ACCEPTED MANUSCRIPT
was refluxed for 2 h. After evaporation of the solvent in vacuo the compound was purified by silica-gel column
chromatography using chloroform as an eluent, then was characterized by IR, NMR spectroscopy, EI, and
identified as 1-((ferrocenyl)methylene)-2-(9H-fluoren-9-ylidene)hydrazine 3a (89% yield) or 1-
((ferrocenyl)methylene)-2-(diphenylmethylene)hydrazine 3b (90% yield), respectively.
-1
1
1
-((Ferrocenyl)methylene)-2-(9H-fluoren-9-ylidene)hydrazine, 3a: IR (cm ): 1625 (C=N). H NMR
(
CDCl ), δ: 4.28 (s, 5H, CH ), 4.58 (s, 2H, CH ), 4.88 (s, 2H, CH ), 7.33-7.37 (m, 2H, CH ), 7.43-7.47 (q,
3 Fc Fc Fc ar
2
7
H, JHH = 6.6 Hz, CH ), 7.64 (d, 1H, JHH = 7.4 Hz, CH ), 7.67 (d, 1H, J = 7.4 Hz, CH ), 7.93 (d, 1H, J
=
ar
ar
HH
ar
HH
1
3
3
.4 Hz, CHar), 8.54 (s, 1H, C(H)=N), 8.59 (d, 1H, JHH = 7.4 Hz, CHar). C NMR (CDCl ), δ: 69.1 (CHFc), 69.6
(CH ), 71.5 (CH ), 78.2 (C ), 119.8 (CH ), 122.6 (CH ), 128.0 (CH ), 128.1 (CH ), 130.6 (CH ), 130.7
Fc Fc Fc ar ar ar ar ar
(
CHar), 131.1 (CHar), 131.9 (Car), 137.1 (Car), 141.5 (Car), 142.2 (Car), 159.2 (C=N), 161.6 (C(H)=N). DEPT-
35 NMR (CDCl ), δ: 69.1 (CH ), 69.6 (CH ), 71.5 (CH ), 119.8 (CH ), 122.6 (CH ), 128.0 (CH ), 128.1
1
3
Fc
Fc
Fc
ar
ar
ar
+
(
CH ), 130.6 (CH ), 130.7 (CH ), 131.1 (CH ). EI: m/z 390.1 [M] .
ar ar ar ar
-1
1
-((Ferrocenyl)methylene)-2-(diphenylmethylene)hydrazine, 3b: IR (cm ): 1679 and 1656 (C=N), 1602
1
(
C=C). H NMR (CDCl
3
), δ: 4.22 (s, 5H, CHFc), 4.43 (s, 2H, CHFc), 4.62 (s, 2H, CHFc), 7.29-7.84 (m, 10H,
1
3
CH ), 8.53 (s, 1H, C(H)=N). C NMR (CDCl ), δ: 68.8 (CH ), 69.3 (CH ), 69.7 (CH ), 70.9 (CH ), 73.2
ar
3
Fc
Fc
Fc
Fc
(
(
(
(
(
(
CH ), 77.3 (CH ), 78.7 (C ), 127.5 (CH ), 127.9 (CH ), 128.0 (CH ), 128.2 (CH ), 128.3 (CH ), 128.7
Fc Fc Fc ar ar ar ar ar
CHar), 128.9 (CHar), 129.0 (CHar), 129.3 (CHar), 129.9 (CHar), 130.1 (CHar), 130.4 (CHar), 132.4 (CHar), 135.7
C ), 138.6 (C ), 159.6 (C=N), 164.2 (C(H)=N). DEPT-135 NMR (CDCl ), δ: 68.8 (CH ), 69.3 (CH ), 69.7
ar
ar
3
Fc
Fc
CHFc), 70.9 (CHFc), 73.2 (CHFc), 77.3 (CHFc), 127.5 (CHar), 127.9 (CHar), 128.0 (CHar), 128.2 (CHar), 128.3
CH ), 128.7 (CH ), 128.9 (CH ), 129.0 (CH ), 129.3 (CH ), 129.9 (CH ), 130.1 (CH ), 130.4 (CH ), 132.4
ar
ar
ar
ar
ar
ar
ar
ar
+
CH ). EI: m/z 392.1 [M] .
ar
2
2
.2. Biology
.2.1. Antimicrobial activities
The antimicrobial activities of 2a, 2b, 3a and 3b were determined against some Gram-positive and Gram-
negative bacteria using paper disc diffusion method [26].
The tested Gram-positive bacteria were Staphylococcus aureus, S. saprophyteus and Streptococcus pyogenes
while, the tested Gram-negative were Acinetobacter baumannii, Escherichia coli, Klebsilla pneumonia,
Pseudomonas earuginosa, Salmonella sp. and Serrratia marcescens. All the previous bacteria were obtained
from KFG hospital, Jeddah, Saudi Arabia. Mueller Hinton agar (MHA) medium was used for bacteria, while
MHA supplemented with 2% dextrose was used for Candida albicans and Aspergillus niger. Each plat was
6
inoculated with 100 µL of the tested microbe (4x10 cfu/mL) and using sterile glass rod, the inoculum was