The effect of roasting, storage temperature, and ethanoic basil (Ocimum basilicum L.) extract on the oxidative stability of crude sesame (Sesamum indicum L.) oil

Abstract Sesame (Sesamum indicum L.) is an annual oilseed crop that is widely used for crude sesame oil (CSO) extraction. CSO is unrefined, thus, may impact the storage stability due to the uncontrolled processing and handling conditions, and absence of preservatives. The purpose of this study intends to analyze the fatty acid profile and oxidative stability of CSO. In a complete randomized (2*2*2) factorial design, clean sesame seeds (3.92% moisture content) were used for oil extraction with/without roasting treatment (200°C). The fatty acid profile and oxidative value of the CSO were analyzed. The roasted sesame seed oil extraction yield was higher (50.9%) and composed of 83.15% unsaturated fatty acids with an omega‐6 to omega‐3 ratio of 95.3. The ratio of polyunsaturated fatty acids (PUFAs) to saturated fatty acids (SFAs) was 2.9, while the moisture content of the CSO was 0.37%, exceeding the tolerable limit. The predominant fatty acids of CSO include palmitic, stearic, oleic, and linoleic acids. Elevated thermal condition during processing and handling speeding up oxidation exceeded the tolerable limit after 25 days of storage, while the ethanoic basil extract was found to inhibit oxidation in the range of 16.38%–90% (p‐value < .05). The peroxide value (PV), para‐anisidine value (p‐AV), and total oxidation (TOTOX) value of CSO with 50 ppm (parts per million) basil extract were detected within the range of 0.29–3.92, 0.75–2.59, and 1.57–8.6 milliequivalents (meq) O2/kg oil, respectively, below the tolerable limit. Nevertheless, basil extract's antioxidant property was declined during prolonged storage, in particular, at elevated temperature. The use of organic extracts of locally available sweet basil herb is capable of mitigating oxidation and substituting inorganic antioxidant for a healthier diet.


| INTRODUC TI ON
Oils, also known as fats, are important structural and functional components of foods (Akoh, 2017). The global oil consumption rose to approximately 600 million tons in 2019/2020, while plant-based edible oil accounted for 203.83 million metric tons by 2018/19 (Statista, 2019). The use of plant-based edible oils raising interest in the consumer market might be due to its nutritional and energy provision in daily diet, organoleptic profile enhancing property, pharmaceutical and cosmetic, health benefits, and nonfood applications (El-hamidi & Zaher, 2018;Kumar et al., 2016;Mensah et al., 2018). Sesame seed is a rich source of protein and essential fatty acids (EFAs), minerals, and antioxidants (Deme et al., 2017). It has medical importance due to the presence of sesamin and sesamolin to lower cholesterol accumulation, prevent high blood pressure, triglyceride linoleate to inhibit malignant melanoma growth and demulcent, used as cosmetics such as mildly laxative, emollient, poultice, a preservative of antimicrobial activity, and antioxidant (lignans) to prevent oxidation (Anilakumar et al., 2010;Gharby et al., 2017). Flavor compounds in sesame-based products include hydrocarbons, aldehydes, ketones, alcohols, acids, esters, furans, sesamol, and pyrazine which significantly changed the sesame oil quality and stability regardless of seed composition and physical quality (Elkhaleefa & Shigidi, 2015) and differences in extraction method (Hou et al., 2019;.
The small-and medium-scale entrepreneurs inspired by supplying edible oil from locally available oil crops due to edible oil shortage and imported oils are expensive and have insufficient control.
Sesame seed is the dominant oilseed and crude sesame oil extraction and consumption is predominant in the community. However, quality degradation, such as discoloration, rancid odor, and unpleasant flavor during storage, confuses regarding oil quality and public health.
Oxidation metabolites are the result of quality degradation such as fatty acid interactions, cleavage of short-chain fatty acids, and physicochemical and structural transformation of fatty acids (Medhujith & Sivaakanthan, 2019;Vaskova & Buckova, 2015), undesirable organoleptic and toxic metabolites' formation (Adibhatla & Hatcher, 2010;Ramana et al., 2017). Studies have reported that the use of oils and oily food with oxidative metabolites composed of reactive oxygen species (ROS) damages various organs in the body by producing cytotoxic and genotoxic compounds that lead to chronic diseases (Jung et al., 2016), disturbance of the redox state of the body, weakening the antioxidant network associated with a higher risk for atherosclerosis, diabetes and obesity (Sies et al., 2005), cancer (Rockenbach et al., 2011;Zhong & Yin, 2015) that increases the pathogenesis of myeloma and lymphoma by damaging DNA (Tandon et al., 2013), chemotherapy in non-Hodgkin's lymphoma (El-mezayen et al., 2015), aging (Liguori et al., 2018), increase in plasma total cholesterol level, larger atherosclerotic lesions (Hao & Friedman, 2014), and atherogenesis that affects the arterial wall leading to heart diseases (Münzel et al., 2017) and cell death (Gaschler & Stockwell, 2017;Stockwell et al., 2017;Wenzel et al., 2017).
The rate of oxidation can, however, be controlled or reduced.
Hence, an understanding of the type and concentration of fatty acids, and valency of ions, processing and handling conditions, and mechanisms of oxidation is important (Thanonkaew et al., 2006). oilseed pretreatments (Kombe & Temu, 2017), direct light exposure (Dalsgaard et al., 2010), presence of enzyme ferrous atoms interacting with the cis-cis-configured fatty acid during extraction in the presence of oxygen at an elevated temperature (Medhujith & Sivaakanthan, 2019;Silvagni et al., 2012;Vaskova & Buckova, 2015), storage duration , and presence of antioxidants (Taghvaei & Jafari, 2015). Oxidation involves three steps: (I) initiation, (II) propagation, and (III) termination reaction, as shown in Figure 1, which produced unstable and stable radicals, compelling to bear in mind the safety and quality of oils and consumer health complications (Choe & Min, 2009).
Sweet basil (Ocimum basilicum L.) is a culinary herb that belongs to the Lamiaceae family, widely produced and used as a fresh vegetable or dried form in food as a preservative and flavor enhancer.

