Phytochemical Constituents of Indonesian Adlay ( Coix lacrima-jobi L.) and Their Potential as Antioxidants and Crop Protection Agents

: Adlay ( Coix lacryma-jobi L.) is a cereal crop that has traditionally been used for medicinal purposes. It is processed into nutritious food in China and Southeast Asian countries. This study assesses the phytochemical constituents of this plant and their potential as antioxidants and crop protection agents. The methanolic extracts from seeds of Indonesian adlay ( C. lacryma-jobi ) varieties including Agrotis , Ma-yuen , and Aquatic , were tested against 2,2-diphnyl-1-picrylhydrazyl (DPPH) and 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) to determine their free radical scavenging activity. The relationship between extraction solvents, phytochemical composition, and antioxidant activity was analyzed statistically using principal component analysis (PCA) to classify them based on the similarities among the components. The potential use of the phytochemicals as crop protection agents was also explored through a review of the literature. The Agrotis variety demonstrated the highest antioxidant activities (IC 50 DPPH = 741.49 and ABTS =152.69 µ g/mL). The ethyl acetate fraction of this variety showed the greatest antioxidant activity (IC 50 DPPH and ABTS = 106.34 and 17.62 µ g/mL, respectively), total phenolic content (275.16 mg GAE/g extract), and ﬂavonoid content (37.41 mg QE/g extract). Fatty acids (FAs) and fatty acid methyl esters (FAMEs) accounted for 47.71 ± 0.02 and 41.73 ± 0.04%, respectively, and they were the major components of the extracts. The principal component analysis (PCA) revealed three different groups of phytochemical components in the seeds of Agrotis variety, including fatty acid methyl esters (FAMEs), such as methyl linoleate, methyl stearate, methyl vaccinates, and methyl palmitate, and fatty acids (FAs), including 7-hexadecanoid acid, bovinic acid, and 15-hydroxipentadecanoic acid. The ﬁnal phytochemical group consisted of minor components, including uncategorized compounds such as decamethyl-tetrasiloxane and cycloalkenes. This study highlights the fact that C. lacrima-jobi is a promising source of natural


Introduction
The search for natural products as crop protection agents is becoming increasingly important due to the growing global demand for organic products and the need for a sustainable food supply. Safe, selective, and cost-effective agrochemicals are needed to replace synthetic ones, which can have negative impacts on the environment [1]. Higher plants, including those that contain phytochemicals such as polyphenols, are promising sources of natural products that can be used as crop protection agents.

Seeds Extraction for Screening Antioxidant Potentials
The antioxidant potential of three varieties of C. lacryma-jobi (Ma-yuen, Agrotis, and Aquatic) was screened using the procedure outlined in Figure 2. The seeds were dried at 40 °C in an oven and then ground into fine powder. After that, 5 g of each variety of pulverized powder was soaked in 170 mL of methanol, stirred for 24 h, filtered, and evaporated using a rotary evaporator at 30 °C (QR 2005-S, Shimadzu, Tokyo, Japan) to obtain the extract. Methanol was chosen due its high polarity, which allows for a high extraction yield and extracts a diverse range of compounds, as reported by Truong et al. [14]. All extracts were then tested for their antioxidant activity against DPPH and ABTS free radicals. The adlay variety with the highest antioxidant activity was selected for further screening in a fractionation procedure using different polar solvents (hexane, chloroform, ethyl acetate, and water).

Fractionation of Agrotis Variety from C. lacryma-jobi
A total of 1365 g of seed powder of the selected variety of adlay was soaked in 2625 mL methanol for nine days under ambient conditions, followed by filtration. The residue was soaked with the same volume of methanol 4 times until a final methanol extract of 10.95 g was obtained. The filtrate was then evaporated under vacuum at 40 °C using a rotary evaporator (QR 2005-S, Shimadzu, Tokyo, Japan. The yields obtained were 0.72,

Seeds Extraction for Screening Antioxidant Potentials
The antioxidant potential of three varieties of C. lacryma-jobi (Ma-yuen, Agrotis, and Aquatic) was screened using the procedure outlined in Figure 2. The seeds were dried at 40 °C in an oven and then ground into fine powder. After that, 5 g of each variety of pulverized powder was soaked in 170 mL of methanol, stirred for 24 h, filtered, and evaporated using a rotary evaporator at 30 °C (QR 2005-S, Shimadzu, Tokyo, Japan) to obtain the extract. Methanol was chosen due its high polarity, which allows for a high extraction yield and extracts a diverse range of compounds, as reported by Truong et al. [14]. All extracts were then tested for their antioxidant activity against DPPH and ABTS free radicals. The adlay variety with the highest antioxidant activity was selected for further screening in a fractionation procedure using different polar solvents (hexane, chloroform, ethyl acetate, and water).

