Preparation of Chalcone-Azo reagents and study of their chemical analysis, thermal studies, and biological effects against fungi

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INTRODUCTION
Chalcone Reagent is an aromatic ketone that forms the central core of various important biological compounds1,2, collectively known as chalcones or chalconoids.Alternate names for chalcone include Benzylideneacetophenone3, phenyl Styryl ketone, benzalacetophenone4, β-phenyl acrylophenone5, γ-oxo-α, γ-diphenyl-α-propylene6 and α-phenyl-β-benzoyl ethylene.7In a narrow sense, it refers to benzal acetophenone8, but in a broader sense, it is collectively called chalcone9, including its derivatives.Although benzalacetophenone is not found in nature, many chalcones exist.10For example, there are plant pigments such as biotin (a yellow pigment that contains a glycoside in flowers such as the yellow varieties of dahlia and kibana cosmos)11-13 and Carthamin (a red pigment found in the crown of safflower, also used as a food coloring in Japan).The closed-loop isomer is flavanone, along with flavonoids14,15 and anthocyanins.These yellow pigment compounds are collectively called flavonoids, a type of polyketide pro-duced16-18 using acetyl CoA as raw materials.Polyketide is a general term for substances that are biosynthesized19,20 via β-polyketide acid represented by the general formula [R (CH2 CO) n CO2 H].There are two isomers of benzalaceto-phenone21, cis and trans, but they usually refer to the trans isomer.22-24It is a pale-yellow crystal with a melting point of 59 °C, but there are many other forms with different melting points25,26 such as 57 °C in ether, 49 °C in carbon dioxide, 48 °C in chloroform27,28, 28 °C in benzene and 18 °C in slightly ethyl alco-hol29,30 dimmed by light irradiation (Renji Okazak).As for azo reagent and azo dye compounds31,32, they have a basic principle of introducing two small water-soluble components inside the fibers and then using certain conditions suitable for the occurrence of duplication, giving a larger, insoluble 33,36 and colored molecule.The last component is one of the diazotized primary amines35-37, or the parent amine, when the dye undergoes dialysis.It is used for dyeing cotton materials.38,39These pigments are characterized by their bright colours40,41 and a high degree of stability.

MATERIALS AND METHODS
Various studies for measurements have been reinforced by maneuvers at the University of Tehran (Iran) and were embodied by Thermo-Curves and Thermal stability in various Temperatures and Fungal-Assay (Tehran University).In Iraq, other analytical studies like solubility, physical properties, infrared measurement, and UV-visible identification.

Formation of Chemical Reagents Formation of Reagent {1}
The chemical Reagent {1} was prepared by a reaction of 1,4-diaminobenzidine (0.1 moles) in (2 ml) of (HCl), and then a solution of sodium nitrite in the ace path was added.After that, it was added to a basic solution of (0.02 mole) of (4methoxy benzaldehyde), then naturalized the medium, after that filtrated, washed and dried, to be responsible for new Reagent {1} supportive to procedures. 26,28

Formation of Reagent {4}
The chemical Reagent {4} was prepared by reaction (0.1 moles) of Reagent {1} with 3-acetoindole (0.2 moles) through a rotation step for (9 hours) in the occurrence of (5% NaOH) at room temperature, the next step embodied by filtration, aeration, then a step of recrystallization to afford new reagent {4} supportive to procedures. 26,28Antifungal Assessment 15,16 The assessment of formatted reagents has been premeditated against some bacteria through the medium (agar) via numerous elegant processes.15,16The assessment of microbial inhibition was tested at three concentrations: 40, 70, and 80 micrograms with blank solvent (DMSO) per fungi (Aspergillus niger and Candida albicans) with incubation for 24 hours at 37 oC.In this research, different types of fungi were used.These organisms were chosen because of their importance in the medical field, as they cause various diseases and differ in their resistance to antibiotics and therapeutic chemicals.The biological activity was measured by calculating the inhibition area and time, and then the biological activity.

RESULTS
Chalcone reagents have several applications in analytical chemistry, especially in estimating the amount of trace elements through forming metal complexes.Spectroscopic and physical methods can estimate the quantity of these elements.In recent years, chalcone reagents have received great attention as medicinal materials due to the biological efficacy that has appeared in some of them.Fungi are also of industrial importance, as they are used as plasticizers and stabilizers for polymers and gasoline and as initiators of the polymerization process.They are also used as antioxidants and catalysts.For this, the present study involved many studies like spectral studies with thermo-curves and thermal stability in various temperatures and Fungal-Assay: Spectral Study of Chemical Reagents UV-Visible -Spectra: The structure of new Chalcone-Azo reagents proved with Uv-Vis-spectra with perfect concentration in Ethanol as a solvent, all spectral information in figures 1-4:

Behaviour of Reagents in Different Solvents
Conferring to the polarity of formatted reagents, results showed that the reagents have good solubility in some of the solvents due to functional groups in our analytical reagents and structure (nature of active groups) in every compound.The effects are shown in

Antifungal Assessment
Results in table (3) showed that the prepared compounds have different biological activities with different concentrated groups on the base compound, as these new compounds work to eliminate the new pathogenic strains of these types of fungi.The table below shows the results of the effect of the prepared chemicals on fungal growth.Some of the tested compounds generally showed medium and close inhibitory activity against these fungi.The assessment of reagents on kinds of fungi produced vibrant data with reagents {3 and 4} more than reagents {2 and 1} due to thiophene ring and indole ring in reagents, which increases the inhibition of fungi-results in Table 3.

DISCUSSION
The literature supports the total explanation of Spectra in reference.Each reagent gave good stability towards various temperatures, allowing study 8.

CONCLUSIONS
Numerous guides tested all the formatted new (Chalcone-Azo)-Reagents using investigative and chemical performances and other instrumental methods to ensure that all reagents have good stability in various temperatures according to their structures and good results as inhibitors against fungi.
PATENTS Funding: Self-funding