Study of Binary Complex formation of Fe (III) with 3-Hydroxy-3-(phenyl)-1- 2, 4Dimethyl Phenyl Triazene

 

Naresh Kumar Choubisa*, Ashok Panwar, Mangilal Regar, G.P. Singh, Rekha Dashora,R.S. Chauhan and A.K. Goswami

Coordination Chemistry Laboratory, Department of Chemistry, University College of Science,

Mohanlal Sukhadia University, Udaipur-313001, Rajasthan, India.

*Corresponding Author E-mail: choubisan@gmail.com

The present paper reports synthesis, characterization, spectrophotometric studies and activity prediction of HPMPT with Fe (III) complexes. It was observed that Fe (III)  forms complex with HPMPT in1:3 ratio at wavelength of 510 nm and between the pH range of 4.3- 4.8. The molar absorptive and Sandell’s sensitivity values are 52175 L mol^-1 cm^-1 and 1.32 ng cm^-2 respectively. The study shows that metal chelates are better in activity as compared to their respective ligands. The results have been interpreted in terms of tables, figures and theories.

 

 

 

INTRODUCTION:

The present work has been centered upon the study of metal complexes spectrophotometrically. HT (HPMPT) is very good chelating agents^1-2. Their metal complexes have been shown excellent biological activities such as anti-inflammatory, insecticidal, antibacterial and antifungal^3-7.

 

EXPERIMENTAL:

HPMPT have been synthesized as per the standard method^8-10. The overall process has been completed in three steps. The compound has been characterized by IR studies, CHN analysis, and m.p. determination (AR grade chemicals were used for synthesis and crystallization).

 

The ligand HPMPT has been screened for biological activity on the basis of PASS^11-12. Computer aided program PASS provides large no. of possible biological activities with their percent activity and percent inactivity, which helps to design drug in huge chemical pharmacological space. Probable activities predicted by PASS are validating by experimental bioassay, which would have been a way to CADD.

 

Materials and Methods

For the synthesis of HPMPT, AR grade chemicals were used. The process is completed in three steps. The general method of synthesis has been given by the following three step reaction procedure.

 

Step-1- Reduction of nitrobenzene

0.25 mole of nitrobenzene, 15 gm. of NH₄Cl were taken in one L beaker containing 400 ml H₂O and 100 ml of rectified spirit. The contents were stirred mechanically and the temp. of the reaction mixture remain between 55-60⁰C. 40gm Zinc dust was added in small lots till one hour. The resulting mixture was filtered under suction and the filtrate was taken into another beaker and kept it in freezer to cool to 0⁰C.

 

Step-2- Diazotization of 2,4 Di-methyl Aniline

0.2 mole of 2,4 Di-methyl Aniline was dissolved in a warm mix. of 50 ml. conc. HCl and 50 ml of H₂O in a 500ml beaker . This mixture was kept in freezer to cool after stirring. In a beaker 13 gm of NaNO₂ was dissolved in H₂O and it was kept in freezer to cool. The beaker containing 2, 4 Di-methyl Aniline solution, was kept in an ice bath and to this NaNO₂ solution was added slowly with continuous stirring. This resulted in the formation of diazotized product, used for coupling in the next step.

 

Step-3- Coupling

Hydroxylamine prepared in step (1) was coupled with the diazotized product obtained in step (2) at 0-5⁰C to maintain the pH between 5-6 by using sodium acetate solution as buffer. When the addition was completed, the resultant mix. was crystallized with appropriate solvent. This result the formation of HPMPT in the form of yellow Shining crystals.

 

 

 

Step-1 Reduction of Nitro benzene

 

 

Step-2 Diazzotization of 2,4 di-Methyl Aniline

 

Step- 3 Coupling

 

Table 1. Physical Characteristics, M.P., C H N Values of the reagent

 

S. No.	Molecular Formula	Melting Point (oC)	Colour and shape of the crystal	% Analysis
				Calculated			Experimental
				C %	H %	N %	C %	H %	N %
1	C14H15N3O	109	Yellow Shining Crystal	69.70	6.22	17.43	69.96	6.38	17.30

 

 

Resultant compound was subjected to four spot tests detection as described by Purohit^13-16. This compound gave positive test with all the four reagents proving that the synthesized compound is a hydroxytriazene.