Fractionation of Agrotis Variety from C. lacryma-jobi
A total of 1365 g of seed powder of the selected variety of adlay was soaked in 2625 mL methanol for nine days under ambient conditions, followed by filtration. The residue was soaked with the same volume of methanol 4 times until a final methanol extract of 10.95 g was obtained. The filtrate was then evaporated under vacuum at 40 °C using a rotary evaporator (QR 2005-S, Shimadzu, Tokyo, Japan. The yields obtained were 0.72, Figure 2. Screening of the antioxidant potential of adlay (C. lacryma-jobi).

Fractionation of Agrotis Variety from C. lacryma-jobi
A total of 1365 g of seed powder of the selected variety of adlay was soaked in 2625 mL methanol for nine days under ambient conditions, followed by filtration. The residue was soaked with the same volume of methanol 4 times until a final methanol extract of 10.95 g was obtained. The filtrate was then evaporated under vacuum at 40 • C using a rotary evaporator (QR 2005-S, Shimadzu, Tokyo, Japan. The yields obtained were 0.72, 0.78, 0.31, and 3.72 g for the hexane, chloroform, ethyl acetate, and water extracts, respectively. After obtaining the dried extracts or fractions, all samples were dissolved in methanol to prepare a stock solution, then stored at 4 • C for future analysis.

DPPH Free Radical Scavenging Activity
The determination of DPPH free radical scavenging activity from adlay extract was performed using the method reported by Andriana et al. [15], with slight modifications. A volume of 1.0 mL of the sample (10-3000 µg/mL in methanol) or quercetin (1-20 µg/mL) as a standard was mixed with 0.5 mL of 0.5 mM DPPH and 1 mL of acetate buffer 0.1 M (pH 5.5), then incubated in the dark at 26 • C for 30 min. The absorbance of the mixture was recorded at a wavelength of 517 nm with the use of a spectrophotometer (Shimadzu 1800 Uv-Vis 115 VAC, Tokyo, Japan). The inhibition level was calculated using the following formula: where A control refers to the absorbance of the reaction without sample and A sample represents the absorbance of the reaction with the sample.

ABTS Free Radical Scavenging Activity
The determination of ABTS free radical scavenging activity was performed using the method described by Andriana et al. [16] with a minor modification. An ABTS + solution was created by mixing 25 mL of a 7 mM ABTS solution and 25 mL of a 2.45 mM potassium persulfate. The mixture was left to incubate at room temperature in the dark for 16 h. Prior to the test, the ABTS+ solution was diluted with methanol to achieve an absorbance of 0.70 ± 0.05 at 734 nm. A total of 2 mL of ABTS+ methanol solution was combined with 0.4 mL of the sample (10-3000 µg/mL in methanol), and the mixture was stored under ambient conditions for 30 min. The absorbance of the mixture was then measured with a spectrophotometer (Shimadzu 1800 Uv-Vis 115 VAC, Tokyo, Japan) at a wavelength of 734 nm. Quercetin solutions (1-20 µg/mL) were used as standard solutions. The level of antioxidant activity was obtained by the following formula: where A control is the absorbance of the reaction without the sample and A sample is the absorbance of the reaction with the sample.

Total Phenolic Contents (TPC)
The total phenolic contents of extracts were evaluated by using the Folin-Ciocalteu's method, following the previous study reported by Iwansyah et al. [17] with some modifications. A volume of 0.2 mL sample (1000 µg/mL) was mixed with 1 mL of 10% Folin-Ciocalteu reagent. The mixture was vortexed; then, 0.8 mL of 5% sodium carbonate solution was added. The reaction mixture was incubated in a dark room for 30 min. The absorbance was measured at a wavelength of 750 nm by using a spectrophotometer (Shimadzu 1800 Uv-Vis 115 VAC, Tokyo, Japan). The total phenolic contents were expressed as mg gallic acid equivalent (GAE)/g of extract.

Total Flavonoid Content (TFC)
The total flavonoid content of the extracts was determined using the method reported previously by Minh et al. [18], with a minor modification. A sample of 1 mL volume (1000 µg/mL) was mixed with 1 mL aluminum chloride-methanol solution (2%). The mixture was then left to incubate for 15 min at room temperature. The absorbance was recorded at a wavelength of 430 nm using a spectrophotometer (Shimadzu 1800 Uv-Vis 115 VAC, Tokyo, Japan). The total flavonoid content was expressed as the equivalent of mg of quercetin in g of extract.

