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Evaluation of antimicrobial activities of Harmine, Harmaline, Nicotine and their

complexes

Article in Pakistan journal of pharmaceutical sciences · August 2016

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SHORT COMMUNICATION

Evaluation of antimicrobial activities of Harmine, Harmaline, Nicotine

and their complexes

Saad Salman1, Fariha Idrees2, Sadia Pervaiz3, Fahad Hassan Shah4, Sareer Badshah2,
Abdullah1, Mohammad Usman1, Syed Muhammad Ashhad Halimi1 and Jawaria Idrees2
1
Department of Pharmacy, Peshawar, Pakistan
2
Islamia College University, Peshawar, Pakistan
3
Department of Pharmacy, Abdul Wali Khan, Mardan, Pakistan
4
Centre of Biotechnology and Microbiology, Peshawar, Pakistan

Abstract: Harmine, Harmaline, Nicotine and its various complexes synthesized have been characterized by physical,
spectral and analytical methods and curtained for in-vitro antimicrobial activity against different bacterial and fungal
species at two different concentrations i.e.100µg/100µl and 200µg/100µl dose level respectively. Analysis showed that
Nicotine, Zinc-Nico, Cd-Nico, Hg-Nico, Ni-Nico, Cu-Nico, Co-Nico, Harmine, and Harmaline having conc. of 100ug/
100ul had antibacterial activity on zero, 5, 4, 10, zero, 5, 7, zero, zero strain of bacteria having an average of zero
(SD=0.0000), 15.2000 (SD=1.30384), 18.2500 (SD=3.30404), 20.2000 (SD=1.39841), zero (SD=0.0000), 14.6000
(SD=0.89443), 15.8571 (SD=1.34519), zero (SD=0.0000), zero (SD=0.0000) respectively. Zinc (II) chloride, Cadmium
(II) Iodide, Mercury (II) chloride, Nickel (II) chloride, Copper (II) chloride, Cobalt (II) chloride, Mercury (II) chloride,
Mercury (II) harmine, Mercury (II) harmaline at 100ug/100ul is valid for 7, 8, 9, 2, 7, 8, 9, 10, 8 strains of bacteria with
an average of 7.1429 (SD=1.06904), 10.0000 (SD=5.01427), 14.8889 (SD=6.00925), 6.0000 (SD=0.0000), 8.5714
(SD=4.27618), 8.2500 (SD=0.88641), 14.8889 (SD=6.00925), 18.6000 (SD=2.45855), 18.5000 (SD=1.85164)
respectively. The above given compounds at the conc. of 200 µg/100ul is valid for 10, 9, 10, 8, 8, 10, 10, 10, 10 strains of
bacteria with an average of 8.1 (SD=1.66333), 11.7778 (SD=5.28625), 16.1000 (SD=6.36745), 6.5000 (SD=0.92582),
9.7500 (SD=4.43203), 9.9000 (SD=2.76687), 16.1000 (SD=6.36745), 22.0000 (SD=2.44949), 20.4000 (SD=2.75681)
respectively. The above given compounds at conc. of 200 µg/100ul showed antibacterial action on 3, 8, 8, 10, 3, 9, 8,
zero, 3 strains of bacteria with an average of 14(SD=0.000), 16.8750 (SD=1.35620), 18.2500 (SD=3.45378), 22.7000
(SD=1.82878), 14.3333 (SD=0.57735), 16.7778 (SD=1.71594), zero (SD=0.000), 12.0000 (SD=1.00000) respectively.
Hence according to the average value of the zone of inhibition, maximum antibacterial activity at 100-200ug/100ul is of
Hg-Nico and Mercury salt; Mercury (II) harmine having an average of 20.2000 (SD=1.39841)-22.7000 (SD=1.82878)
and 18.6000 (SD=2.45855)-22.0000 (SD=2.44949). Minimum antibacterial activity at 100-200ug/100ul is Nicotine100,
Nicotine-Nico100, Harmine 100,Harmaline 100, Harmine 200 having zero average (SD=0.000).

