INTRODUCTIONFood preservation has remained a problemthroughout the human history. It is caused by the activityof environmental microorganisms and enzymaticreactions in the products during their productionand storage [1, 2]. About a third of the populationin the developed countries are estimated to sufferfrom diseases transmitted through food especiallyfalsified [3]. Food safety is directly related to thedevelopment of chemicals that prevent or slow down thespoilage of these products.Sorbic and benzoic acids, as well as their salts, areknown to be widely used as food preservatives. Theirproduction is steadily increasing. These acids arecontained in some fruits, berries, dairy products. Sorbicacid is an unsaturated fatty acid and is used only as apreservative in food, animal feed, tobacco, cosmeticsand pharmaceuticals. It is metabolized like normalfatty acids, so this acid was assumed to have no sideeffects. Benzoic acid is a synthetic additive, used as apreservative and antioxidant. It is excreted by the humanbody through the kidneys.126Samoylov A.V. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 125–133There are numerous data on the health safety ofthese compounds in regulated food products. Recently,however, there are more discussions on the necessity todevelop scientific approaches to studying mechanisms oftheir toxicity [4, 5]. The interest in this problem is dueto by the detected adverse effects of the chronic andsubchronic benzoate and sorbate intake by both animalsand humans. Thus, adding benzoic acid to pig feedincreased the liver enzymes activity and changed theblood formula negatively, eventually damaging the liverand spleen, respectively [6].In vitro studies of human erythrocytes demonstratedthat sodium benzoate reduced the level of keymetabolic enzymes of amino acids (aspartate andalanine aminotransferase) and alkaline phosphatasesignificantly [7]. There is strong evidence that attentiondeficit and hyperactivity syndrome in children andanxiety conditions in rats could be associated withhigh doses of sodium benzoate [8–9]. Other researchersshowed that sodium benzoate caused numerous negativephysiological and biochemical changes in mice and rats.The changes included reducing the mass of reproductiveorgans and embryos and the level of sex hormones inmice [10]. As for human blood cell culture, sorbic aciddemonstrated the inhibitory effect on biochemicalreactions in the activated immune response [11].However, the mechanism of toxicity for thesepreservatives is still unclear. In addition, creating a newalgorithm for assessing food safety is debated a lot. Thealgorithm especially concerns foods containing severalfood additives because of their potential additive andsynergetic effect of toxicity [12]. It is yet to be found outif the food preservatives may exert increased activity inpeople with specific diseases or genetic defects.To rise up to the challenge, it is necessary to gobeyond standard toxicity tests to identify molecularbiological protection mechanisms and to identifybiomarkers responsible for the body reaction to theeffects of chemical compounds. All the more so, asmodern methodology and instrumentation system areable to tackle these complex problems. New approachesshould not only monitor and evaluate toxic effects, butalso result in the adequate test systems for modellingdetoxification and metabolism of food preservatives inthe human body. It is important to develop new modelsystems. They should be simple to execute, cheap, andable to simulate the reactions of the human body, bothon the physiological and molecular levels, with themaximum available accuracy.In this aspect, the special interest is given to thework on a comprehensive assessment of biomarkersof neurotoxicity and antioxidant enzymes activity indaphnia under the influence of food sweetener sucralose.It is due to the evidence that Gammarus zadachi andDaphnia magna crustaceans exposed to this sweeteneraltered their swimming behavior [13]. The tests werecarried out on these organisms to compare the activityof acetylcholinesterase (AChE), lipid peroxidationenzymes, and the ability to absorb oxygen radicals(ORAC assay) in them. The authors observed thestimulating effect of sucralose on the activity of AChEand lipid peroxidation, but not on the antioxidantcapacity (ORAC). In humans, an increased AChEactivity was also associated with neurodegenerativediseases such as Alzheimer’s disease, Parkinson’sdisease, multiple sclerosis, and restless legs syndrome.It is important to note that the data obtained in this workare consistent with those in other experimental