Identification of Chemicals Constituent of C. lacrima-joby var. Agrotis by Gas Chromatography-Mass Spectrometry (GC-MS)
The Agilent GC-MS system (Agilent 7890B/MSD 5977 A, Agilent Technology, Inc., Santa Clara, CA, USA) was employed to detect chemical constituents of ethyl acetate extract of the Agrotis variety. The volume of the injected sample was 1 µL. The Agilent HP-5MS (19091S-433:93.92873) (Agilent Technology, Inc., J&W Scientific Products, Santa Clara, CA, USA), with dimensions of 30 m in length, 250 µm internal diameter, and 0.25 µm in thickness, was used as the column. Helium was chosen as the carrier gas. The mode of the helium inlet was spitless, with a heater temperature of 250 • C, a pressure of 7.0699 psi, and a total flow of 104 mL/min. The operating conditions of the GC oven temperature were maintained as follows: The initial temperature = 40 • C with 1 min of hold time, which increased by the programmed rate = 10 • C/min up to a final temperature of 325 • C, with 4 min of hold time. The mass range scanned from 122-1021 amu. Electron ionization (EI) was employed as an ion source, and positive ions were collected in the MS analyses. The control of the GC-MS system and the data peak processing were carried out using the Agilent ChemStation software featured with the NIST mass spectral library (Agilent Technology, Inc., J&W Scientific Products, Santa Clara, CA, USA).

Statistical Analysis
Data analysis was performed using the one-way ANOVA and R Software (R Core Team, Vienna, Austria, 2020), and each assay was conducted in triplicate. Data were presented as mean ± standard deviation. A Duncan's test was used to determine the significant difference between the treated samples and the controls at a confidence level of 95% (p < 0.05). A multivariate analysis was conducted by performing principal component analysis (PCA) using R software [19] to determine the relationship between the extracting solvents and the concentration of compounds contained in the extracts.

Screening Antioxidant Potentials from Adlay (C. lacryma-jobi) Varieties
The DPPH and ABTS radical scavenging activity assays revealed that the antioxidant potential of three varieties of adlay (C. lacryma-jobi) were significantly different, according to Duncan's test (p < 0.05). The Agrotis variety exhibited the highest antioxidant activity (IC 50 DPPH and ABTS = 741.39 and 152.69 µg/mL, respectively) followed by the Aquatic and Mayuen cultivars (Table 1). However, when compared to the positive control, quercetin, these results were still lower. The Agrotis variety appeared to have higher antioxidant capacities to compare to other varieties, making it the preferred option for further fractionation.

Antioxidant Potentials from C. lacryma-jobi var. Agrotis Extracts
Since the Agrotis variety demonstrated the highest antioxidant activity, its methanol extract was fractionated using solvents of varying polarities. Table 2 displays the antioxidant activity of the extracts obtained from the Agrotis variety. The type of solvent influenced the IC 50 of the DPPH and ABTS assays of the Agrotis variety of adlay, and the results were found to be significantly different according to Duncan's test (p < 0.05). Ethyl acetate extract displayed the strongest antioxidant activity (IC 50 DPPH and ABTS= 106.34 and 38.20 µg/mL, respectively) followed by chloroform, water, and hexane extracts. However, when compared to quercetin as the positive control (IC 50 DPPH and ABTS = 6.55 and 9.24 µg/mL, respectively), the extract still showed a lower level of activity.

Total Phenolic and Flavonoid Contents (TPC and TFC)
Figures 3 and 4 indicate the total phenolic and flavonoid content of several seed extracts of the Agrotis variety. The results revealed that the polarities of the solvents significantly impacted the total phenolic and flavonoid content of the Agrotis variety, according to Duncan's test (p < 0.05). When ethyl acetate was used as the extracting solvent, the highest quantities of phenolic compounds (275.18 ± 16.71 mg GAE/g extract) and flavonoids (37.91 ± 0.68 mg QE/g extract) were found. On the other hand, the hexane fraction yielded the lowest amounts of phenols and flavonoids (35.18 ± 2.19 mg GAE/g extract and 3.96 ± 0.01 mg QE/g extract, respectively).