Keywords: Nicotine, Harmaline, Harmine, zone of inhibition, anti-microbial activity, complexes.

INTRODUCTION towards all the selected species of fungi except

Fusariummoniliforme but its metal complexes showed
Harmine and its metal complexes were isolated from good antimicrobial properties (Chen et al., 2015). One
seeds of Peganum harmala. Inhalation of its smoke study showed that harmaline inhibited the dioxin-
relieves pain in the liver. It is used for the treatment of mediated induction of CYP1A1 at the transcriptional and
jaundice, asthma and colic as well. (Niroumand et al., posttranslational levels (Gendy et al., 2012) and showed
2015) Usually powder of the seeds and watery infusion effective antimicrobial activity against suspensions and
are given for treatment of these disease. The active biofilm cultures of S. aureus. It also shows anti-HIV and
alkaloids of Harmal seeds are the MAOI-A (monoamine anti-parasitic properties (Khan et al., 2013).
oxidase inhibitor A) compounds. Pegnumharmala extract
have antifungal and antimicrobial effects (Sassoui et al., Nicotine, 3-(1-methyl-2-pyrrolidinyl) pyridine is present
2015). Psychoactive effects of Esfand seeds are rooted in in the leaves of Nicotiana tabacum. Nicotine is an
inhibitory effect of monoamine oxidase (MAO) of important bioligand and has good chelating sites for
betacarboline alkaloids effects the cardiovascular system coordination with numerous metals (Rajesh Arora et al.,
(Zhang et al., 2014). Harmine alone was ineffective 2010, Zaidi et al., 2012). The novel copper complexes
with nicotinic acid and carboxylic acids were shown to
*Corresponding author: e-mail: [email protected] exhibit super oxide scavenging activity as well as
Pak. J. Pharm. Sci., Vol.29, No.4, July 2016, pp.197-200 197
Evaluation of antimicrobial activities of Harmine, Harmaline, Nicotine and their complexes

antimicrobial activities against B. subtilis (Malczewska-

Jaskóła et al., 2015).

Nicotinic acid derived Schiff bases and their transition

metal complexes showed antibacterial activity against
pathogenic strains of Escherichia coli, Staphylococcus
aureus and Pseudomonas aeruginosa (Patel et al., 2012,
Xing et al., 2012). Siverbidentateligands based on
oligomers of polyethylene glycol, functionalized at both
ends with either nicotinic or iso-nicotinic acid showed
good antimicrobial properties(Suksrichavalit et al., 2008).

