INTRODUCTION:

Green chemistry involves design and redesign of chemical synthesis¹ and chemical products to prevent pollution and thereby solve environmental problems. Clearly there will be a continuing need for the definition of novel reaction routes to both multifunctional scaffolds for lead generation and to unique drug like heterocyclic structures Microwave irradiation has proved to be a powerful tool for both speeding up chemical optimizations and for efficient preparation of new target compounds.

Pyrazole derivatives have a long history of application in agrochemicals and pharmaceutical industry as herbicides and active pharmaceuticals. The recent success of pyrazole COX-2 inhibitor has further highlighted the importance of these heterocycles in medicinal chemistry. The prevalence of pyrazole cores in biologically active molecules has stimulated the need for elegant and efficient ways to make these heterocyclic lead. The treatment of pain continues to be the subject of considerable pharmaceutical and clinical research. Microbial infections often produce pain and inflammation.

Chemotherapeutic, analgesic and anti-inflammatory drugs are prescribed simultaneously in normal practice. It has been reported that pyrazoline possess analgesic, anti-inflammatory^2-4 and antimicrobia^5-7 activities. In view of these above, an attempt has been undertaken for the synthesis of the some 1-isonicotinoylpyrazolidine-3,5-dione. The synthesized compounds were tested for their possible anti-microbial activities.

Pyrazoline also shows various biological and Pharmacological activities such as, antitumor⁸, anticonvulsant⁹, antiviral¹⁰, diurectic¹¹, antidiabetic¹², anticancer¹³ activities. Synthesis of some pyrazolines derivatives have been carried under microwave irradiation as well as by conventional procedure. In conventional method, it took 6-12 hours for completing of reaction whereas under microwave irradiation, only 2-4 min required. The use of microwave irradiation^14-17 in chemical synthesis is eco-friendly and free from pollution hazard. The microwave-assisted reactions are rapid, safe, high yielding¹⁸ and superior to conventional methods.

Experimental Section:

Chemicals were purchased from commercial suppliers and were used without any further purification. All the reactions were carried out in a Modified Microwave oven (Kenstar, model no: OM26.EGO). The progress of the reaction was monitored on percolated silica gel 60 F254 plates (Merck) using ethyl acetate: n-hexane (7:3) as an eluent and spot was detected by using iodine vapors. Melting points are determined by open capillary tube method and are uncorrected. The IR (KBr pellets) spectra were recorded on a Perkin Elimer-1800- spectrophotometer and H¹NMR spectra were recorded on BRUKER DRX-300MHz spectrophotometer, (TMS as a internal reference) and chemical shifts are expressed in δ. MASS spectra were recorded on Jeol D30 spectrophotometer. Elemental analyses for C, H and N were conducted using a Perkin -Elmer C, H, and N analyzer.

Synthesis of 1-isonicotinoylpyrazolidine-3, 5-dione, 1

Method A (Conventional): Isonicotinohydrazide (0.01 mol), diethyl malonate (0. 01mol), two to three drops of glacial acetic acid and absolute ethanol (20mL) were taken in a round bottom flask. The mixture was well stirred and refluxed on a water-bath for about 12 hr. The TLC monitored the completion of the reaction. After then the reaction mixture was cooled at room temperature and pour into crushed ice. Solid separated was recrystallized from ethanol.

Method B (Microwave): Isonicotinohydrazid (0.01 mol), diethyl malonate (0.01mol) and two to three drops of glacial acetic acid were taken in a Erlenmeyer flask. Then the well-stirred mixture was irradiated in microwave oven for 4 min at 480 W (i.e. 40% microwave power). The completion of the reaction was monitored by TLC. The brown colour oily mass obtained was cooled and the crud product was recrystallized from ethanol to give compound 1. Spectral and analytical data were found to similar as reported for conventional method. The physical data and R_fvalueare recorded in Table-1

General procedure for the synthesis of Mannich bases of 1- isonicotinoylpyrazolidine-3, 5-dione (2a-j)

Method A (Conventional): 1-isonicotinoylpyrazolidine-3, 5-dione derivatives were synthesis under Mannich reaction. Mixtures of 1 (0.01 mole) and various types of secondary or primary amines (0.015 mol), catalytic amount of hydrochloric acid were dissolved in alcohol (30 mL). To this well stirred cooled solution, formaldehyde (0.02 mol) was added drop wise drop during a period 20 min. The reaction-mixture was then stirred for 1hr and refluxing for 12 hr. After completion of the reaction, as indicated by TLC, the solvent was removed under reduced pressure then poured onto crushed ice. The product obtained was filtered, washed with water and recrystallized from methanol.