Total Phenolic and Flavonoid Contents (TPC and TFC)
Figures 3 and 4 indicate the total phenolic and flavonoid content of several seed extracts of the Agrotis variety. The results revealed that the polarities of the solvents significantly impacted the total phenolic and flavonoid content of the Agrotis variety, according to Duncan's test (p < 0.05). When ethyl acetate was used as the extracting solvent, the highest quantities of phenolic compounds (275.18 ± 16.71 mg GAE/g extract) and flavonoids (37.91 ± 0.68 mg QE/g extract) were found. On the other hand, the hexane fraction yielded the lowest amounts of phenols and flavonoids (35.18 ± 2.19 mg GAE/g extract and 3.96 ± 0.01 mg QE/g extract, respectively).

Identification of Phytochemical Constituents Adlay (C. lacryma-jobi) var. Agrotis by Gas Chromatography-Mass Spectrometry (GC-MS)
GC-MS analysis was performed to identify and quantify the phytochemical constituents of adlay (C. lacryma-jobi) var. Agotis. The GC-MS chromatograms of its extracts are presented in Figure 5, and the chemical constituents are listed in Table 3. A total of 20 compounds in various categories, such as cycloalkane, amides, alkanes, fatty acid methyl esters (FAMEs), fatty acids, polycyclic aromatic hydrocarbons (PAHs), alcohols, aldehydes, cyclosiloxanes, and others, were identified. The highest number of detected compounds was found in hexane, followed by the water, ethyl acetate, and chloroform extracts. FAME was determined to be the most dominant chemical group. The highest concentration of a component was found for linoelaidic acid (38.27%), followed by methyl linoleate (14.07%), palmitic acid (9.66%), methyl elaidate (9.51%), and the rest (<9.5%).

Identification of Phytochemical Constituents Adlay (C. lacryma-jobi) var. Agrotis by Gas Chromatography-Mass Spectrometry (GC-MS)
GC-MS analysis was performed to identify and quantify the phytochemical constituents of adlay (C. lacryma-jobi) var. Agotis. The GC-MS chromatograms of its extracts are presented in Figure 5, and the chemical constituents are listed in Table 3. A total of 20 compounds in various categories, such as cycloalkane, amides, alkanes, fatty acid methyl esters (FAMEs), fatty acids, polycyclic aromatic hydrocarbons (PAHs), alcohols, aldehydes, cyclosiloxanes, and others, were identified. The highest number of detected compounds was found in hexane, followed by the water, ethyl acetate, and chloroform extracts. FAME was determined to be the most dominant chemical group. The highest concentration of a component was found for linoelaidic acid (38.27%), followed by methyl linoleate (14.07%), palmitic acid (9.66%), methyl elaidate (9.51%), and the rest (<9.5%). Agrochemicals 2023, 2, FOR PEER REVIEW 8

Relationship of Extracting Solvents to Antioxidant Activity and Phytochemical Constituents of Adlay (C. lacryma-jobi) var. Agrotis by Principal Component Analysis (PCA)
The score plot of concentrations of phytochemicals from the Agrotis variety was created using PCA, as illustrated in Figure 6. The detected compounds were classified into three different groups. The compounds extracted from chloroform and ethyl acetate were grouped together in a similar quadrant, as they were largely similar. Figure 6 also shows the differentiation between the identified compounds present in the C. lacryma-jobi var. Agrotis extracts along the first two principal components, PC1 and PC2, in the scatter plot. The explained variances for PC1 and PC2 were 41.7% and 38.0%, respectively. The cumulative contribution rates of PC1 and PC2 reached a high level, which was enough to explain the maximum variation of compounds and to classify them in PCA. As shown in the scatterplot of PC1 and PC2, different extracts of C. lacryma-jobi seeds were clearly distributed in distinct regions of the PCA plot (quadrants I, II, and IV). The first group (quadrant I) consisted of six compounds (3−acetoxypentadecane, 7−hexadecenoic acid, bovinic acid, 2−methyl−Z, Z−3,13−octadecadienol, 15−hydroxypentadecanoic acid, and linoleyl aldehyde). The second group (quadrat II) included nine compounds (methyl palmitate, palmitic acid, methyl linoleate, methyl vaccinates, methyl stearate, cis−13−octadecenoic acid, linoelaidic acid, elaidolinoleyl alcohol, and 9,17−octadecadienal). The third group (quadrant IV) comprised five compounds (cyclopentene, 1,4−bis(methoxyethoxymethoxy) −, cis−pyrene, acetothioamide, cyclotrisiloxane, tetrasiloxane, and decamethyl−).