studieson human cell cultures and vertebrates. However, planttest systems are also of interest, in particular Alliumcepa L. onion roots (Allium test).Traditionally, the Allium test has been usedas a bioindicator in numerous studies on toxicity,cytotoxicity, and genotoxicity of various chemicalcompounds. It is recommended by WHO expertsas a standard for the cytogenetic monitoring of theenvironment. Recently, it has been increasingly usedto assess the genotoxic potential of medicinal plants,food additives, and even ionizing radiation [14–17]. TheAllium test was an excellent eukaryot model in vivo.It was one of the few direct methods for measuringdamages in biological systems after exposure tovarious toxicants and mutagenes. Its main advantagesinclude the following characteristics. First, the apicalmeristematic root cells can show constant mitoticdivision. Second, the roots may incubate directlywith the object being tested. Third, these cells havelarge chromosomes, which allows a comprehensiveanalysis of DNA damage. In addition, the test indicatorswere shown to be more sensitive than the models onmicroorganisms, cell cultures, and even animals [15, 18].Allium cepa was also presented as an effective testobject in studying the reaction of plant cell biomarkersto chemical toxicants of different nature. It is knownthat chemical pollutants can induce the formation ofactive forms of oxygen. In its turn, oxygen can activateenzymes of peroxidation and result in damagingvarious biological molecules, including lipids. Thus,it was found that herbicide glyphosate and coppersalts significantly increased lipid peroxidation in plantcells [19, 20]. In our opinion, the Allium test can helpsignificantly expand our knowledge of the mechanismsof damage to biological systems of eukaryotes, includingthe damage after exposure to sorbic and benzoic acids.Moreover, no information was found on an effect ofthese preservatives on physiological and biochemicalparameters in the meristematic cells of onion roots.The aim of the research was to compare changes in themass growth, activity of lipid peroxidation enzymes,cytological and cytogenetic parameters of Allium ceparoots after treatment with sorbic and benzoic acids.STUDY OBJECTS AND METHODSIn the research we used such preservatives as sorbicacid (Alfa Aesar by Thermo Fisher Scientific) andbenzoic acid (Alfa Aesar by Thermo Fisher Scientific).127Samoylov A.V. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 125–133Allium cepa onion bulbs (Stuttgarter sort) of the samesize (2.5–3 cm in diameter) and mass (5–7 g) wereselected as a test organism. Dry scales were removedfrom the bulbs before incubation. Preliminarily, thegermination was conducted in 15 mL test tubes withbottled water for 2 days in the dark at 25°С.The bulbs with roots over 1 cm long were selectedfor further studies. Before treatment with benzoic andsorbic acid solutions, the average mass of the roots wasdetermined in a separate group of the control bulbs.Then the bulbs were transferred to the solutions ofthese acids in the bottled water and incubated for 2 or3 days depending on the purpose of the experiment.After the incubation, the roots were cut off, dried withfilter paper, and weighed [21]. The EC50 value wasdetermined by the concentration of the preservative,which retarded the root mass growth by 50% comparedto control, considering the average mass of the rootsbefore treatment with acids. For cytogenetic analysis,the apical meristematic cells of the roots were stainedwith acetoorcein (1 g of orcein dye was diluted in50 mL of 45% CH3COOH). The roots were placed in a70% solution of ethyl alcohol for the long-term storage.Next, instant squash preparations were obtained, theanalysis of which was carried out with the help of a lightmicroscope Axioskop 40, Zeiss.The lipid oxidation level was determined by theconcentration of malon dialdehyde (MDA) in the onionroots [22]. The sample weight of approximately 0.25 g tothe fourth decimal place was measured in a 15 mL testtube. Then 1 mL of trichloroacetic acid solution (Merck,Germany), concentration of 200 g/dm3, was added. Themixture was thoroughly stirred with a glass stick. Thenthe stick was washed with 3 mL of the same solution oftrichloroacetic acid. The tubes were tightly corked andcentrifuged at 1000 g and 4°C for 15 min. One milliliterof supernatant was transferred to a clean 15 mL testtube. Four milliliters of thiobarbituric acid solution(0.5 g of thiobarbituric acid (Diaem, Russia)) was addedto 100 mL of trichloroacetic acid solution (200 g/dm3).The test tubes were closed and placed in a water bathat 