Synthesis, Characterization and Evaluation of Biological Activity of Corn Oil -Based Difatty Acyl Carbamodithioic Acid

Wisam H. Hoidy¹*, Shaimaa M. Essa¹, Fouad A. Al-Saady², Emad A. Jaffar Al-Mulla³

¹Chemistry Department, College of Education, University of Al-Qadisiyah, Al-Qadisiyah, Iraq.

²College of Pharmacy, Al-Mustansiriyh University, Baghdad, Iraq

³College of Health and Medical Techniques, Al-Furat Al-Awsat Technical University, Kufa, Iraq

*Corresponding Author E-mail: wisam.hoidy@qu.edu.iq

ABSTRACT:

Carbamodithioic acid is a compound derived from thiourea. Thioureas have a significant role in medical chemistry and agriculture because of their biological action as herbicides and rodenticides, also antiviral antibacterial, and antifungal, in addition, anti-HIV and anti-tumor agents and phenoloxidase enzymatic inhibitors. The paper sheds light on Difatty acyl carbamodithioic acid (DFACA) that has antibiotics and antifungal actions that have been made out of corn oil and carbamodithioic acid employing sodium ethoxide as a catalyst. glycerol and Ethyl fatty ester (EFE) are manufactured as secondary products. The synthesis is achieved through refluxing corn oil with carbamodithioic acid in ethanol. In this method, corn oil transformed into about 83% pure DFACA after 10 hrs and a molar ratio of carbamodithioic acid compared to corn oil was 6.0: 1 at 80C. Analysis of elements, 1H nuclear magnetic resonance (NMR) technique and Fourier transform infrared (FTIR) spectroscopy are employed to portray DFACA and EFE. The new compounds (DFACA) was filtered for antimicrobial actions using disc diffusion method for antimicrobial activity against two samples of Gram-positive bacteria and two samples of Gram-negative bacteria represented by "Staphylococcus Aureus and Bacillus Subtilis" and "Escherichia Coli and Klebsiella pneumonia", respectively. Also, examine the effect of DFACA on fungus Candida albicans. The result disclosed that the (DFACA) has high biological activity. The result disclosed that the (DFACA) has high biological action.

KEYWORDS: Corn oil, Carbamodithioic acid, ethyl fatty ester, antimicrobial, thiourea.

INTRODUCTION:

Carbamodithioic acid is a type of organic compound containing two sulfur atoms, namely organic sulfur compounds, which contain a single and double bond between sulfur and carbon, this acid is derived from thiourea where nitrogen atom has been replaced with sulfur¹. Much attention has been given to Thiourea and its derivatives because of antifungal resistance, insecticidal and antibacterial characteristics especially against tubercle bacillus^2,3. Moreover, they are employed as detergents, lubricants, surfactants and antifoams in industrialized uses⁴.

Also, attention has been given to compounds of fatty nitrogen because of their biological activities and industrialized uses as detergents, antifoams, lubricants surfactants, and other utilizations. Fatty amides are among important fatty nitrogen compounds that are made in great number of metric tons annually from the fatty acids, which depends on its reaction with anhydrous ammonia about 200˚C and 345–690 kPa4). Number of reports of synthesizing fatty amides out of fatty acids have been issued or esters of different amine compounds through pressure with high temperature^5,6 or by synthesizing enzymes^7–9.

Some agents, antimicrobial, has been the focus of several reports lately because of The rapid increase of fungal and bacterial infections diseases. Since 1940, when penicillin was presented as an antibiotic, it was among the first medicines used effectively to fight many bacterial infections. Antibiotics were the weapon that employed commonly to resist infective diseases due to bacteria and fungi. Nevertheless, the use and misapplication of antibiotics resulted in the rise of pathogens resisting antibiotic as multidrug-resistant strains^10,11. The result is that not all infectious diseases are treated successfully. In this regard, there is an increasing tendency to open new horizons of new antibacterial medicines¹².

In the current paper, difatty acyl carbamodithioic acid is manufactured straightly from corn oil through refluxing it with carbamodithioic acid by sodium ethoxide as a catalyst. Difatty acyl carbamodithioic acid gives the probable application as antimicrobial and antifungal due to the nitrogen and sulphur in its structure. Furthermore, the existence of long chains fatty acids from corn oil encompassing O and N atoms suggest difatty acyl carbamodithioic acid could be beneficial as organic reagents for taking out and split-up metal ions from aqueous solutionEthyl fatty ester is made as a byproduct. EFE can be used as a primary component of biodiesel, which is an important substitute energy for petroleum substitution^13-15.

METHODS AND MATERIALS:

Materials:

The researcher bought corn oil from the marketplace, Iraq. Carbamodithioic acid, sodium metal and ethanol were bought by local providers from Sigma-Aldrich, united state.