In our study: Nicotine, Zinc-Nico, Cd-Nico, Hg-Nico, Ni-

Nico, Cu-Nico, Co-Nico, Harmine and Harmaline, Zinc Fig 3: Activity of Harmine, Harmaline, Nicotine and its
(II) chloride, Cadmium (II) Iodide, Mercury (II) chloride, various complexes at 100 µg/100µl.
Nickel (II) chloride, Copper (II) chloride, Cobalt (II)
chloride, Mercury (II) chloride, Mercury (II) harmine and Zinc (II) chloride, Cadmium (II) Iodide, Mercury (II)
Mercury (II) harmaline were prepared and analyzed at chloride, Nickel (II) chloride, Copper (II) chloride, Cobalt
different concentrations for antimicrobial properties. (II) chloride, Mercury (II) chloride, Mercury (II) harmine,
Mercury (II) harmaline at 100ug/100ul is valid for 7, 8, 9,
MATERIALS AND METHODS 2, 7, 8, 9, 10, 8 strains of bacteria with an average of
7.1429 (SD=1.06904), 10.0000 (SD=5.01427), 14.8889
The study was conducted at HEJ Karachi, Pakistan. The (SD=6.00925), 6.0000 (SD=0.0000), 8.5714
antimicrobial sensitivity tests of the compounds were (SD=4.27618), 8.2500 (SD=0.88641), 14.8889
tested against ten different species of gram positive and (SD=6.00925), 18.6000 (SD=2.45855), 18.5000
gram negative bacteria including Escherichia coli, (SD=1.85164) respectively.
Streptococcus pyogenes, Klebsiellapneumoniae, Bacillus
cereus, Pseudomonas aeruginosa, Proteus mirabilis, At 200ug/100ul
Shigelladysenteriae, Salmonella typhi, Staphylococcus 5. The above given compounds at the conc. of 200
aureus and Corynebacterium diphtheriae bacterial strains ug/100ul was valid for 10, 9, 10, 8, 8, 10, 10, 10, 10
and for in vitro antifungal activity against Candida strains of bacteria with an average of 8.1(SD=1.66333),
albicans, Epidermophyton floccosum, Microsporumcanis, 11.7778 (SD=5.28625), 16.1000 (SD=6.36745), 6.5000
Fusariummonoliformi, Alternariainfectoria, (SD=0.92582), 9.7500 (SD=4.43203), 9.9000
Fusariumsolani, Aspergillusnigar, Pseudallescherisboydii, (SD=2.76687), 16.1000 (SD=6.36745), 22.0000
Candida solani, Penicillium, Penicilliumnotatum, (SD=2.44949), 20.4000 (SD=2.75681) respectively. The
Saccharomyces ceravaceae, Candia tropicalis, Candia above given compounds at conc. of 200 ug/100ul showed
lusitaniae. The agar medium for testing of both anti- antibacterial action on 3, 8, 8, 10, 3, 9, 8, zero, 3 strains of
bacterial and anti-fungal activities is given (table 1). The bacteria with an average of 14 (SD=0.000), 16.8750
compounds were used in two concentrations (SD=1.35620), 18.2500 (SD=3.45378), 22.7000
i.e.100µg/100µl (first dose level) and 200µg/100µl, the (SD=1.82878), 14.3333 (SD=0.57735), 16.7778
second dose level. (SD=1.71594), zero (SD=0.000), 12.0000 (SD=1.00000)
respectively.
RESULTS

Analysis of the zone of inhibition of Harmine,

Harmaline, Nicotine and their complexes in millimeters
At 100ug/100ul
Analysis showed that Nicotine, Zinc-Nico, Cd-Nico, Hg-
Nico, Ni-Nico, Cu-Nico, Co-Nico, Harmine, and
Harmaline having conc. of 100ug/100ul had antibacterial
activity on zero, 5, 4, 10, zero, 5, 7, zero, zero strain of
bacteria having an average of zero (SD=0.0000), 15.2000
(SD=1.30384), 18.2500 (SD=3.30404), 20.2000
(SD=1.39841), zero (SD=0.0000), 14.6000
(SD=0.89443), 15.8571 (SD=1.34519), zero
(SD=0.0000), zero (SD=0.0000) respectively. Fig 4: Activity of Harmine, Harmaline, Nicotine and its
various complexes at 200µg/200µl.

198 Pak. J. Pharm. Sci., Vol.29, No.4, July 2016, pp.197-200

 Saad Salman et al

Table 1: Ingredients of the medium and preparation of the stock solutions.

Preparation of Mueller-Hinton Medium (DIFCO Sabouraud Dextrose Agar (Oxoid Manual, 1976,
laboratories, 1984) Sabouraud, 1910)
Constituents Strength, g/l Ingredients Strength,g, g/l
Beef Infusion 300.0 Mycological peptone1 10g
Casamino acids 17.5 g/l Dextrose 40
Starch 1.5
Agar No.12 15
Bacto Agar 17.0
pH at 25°C 7.3± 0.1 pH at 25°C 5.6
Stock solution 1mg/ml DMSO* Stock solution 5mg/ml DMSO**
1
(oxoid L40) *diluted to 100µg and 200µg/mldimethylsulphoxide (DMSO) 2(oxoid LII)**diluted to 300 µg/ml

Table 2: Anti-microbial activity of Harmine, Harmaline, Nicotine and their complexes.