 

To check the purity of the compound CHN-analysis and IR studies have been done. CHN data has been given in table-1. IR-data of this compound has been given in table 2, showing the characteristic peaks of (O-H str.) (C-H str. Ar)(C-H str. CH₃) (N=N str.) (N-N str.) and (C-CH₃str.)vibrations.

 

 

 

Table 1.2 IR data of the reagent(Cm^-1)

 

S. No.	O-H str.	C-H str. Ar	C-H str. CH3	N=N str.	N-N str.
1	3618	3028	2920	1581	1454

 

 

Spectrophotometric Study of the Complex-

(A) Preparation of Solutions

(i)Reagent solution

A fresh stock solution of 1.0*10^-2 M of the reagent 3-hydroxy-3-phenyl-1-(2,4dimethyl phenyl) triaz-1-ene was prepared by dissolving requisite quantity of thereagent in ethanol. Dilute solution were prepared from thisstock solution as and when required.

 

(ii)Standard solution of Fe(III)

A 1.0*10^-2 M stock solution of Fe (III) was prepared by dissolving the requisite quantity of A.R. grade Ferric nitrate and making it up to the required volume with double distilled water. It was standardized with standard1.0*10^-2 EDTA solutions at pH 2.5-3.0 using sulphosalicylic acid as an indicator. Dilute solution of different concentration were prepared from stock solution by proper dilution with double distilled water.

 

(iii) Solutions for pH adjustment

(a)Trisbuffer solution

A 1.0% of tris buffer solution was prepared by dissolving 1.0 gm of the tris buffer in minimum quantity of double distilled water and then making it up to 100 ml with distilled water.

 

(b) Perchloric acid solution

A 1.0% Perchloric acid solution was prepared by dissolving1.0 ml of the Perchloric acid in minimum quantity of double distilled water and then making up to 100 ml with distilled water.

 

(iv)Instruments

The spectrophotometric studies have been carried out onSystronic-108 UV-VIS spectrophotometer and Systronic-324 was used for pH measurement.

 

(B) Selection of suitable working wavelength

3ml (1*10^-5M) Fe(III)solution and 3ml (1*10^-4M) reagent solution was taken in 10ml volumetric flask and then makeup to the mark with alcohol. Absorbance of solution against its reagent blank was measured selected in a region where the absorption of Fe (III) complex was maximum. For working wavelength, maximum absorbance was found at 510 nm.

 

(C) Effect of pH on absorbance

Absorbance of the solutions at various pH values containing Fe (III) and reagent solutions in the ratio of 1:10 was taken at working wavelength 510 nm against reagent blank. The optimum pH range for constant maximum absorption was selected.

 

(D) Composition of the Fe (III)complex

The composition of the Fe(III) complex was determine dusing Job’s method, mole ratio method of Yoe and Jone’s and slope ratio method.

 

(a) Job’s method

The composition of Fe (III) complex was determined at two different concentrations with Job’s method. For each concentration, set of solution was prepared by varying the volume of equimolar Fe (III) and reagent solution from 0 to6 ml. After pH adjustment, the solutions were marked (10ml) with ethanol. The absorbance of solution was measured at suitable working wavelength against reagent blank. The second set of this method differed from the first set only in the concentration used. By this method the composition was found to be 1:3 [Fe: R].

 

(b) Mole ratio method of Yoe and Jone’s

In this method Fe(III)concentration was kept constant and reagent concentration was varied. A series of solutions having Fe(III)to regent ratio 1:1 to 1:10 were prepared with maintaining the pH of constant absorbance. Absorbance of each solution of a set was measured at working wavelength against the reagent blank. By this method the composition was found 1:3[Fe: R].

 

(E) Beer’s Law

A set of solution, having to ligand ratio 1:10 was prepared. The studies were performed under optimum condition of pH, concentration and solvent at corresponding working wavelength. The absorbance was measured for the complex against the reagent blank. The results are shown in Table 2.

 

(F) Sandell’s sensitivity

The molar absorptivity of the Fe (III) complex was calculated from the Beer’s law graph and it was found to be €= 52,175 L/mol.cm. The value thus obtained was used for determining Sandell’s sensitivity of the complex that are obtained 1.32 ng/cm². This value shows that the method used is quite sensitive and satisfactory for the determination of Fe(III).