Synthesis of DFACA:

By using A round-bottomed flask of 250 mL fixed with a refluxing and use a magnetic stirrer. Pieces of sodium cut precisely were melted in 150 ml of dry ethanol. When all the sodium in the dry ethanol is reacted, the corn oil is then added to the solution followed by carbamodithioic acid melted in 60 ml of hot ethanol(68°C). After mingling by stirring, the blend was refluxed for 10 hrs in an oil bath at 75°C. To the room temperature, flask materials were cooled. They were relocated to a separator funnel, permitted to calm down for a night. The lowest level included of glycerol was detached. The highest level that includes the products are transferred to a beaker, mixed with 150 mL of hot distilled water (70°C) and 15 mL of condensed HCl, and shacked for 20 minute. Difatty acyl carbamodithioic acid (DFACA) solution was isolated out of white particles of ethyl fatty ester through filtering. The pure solution contains difatty acyl carbamodithioic acid and in ice bath was used to cool it. The yellow product was collected on the Buchner funnel and was washed by 150 ml of cold water and then dried by a vacuum desiccator. Corn oil transformation to DFACA was nearly 80%. Preparation method is displayed in the Scheme 1.

The suggested process of transforming corn oil to DFACA and EFE are displayed in Scheme 2. Sodium ethoxide is used as a, Stage No.1 directs to proton retreating from (ــNH2) in carbamodithioic acid molecule, as suggested in the system, (the Scheme 2A). As per demonstrated through Scheme 2B, carbamodithioic acid ion attacks one carbonyl of the triglyceride set and shapes N-carbamoyl fattythioamide that attacks the second carbon thioamide in the glycerides set after proton retreating from it by sodium ethoxide. When two successive reactions, "2,3-dihydroxypropyl fatty ester, 1,3 dihydroxypropan-2-fatty ester (as intermediates) and difatty acyl carbamodithioic acid" are made ¹⁶. The final stage (the Scheme. 2C), the alkoxy set attacks the third carbonyl of the triglyceride set and produces ethyl fatty ester and glycerol¹⁷.

Antimicrobial Study:

The antibacterial tests were performed according to the disc diffusion method. Difatty acyl carbamodithioic acid was examined for their antimicrobial action in vitro against four strains of bacteria, two of them were gram-positive and two gram-negative represented by "Staphylococcus aureus and Bacillus Subtilis" and "Escherichia coli and Klebsiella pneumoniae", respectively. In addition, single fungal Candida albicans was used, Amoxicillin and Cefalexin were utilized as positive control and DMSO was employed as negative control, nystatin was employed as antifungal reference drug.

Bacteria were incubated and cultured at 37°C for 24 hrs. To prepare media, agar was poured into the sterilized petri discs. After that, the cultures were scattered on the surface of Nutrient Agar (NA). Antimicrobial action was defined by measuring the diameter of inhibition zone (IZ) in comparison with positive control. concentration of all compounds where was 30 μg/ml

RESULTS AND DISCUSSION:

Identification of DFACA and EFE:

Test of complex formation:

DFACA Complexes with copper (II) gives a green color that is a usual color of the compound detected as copper ion is reacted with amide group¹⁸.

Spectra of FTIR:

Distinguished bands of corn oil are seen at "3011, 2923, 2856, 1742, 1464, 1162, and 723 cm^-1" resulted from "C–H stretching of CH=CH, C–H asymmetric stretching of CH₂, C–H symmetric stretching of CH₂¹⁹, C=O stretching of ester (glyceride), CH₂ scissoring, O–C–C stretching, and CH2 rocking", respectively 11 . DFACA spectra display new bands at "3349, 1626, 1049, 1030 cm-1 and 570 cm^-1"attributed to "N-H stretching, C=O stretching, C-N stretching of amide, C=S stretching and C-S stretching", respectively. EFEs spectra display the existence of main absorbing groups of corn oil with vanishing of peaks that parallel to amide

¹H NMR Spectra of DFACA:

"(400MHz) (CDCl₃): δ 0.88 (t, J = 8.8 Hz, 6H, 2 x CH₃), 1.24 (m, H, CH₂), 1.53 (4H, 2 x CH₂ CH₂C=O NH), 2.19 (4H, 2 x CH₂ CH = CH), 2.33 (t, J = 10.9 Hz, 2H, 2 x CH₂ CO NH), 5.87 (2H, CH = CH), 10.8 (br, s, 2H, CO NH)"

1H NMR Spectra of EFEs:

"(400MHz) (CDCl₃): δ 0.88 (t, 3H CH₃), 1.27 (m, H, CH₂), 1.28 (t, 3H CH₃), 1.64 (2H, CH₂ CH₂C=O), 2.19 (4H, 2 x CH₂ CH = CH), 2.34 (t, 2H, CH₂C=O), 4.14 (q, 2H, CH₂ – O), 5.83 (2H, CH = CH)".