Relationship of Extracting Solvents to Antioxidant Activity and Phytochemical Constituents of Adlay (C. lacryma-jobi) var. Agrotis by Principal Component Analysis (PCA)
The score plot of concentrations of phytochemicals from the Agrotis variety was created using PCA, as illustrated in Figure 6. The detected compounds were classified into three different groups. The compounds extracted from chloroform and ethyl acetate were grouped together in a similar quadrant, as they were largely similar. Figure 6 also shows the differentiation between the identified compounds present in the C. lacryma-jobi var. Agrotis extracts along the first two principal components, PC1 and PC2, in the scatter plot. The explained variances for PC1 and PC2 were 41.7% and 38.0%, respectively. The cumulative contribution rates of PC1 and PC2 reached a high level, which was enough to explain the maximum variation of compounds and to classify them in PCA. As shown in the scatterplot of PC1 and PC2, different extracts of C. lacryma-jobi seeds were clearly distributed in distinct regions of the PCA plot (quadrants I, II, and IV). The first group (quadrant I) consisted of six compounds (3−acetoxypentadecane, 7−hexadecenoic acid, bovinic acid, 2−methyl−Z, Z−3,13−octadecadienol, 15−hydroxypentadecanoic acid, and linoleyl aldehyde). The second group (quadrat II) included nine compounds (methyl palmitate, palmitic acid, methyl linoleate, methyl vaccinates, methyl stearate, cis−13−octadecenoic acid, linoelaidic acid, elaidolinoleyl alcohol, and 9,17−octadecadienal). The third group (quadrant IV) comprised five compounds (cyclopentene, 1,4−bis(methoxyethoxymethoxy) −, cis−pyrene, acetothioamide, cyclotrisiloxane, tetrasiloxane, and decamethyl−).  Figure 7 shows the similarities among the different solvents used to extract C. lacryma-jobi var. Agrotis seeds. It can be observed that chloroform (2) and ethyl acetate (3) extracts were placed into the second group (quadrant II), meaning these solvents possess similarities in terms of the yield of the extracted compounds. Moreover, the hexane extract was arranged into the second group (quadrant II), while the water extract was arranged into the fourth group (quadrant IV), indicating that these solvents produced distinct results.  Figure 7 shows the similarities among the different solvents used to extract C. lacrymajobi var. Agrotis seeds. It can be observed that chloroform (2) and ethyl acetate (3) extracts were placed into the second group (quadrant II), meaning these solvents possess similarities in terms of the yield of the extracted compounds. Moreover, the hexane extract was arranged into the second group (quadrant II), while the water extract was arranged into the fourth group (quadrant IV), indicating that these solvents produced distinct results.

Potential Use of Adlay Phytochemicals for Agrochemicals
A potential use of phytochemicals from C. lacrima-joby, based on a literature review, is shown in Table 4. The major constituents of this plant, such as bovinic acid and methyl linoleate in hexane and chloroform extracts, have been reported to possess insecticidal activity against the potato bug (Leptinotarsa decemlineata) [20] and herbicidal activity against wild oat (Avena fatua), sun spurge (Euphorbia helioscopia), goosefoot (Chenopodium album), canary grass (Phalaris minor), and knotweed (Rumex dentatus), respectively [21]. On the other hand, in ethyl acetate extract, some chemical constituents such as palmitic and linoelaidic acids, were found to show larvicidal and insecticidal activities against Aedes aegypti Linn and cotton leafworm (Spodoptera littoralis), respectively [22,23]. Moreover, methyl palmitate presents in the water extract exhibited acaricidal activity against Tetranychus cinnabarinus (Boisduval) [24]. Most of the chemical constituents detected in C. lacryma-jobi have been reported to exhibit potential activity against various plant pests, weeds, fungi, and bacteria. These phytochemicals have the potential to be developed as natural pesticides, but further research is necessary to fully understand their biological activities.

Potential Use of Adlay Phytochemicals for Agrochemicals
A potential use of phytochemicals from C. lacrima-joby, based on a literature review, is shown in Table 4. The major constituents of this plant, such as bovinic acid and methyl linoleate in hexane and chloroform extracts, have been reported to possess insecticidal activity against the potato bug (Leptinotarsa decemlineata) [20] and herbicidal activity against wild oat (Avena fatua), sun spurge (Euphorbia helioscopia), goosefoot (Chenopodium album), canary grass (Phalaris minor), and knotweed (Rumex dentatus), respectively [21]. On the other hand, in ethyl acetate extract, some chemical constituents such as palmitic and linoelaidic acids, were found to show larvicidal and insecticidal activities against Aedes aegypti Linn and cotton leafworm (Spodoptera littoralis), respectively [22,23]. Moreover, methyl palmitate presents in the water extract exhibited acaricidal activity against Tetranychus cinnabarinus (Boisduval) [24]. Most of the chemical constituents detected in C. lacryma-jobi have been reported to exhibit potential activity against various plant pests, weeds, fungi, and bacteria. These phytochemicals have the potential to be developed as natural pesticides, but further research is necessary to fully understand their biological activities. 8. Methyl vaccinates n/a n/a n/a n/a 9. Methyl