Practical Application
The crude sesame oil (CSO) oxidation contributor factors identified were: • Elevated time/temperature roasting conditions, extraction machine exposure to environmental air, longer holding time during extraction, and a higher ambient temperature during storage.
The aim of this study was designed to determine the CSO oxidation with roasting treatment and application of ethanoic basil extract during 35 days of storage at ambient condition and 65°C.

| Sample preparation
Sesame seeds were collected from Western Tigray, Ethiopia  Table 1. The CSO sample was drawn during 0, 3, 7, 12, 17, 21, 25, 30, and 35 days for F I G U R E 1 Summary of oil oxidation mechanisms involves the initiation, propagation, and termination reaction to produce stable and unstable radicals and other derived compounds responsible for oil rancidity and application of natural and antioxidants donating a hydrogen atom or transform metallic ions to prevent oxidation and neutralize radical formation F I G U R E 2 Crude sesame oil extraction flow diagram using a mechanical milling and cold pressing method

| Crude basil extract preparation
The ethanoic basil extract was extracted according to Sholichah () procedure. Basil powder (gram) and petroleum ether (

| Fatty acid composition analysis
The moisture content of sesame seed and sesame oil was determined Saturated fat is the summation of fatty acids without double bonds in the chain of fatty acids and calculated as; � * 100 and monounsaturated fat, the summation of fatty acids, contains single and double bonds in their chain The polyunsaturated fat, the summation of fatty acids, contains more than one double bond in their chain structure.

Peroxide value
The peroxide value (PV) of the CSO was determined according to the International Dairy Federation (AOAC 920.160), as described by Shantha and Decker, (1994). All the reagents used (barium chloride dihydrate, iron (II) sulfate solution (0.5 g of FeSO 4 .7H 2 O), hydrochloric acid, ammonium thiocyanate, and chloroform-methanol) were of analytical grade. Iron (II) chloride solution and ammonium thiocyanate solution were prepared in the laboratory. About 0.2 g of sesame oil was put into a conical flask (250 ml) and vortexed with 9.8 ml chloroform-methanol (7:3V/V) for 5 s. A 50 µl ammonium thiocyanate solution was added and vortexed for 4 s, followed by the addition of 50 µl iron (II) solution. The mixture was vortexed for 2-4 s, absorbance reading at 500 nm performed immediately but no longer than 10 min. The absorbance of the blank cell containing all reagents, except the oil sample, was measured at a wavelength of 500 nm. The peroxide value (PV) was computed as; Where: A s is the absorbance of the sample solution, A b is the absorbance of blank, S is the slope, and mo is the sample weight.