95°С for 30 min. Then the test tubes were pulled outand cooled in an ice bath. The cooled solutions werecentrifuged at 1000 g and 20°C for 10 min.The spectrophotometric detection was performedwith the obtained solutions at 600 and 532 nm. TheMDA content was calculated according to the formula:where ABS532 is the absorption value at 532 nm;ABS600 is the absorption value at 600 nm;K is the dilution factor;Ke is the molar coefficient of extinction;l is a beam path length, cm;mwt is the weight of the sample, g.The statistical processing of the results was carriedout in Microsoft Excel and Statistica programs (v. 12).In the paper, the analysis of average values by Student’scriterion with Fisher’s angular transformation was usedfor comparative estimation of percentages.RESULTS AND DISCUSSIONThe macroscopic parameters were studied andcomparatively evaluated, particularly, for the levelsof mass growth in the onion roots after treatment withbenzoic and sorbic acid solutions. According to theliterature review, the macroscopic parameters appearedmore sensitive in comparison with the cytological andcytogenic parameters [23]. This conclusion seemedlogical because these parameters reflected the finaleffect of all disorders in the plant cells. In this work,when calculating the growth of root mass, the averageweight of roots was subtracted both in control andexperimental samples before their treatment withpreservatives solutions. Thus, the EC50 overstatementerror was eliminated in these samples. In thepreliminary experiments the solutions of preservativeswere used with the concentrations not exceeding thepermissible levels for some food products, namely,1 g/L and 2 g/L. Death of the roots was observed after2 days of incubation. Therefore, we reduced the rangeof acid concentrations significantly. As a result, the rootgrowth and dose-dependent toxic effects were observedduring the same incubation period (Figs. 1 and 2). Theroots in the samples remained white and unchanged inshape throughout the incubation. However, there werestatistically significant differences between the controland test samples, namely, when treated with benzoicacid at concentrations of 0.01 (P < 0.1); 0.05 (P < 0.05);0.1 (P < 0.05) and 0.2 g/L (P < 0.05) and with sorbicacid at concentrations of 0.02 (P < 0.1); 0.1 (P < 0.05);0.2 (P < 0.05) and 0.3 g/L (P < 0.05). EC50 was 10 mg/Lfor benzoic acid and 110 mg/L for sorbic acid. Thus,these values differed significantly from the domesticregulatory norms on the content of these food additivesin certain types of food.As far as we know, this is the first study in whichEC50 values were identified for these preservatives inthe Allium test. At the same time, cyanobacteria withFigure 1 Root mass growth inhibition after treatment withsorbic acid (n = 10). * P < 0.05, ** P < 0.1Sorbic acid concentration, mg/L𝑥𝑥 􀵌𝐴𝐴𝐵𝐵𝑆𝑆􀍷􀍵􀍴 − 𝐴𝐴𝐵𝐵𝑆𝑆􀍸􀍲􀍲 ∗ 𝐾𝐾𝐾𝐾𝑒𝑒 ∗ 𝑙𝑙 ∗ 𝑚𝑚𝑤𝑤𝑡𝑡03060900 50 100 150 200 250 300 350Root mass growth, %Sorbic mg/l0204060801000 Root mass growth, %50100150200250300MDA concentration, μmol/gof wet weightEC50 *******× × ×128Samoylov A.V. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 125–133EC50 from 9 mg/L were the most sensitive to benzoicacid in similar studies using different organisms livingin water when treated for 14 days. In molluscs, fishand amphibians, EC50/LC50 values were determinedwithin 100–1291 mg/L for 24–96 h [24]. The resultsof these studies confirmed the high sensitivity of themacroscopic parameters in the Allium test.However, the studies on the toxicity of benzoic andsorbic acids using the Allium test focused mainly onthe microscopic indicators reflecting the peculiarities ofcell division and chromosomal aberrations occurring inits process, and EC50 was not determined. At the sametime, the tested concentrations of preservatives wereusually much higher than the EC50 values we found. Sothe exposure of the roots to the preservative solutions inthese studies usually did not exceed several hours [25,26]. We believe that such experimental conditions aresuitable only for acute toxicity testing. They are totallyunacceptable for the study of subchronic and chronicconsequences of negative effects, especially at thebiochemical level. The last aspect should be consideredthe most interesting in the case of food preservatives.Therefore we believe that the Allium test schemepreviously proposed for environmental monitoringdid not lose its relevance for studying the toxic effectsof these preservatives. The Allium test included acomparative analysis of macro- and microindicators atconcentrations of toxicants within their EC50 [23].The mitotic