Method B (Microwave): A mixture of 1-isonicotinoylpyrazolidine-3, 5-dione, various types of secondary or primary amines (0.015 mol), formaldehyde (0.02 mol), a catalytic amount of hydrochloric acid and DMF (20 mL) was taken in Erlenmeyer flask and mixed thoroughly. The resulting reaction mixture was then irradiated under microwave irradiation at intermittently at 30 sec intervals for 3-to5 min (30 power level), until completion of reaction, which was monitored by using silica gel TLC. The reaction mixture was cooled and poured onto crushed ice. The precipitates thus obtained were filtered, washed with water and purified by recrystallization from methanol. The physical data and R_fvalueare recorded in Table-1.

Antimicrobial activity:

All the newly synthesized compounds were tested in vitro for antimicrobial activity against four bacterial strains two-gram positive bacteria (Bacillus subtilis and staphylococcus aureus) and two-gram negative bacteria (Escherichia coli and P.aeruginosa) and antifungal activity against Candida albicans and Asperigillus niger at the concentration of 200 µg/ml by using the disk diffusion technique^[19]. The dimethylsulfoxide (DMSO) was used as a solvent. For this method, Mueller-Hinton agar was used as the growth medium for the bacterial strains and Sabouraud agar was growth medium for fungal species. The sterilized agar media were poured in to petridishes and allow solidifying. On the surface of the media microbial suspension were spread with the help of sterilized triangular loop. (Inoculums of standard suspension 0.1 mL of the test organism strain which contains 106 bacilli / mL). The petridishes used for antibacterial screening were incubated at 37 ±1°C for 24 h, while those for antifungal activity were incubated at 28 ±1°C for 48-72 h. DMSO was used as a control for all the type of microorganisms. The control showed no activity against the strains of microorganisms used. Antimicrobial activity and antifungal activity were measured as a function of diameter of zone of inhibition (mm). The result was compared with standard drugs Ciprofloxacin for antibacterial activity and Clotrimazole for antifungal activity by measuring the zone of inhibition in mm. The results are given in Table 2.

The minimum inhibitory concentration (MIC) was determined using tube dilution method according to the standard procedure^[20]. Mueller Hinton broth was used as a culture medium. Sterilized medium was dispensed in each borosilicate glass test tube. The synthesized compounds solution was added in order to attain final drug concentrations of 400, 200, 100, 50, 25, 12.5 µg/mL. Inoculums of standard suspension (0.1 mL of the test organism strain which contains 106 bacilli / mL) were added. The tubes were incubated at 37 ºC for 48 hr and then examined for the presence or absence of growth of the organism. The lowest concentration, which showed no visible growth, was taken as an end point minimum inhibitory concentration (MIC). The results are given in Table 3.

The antifungal activity of compounds has been assayed in vitro at a concentration of 100 µg/mL against Candida albicans and Asperigillus niger. Which were maintained on nutrient agar slants, stored at 4 ºC. The testing results are given in Table 2 and MIC on Table 3.

Table.1: The physical data and R_fvalueof the synthesized compounds1 and 2a –2j.

Compound.	Molecular Formula.	MolWt.	Conventional Method		Microwave Method		m.p °C	R_f
Compound.	Molecular Formula.	MolWt.	Yield (%)	Reaction time (Hours)	Yield %	Reaction time (min).	m.p °C	R_f
1	C₉H₇N₃O₃	205	51	12	88	4.0	116	0.72
2a	C₁₄H₁₆N₄O₄	304	47	8	87	3.0	122	0.77
2b	C₁₅H₁₉N₅O₃	317	52	6	88	2.5	124	0.72
2c	C₁₆H₁₂N₆O₃	336	53	11	86	3.5	136	0.74
2d	C₁₇H₁₃N₅O₃	335	50	9	87	3.0	145	0.75
2e	C₁₃H₁₁N₅O₃	285	49	12	88	3.0	134	0.79
2f	C₁₄H₁₈N₄O₃	290	50	13	83	3.5	189	0.71
2g	C₁₂H₁₄N₄O₃	262	48	12	84	4.0	169	0.78
2h	C₁₃H₁₃N₅O₃S	319	45	10	80	3.5	187	0.73
2i	C₁₆H₁₃FN₄O₃	328	45	12	84	4.0	156	0.76
2j	C₁₆H₁₃ClN₄O₃	334	45	13	87	4.5	169	0.71

