ANKO, Poland) was used for all voltammetric measurements. The classical three-electrode quartz cell of 10 mL volume was applied. Various carbon sensors were utilized as the working electrodes, that is, glassy carbon electrode (BASi, ϕ = 3 mm and home-made, ϕ = 1.5 mm), boron diamond-doped electrode (Windsor Scientific, ϕ = 3 mm), carbon paste electrode, carbon paste electrode doped by cerium(IV) oxide, and carbon paste electrode doped by lanthanum(III) oxide. Carbon paste electrodes were prepared in our laboratory. Also a double-junction reference electrode Ag/AgCl/KCl (3 M) with replaceable outer junction (2.5M KNO3) and a platinum wire as an auxiliary electrode were used. The ambient temperature was ca. 23°C. The MTM-Anko EAPro 1.0 software enabled electrochemical measurements, data acquisition, and processing of the results.

Electrochemical impedance spectroscopy measurements were performed using a frequency analyzer (Solartron model FRA 1260) coupled with dielectric interface (model 1296). The surface morphology of electrode material was observed using ultrahigh-resolution scanning electron microscope with field emission (FEG-Schottky emitter; Nova NanoSEM 200, FEI Europe BV) cooperating with EDAX EDS analyzer.

2.2. Carbon Paste Electrodes Doped by La2O3 and CeO2

Carbon-based electrodes are useful for voltammetric determination of wide range of analytes in liquid solutions. Moreover, their applicative properties can be improved by doping with metal oxide modifiers. The usage of different doping oxides was reported for modification of carbon-based electrodes so far [1822]. According to the literature, addition of controlled amounts of cerium(IV) oxide to glassy carbon electrodes material led to the significant enhancement of sensitivity, selectivity, reproducibility, and response time in the amperometric quantification of eugenol [22].

In this work, the experimental data obtained by use of lanthanum oxide-doped graphite paste electrodes applied in voltammetric analysis are presented for the first time. The carbon pastes were prepared by hand mixing an adequate amount of graphite powder and rare earth oxide powder with paraffin oil using a pestle and mortar for at least 30 minutes in the case of each batch. Nanopowders of lanthanum(III) oxide (99.99%) and cerium(IV) oxide (99.9%) were provided by Acros Organics. The ratio of used paraffin oil and graphite powder was determined based on literature repots as well as our experience in order to get electrodes characterized by high chemical and mechanical stability during performance in liquid solutions. After standing overnight, the resulting homogenous pastes were packed into the well of the working electrodes to depth of 2 mm with two different diameters (1.5 and 3 mm). The body of working electrode was a Teflon tube with stainless steel rode of 1.5 mm diameter serving as electric contact. To provide the required smoothness of electrodes, working surfaces the forehead of electrodes were polished on a print paper or tissue paper.

The amounts of used reagents, details of prepared electrodes, and pastes are presented in Table 3.

ElectrodeElectrode diameter (mm)IngredientsPaste labelWeight ratio of powder ingredients

CeO2/CPE 20%1.52 g graphite, 1.5 ml paraffin oil, 0.5 g CeO2PCe20-50.2 CeO2–0.8 graphite
La2O3/CPE 20%1.52 g graphite, 1.5 ml paraffin oil, 0.5 g La2O3PLa20-20.2 La2O3–0.8 graphite
La2O3/CPE 30%1.52 g graphite, 1.5 ml paraffin oil, 0.857 g La2O3PLa30-30.3 La2O3–0.7 graphite
La2O3/CPE 40%1.52 g graphite, 1 ml paraffin oil, 1.333 g La2O3PLa40-40.4 La2O3–0.6 graphite

2.3. Chemicals and Glassware

As a supporting electrolyte, buffers of a different pH were prepared in our laboratory (from reagents pure for analysis, POCH, Poland): acetate buffer—mix acetic acid and sodium acetate; Britton–Robinson buffer—mix boric acid, phosphoric acid, acetic acid, and sodium hydroxide; Sørensen phosphate buffer—mix sodium hydrogen phosphate and sodium dihydrogen phosphate; ammonia buffer—mix ammonia and ammonium chloride. As a standard solution, trans-anethole (analytical standard, Sigma-Aldrich) was used. 1 µL of solution contains 3.48 µg of trans-anethole. Reagents used to determine the impact of interferents are 99% eugenol (Reagent Plus, Sigma-Aldrich), 99% carvacrol (food grade, Sigma-Aldrich), ≥98.5% thymol (pure, Sigma-Aldrich), and zinc, lead, cadmium, bismuth, aluminum, thallium, chromium, and vanadium (all metals from Certipur, Merck). The other chemicals were 95% ethanol (food grade, Polmos, Poland) and 0.1 mol L−1 solution of sulfuric acid (pure for analysis, POCH, Poland) for activation of BDD electrode. All reagents used were prepared using quadruply distilled water (two last stages from quartz). Glassware was first immersed in 6 M nitric acid and then rinsed repeatedly with distilled water.