index is one of the microindicators in theAllium test which is used as an indicator of the level ofcell proliferation. It is known that the dose-dependentdeviation of the mitotic index in the experimentalsamples compared to the control values, both increasingand decreasing, indicates cytotoxicity of the testedchemical. In our previous study, the mitotic index ofcells of their meristem was decreasing significantly withan increase in concentrations of preservatives (Table 1).In testing highly toxic doses of sorbic acid (from 1 to2 g/L), the mitotic index decreased only slightly whenthe concentration of this acid increased [26]. This dataconfirmed the previous assumption that there may bedifficulties in interpreting the research results due to thehigh concentration of preservative.The cytogenetic analysis was carried out on thesquash preparations of the apical meristematic cells ofonion roots obtained in the previous study. The analysisdetermined the accumulation dynamics of chromosomalaberrations when the concentrations of sorbic andbenzoic acids were increased. According to Table 1,when acid concentrations increased, the proportion ofmitosis pathologies also increased, peaked, and thendecreased. It is noteworthy that the highest percentageof chromosomal aberrations coincided with acidconcentrations close to the corresponding EC50 of thesepreservatives. The drop in chromosomal aberrations athigh concentrations of acids is probably associated witha significant decrease in the number of divisible cells inthe meristematic cells of roots.The types of major chromosomal aberrationsdetected in the experiment are shown in Fig. 3. Theanalysis of the data allows us to conclude that sticknessof chromosomes in metaphase and chromosomes withlaggard in anaphase make the main contribution to thespectrum of chromosomal aberrations. These anomaliesaccount respectively for aberrations ranging from 23.8%to 70% (for sticky metaphase) and from 13.6% to 45.2%(for chromosome with laggard). Also, there were thefollowing aberrations of the mitosis process detected inmicropreparations: C-mitosis, multiple fragmentationof chromosomes, change in the spatial orientation ofchromosomes at the metaphase stage in cells. The leastobserved anomalies included bridges and fragments(about 2%, depending on the concentration of the testedsubstances).It seems remarkable to consider the whole spectrumof aberrations. The most numerous anomalies foundwhile analyzing biomaterial can be due to the effectsFigure 2 Root mass growth inhibition after treatment withbenzoic acid (n = 10). * P < 0.05, ** P < 0.1Table 1 Mitotic index and chromosomal aberrationsin meristematic cells of onion roots after exposure to benzoicand sorbic acids (n = 10)Acid concentration,g/LMitoticindex, %Chromosomal aberrationsbased on the totalnumber of cells, %Benzoic acid0.01 12.02 ± 0.48c 0.88 ± 0.14a0.02 10.20 ± 0.49a 1.00 ± 0.16a0.1 6.89 ± 0.33a 0.50 ± 0.09b0.2 0.75 ± 0.42a 0.05 ± 0.04aSorbic acid0.02 9.63 ± 0.42a 0.95 ± 0.14a0.1 6.81 ± 0.38a 0.79 ± 0.12a0.2 1.52 ± 0.16a 0.10 ± 0.04a0.3 0.00 ± 0.00a* 0.00 ± 0.00a*Control 12.97 ± 0.48 0.30 ± 0.08а P < 0.05, b P < 0.1, c P < 0.15, * 6503 cells were observedBenzoic acid concentration, mg/L􀍷􀍵􀍴 − 𝐴𝐴𝐵𝐵𝑆𝑆􀍸􀍲􀍲 ∗ 𝐾𝐾𝑒𝑒 ∗ 𝑙𝑙 ∗ 𝑚𝑚𝑤𝑤𝑡𝑡300 350mg/l0204060801000 50 100 150 200 250Root mass growth, %Benzoic acid concentration, mg/l0481216Контроль 0,01 0,05 0,1 0,2MDA concentration,μmol/g of wet weightBenzoic acid concentration, g/l020406080100120Контроль MDA concentration, μmol/gof wet weight0.2 0.3 0.4 0.5 0.6acid concentration, g/l0100200300Контроль 0,02 0,2MDA concentration, μmol/gof wet weightSorbic acid concentration, g/lEC50****** ** **** *129Samoylov A.V. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 125–133of mitotic spindle disorder and changes in the surfaceof chromosomes. The aberrations occur in this groupprobably due to the influence of the tested substance onthe proteins regulating the work of the mitotic spindle inthe cell [27]. On the other hand, bridges, fragments, andmicronuclei are associated with clastogenic aberrations(arising from the fracture of the chromosome andanomalies of the further molecular genetic processes,unequal translocation or inversion of the chromosomesegments). In the study [28], the analysis of genotoxicityof sodium benzoate (in concentrations from 20 to100 mg/kg) discovered the prevalence of aberrationsrelated to mitotic spindle disorders and changes in thesurface of chromosomes. These are the main types ofagglutination and C mitosis disorders. The clastogeniceffect of the factor was not recorded at all for thisindicator.On the