Table 2. Zone of Inhibition in mm

Zone of Inhibition in mm (MIC in µg/mL)
Compd	Conc. (μg\ mL)	Antibacterial Activity				Antifungal Activity
Compd	Conc. (μg\ mL)	*E.Coli*	*P. aeriginosa*	S. aureus	B. subtilis	*C. albicans*	A. niger
2a	100	15	13	11	14	15	12
2b	100	16	14	14	20	18	16
2c	100	18	12	23	14	23	22
2d	100	14	12	18	14	24	22
2e	100	19	18	13	14	23	22
2f	100	20	12	23	14	12	13
2g	100	20	12	13	24	13	12
2h	100	28	25	20	16	16	18
2i	100	26	24	20	14	16	15
2j	100	24	26	21	21	16	16
Ciprofloxacin	10	32	31	30	28	-	-
Clotrimazole	20	-	-	-	-	28.	27.

Table-3:- Minimum Inhibitory Concentration (MIC) of the synthesized compounds (3a-fand 4a-f)

	Minimum Inhibitory Concentration (MIC) in µg/mL
Compd.	Antibacterial Activity				Antifungal Activity
Compd.	*E. .coli*	*P. eruginosa*	*S. aureus*	*B .subtilis*	*C. albicans*	*A. niger*
2a	50	50	50	50	50	50
2b	25	25	50	50	50	50
2c	50	50	50	50	12.5	12.5
2d	25	50	25	25	12.5	12.5
2e	25	50	50	50	12.5	12.5
2f	25	25	25	12.5	50	50
2g	25	25	25	12.5	100	100
2h	12.5	12.5	25	25	50	50
2i	12.5	12.5	25	25	50	50
2j	12.5	12.5	25	25	50	50
Ciprofloxacin	3.125	3.125	3.125	3.125	--	--
Clotrimazole	--	--	--	--	6.25	6.25

1-isonicotinoylpyrazolidine-3, 5-dione.1

Elemental analysis (Found, %): C, 52.35; H, 3.24; N, 20.77; (Calculated, %): C, 52.69 H, 3.44; N, 20.48; Mol. Formula: C₉H₇N₃O₃; IR (KBr, cm-¹): 3230 (-N H), 3100 (pyridine-H), 2325 (-CH₂, pyrazolidine), 1720 (C=O, isonicotinohydrazide), 1690, 1670, (C=O, pyrazolidine.), 1630, (C=N), 1345 (N-N); ¹HNMR (δ, DMSO): 7.65-8.40 (m, 4H, pyridine), 8.70 (s, H, HN), 3.35 (s, 2H, -CH₂ -); MS: m/z [M]⁺ 205.

1-Isonicotinoyl-2- (morpholinomethyl) pyrazolidine-3, 5-dione, 2a

Elemental analysis (Found, %): C, 55.40; H, 5.50; N, 18.20; (Calculated, %): C, 55.26; H, 5.30; N, 18.41; Mol. Fofmula: C₁₄H₁₆N₄O₄;IR (KBr, cm-¹): 3110 (pyridine-H), 2925 (alipatic-CH₂-.), 2325 (CH_2, pyrazolidine), 1728 (C=O, isonicotinohydrazide), 1685, 1675 (C=O pyrazolidine), 1630 (C=N), 1340 (N-N); 1280 (-C=N- tertiary amine); ¹HNMR (δ, DMSO): 7.70-8.38 (m, 4H, pyridine), 4.90 (s, 2H, -N-CH₂ -N-), 4.07 (s 2H -CH₂ - Pyrazolidine), 3.57 (t, 4H, CH₂-O-CH₂-), 2.80 (t, 4H, CH₂-N-CH₂-); MS: m/z [M]⁺ 304

1-Isonicotinoyl-2- ((4-methylpiperazin-1-yl) methyl) pyrazolidine-3, 5-dione, 2b

Elemental analysis (Found, %): C, 57.00; H, 5.80; N, 21.80; (Calculated, %): C, 56.77; H, 6.03; N, 22.07: Mol. Formula: C₁₅H₁₉N₅O₃; IR (KBr, cm-¹): 3115 (pyridine-H), 2935 (alipatic-CH₂-.), 2322 (-CH₂- pyrazolidine), 1735 (C=O, isonicotinohydrazide), 1680, 1672 (C=O pyrazolidine), 1632 (C=N), 1343 (-N-N-), 1282 (-C-N, tertiary amine); ¹HNMR (δ, DMSO): 7.92-9.04 (m, 4H, pyridine), 5.10 (s, 2H, -N-CH₂ -N-) 3.10 (s, 2H-CH₂pyrazolidine), 2.40 (m, 8H, -CH₂-, piperazin) 1.80 (s, 3H, CH₃); MS: m/z [M]⁺ 317