 

 

Table. 2: Spectrophotometric determination of Fe(III)with HPMPT-1

Fe(III)complex with reagent	Composition of the complex [Fe (III): R]	Working Wavelength (nm)	Optimum pH range	Beer’s Law range (M)	Molar absorptivity [mol-1cm-1]	Sendell’s Sensitivity Ng cm-2
3-hydroxy-3- phenyl-1-(2,4 di methyl phenyl) triazene	1:3	510	4.3-4.8	1.0 x10-5to 4.5 x10-5	52175	1.32

 

 

Prediction of the spectrum of biological activities:

Brief description of PASS-The computer system for the prediction of the spectrum of biological activity according to the structural formula (PASS) was used to predict the spectrum of biological activity of the test selection compounds. The functioning of the PASS system is based on a training selection of chemical substances with known biological activities. The PASS system involves the following basic elements: description of the chemical structure, representation of the biological activity, training selection, and mathematical search algorithm for the structure-activity relationship. The biological activity in the PASS system is predicted qualitatively (presence or absence). The general number of the predicted activities (the spectrum of activity) involves more than 400 pharmacological effects and mechanisms of action, as well as carcinogenicity, probabilities that a definite activity would be or would not be exhibited (Pa and Pi, respectively) are calculated for each predicted type of activity with the use of a specially developed mathematical algorithm.

 

Activity prediction

The biological activity spectra of the 3-hydroxy-3- phenyl-1-(2,4 di methyl phenyl) triaz-1-enewere obtained by PASS software. The predictions were carried out on the basis of analysis of training set containing about 10000 drugs and biologically active compounds.

 

This set consider as reference compounds for known chemical compounds as well as different biological activities. Percent activity (Pa) and inactivity (Pi) of compound have been represented in Table- 3.

 

RESULT AND DISCUSSION:

As described in the Table-2, 3-hydroxy-3-phenyl-1-(2,4 di methyl  phenyl) triaz-1-eneforms complex in the ratio of 1:3.  The conditional stability constants have also been given in the table, determined by using two different methods. Hydroxytriazenes act as, bidentate ligand and in the present case, the reagent has been found to form a 1:3 complex with 3-hydroxy-3-phenyl-1-(2,4 di methyl  phenyl) triaz-1-ene, which indicates hexa coordinated Fe (III) complex with a probable geometry being Octahehral. Biological activity spectra were predicted for title compound with PASS computer program. The result of prediction is presented in table 3 as the list of activities with appropriate Pa and Pi (Pa-Pi) > 0.PASS is based on a robust analysis of structure–activity relationship in a heterogeneous training set currently including about sixty thousand of biologically active compounds from different chemical series with about four thousand five hundred types of biological activity. It can be seen from the results of PASS that most probable activities are Hematotoxic, Antineurotoxic, Anti-inflammatory and Analgesic.

 

Table 3: Predictions of Percent activity (Pa) and inactivity (Pi) of compound

Pa	Pi	Activity
0.744	0.018	Glutamylendo peptidase II inhibitor
0.686	0.026	5-O-(4-coumaroyl)-D-quinate 3'-monooxygenase inhibitor
0.673	0.060	CYP2J substrate
0.669	0.074	Aspulvinonedimethylallyltransferase inhibitor
0.598	0.004	Thiosulfate dehydrogenase inhibitor
0.626	0.037	Omptin inhibitor
0.636	0.057	Polyporopepsin inhibitor
0.575	0.003	AICAR transformylase inhibitor
0.645	0.074	Testosterone 17beta-dehydrogenase (NADP+) inhibitor
0.579	0.011	Plastoquinol-plastocyanin reductase inhibitor
0.600	0.032	Venombin AB inhibitor
0.582	0.032	Alkane 1-monooxygenase inhibitor
0.576	0.033	Phosphatidylcholine-retinol O-acyltransferase inhibitor
0.573	0.032	Pterindeaminase inhibitor
0.630	0.095	Ubiquinol-cytochrome-c reductase inhibitor
0.557	0.027	Aminobutyraldehyde dehydrogenase inhibitor
0.579	0.056	Fusarinine-C ornithinesterase inhibitor
0.572	0.054	Kidney function stimulant
0.577	0.061	Sugar-phosphatase inhibitor
0.552	0.043	Phospholipid-translocating ATPase inhibitor
0.563	0.060s	Pseudolysin inhibitor

 

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Received on 18.12.2014         Modified on 30.12.2014

Accepted on 06.01.2015         © AJRC All right reserved