2.4. Samples

To verify the possibility to determine anethole in herbal matrices A, B, and D, three solutions were prepared. Also matrix C was tested, which did not contain anethole. The composition and preparation of the matrices imitated different anethole-containing beverages. Each matrix was prepared by pouring with the ethanol (95%, food grade) the appropriate herbal composition and the five-day maceration of the mixture. After this time, each matrix was rectified once.

Anethole was also determined in commercially available products such as anise oil (KEJ, Poland), Efe Rakija (Turkey), Ouzo Typnaboy (Greece), and Metaxa (Greece).

Three independent samples of the same type were tested.

2.5. Standard Procedure of Voltammetric Measurements

Measurements were performed using differential pulse voltammetry (DPV). Before each series of measurements, surface of the BDD electrode was activated 15 minutes in 0.1 mol L−1 sulfuric acid solution by the potential of 2400 mV. Before each calibration, the BDDE surface was additionally renewed by the potential 1500 mV and time of 30 s in supporting electrolyte. GCE was activated by polishing with polishing powder MicroPolish Alumina 0.05 μm (Buehler, USA).

The investigation of anethole was performed in different supporting electrolytes depending on the working electrode used, that is, 0.1 M acetate buffer with pH 3, 4, 5, or 6; Britton–Robinson buffer with pH 2 and 3; 0.1 M Sørensen’s phosphate buffer with pH 6, 7, and 8; or 0.1 M ammonia buffer with pH 9 and distilled water, giving total volume of 5 mL filling the quartz voltammetric cell. The best results were obtained in supporting electrolyte consisting 5 mL of 0.1 M acetate buffer with pH 6. The volume of added standard solution of anethole was of 1–5 µL.

The solution in cell was stirred (ca. 500 rpm) using a magnetic stirring bar. Then, after a rest period of 5 s, differential pulse voltammograms were recorded in the potential window: 0–1200 mV (BDDE), 500–1300 mV (CPE, La2O3/CPE, CeO2/CPE), and 600–1200 mV (GCE). The other standard experimental parameters were as follows: potential step Es = 5 mV; pulse potential dE = 50 mV; and time of potential step = 40 ms (20 ms waiting time + 20 ms sampling time).

All experiments were performed at 23°C. All experiments were carried out in triplicate.

3. Results and Discussion

3.1. Carbon Electrodes in Determination of Anethole

The purpose of the study was to investigate whether carbon paste electrodes doped by two new rare earth element oxides may be useful in voltammetric determination of the anethole. Commercially available and popular sensors were used as a comparison. The well-defined DPV anethole peak (Figure 2) was obtained on the various carbon electrodes, that is, glassy carbon, boron-doped diamond, carbon paste, and carbon paste doped by lanthanum(III) oxide and cerium(IV) oxide, which were considered in this work. The peak position was observed between 965 and 1155 mV (Table 4, second column). Anodic shift (of 150–200 mV) of anethole oxidation potential has been obtained for carbon paste and two nanoparticles-modified electrodes compared to GCE and BDDE, confirming the lower transfer rate on CPE and nanoparticles/CPE.

ElectrodeAnethole peak position (mV)a ± SDa (µA/mg L−1)b ± SDb (µA)rLOD (mg L−1)

Electrode diameter 3 mm, area 7.07 mm2
BDDE9650.135 ± 0.0030.050 ± 0.0130.99900.024
GCE9900.306 ± 0.0050.031 ± 0.0460.99950.011
CPE11550.546 ± 0.036−0.068 ± 0.0830.99360.006
CeO2/CPE 20%11250.341 ± 0.009−0.060 ± 0.0210.99890.010
La2O3/CPE 20%10950.885 ± 0.0160.076 ± 0.0690.99930.004
Electrode diameter 1.5 mm, area 1.77 mm2
GCE9700.022 ± 0.0010.0035 ± 0.00510.99890.148
CPE11100.111 ± 0.004−0.005 ± 0.0430.99830.030
CeO2/CPE 20%10750.227 ± 0.008−0.054 ± 0.0800.99780.014
La2O3/CPE 20%10800.449 ± 0.021−0.058 ± 0.0440.99590.007
La2O3/CPE 30%10950.350 ± 0.016−0.039 ± 0.0350.99580.009
La2O3/CPE 40%11000.332 ± 0.0100.029 ± 0.1100.99800.010

Quantitative analysis was preceded by especially projected procedure of baseline modeling and subtraction (Figure 3). The first step of the proposed approach was subtraction of the experimental baseline obtained for the supporting electrolyte. Next, the typical approximation by the polynomial of the 2nd degree was utilized. These two steps were necessary, because the background shape was very different from the polynomial function.