Para-anisidine value
The p-anisidine value (p-AV) was determined according to the AOCS method (1995) described by Abdelazim et al. (2013). Analyticalgrade chemicals isooctane and p-anisidine in glacial acetic acid (1 ml of 0.25%, w/v) were used for analysis. Oil samples (~0.5 L) in a flask were dissolved with isooctane (25 ml) to measure the absorbance at 350 nm using a ultraviolet-visible (UV-vis) spectrophotometer.
Five milliliters of the above mixture was mixed with 1 ml of 0.25% panisidine in glacial acetic acid (w/v), allowed to stand for 10 min, and the absorbance was taken at 350 nm using a UV-vis spectrophotometer against the blank. The total oxidation (TOTOX) value was taken as the summation of the para-anisidine (p-AV) value and twice that of the peroxide value (pV). The AV and TOTOX value were computed as follows; Where: A s is the absorbance of the sample solution after reaction with the p-anisidine reagent; A b is the absorbance of the fat solution without the p-anisidine reagent and m is the mass of oil sample (g).

| Data analysis
A complete randomized design with a factorial arrangement was used. The mean ±standard deviation (SD) was generated by a JMP ® Pro 13.0.0 statistical software at 95% confidence interval (CI). Tukey for statistical significance test was used, whereby the predictor variables were roasting temperature, storage temperature, basil extract, and storage time. The regression function was generated.

| Crude sesame oil
The oil extraction yield of white sesame seed was obtained as 50.9% and 45.8% with and without roasting treatment, respectively.
Increased extraction temperature, pressure, and smaller particles similarly increase oil extraction yield and reduce extraction holding time (Döker et al., 2010;Elkhaleefa & Shigidi, 2015). In the meantime, extraction yield depends on the extraction method (Junpeng et al., 2019), operating conditions, and solvent type/solvent:seed ratio (Osmanet al., 2019). Sunflower seeds' oil expression efficiency was improved with increased thermal treatment, explaining why the seeds' mild thermal treatment increases cell porosity and weakens the cell membrane resulting in increased oil extraction efficiency (Guradil et al., 2020;Sánchez Chino et al., 2019). Fatty acids identified in CSO were palmitic, stearic, oleic, linoleic, linolenic, arachidic, and eicosanoid acids which are presented in Table 2. The amount of unsaturated fatty acids was 83.15%, omega-6 to omega-3 ratio of 95.3 within the recommended limit for a healthier diet (3:1-4:1) (Simopoulos, 2010). The unsaturated fatty acids cause an upsurge in oxidation sensitivity and storage instability due to the unstable double bonds in the chains of fatty acids (Maszewska et al., 2018;Syed et al., 2016). The lower level of linolenic acid (omega-3 fatty acid) in CSO might be due to oxidation (Maszewska et al., 2018). According to Yun and Surh (2012) investigation, fatty acid composition of edible oils is a predictor of oxidation and oxidative stress, besides the impact of seed pretreatments and processing Abou-gharbia et al., 1997Abou-gharbia et al., , 2000.
The ratio of omega-6 to omega-3 essential fatty acids is an indicator of healthier diet and the lessened saturated fatty acid intake.
For example, increasing the ratio of omega-6 to omega-3 essential fatty acids prevents up to 70% health risks of cardiovascular diseases, infection, and metabolic disorders (Hashempour-baltork et al., 2017; and promote a healthy development (Candela et al., 2011). On the other hand, the total fatty acid content, individual fatty acid level, saturated and unsaturated fatty acids (Tabee et al., 2008), and the ratio of omega-6 to omega-3 were significantly affected by the soil composition and climatic condition, season and year of harvest (Wacal et al., 2019), and processing (Dar et al., 2019;Hama, 2017). The CSO moisture content (0.37%) exceeded the recommended limit for storage and overall oil quality (0.2%) according to the CODEX specification which contributes to hasten oxidation.
The higher moisture content might be due to lack of refining and conditioning effect during extraction. Choe and Oh (2013) and Gumus and Decker (2021) investigated high moisture of oils and oily foods which were sensitive to oxidation.