1-((1H-benzo[d][1,2,3] triazol-1-yl) methyl)-2-isonicotinoylpyrazolidine-3, 5-dione, 2c

Elemental analysis (Found, %): C, 57.44; H, 3.89; N, 25.15; (Calculated, %): C, 57.14; H, 3.60; N, 24.99; Mol. Formula: C₁₆H₁₂N₆O₃;IR (KBr, cm-¹): 3095 (pyridine-H), 2925 (alipatic-CH₂-), 2300 (-CH₂-pyrazolidine), 1745 (C=O, isonicotinohydrazide), 1678, 1662 (C=O, pyrazolidine.), 1632 (C=N), 974 (N=N-N); ¹HNMR (δ, DMSO): 7.92-9.04 (m, 4H, pyridine), 7.16-7.60 (m, 4H, Ar-H), 5.30 (s, 2H, -N-CH₂ -N-), 3.15 (s 2H-CH₂) pyrazolidine, MS: m/z [M]⁺ 336.

1-((1H-benzo[d] imidazol-1-yl) methyl)-2-isonicotinoylpyrazolidine-3, 5-dione, 2d

Elemental analysis: (Found, %), C, 61.10; H, 4.20; N, 21.20, (Calculated, %), C, 60.89; H, 3.91; N, 20.89; Mol. Formula: C₁₇H₁₃N₅O₃; IR (KBr, cm-¹), 2930 (alipatic-CH₂-.), 2332 (CH₂ pyrazolidine), 1740 (C=O, isonicotinohydrazide), 1675, 1660, (C=O pyrazolidine.), 1628,1606, (C=N), 1340 (N-N); 1267 (-C-N tertiary amine); ¹HNMR: (δ, DMSO): 7.16-7.50 (m, 4H, Ar-H), 7.70-8.12 (m, 4H, pyridine), 8.24 (s, H, H-C=N-) 5.00 (s, 2H, -N-CH₂-N-), 3.80 (s, 2H-CH₂-Pyrazolidine); MS: m/z [M]⁺ 335

1-((1H-imidazol-1-yl) methyl)-2-isonicotinoylpyrazolidine-3, 5-dione, 2e

Elemental analysis: (Found, %): C, 54.89; H, 4.10; N, 24.80 (Calculated, %): C, 54.74; H, 3.89; N, 24.55: Mol. Formula: C₁₃H₁₁N₅O₃; IR (KBr, cm-¹), 2936 (alipatic-CH₂-.), 2338 (-CH₂-pyrazolidine), 1745 (C=O, isonicotinohydrazide), 1665, 1650 (C=O pyrazolidine), 1632, 1612 (C=N), 1343 (N-N); ¹HNMR: (δ, DMSO), 7.70-8.68 (m, 4H, pyridine), 6.88-7.45 (m, 3H, imidazole), 4.25 (s, 2H-CH₂- pyrazolidine), 5.64 (-N-CH₂-N-) MS: m/z [M]⁺ 285

1-((diethylamino) methyl)-2-isonicotinoylpyrazolidine-3, 5-dione, 2f

Elemental analysis (Found, %): C, 58.24; H, 5.92; N, 19.10 (Calculated, %): C, 57.92; H, 6.25; N, 19.30; Mol. Formula: C₁₄H₁₈N₄O₃; IR (KBr, cm-¹): 3090 (pyridine-H), 2928 (alipatic-CH₂-.), 2310 (-CH₂-pyrazolidine), 1742 (C=O, isonicotinohydrazide), 1675, 1667 (C=O, pyrazolidine.), 1634 (C=N), 1278 (-C-N, tertiary amine);¹HNMR (δ, DMSO): 7.88-9.10 (m, 4H, pyridine), 5.09 (s, 2H, -N-CH₂ -N-), 3.25 (s, 2H, -CH₂- pyrazolidine), 2.84 (q, 4H, N-CH₂-) 1.28 (t, 6H -CH₃); MS: m/z [M]⁺ 290;