Analytical parameters were determined and tested for two groups of electrodes (Table 4), that is, of the diameter of 3 mm (geometric area of 7.07 mm2) and of the diameter of 1.5 mm (geometric area of 1.77 mm2). After signal processing, the linear relation between peak current and concentration of the anethole in the range of 0.7–17.5 mg L−1 was noticed. Generally, paste electrodes were characterized by the greater sensitivity and lower detection limit in comparison to BDDE and GCE. However, the repeatability of the signal for successive analyte concentration was excellent for the latter (CV < 1%).

The highest sensitivity of 0.89 µA/mg L−1 among the considered electrodes was obtained on the carbon paste doped by the 20% of lanthanum(III) oxide nanoparticles, with the correlation coefficient r of  0.9993 (for averaged signals for each concentration) and the lowest detection limit of 0.004 mg L−1. The sensitivity for the anethole on the electrode doped by the 20% of cerium(IV) oxide nanoparticles (ϕ = 3 mm) was even lower (0.34 µA/mg L−1) than the reference value obtained on CPE (0.55 µA/mg L−1). The lowest sensitivity in the group of sensors with a diameter of 3 mm was obtained on BDDE (0.14 µA/mg L−1) what was ca. 6 times less than on La2O3/CPE. Considering the sensors with a diameter of 1.5 mm, the highest sensitivity of 0.45 µA/mg L−1 was obtained on CeO2/CPE. For CPE- and La2O3-doped CPE, the repeatability of the signal relied on the percent (w/w) of the added nanoparticles and was on the level of 5–8% (CV) when the nanopowder addition was lower than 20%. For the addition greater than 20%, the repeatability of the signal rapidly deteriorated (CV > 10%), and therefore these electrodes were not considered in further tests. CP electrodes doped by CeO2 did not also show the satisfactory repeatability of the signals recorded for each concentration. It was also observed that increasing addition of the lanthanum(III) oxide nanoparticles decreased sensitivity for the anethole.

For further detailed analysis, La2O3/CPE (ϕ = 3 mm) as a sensor of the greatest sensitivity for the anethole was chosen and for comparison BDDE, which is reliable after appropriate activation. Voltammograms and calibration lines for the anethole in the concentration range from 1.39 to 6.96 mg L−1 prepared on the mentioned two sensors are presented in Figure 2(b).

3.2. Supporting Electrolyte Effects

There are several ways in which the supporting electrolytes solvent system can influence mass transfer, the electron reaction (electron transfer), and the chemical reactions which are coupled to the electron transfer. As a supporting electrolyte, 4 different buffers (acetate, Britton–Robinson, phosphate, and ammonia) were applied in examination of the analyte behavior in pH range from 2.0 (BR buffer) to pH 9.0 (ammonia buffer). The best parameters—repeatability, sensitivity, limit of detection, and the favorable relation between signal and baseline—were obtained using acetate buffer; therefore, the pH effect was tested carefully in the pH range typical for this electrolyte, that is, from 3.0 to 6.0. In the considered supporting electrolytes at strongly acidic pH (Britton–Robinson buffer, pH 2.0), neutral pH (phosphate buffer, pH 7.0), and basic pH (phosphate buffer, 8.0; ammonia buffer, pH 9.0), the investigated analyte did not show adequate analytical sensitivity and repeatability.

Figure 4 presents the influence of the acetate buffer pH on the anethole voltammetric signal. The well-defined DPV peak was observed in the whole range of the considered pH, that is, 3.0–6.0. For BDDE, the peak position changed in the range from 950 to 990 mV, without a distinct maximum current change. For La2O3/CPE, the oxidation peak currents decreased to pH 5.0 and then increased. Anethole oxidation potential decreased from 1180 mV to 1080 mV as pH increased. Further experiments were done by pH 6, because less positive peak position equal to 1080 mV is more suitable for oxidation. Sensitivity in this case was also ca. 44% greater in comparison with the best variant obtained for the other pH.

3.3. Parameters of Anethole Electrooxidation on BDDE and La2O3/CPE

The voltammetric behavior of anethole on two carbon electrodes, that is, BDD and La2O3-modified electrodes, in 0.1 mol L−1 acetate buffer of pH 6.0 has been investigated by recording cyclic voltammograms (CV) using the scan rates of 0.025, 0.05, 0.1, 0.2, 0.25, and 0.5 V s−1. It was observed that anethole is irreversibly oxidized on these electrodes (Figure 5), what was confirmed by the absence of cathodic step on the backward branch of the CV. The CP electrode modification with La2O3 nanoparticles leads to the anodic shift of anethole oxidation potential on ca. 200 mV. The effect of potential scan rate in the range of 0.025–0.5 V s−1 on the voltammetric behavior of anethole is also presented in Figure 5. The anethole oxidation currents were proportional to the square root of the potential scan rate (1), confirming that the electrochemical process is diffusion controlled [23].