other hand, studies with high concentrationsof sodium benzoate exposed a much wider spectrumof chromosomal aberrations. The aberrations includedagglutination and fragmentation of chromosomes, theirreduction, the formation of binuclear cells, chromosomalbridges and other disorders [29]. According to data [30],treating cells with sorbic acid resulted in thechromosomal aberrations associated with mitotic spindledisorder. Clastogenic aberrations were not detected.Similar data were obtained in the study of the effects ofsorbic acid on the formation of micronuclei in cells [31].This study recorded reliable mitodepressiveand genotoxic effects at very low concentrations ofpreservatives (10 and 20 mg/L for benzoic and sorbicacids, respectively). It is important to note that the dataobtained were consistent with the results on genotoxicityof these acids and their salts for human and animalcell culture. The results were published in a number ofpapers, describing the exposure to both low and highdoses of these preservatives. Thus benzoic acid causedsister chromatid exchange, chromosomal aberrations,and micronuclei formation in human lymphocytecells [32]. Other researchers demonstrated the genotoxiceffect of sodium sorbate on Chinese hamster cells, aswell as clastogenic, mutagenic and cytotoxic effectsof sodium benzoate on the cell culture of humanlymphocytes [29, 33].MDA concentration is commonly used as anindicator of lipid peroxidation when the tissues areexposed to chemical toxicants. MDA was measured inthe onion roots of the control and experimental groupsobtained in our previous study. In the experimentalgroups of onion roots this biomarker analysis showedFigure 3 Stained preparations of meristemic cells of oninonroots: (a, b) fragmentation in anaphase; (c, d) fragmentsof chromosomes in anaphase; (e) C-mitosis; (f) anaphasewith laggards; (g) fragmentation in metaphase; (h) stickymetaphase, without pathologies: (i) prophase;(j) metaphase; (k) anaphase; (l) telophaseFigure 5 Effects of different doses of sorbic acid on MDAlevel in roots after 2 days of incubation (n = 10). * P < 0.05(a) (b) (c)(d) (e) (f)(g) (h) (i)(j) (k) (l)Figure 4 Effects of different doses of benzoic acid on MDAlevel in roots after 2 days of incubation. * P < 0.05, ** P < 0.1ControlControlg/Lg/L130Samoylov A.V. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 125–133a significant dose-dependent increase in the MDAlevels (by 760%) compared to the control samples. Atthe same time benzoic acid did not have a significanteffect on the process (Figs. 4 and 5). Since sorbic acidcan be subjected to partial hydrolysis, MDA wasmeasured in a solution of sorbic acid (0.2 g/L) after4 days of incubation, but its content exceeded only by57% compared to the values in the control samples ofonion roots.Since the biomarkers of oxidative stress usuallyshowed a two-phase response rather than a linearresponse, we expanded the range of sorbic acid dilutionsand increased the period of incubation with acids up to3 days to identify the dynamics of MDA biosynthesis.Indeed, this pattern of the two-phase response wasconfirmed again [13]. With an increase in sorbic acidconcentration, the level of this biomarker increasedevenly at first, reached its peak, and then droppeddose-dependent until it reached MDA value in thecontrol samples (Fig. 6). Like in the previous study,the maximum concentration of MDA was recorded forsorbic acid at 200 mg/L. The concentration was abovethat in the control group by almost 2000%, i.e. lipidoxidation level also increased with exposure time.As far as we know, these are the first experimentsto study the dynamics of lipid oxidation biomarkergeneration in the Allium test after exposure to benzoicand sorbic acids. Both acids reduced root growth andthe mitotic index of apical meristematic cells. However,these negative phenomena were accompanied by thesimultaneous increase in MDA only in the case of sorbicacid. These results are consistent with the data on thetreatment of wheat seeds with benzoic acid [34]. Thisstudy did not detect any change in lipid peroxidationactivity different from control when treated with lowconcentrations of this preservative.In the case of benzoic acid, its toxic effects wereprobably not associated with oxidative damage to lipids.In addition, the study showed the protective reaction inthe plant cell to benzoic acid in concentrations of 1 to10 mM. The reaction was accompanied by an increasedactivity of glutamate and malate dehydrogenase,enzymes activating catabolic and metabolic processes[35]. In the current study, both the MDA level androot mass growth