1-((dimethylamino) methyl)-2-isonicotinoylpyrazolidine-3, 5-dione, 2g

Elemental analysis (Found, %): C, 55.24; H, 5.59; N, 21.15; (Calculated, %): C, 54.96; H, 5.38; N, 21.36; Mol. Formula: C₁₂H₁₄N₄O₃; IR (KBr, cm-¹): 3105 (pyridine-H), 2928 (alipatic-CH₂-.), 2310 (-CH₂- pyrazolidine), 1738 (C=O, isonicotinohydrazide), 1672, 1660 (C=O, pyrazolidine), 1642 (C=N), 1282 (-C-N, tertiary amine); ¹HNMR (δ, DMSO): 7.87-9.02 (m, 4H, pyridine), 4.78 (s, 2H, -N-CH₂ -N-), 3.15 (s 2H, -CH₂- pyrazolidine), 2.84 [s, 6H, -(CH₃)₂]; MS: m/z [M]⁺ 262

1-isonicotinoyl-2- ((thiazol-2-ylamino) methyl) pyrazolidine-3, 5-dione, 2h

Elemental analysis (Found, %): C, 49.54; H, 3.12; N, 22.25; (Calculated, %): C, 49.21; H, 3.49; N, 22.07; Mol. Formula: C₁₃H₁₁N₅O₃S; IR (KBr, cm^-1): 3340 (N-H), 3115 (pyridine-H), 2922 (alipatic-CH₂-), 2318 (-CH_2, pyrazolidine), 1748 (C=O, isonicotinohydrazide), 1672, 1662 (C=O, pyrazolidine.), 1630 (C=N); ¹HNMR (δ, DMSO): 7.88-9.10 (m, 4H, pyridine), 4.56 (t, 1H, N-H), 5.09 (s, 2H, -N-CH2 -N-), 3.25 (s, 2H, -CH₂- pyrazolidine) 7.45 -6.56 (m, 2H,thiazol ring); MS: m/z [M]⁺ 317;

1-((4-fluorophenylamino) methyl)-2-isonicotinoylpyrazolidine-3, 5-dione, 2i

Elemental analysis (Found, %): C, 58.34; H, 5.52; N, 16.85; (Calculated, %): C, 58.54; H, 3.99; F, 5.79; N, 17.07; Mol. Formula: C₁₆H₁₃FN₄O_3; IR (KBr, cm-¹): 3345 (N-H), 3115 (pyridine-H), 2922 (alipatic-CH₂-), 2318 (-CH₂- pyrazolidine), 1748 (C=O, isonicotinohydrazide), 1672, 1662 (C=O, pyrazolidine.), 1630 (C=N), 1150 (C-F);¹HNMR (δ, DMSO): 7.88-9.10 (m, 4H, pyridine), 6.87-7.15 (m, 4H, Ar-H), 6.30 (s, 1H, N-H), 5.09 (s, 2H, -N-CH₂ -N-), 3.25 (s, 2H, -CH₂- pyrazolidine); MS: m/z [M]⁺ 328;

1-((4-chlorophenylamino) methyl)-2-isonicotinoylpyrazolidine-3, 5-dione, 2j

Elemental analysis (Found, %): C, 55.44; H, 4.12; N, 16.45; (Calculated, %): C, 55.74; H, 3.80; N, 16.25; Mol. Formula: C₁₆H₁₃ClN₄O_3; IR (KBr, cm-¹): 3340 (N-H), 3115 (pyridine-H), 2922 (alipatic-CH₂-), 2318 (-CH₂- pyrazolidine), 1748 (C=O, isonicotinohydrazide), 1672, 1662 (C=O, pyrazolidine), 1630 (C=N), 1050 (C-Cl);¹HNMR (δ, DMSO): 7.88-9.10 (m, 4H, pyridine), 6.87-7.15 (m, 4H, Ar-H), 6.30 (s, 1H, N-H), 5.09 (s, 2H, -N-CH₂ -N-), 3.25 (s, 2H, -CH₂- pyrazolidine); MS: m/z [M]⁺ 344;

RESULT AND DISCUSSION:

The starting materials obtained commercially and were used after purification. 1-isonicotinoylpyrazolidine-3, 5-dione (1) has been synthesized by the condensation of isonicotinohydrazide, diethyl malonate and a catalytic amount of glacial acetic acid. The mannich bases have been synthesized by the reaction of (1) with formaldehyde and various primary or secondary amines in the presence of catalyst hydrochloric acid under microwave irradiation and also by the conventional method. It is noteworthy that the reaction which required 9-12 hr in conventional method, was completed within 3-5 min in Microwave system power level of 300 W. Yield has been remarkably improved from 45- 53 % to 80- 87 %. The yield and the physical constants of the compounds synthesized by the conventional and microwave irradiation methods are given (Table I).