Moreover, the natural logarithm of anethole peak current (lnip) increases linearly with the natural logarithm of scan rate (lnν) in the range of potential scan rate under investigation, and the regression equation is described by

The value of the slope is below the theoretical value of 0.5, what proves the diffusion nature of anethole oxidation peak once again [23]. A linear relationship between the oxidation potential Ep and lnν has been observed, confirming totally irreversible electrode processes:

In this case, the number of electrons participating in the reaction can be calculated according to [24]where α is assumed to be 0.5 for a totally irreversible electrode process. The Ep − Ep/2 is 53 mV for BDDE and 59 mV for La2O3/CPE. Hence, the number of electrons participating in the anethole oxidation process equals nα to 2.22 for BDDE and 2.47 for La2O3/CPE, what agree well with the values reported earlier [25]. Figure 6 presents the proposition of the electrode reaction.

3.4. Investigation by Energy-Dispersive X-Ray Spectroscopy and Scanning Electron Microscopy

The chemical composition of the pastes used for construction of CPE and La2O3/CPE was analyzed by EDX. The EDX spectrum for the nondoped carbon paste (Figure 7(a)) confirmed the presence of carbon, as the dominant element, and a small amount of oxygen. The EDX spectrum for carbon paste doped by lanthanum(III) oxide (Figure 7(b)) confirmed the presence of the elements carbon, lanthanum, and oxygen.

The surface morphology of the CPE and La2O3/CPE was observed by SEM. The SEM test showed that the nondoped carbon paste was characterized by a surface formed by irregular graphite flakes (Figures 8(a) and 8(b)). The surface of the La2O3-doped carbon paste is more porous, heterogeneous, and irregular than the surface of nondoped carbon paste (Figures 8(c) and 8(d)). This suggests that the presence of La2O3 molecules in carbon paste significantly increases the morphological structure of the material, which facilitates the electron transfer process in the electrode–solution interface, giving better sensitivity and higher repeatability of the voltammetric signal.

3.5. Application of Electrochemical Impedance Spectroscopy

Electrical properties of experimental set comprised of the studied carbon paste electrode and Ag/AgCl/KCl reference electrode immersed in solution containing analyte were determined by Electrochemical Impedance Spectroscopy (EIS) method. The measurements were performed in room temperature, with the frequency range of 0.1–10 MHz and the amplitude of sinusoidal voltage signal of 20 mV. The experimental data were analyzed using the ZView software (version 2.2, Scribner Associates, Inc.), which helped in determination of equivalent circuits’ optimal parameters.

The comparison of Nyquist’s spectra obtained for three different carbon paste electrodes is given in Figure 9. In each case, spectrum was comprised of semicircle visible in high-frequency range and the spur in middle and low frequencies. Electrical equivalent circuits were fitted to the experimental data sets. Spectra were analyzed by connected in series two parallel equivalent circuits consisted of resistors (R) and constant phase elements (CE) and additional constant phase element indispensable to model the spur in low frequencies. The scheme of the used equivalent model is depicted in inset of Figure 9. The simulated spectra are plotted by solid black line and exhibit good agreement with experimental data presented by points. The semicircle parts of the spectra in high frequencies look similar in each case. The course of above mentioned part of the spectrum depends on the reference electrode and solution used during the measurements. Therefore, the parameters of R1 and CE1 are of similar value (Table 5). The differences in course of spectrum in the middle- and low-frequency parts indicate that it is attributable to carbon paste electrode properties. On the basis of conducted analysis, there is a strong relation between the applicable properties of carbon paste electrode and the value of resistance exhibited in middle-frequency fragments of spectra. In particular, the highest value of resistance R2 shows the electrode modified by lanthanum(III) oxide, while the electrode without rare earth oxide addition is characterized by the lowest value of this parameter. Concomitantly, the lower CE-T-2 value, the better performance of electrode. The most significant differences between behaviors of studied electrodes are visible in low frequencies part of the spectra. It is reflected in particular in CE3 element. CE-T-3 value determined for undoped electrode is of order higher than for electrodes modified by rare earth metal oxides. Moreover, parameter n3 for CPE is somehow higher than for doped electrodes and close to 1, indicating stronger capacitive properties of undoped graphite than in the case of electrodes modified modified by lanthanum and cerium oxides.