| Oxidation
The residuals are the differences with the observed and predicted values of oxidation data presented in a scatter plot, Figure 3, which explains that the residual oxidative data were inclined to one side, in particular, the peroxide and TOTOX residual with nearly normal distribution with nonlinear association. The para-anisidine residual data are, however, scattered and normally distributed which explains that the oxidative data were defined to predict the oxidative quality of CSO. CSO oxidation was linearly associated and continued to build up during storage, as shown in Figure 4. Tabee  reported that the primary, secondary, and overall oxidation indicators, including with antioxidants, continued to increase with time.
The overall oxidation of CSO exceeded the tolerable limit according to the CODEX recommendation for edible vegetable oil after 25 days of storage at ambient condition. This explains that the shorter CSO shelf life might be due to inadequate processing and handling conditions. When CSO is unrefined, an enzyme-induced biochemical reaction might be introduced, causing a high CSO moisture content, exposure of the extraction process, and filling of oxygen and light that catalyze oxidation. Vidrih et al. (2010) reported that unrefined oils are vulnerable to oxidation than refined oil. In the meantime, pretreatment, processing, packaging, storage condition, and preservatives affect the oil quality, safety, and stability (Ali  was estimated to be no more than 3 weeks at ambient condition storage, unless the storage condition such as hotness and CSO moisture content reduced, refining process introduced and preservatives were applied, Figure 6.

| Effect of roasting
The effect of roasting treatment on CSO oxidation is presented in  (Durmaz & Gökmen, 2010;Suri et al., 2019). Elkhaleefa & Shigidi (2015) investigated the seed quality, operating conditions, and processing techniques such as increased holding time (increase yield), roasting temperature (150-250°C), pressing and stirring speed (≥350 rpm), and extraction temperature (≥40°C). According to Xu-yan et al. (2012) investigation, the flavor compounds of different sesame seed varieties were increased with increased roasting temperature. A similar finding was reported by Suri et al. (2019) where peanut oil extraction yield was increased after roasting pretreatment. Roasting also improves the flavor/aroma profile, imparts color change and bioactive compounds, and negatively affects the oxidative stability and nutritional value. CSO from roasted sesame seed develops amber brown color, deep flavored, and thicker in viscosity which might be due to the roasting-induced Maillard reaction, oxidation, and degradation/hydrolysis of the major constituents that include fats, proteins, carbohydrates, and cleavage of short-chained fatty acids. This explains that roasting and conditioning affect oil quality. Ribeiro, Eduardo, et al. (2016), Ribeiro, Nicacio, et al. (2016) and Yan et al. (2016) reported that roasting impacts physical quality, flavor compounds, antioxidants, essential fatty and amino acids.
Unsaturated fatty acids were largely transformed as a result of heatinduced oxidation and fatty acid modification (Hama, 2017;Hou et al., 2019).
During sesame oil extraction, sesame seed was conditioned (25 kg seed:1 L water) for the purpose of ease of seed breakage and increased oil release. Udoh et al. (2017) and Junpeng et al. (2019) described that increase in moisture content of oilseeds improves oil extraction yield and reduces energy expenditure while affecting oil quality. However, the higher moisture content of the seed results in increased moisture content of the unrefined oil (0.37%) which might be responsible for initiating an enzyme-induced oxidation and microbial growth (Choe & Oh, 2013). According to Dymińska et al. (2017) reports, oil stability was influenced by the process condition and composition. Pumpkin and brown sesame seeds' oil yield, saturated and monounsaturated fatty acids were increased, while the moisture content and polyunsaturated fatty acids were decreased with roasted seeds Hama, 2017).
The PV of CSO measured immediately after extraction was as high as 0.13 and 0.49 meq O 2 /kg oil and p-AV was 0.02 and 1.10 meq O 2 /kg oil with and without roasting treatment, respectively. The higher PV and p-AV of CSO without roasting treatment after extraction explain that enzymes were lively to induce oxidation and other side reactions while roasting represses enzymes.
According to Fu et al. (2009) andSilvagni et al. (2010) reports, endogenous enzymes enhance biochemical reactions including oxidation and rancidity development. Yet, PV and p-AV detected F I G U R E 4 Crude sesame oil oxidation (milligram equivalents (meq) O2/kg oil) pattern during 35 days of storage at ambient condition in a blue-brown glass bottle package