Discussion
This study evaluated the phytochemical constituents of Indonesian adlay (C. lacrymajobi) by screening its three varieties, namely Agrotis, Ma-yuen, and Aquatic, and assessed their potential for use as crop protection agents. Based on DPPH and ABTS assays, the Agrotis variety was chosen for further fractionation due its high antioxidant activity. Total phenolic and flavonoid evaluations in the Agrotis variety using different extraction solvents (Figures 1 and 2) revealed that ethyl acetate extract had the highest content (275.18 ± 16.71 mg GAE/g extract and 37.91 ± 0.68 mg QE/g extract, respectively). This demonstrates that phenolics are not always found in polar extracts, depending on the structure of the phenolic compounds present [40]. According to Woo [41], the darker the color of the extract, the more flavonoids it contains. The yellow color of the hexane fraction is due to the presence of non-polar active chemicals dispersed in the extract [42]. In this study, the total phenol and flavonoid contents and antioxidant activity of adlay extracts were investigated. According to Tuyen et al. [43], phenols and flavonoids are well-known natural antioxidants. Phenolic and flavonoid compounds have been reported to have potential use as natural agrochemicals such as insecticide [44] and herbicide [45].
According to GC-MS analysis, a total of 20 compounds from different chemical groups were identified and quantified ( Table 4). The main components, which were mainly volatile compounds, were linoelaidic acid (38.27%), methyl linoleate (14.07%), palmitic acid (9.66%), methyl elaidate (9.51%), and the rest (<9.5%). Based on their similarity and abundance in the extracts, particle component analysis (PCA) clustered them into three different groups ( Figure 6). Hexane and water were classified as different groups due to hexane being a nonpolar solvent, while water is a polar one. Additionally, chloroform and ethyl acetate were placed in the same group because they are both are semi-polar solvents, which agrees with the study of Andriana et al. [46], which found ethyl acetate to be an effective solvent for extracting phenolic compounds.
Based on a study of the literature, it has been shown that most of the major chemical constituents from C. lacryma-jobi have potent pesticidal effects against plant pests and weeds, as well as against bacteria and fungi. Generally, certain fatty acids and their derivatives, such as linoleic acid, oleic acid, methyl linoleate, and methyl stearate, have been shown to possess antioxidant and fungicidal properties due to their unsaturated bonds [47]. Furthermore, linoleic acid has demonstrated anti-histaminic, anti-coronary, anti-acne, anti-bacterial, anti-fungal, anti-arthritic, and anti-inflammatory potential [48]. It was also found predominantly in green, black, and white adlay seeds in the range of 66.94-88.29% [49]. Palmitic acid was detected in ethyl acetate extract at 9.66%, and has been reported to possess antioxidant, larvicidal, and insecticidal activities [50]. Mohadjerani [51] also reported that fatty acids and their derivatives have antioxidant activities. Based on the study conducted by Andriana et al. [52], methyl palmitate was also detected in the extract with the highest levels of antioxidants. Therefore, it can be assumed that fatty acids and their derivatives might contribute to antioxidant activity. In addition, there are some saturated fatty acids that show activity as antioxidants as well as antifungal, bactericidal, and pesticidal agents [53]. Table 3 summarizes the pesticidal effects of phytochemical constituents from C. lacryma-jobi. It shows a wide range of inhibitory effects against several weeds, insects, bacteria, and fungi. Further research is required to investigate the phytochemical constituents of this plant in order to explore its potential as a source of natural agrochemicals and to better understand its pesticidal activity.

Conclusions
In this study, the antioxidant properties and phytochemical contents of different seed extracts of Indonesian adlay (C. lacryma-jobi) were successfully screened using a GC-MS analysis, with the aim of exploring their potential use as agrochemicals. The Agrotis variety was found to have the highest total phenol and flavonoid content, and its antioxidant capacities were comparable to other studied varieties. Through GC-MS analysis, a total of 20 compounds were identified and quantified, with the majority consisting of fatty