Frequency rangeParameterElectrode

High (106–107 Hz)R1/Ω18,24815,68016,751
CE-T-1/Ssn7.850 × 10−106.262 × 10−105.009 × 10−10
Medium (106–102 Hz)R2/Ω1.857 × 1062.272 × 1066.027 × 106
CE-T-2/Ssn7.05 × 10−72.65 × 10−71.21 × 10−7
Low (102–10−1 Hz)CE-T-3/Ssn1.03 × 10−66.66 × 10−75.81 × 10−7

3.6. Interferences

Such parameters as potential window, potential step, potential pulse, and time of potential step were tested to optimize the procedure of the anethole determination. The criteria of optimization were repeatability, sensitivity of the method, and the favorable relation between signal and baseline. It was observed that starting potential does not have influence on the anethole peak. Taking into account all the criteria selected experiment conditions are potential step 5 mV, potential pulse 50 mV, and time of the potential step 40 ms (i.e., waiting time 20 ms + sampling time 20 ms).

As possible interferences, metal ions such as Zn(II), Pb(II), Cd(II), V(III), Bi(III), Al(III), Tl(I), and Cr(III) and organic compounds such as eugenol, carvacrol, and thymol were tested, which may be present in plants and products of biological origin, in which anethole also occurs. The concentration of the metal ions was in the range of 1.4–14 mg L−1 by the 13.92 mg L−1 of anethole, which was present in the measured solution. The anethole peak position was not moved, and also no additional peaks were observed. However, the impact of the analyzed ions on the height of the anethole peak after addition of metals was noticeable: change for BDDE anethole signal was in the range of 93–99%, and change for La2O3/CPE was in the range from 82 to 127% (Table 6). The greater sensitivity variation in the last case may be connected with the chemical reactions between metal ions and active lanthanum(III) oxide nanoparticles, what could cause the change of the number of active centres on the electrode surface.

InterferentProportion anethole: interferentianethole+interferent/ianethole

Metal ions
Zn2+20  :  01.001.00
20  :  20.991.02
20  :  100.991.21
20  :  200.991.27
Cd2+20  :  01.001.00
20  :  20.991.07
20  :  100.981.12
20  :  200.961.15
Pb2+20  :  01.001.00
20  :  20.991.04
20  :  100.991.10
20  :  200.981.13
V3+20  :  01.001.00
20  :  21.000.95
20  :  100.990.88
20  :  200.990.82
Bi3+20  :  01.001.00
20  :  20.991.02
20  :  100.981.04
20  :  200.961.22
Al3+20  :  01.001.00
20  :  21.001.02
20  :  100.991.02
20  :  200.991.03
Tl+20  :  01.001.00
20  :  20.980.99
20  :  100.950.91
20  :  200.930.90
Cr3+20  :  01.001.00
20  :  20.991.12
20  :  100.991.22
20  :  200.951.23
Organic compounds
Eugenol20  :  01.001.00
20  :  11.191.14
20  :  21.461.14
20  :  51.900.84
Thymol20  :  01.001.00
20  :  11.111.11
20  :  21.161.21
20  :  51.321.26
Carvacrol20  :  01.001.00
20  :  11.231.09
20  :  21.551.07
20  :  52.001.04

No additional current peaks coming from eugenol, carvacrol, and thymol were observed in the considered acetate buffer (pH 6.0) and potential area where anethole peak was recorded. The concentration of the added substances was in the range of 0.7–3.5 mg L−1 by the 13.92 mg L−1 of anethole. The presence of biological compounds in the solution caused, in the experiments with BDDE, increase of the sensitivity up to 100%. This value is related to the study of the carvacrol effect at the 4 times excess of anethole. The mentioned interferents may facilitate the charge transfer between the analyte and the electrode. In the case of measurements on La2O3/CPE, an addition of 3 biological compounds resulted in the change of the signal amplitude in the range of 84–126%.

3.7. Determination of Anethole in Herbal Matrices and Commercial Products

Because anethole occurs in food products (beverages, herbal oils, and tinctures) and herbs such as anise, star anise, fennel, liquorice, and caraway, the problem of anethole determination of specially prepared herbal matrices was considered. The composition of these mixtures which mimics the real commercially available products is given in Table 7. It is important that some matrices contain anethole, while the other did not contain this analyte, and it was added at the stage of recovery studies. The concentration of the anethole in matrices A, B, and D was on the level 0.1–1.6 g L−1 (Table 7). The highest concentration was in the most complex mixture B, while the lowest in D, where only one component contained anethole. The significant decrease of the sensitivity of the method was observed in comparison to the measurements in only supporting electrolyte. The decrease was to 73% (matrix B) in the case of BDDE and to 66% (matrix D) in the case of La2O3/CPE. Exemplary voltammograms recorded on La2O3/CPE in the case of anethole determination in matrix D are presented in Figure 10(a).