Conditions for optimal reaction:

Difatty acyl carbamodithioic acid was set with diverse molar ratios of carbamodithioic acid and corn oil. Table 1. shows the molar ratio effect in the carbamodithioic acid molar ratio to corn oil as 5.0:1.0 was regarded optimal. The reaction duration was increased from an hour to 15 hrs. Reaction time effect is revealed in Figure 1, in this figure, it is proven that 10 hrs is the appropriate duration for the highest transformation of corn oil into difatty acyl carbamodithioic acid. In this reaction the catalyst has a very important role, the catalyst percentage (sodium ethoxide) to corn oil were taken into consideration. Table 2 displays the effect of catalyst amount to corn oil. Molar ratio of catalyst to corn oil as 6.0: 1.0 gave the full conversion²⁰.

Table 1. Transformation of corn oil as a purpose of carbamodithioic acid/corn oil molar ratio. Circumstances of reaction: Reaction time = 10 hrs

Carbamodithioic acid /corn oil (mmol: mmol)

Transformation of corn oil into difatty acyl carbamodithioic acid (%)

1:1

2:1

3:1

4:1

5:1

6:1

7:1

8:1

12.63

23.91

42.59

66.23

80.42

80.16

79.44

78.22

Table 2. Influence of the catalyst on corn oil transformation

Carbamodithioic acid /corn oil (mmol:mmol)

Transformation of corn oil into DFACA (%)

1:1

2:1

3:1

4:1

5:1

6:1

7:1

8:1

9.18

21.42

46.13

68.36

80.12

80.41

80.10

79,63

The conditions of the highest reaction for full transformation of corn oil into DFACA are displayed in Table 3.

Table 3. The conditions of the highest reaction of transformation corn oil into DFAT through catalest (sodium ethoxide)

Parameters	Conditions
Time of reaction	10 hours
Temperature of reaction	79 C
Ratio of carbamodithioic acid: corn oil	5:00 (mmol) to 1 (mmol)
Ratio of catalyst: corn oil	6:00 (mmol) to 1 (mmol)

Antimicrobial actions:

The antibacterial and antifungal actions of difatty acyl carbamodithioic acid was examined against two samples of Gram-positive bacteria’s and two samples of Gram-negative bacteria’s represented by "Staphylococcus aureus and Bacillus subtilis" and "Escherichia coli and Klebsiella pneumoniae", respectively. Also single fungal Candida albicans utilizing disc diffusion method. Amoxicillin and cefalexin were used as positive controls, Nistatin was used as positive control for fungal (30 μg/ml concentration for all of these compounds) The outcomes are itemized in Table (4) and Table (5)

Table 4. Antimicrobial result for DFACA compound

Compounds (30 mg/ml)	Inhibition Zone (mm)
	Gram positive		Gram negative
	*S. aureus*	B. *subtilis*	*E. coli*	*K. pneumoniae*
DFACA	17.3	16.7	17	18.7
Amoxicillin	14.4	13.4	15.1	12.9
Cefalexin	18.2	16.3	18.1	18.6
DMSO	-	-	-	-

Table 5. Inhibition zones of DFACA and the references antifungal

Compounds (30mg/ml)	Inhibition Zone(mm)
Compounds (30mg/ml)	*Candida albicans*
DFACA	19.6
Nystatin	17.4
DMSO	-

The antibacterial activity of difatty acyl carbamodithioic acid was higher than amoxicillin and cepxin because DFACA contained two atom of sulfur which increase the lipophilicity of this compound. while amoxicillin and cefalexin have one atom only in their structure. These remarks displayed that the compounds were commonly had enhanced permeability towards gram-Positive and gram-negative bacteria cell tissue. It is identified that DFACA compounds were commonly show good antimicrobial actions.

CONCLUSION:

The paper reveals that difatty acyl carbamodithioic acid was positively produced by refluxing corn oil, ethanol and carbamodithioic acid with the company of sodium ethoxide as catalyst. Moreover, the glycerol and ethyl fatty esters are made as byproducts. The most appropriate circumstances of the reaction examined. It is denoted that at 79 ºC temperature, 5.0: 1.0 molar ratio of carbamodithioic acid to corn oil, 6.0: 1.0 molar ratio of catalyst (sodium ethoxide) to corn oil and 10 hrs reaction time were the appropriate circumstances of reaction. The results of FTIR spectroscopy, ¹H NMR technique, elemental analysis and Qualitative analysis of complex formation are affirm the formation of difatty acyl carbamodithioic acid. From the result it can be concluded that the DFACA has a good biological activities against these microorganisms. DFACA showed the highest biological activity against four bacteria higher than the references Antibiotics (Amoxicillin and Cefalexin), this activity may Attributed to C-S and C═ S group'.

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

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Received on 18.07.2019 Modified on 30.07.2019

Research J. Pharm. and Tech. 2020; 13(4):1931-1935.

DOI: 10.5958/0974-360X.2020.00348.0