The molecular formulae of the compounds were confirmed by the elemental analysis and their structures were determined from IR, ¹H NMR, and mass spectral data. The IR spectra of the compound 1 displayed the characteristic N-H stretching vibration at 3230cm^-1. The ¹H NMR spectra of this compound gave characteristic peaks in the expected regions. MS of this compound showed the molecular ion peak (M⁺) at 205 corresponding to molecular formula C₉H₇N₃O₃. The structures of all the Mannich bases (2a-j) showed ¹H NMR singlet at 4.78 to 5.64 due to CH₂ protons of N-CH₂-N group and gave characteristic peaks in the expected regions.

The screening results of antibacterial activities showed that the compounds 2h, 2i, 2j, showed significant activity at 100 µg/mL concentration against E. coli and P.aeruginosa gram positive bacteria strains. Compound 2f showed good activity against B. subtilis at 100 µg/mL concentrations.

Compound 2g showed good activity against S. aureus, compared with the standard drug Ciprofloxacin.

For antifungal activity, compounds 2c, 2d, 2e showed significant activity against C. albicans and A. niger at 100 µg/ml concentration, compared with the standard drug Clotrimazole.

Antimicrobial activity:

All the newly synthesized compounds were tested in vitro for antimicrobial activity against four bacterial strains two-gram positive bacteria (Bacillus subtilis and staphylococcus aureus) and two-gram negative bacteria (Escherichia coli and P.aeruginosa) and antifungal activity against Candida albicans and Asperigillus niger at the concentration of 100 µg/ml by using the disk diffusion technique²⁰. The dimethylsulfoxide (DMSO) was used as a solvent. For this method, Mueller-Hinton agar was used as the growth medium for the bacterial strains and Sabouraud agar was growth medium for fungal species. The sterilized agar media were poured in to petridishes and allow solidifying. On the surface of the media microbial suspension were spread with the help of sterilized triangular loop. (Inoculums of standard suspension 0.1 mL of the test organism strain which contains 106 bacilli / mL). The petridishes used for antibacterial screening were incubated at 37 ±1°C for 24 h, while those for antifungal activity were incubated at 28 ±1°C for 48-72 h. DMSO was used as a control for all the type of microorganisms. The control showed no activity against the strains of microorganisms used. Antimicrobial activity and antifungal activity were measured as a function of diameter of zone of inhibition (mm). The result was compared with standard drugs Ciprofloxacin for antibacterial activity and Clotrimazole for antifungal activity by measuring the zone of inhibition in mm. The results are given in Table 2.

The minimum inhibitory concentration (MIC) was determined using tube dilution method according to the standard procedure²¹. Mueller Hinton broth was used as a culture medium. Sterilized medium was dispensed in each borosilicate glass test tube. The synthesized compounds solution was added in order to attain final drug concentrations of, 100, 50, 25, 12.5 and 6.25 µg/mL. Inoculums of standard suspension (0.1 mL of the test organism strain which contains 106 bacilli / mL) were added. The tubes were incubated at 37 ºC for 48 hr and then examined for the presence or absence of growth of the organism. The lowest concentration, which showed no visible growth, was taken as an end point minimum inhibitory concentration (MIC). The antifungal activity of compounds has been assayed in vitro at a concentration of 100,50,25,12.5 and 6.25 µg/mL against Candida albicans and Asperigillus niger. Which were maintained on nutrient agar slants, stored at 4 ºC. The testing results are given in Table 2.

ACKNOWLEDGMENT:

The authors are thankful to HOD, Department of Chemistry, University College of Science, M.L.Sukhadia University Udaipur (Rajasthan) for providing laboratory facilities authors are also thankful to Director, CDRI Lucknow, India for providing spectral and analytical data.

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Received on 01.12.2010 Modified on 15.12.2010

Asian J. Research Chem. 4(3): March 2011; Page 455-460

Sudhir Bharadwaj1, Bharat Parashar2, Dimple K. Rathore3* and V.K. Sharma3

INTRODUCTION:

ACKNOWLEDGMENT:

The authors are thankful to HOD, Department of Chemistry, University College of Science, M.L.Sukhadia University Udaipur (Rajasthan) for providing laboratory facilities authors are also thankful to Director, CDRI Lucknow, India for providing spectral and analytical data.

Sudhir Bharadwaj¹, Bharat Parashar², Dimple K. Rathore³*and V.K. Sharma³