Matrix labelComposition of herbal mixtureAmount of ethanol for macerationIn the matrix may be anetholeAnethole conc. ± SD (g L−1)

AFruit of anise7.5 g25 mLYesBDDE: 0.143 ± 0.005
La2O3/CPE: 0.102 ± 0.024
Fruit of star anise2.5 g
BHyssop leaves2.125 g50 mLYesBDDE: 1.63 ± 0.07
La2O3/CPE: 1.61 ± 0.19
Root of the sweet flag0.45 g
Lemon balm leaves1.5 g
Fruit of anise7.5 g
Fruit of star anise2.5 g
Fruit of fennel6.25 g
Fruit of coriander0.75 g
CWormwood leaves7.5 g50 mLNoBDDE < LOD
DWormwood leaves7.5 g50 mLYesBDDE: 0.500 ± 0.005
La2O3/CPE: 0.475 ± 0.033
Hyssop leaves2.125 g
Root of the sweet flag0.45 g
Lemon balm leaves1.5 g
Fruit of fennel6.25 g
Fruit of coriander0.75 g

Italic signs herbs from which it is possible to extract anethole.

Matrix C did not contain a detectable concentration of the anethole; therefore, this analyte was added to the herbal extract, and percent of recovery was studied (Table 8). Using BDDE, the concentration of anethole of 3.5–10.5 mg L−1 was successfully determined with recovery 101–108%. La2O3/CPE enabled determination of 1.4–4.2 mg L−1 of anethole with recovery 95–100%. The correlation coefficient r in each case was greater than 0.995. The presence of herbal matrix C caused a significant decrease of the method sensitivity, that is, ca. 30% in the case of BDDE and even 50% in the case of La2O3/CPE.

ElectrodeAdded (mg L−1)a ± SDa (µA/mg L−1)b ± SDb (µA)Found ± SD (mg L−1)Recovery (%)r

BDDE 3 mm3.480.0939 ± 0.040.3460 ± 0.053.68 ± 0.70105.870.9951
6.960.0973 ± 0.030.6859 ± 0.037.05 ± 0.51101.300.9982
10.440.0963 ± 0.041.0814 ± 0.0211.22 ± 0.61107.510.9987
La2O3/CPE 3 mm1.390.4306 ± 0.020.5995 ± 0.111.39 ± 0.19100.020.9951
2.780.435 ± 0.0031.186 ± 0.0012.728 ± 0.03995.790.9999
4.180.4409 ± 0.011.8124 ± 0.014.11 ± 0.0798.450.9999

Further, anethole was determined in commercially available products. Some research objects were chosen in which the mentioned analyte is present on very low and very high concentration level. The measurements were done without sample preparation or anethole extraction. The adequate sample volume was added directly to the electrochemical cell. It was observed (Tables 9 and 10) that, in anise essential oil, the concentration of anethole was ca. 570 g L−1, while in alcohol drinks, like Metaxa, Ouzo, and Rakija, it was ca. 0.13–0.21 g L−1. The results obtained using both electrodes were compatible. At 95% confidence level, the calculated Student’s t-values for the replicate measurements of each sample (Table 10) using both fabricated sensors did not exceed the theoretical value (2.7765), indicating that the results obtained are not significantly different. An F-test revealed no significant difference between the standard deviations of the two sets of replicate measurements for each sample. Exemplary voltammograms recorded on La2O3/CPE in the case of anethole determination in Ouzo and Raki are presented in Figures 10(b) and 10(c).

Samplesa ± SDa (µA/mg L−1)b ± SDb (µA)r

BDDE, electrode diameter 3 mm, area 7.07 mm2
Anise essential oil0.0293 ± 0.0010.0667 ± 0.0050.9967
Metaxa 0.0535 ± 0.0020.0691 ± 0.0080.9975
Ouzo0.0905 ± 0.0040.1699 ± 0.0140.9971
Raki0.1068 ± 0.0040.2216 ± 0.0180.9967
La2O3/CPE, electrode diameter 3 mm, area 7.07 mm2
Anise essential oil0.3682 ± 0.0180.8544 ± 0.0780.9951
Metaxa 0.8061 ± 0.0321.1490 ± 0.0680.9968
Ouzo0.7177 ± 0.0201.4659 ± 0.0670.9989
Raki0.4497 ± 0.0100.9507 ± 0.0340.9993

SamplesAnethole conc. ± SD/g L−1F-testt-test

Anise essential oil569.0 ± 62.5580.00 ± 77.51.53760.1914
Metaxa 0.129 ± 0.0190.143 ± 0.0141.84201.0275
Ouzo0.188 ± 0.0220.204 ± 0.0142.46911.0627
Raki0.208 ± 0.0250.211 ± 0.0124.34030.1874

4. Conclusions

In this work, a sensitive, rapid, and convenient DPV procedure of anethole determination was proposed, which does not require sample preparation and separation of the analyte, even in the case of complex matrices. Additionally, it was proved that various carbon electrodes, that is, BDD, GC, CP, and CP doped by La2O3, and CeO2 nanoparticles, are sensitive for anethole, and the proposed analytical strategies fulfill typical validation criteria. Recording cyclic voltammograms, it was noticed that electrooxidation process has totally irreversible character, controlled by diffusion, in which two electrons take part.