TA B L E 3
The oxidative value of crude sesame oil (meqO 2 /kg oil) (n = 6) during 35 days of storage at ambient condition in a blue-brown glass before and after roasting treatment after roasting treatment explain that environmental exposure, longer holding time, and reaction-induced oxidation were prominent. The exposure of extraction machine to environmental air with longer holding time during milling and pressing significantly affects the oxidative value. Silvagni et al. (2012) illustrate that the use of extraction machine and machine type responds differently to oil oxidation. In addition, other studies have reported that agroecological condition, environmental factors (water stress, or salinity and coldness) and botanical families, extraction method, and types of fatty acids contribute to oxidation differently (Gharby et al., 2017). Oilseed composition includes moisture content, types of fatty acids and antioxidants (Carvajal et al., 2009;Silvagni et al., 2010Silvagni et al., , 2012, seed maturity and quality, storage and handling condition (light, temperature, air) (Sanaeifar & Jafari, 2019;Silvagni et al., 2010) which affect the oxidative stability of edible oils.

| Effect of storage temperature
Effects of storage temperature on the PV of CSO during 35 days of storage at 65°C temperature and ambient condition, presented in

| Effect of basil extract
Oxidation cannot be avoided, yet it can be minimized as the rate of oxidation is determined considering processing and handling conditions. The use of basil extract elucidates a prominent potential to prevent/reduce oxidation and extend oil shelf life, presented in

| CON CLUS ION
Vegetable oils are basic daily human diets. However, oxidationinduced quality degradation and carcinogenic metabolite accumulation remain the challenges in the edible oil value chain. CSO is a nutritionally rich unrefined cooking oil. Oxidation is continuous, irrespective of the processing and handling conditions and composition.
Regardless of the extraction yield increment, roasting accelerates oxidation, discoloration due to Maillard reaction, unpleasant flavor development, and thick oil viscosity due to the cleavage of proteins and carbohydrates. Thermal treatment of oil seeds, oil, and oil storage also promotes the enhanced rate of polymerization and quality degradation. Oxidation metabolites are carcinogenic to humans due to the abstraction of hydrogen and formation of unstable radical compounds. However, oxidation is desired to some extent and cannot be completely avoided and yet it can be minimized. Process optimization and reduced thermal storage in a light-protected packaging reduce oxidation. Basil extract is an organic preservative that shows a prominent potential to mitigate CSO oxidation. The PV, p-AV, and TOTOX value of CSO with basil extract were measured within the range of 0.29-3.92, 0.75-2.59 and 1.57-8.6 meq O 2 /kg oil, respectively, during storage period. However, the oxidation value of CSO without basil extract increases more than threefold. Yet, research should be done to explore the bioactive compounds of sweet basil and commercialize in the food processing as potential preservatives and functional foods for a healthier diet.

ACK N OWLED G EM ENTS
We acknowledge the Dutch organization for internationalization in education (Nuffic), Organization for Women in Science for the Developing World (OWSD), and the World Academy of Sciences (TWAS) for financial support. We also thank the Tigray Sesame oil extraction plant and Addis Adigrat pharmaceutical industry for allowing us to use their facilities and technical support.

CO N FLI C T O F I NTE R E S T
The authors declare there is no conflict of interest.

AUTH O R CO NTR I B UTI O N S
Authors equally contributed to conceptualization and writing.

PA RTI C I PATI O N
Not applicable.

CO N S E NT FO R PU B LI C ATI O N
Not applicable.

DATAT AVA I L A B I L I T Y S TAT E M E N T
Under the caption of each figure/result, data has been provided.