increased with an increase in theconcentration of sorbic acid from 20 to 200 mg/L(Figs. 5 and 6). Thus, there was a clear correlationbetween the physiological index and MDA, the latterbeing even more sensitive.According to the literature, the meristematic cellmitotic index in the control samples when using theAllium test is both close to our result (12.97 ± 0.48) andwell below it [20, 36, 37]. This indicator could changedepending on the quality of the batch of onions, itsvariety, and storage conditions. However, the questionremained whether there was a dependence between theFigure 6 Effect of different doses of sorbic acid on MDA level in roots after 3 days of incubation (n = 10). * P < 0.05, ** P < 0.1ControlTable 2 Inhibition of the root mass growth and the mitoticindex after treatment with sorbic acid (n = 9)Sorbic acidconcentration,g/LRoot massgrowth, gMitoticindex, %Chromosomalaberrations basedon the total numberof cells, %0.02 0.19 ± 0.07 6.84 ± 0.40a 0.29 ± 0.090.2 0.05 ± 0.02a 0.00 ± 0.00a* 0.00 ± 0.00a*Control 0.27 ± 0.04 7.97 ± 0.89 0.15 ± 0.06a P < 0.05, *1748 cells were observedFigure 7 Effect of different doses of sorbic acid on MDA levelin roots after 2 days of incubation (n = 9). * P < 0.05, ** P < 0.1Controlg/Lg/L131Samoylov A.V. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 125–133initial level of the mitotic index and the ability of rootcells to fully respond to the effects of toxicants.To this end, another study was conducted toexamine the toxic effect of sorbic acid solutions at lowand high concentration on onions with a small part ofmeristematic dividing cells in control. In this study, themiotic index in the control samples (Table 2) was 40%lower than that obtained in the previous study (Table 1).The conditions of the Allium test in the previous and thecurrent study did not differ. The comparative analysisshowed the following negative trends in the results.First, the roots in the experimental groups became softand acquired a yellowish hue after 2 days of incubationwith the acid. Second, there was no gain in the rootmass compared to control (Table 2) when treated witha high-concentration acid solution (0.2 g/L), whereas inthe previous study the gain was 25%, and roots did notchange the color (Fig. 1). Similar negative changes wererecorded at the biochemical level in MDA measurement(Fig. 7). The lipid oxidation activity in these samplescompared to the previous study was significantly higherboth in the test and control samples.According to the obtained results, it seems advisableto select batches of bulbs before the Allium test. Thisselection is necessary as the low values of the mitoticindex may result in false positive results, both in termsof EC50 estimates and biochemical indicators.CONCLUSIONThe results of this study showed that sorbic andbenzoic acids caused toxic effects in the roots of Alliumcepa. These preservatives affected the physiological,biochemical, cytological, and genetic characteristics ofthe plant system. Treating onion roots with these acidsin concentrations of 1 and 2 g/L, which are acceptablefor some food products, was so highly toxic as to leadto their death. When concentrations of these acidsdecreased, EC50 limits for benzoic and sorbic acids wereshown to be 10–20 and 20–100 mg/L, respectively.These concentrations of preservative solutions induced a50% retardation in root growth, a significant decrease inthe mitotic index, especially in the case of sorbic acid,and almost a triple increase in chromosomal disorders.Thus, these preservatives at very low concentrationsgave a chronic and subchronic toxic effect. Basedon the conducted studies, it is necessary to use theconcentration of food preservatives within their detectedEC50 values to assess these toxicity indicators in theAllium test. If these conditions are met, it is possible tosimulate the processes of detoxification and metabolismfor these compounds, both at the cellular level and thewhole organism.Therefore, it can help gain a better understandingof the biological actions of these agents. Indeed, thenegative effects found under these conditions for sorbicacid, but not benzoic acid, were correlated with the lipidoxidation biomarker. In this regard, we believe that thestudy of this biomarker can provide valuable informationfor monitoring and predicting early effects of sorbic acidon animal and human cells. Yet, it is probably necessaryto study the role of catabolic processes to determine themolecular mechanisms of activation of enzymes withbenzoic acid [35].CONFLICT OF INTERESTThe authors state that there is no conflict of interest.