The most sensitive electrode turned out to be La2O3/CPE with 20% of nanoparticles in graphite paste (w/w). According to our knowledge, it is the first literature report about application of such a sensor. Sensitivity obtained in DPV experiments realized by optimized parameters in 0.1 mol L−1 acetate buffer was for La2O3/CPE of 3 mm diameter equal to 0.885 ± 0.016 µA/mg L−1 with the correlation coefficient r of 0.9993 and the detection limit of 0.004 mg L−1, while for commercially available sensor BDDE it was 0.135 ± 0.003 µA/mg L−1 with r of 0.9990.

Operation of the selected electrodes was verified using especially prepared herbal ethanol extracts which contained and did not contain anethole. In the last case, recovery was tested applying standard addition method. Anethole was also successfully determined in commercially available products, such as anise essential oil, which contains a large concentration of anethole, and in alcohol drinks like Metaxa, Ouzo, and Rakija, in which the considered analyte occurs on trace levels. The results obtained on La2O3/CPE and BDDE were statistically consistent, at 95% confidence level.

Conflicts of Interest

The authors declare that there are no conflicts of interest regarding the publication of this article.


This work was supported by the National Science Centre, Poland (Project no. 2015/19/B/ST5/01380).


  1. L. S. Moura, R. N. Carvalho Jr., M. B. Stefanini, L. C. Ming, and M. A. A. Meireles, “Supercritical fluid extraction from fennel (Foeniculum vulgare): global yield, composition and kinetic data,” Journal of Supercritical Fluids, vol. 35, no. 3, pp. 212–219, 2005. View at: Publisher Site | Google Scholar
  2. G.-W. Wang, W.-T. Hu, B.-K. Huang, and L.-P. Qin, “Illicium verum: a review on its botany, traditional use, chemistry and pharmacology,” Journal of Ethnopharmacology, vol. 136, no. 1, pp. 10–20, 2011. View at: Publisher Site | Google Scholar
  3. M. Wichtl, Herbal Drugs and Phytopharmaceuticals, MedPharm GmbH Scientific Publishers, Stuttgart, Germany, 2004.
  4. A. A. Shahat, A. Y. Ibrahim, S. F. Hendawy et al., “Chemical composition, antimicrobial and antioxidant activities of essential oils from organically cultivated fennel cultivars,” Molecules, vol. 16, no. 12, pp. 1366–1377, 2011. View at: Publisher Site | Google Scholar
  5. R. S. Freire, S. M. Morais, F. E. A. Catunda-Juniora, and D. C. S. N. Pinheiro, “Synthesis and antioxidant, anti-inflammatory and gastroprotector activities of anethole and related compounds,” Bioorganic and Medicinal Chemistry, vol. 13, no. 13, pp. 4353–4358, 2005. View at: Publisher Site | Google Scholar
  6. A. Kołodziejczyk, Natural Organic Compounds, PWN, Warsaw, Poland, 2013.
  7. S. Ariamuthu, V. Balakrishnan, and M. L. Srinivasan, “Chemical composition and antibacterial activity of essential oil from fruits of Illicium verum Hook. f.,” International Journal of Research in Pharmacology and Phytochemistry, vol. 3, pp. 85–89, 2013. View at: Google Scholar
  8. H. Ullah, A. Mahmood, and B. Honermeier, “Essential oil and composition of anise (Pimpinella anisum L.) with varying seed rates and row spacing,” Pakistan Journal of Botany, vol. 46, no. 5, pp. 1859–1864, 2014. View at: Google Scholar
  9. A. Raal, E. Arak, and A. Orav, “The content and composition of the essential oil Found in Carum carvi L. commercial fruits obtained from different countries,” Journal of Essential Oil Research, vol. 24, no. 1, pp. 53–59, 2012. View at: Publisher Site | Google Scholar
  10. M. K. Abderrezak, I. Abaza, T. Aburjai, A. Kabouche, and Z. Kabouche, “Comparative compositions of essential oils of Citrus aurantium growing in differents oils,” ournal of Materials and Environmental Science, vol. 5, no. 6, pp. 1913–1918, 2014. View at: Google Scholar
  11. M. K. Ibrahim, Z. A. Mattar, H. H. Abdel-Khalek, and Y. M. Azzam, “Evaluation of antibacterial efficacy of anise wastes against some multidrug resistant bacterial isolates,” Journal of Radiation Research and Applied Sciences, vol. 10, no. 1, pp. 34–43, 2017. View at: Publisher Site | Google Scholar
  12. P. Brereton, S. Hasnip, S. Bertrand, R. Wittkowski, and C. Guillou, “Analytical methods for the determination of spirit drinks,” TrAC Trends in Analytical Chemistry, vol. 22, no. 1, pp. 19–25, 2003. View at: Publisher Site | Google Scholar
  13. M. Przygodzka, D. Zielińska, Z. Ciesarová, K. Kukurová, and H. Zieliński, “Comparison of methods for evaluation of the antioxidant capacity and phenolic compounds in common spices,” LWT–Food Science and Technology, vol. 58, no. 2, pp. 321–326, 2014. View at: Publisher Site | Google Scholar
  14. M. Kowalcze and M. Jakubowska, “Voltammetric profiling of absinthes,” Journal of Electroanalytical Chemistry, vol. 776, pp. 114–119, 2016. View at: Publisher Site | Google Scholar
  15. J. Fiori, M. Hudaib, L. Valgimigli, S. Gabbanini, and V. Cavrini, “Determination of trans-anethole in Salvia sclarea essential oil by liquid chromatography and GC-MS,” Journal of Separation Science, vol. 25, no. 10-11, pp. 703–709, 2002. View at: Publisher Site | Google Scholar
  16. J. M. Murado, A. Alcazar, F. Pablos, and M. J. Martin, “LC determination of anethole in aniseed drinks,” Chromatographia, vol. 64, no. 3-4, pp. 223–226, 2006. View at: Publisher Site | Google Scholar
  17. K. Schulz, K. Schlenzc, R. Metaschd, S. Malt, W. Romhild, and J. Dressler, “Determination of anethole in serum samples by headspace solid-phase microextraction-gas chromatography–mass spectrometry for congener analysis,” Journal of Chromatography. A., vol. 1200, no. 2, pp. 235–241, 2008. View at: Publisher Site | Google Scholar
  18. E. Mehmeti, D. M. Stankovic, S. Chaiyo, L. Svorc, and K. Kalcher, “Manganese dioxide-modyfied carbon paste electrode for voltammetric determination of riboflavin,” Microchimica Acta, vol. 183, no. 5, pp. 1619–1624, 2016. View at: Publisher Site | Google Scholar
  19. N. Al-Qasmi, M. T. Soomro, M. Aslam et al., “The efficacy of the ZnO: α-Fe2O3 composites modified carbon paste electrode for the sensitive electrochemical detection of loperamide: a detailed investigation,” Journal of Electroanalytical Chemistry, vol. 783, pp. 112–124, 2016. View at: Publisher Site | Google Scholar
  20. P. Khanh Quoc Nguyen and S. K. Lunsford, “Square wave anodic stripping voltammetric analysis of lead and cadmium utilizing titanium dioxide/zirconium dioxide carbon paste composite electrode,” Journal of Electroanalytical Chemistry, vol. 711, pp. 45–52, 2013. View at: Publisher Site | Google Scholar
  21. V. Arun and K. R. Sankaran, “Nafion coated TiO2 and CuO doped TiO2 modified glassy carbon and platinum electrodes: preparation characterization and application of enhancement of electrochemical sensitivity of azines,” Journal of Electroanalytical Chemistry, vol. 769, pp. 35–41, 2016. View at: Publisher Site | Google Scholar
  22. G. Ziyatdinova, E. Ziganshina, S. Romashkina, and H. Budnikov, “Highly sensitive amperometric sensor for eugenol quantification based on CeO2 nanoparticles and surfactants,” Electroanalysis, vol. 29, no. 4, pp. 1197–1204, 2017. View at: Publisher Site | Google Scholar
  23. R. S. Nicholson and I. Shain, “Theory of stationary electrode polarography. Single scan and cyclic methods applied to reversible, irreversible, and kinetic systems,” Analytical Chemistry, vol. 36, no. 4, pp. 706–723, 1964. View at: Publisher Site | Google Scholar
  24. A. J. Bard and L. R. Faulkner, in Electrochemical Methods: Fundamentals and Applications, John Wiley & Sons, New York, NY, USA, 2nd edition, 2001.
  25. L.-H. Wang, C.-L. Chang, and Y.-C. Hu, “Electrochemical oxidation of fragrances 4-allyl and 4-propenylbenzenes on platinum and carbon paste electrodes,” Croatica Chemica Acta, vol. 88, no. 1, pp. 35–42, 2015. View at: Publisher Site | Google Scholar

Copyright © 2018 Mateusz Kowalcze et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

More related articles

 PDF Download Citation Citation
 Download other formatsMore
 Order printed copiesOrder

Related articles

Article of the Year Award: Outstanding research contributions of 2020, as selected by our Chief Editors. Read the winning articles.