Anti-micro bialactivityat 100 µg/100ul Anti-microbialactivityat 200µg/100ul

Complexes Strains Mean SD Complexes Strains Mean SD
Mercury (II) harmine 10 18.6000 2.45855 Mercury (II) harmaline 10 12.0000 1.0000
Hg-Nico 7 6.5000 0.92582 Hg-Nico 9 20.2000 1.39841
Cadmium (II) Iodide 8 10.0000 5.01427 Cadmium (II) Iodide 8 16.8750 1.35620
Cobalt (II) Chloride 8 8.2500 0.88641 Cobalt (II) Chloride 9 16.7778 1.71594
Mercury (II) Chloride 8 14.8889 6.00925 Mercury (II) Chloride 9 22.7000 1.82878
Mercury (II) Harmaline 8 18.5000 1.85164 Mercury (II) harmine 10 12.0000 1.0000
Co-Nico 7 15.8571 1.34519 Co-Nico 8 16.1000 6.36745
Zinc (II) Chloride 7 7.1429 1.06904 Zinc (II) Chloride 9 14 0.000
Copper (II) Chloride 7 8.5714 4.27618 Copper (II) Chloride 8 14.3333 0.57735
Zn-Nico 5 15.2000 1.30384 Zn-Nico 8 11.7778 5.28625
Cu-Nico 5 14.6000 0.89443 Cu-Nico 8 9.9000 2.76687
Cd-Nico 4 18.2500 3.30404 Cd-Nico 8 16.1000 6.36745
Ni-Nico 0 0 0.000 Ni-Nico 3 9.7500 4.43203
Nickel (II) Chloride 2 8.5714 4.27618 Ni (II) Chloride 10 14.3333 0.57735
Nicotine 0 0 0.000 Nicotine 3 8.1 1.66333
Harmine 0 0 0.000 Harmine 10 22.0000 2.44949
Harmaline 0 0 0.000 Harmaline 10 20.4000 2.75681
DISCUSSION Compounds that showed high antibacterial activity were:
(1) Compounds showing activity on 8 strains of bacteria:
The compounds that showed minimum or low activities Zn-Nico 200, Cadmium (II) Iodide 100, Cd-Nico 200,
were: (1) Compounds presenting zero activity: Nicotine Nickel (II) Chloride, Cu (II) Cl 200, Co (II) Cl 100,Co-
100, Nicotine-Nico 100, Harmine 100, Harmine 200, Nico 200, Mercury (II) Harmaline 100,Cadmium(II)
Harmaline 100. (2) Compound displaying activity on 1 Iodide 200, Mercury (II) Chloride 100,Cu-Nico 200.. (2)
strain of bacteria: Nil. (3) Compounds demonstrating Compounds showing activity on 9 strains of bacteria:
activity on 2 strains of bacteria was just Nickel (II) Zinc (II) Chloride 200, Mercury (II) Chloride 200, Hg-
Chloride. (4) Compound exhibiting activity on 3 strains of Nico 100, Hg-Nico 200, Cobalt (II) Chloride 200. (3)
bacteria were: Nicotine 200, Ni-Nico 200 and Harmaline Compounds showing activity on 10 strains of bacteria:
200. Mercury (II) harmine 100, Mercury (II) harmine 200 and
Mercury (II) harmaline.
Compounds that displayed moderate antibacterial activity
were: (1) Compounds exhibiting activity on 4 strains of Further research is underway on the antimicrobial
bacteria: Cd-Nico 100. (2) Compounds showing activity mechanism of this nicotine, harmaline and harmine metal
on 5 strains of bacteria: Zn-Nico 100, Cu-Nico 100. (3) complexes that either this is cell wall inhibitors or
Compounds displaying activity on 6 strains of bacteria: bactericidal or bacteriostatic. However, some researchers
Nil. (4) Compounds showing activity on 7 strains of studied considerable changes in the bacterial cell
bacteria: Zinc (II) Chloride 100, Copper (II) Chloride, 100 membranes upon metal ion treatment, which might be one
and Co-Nico 100. of the cause or consequence of cell death (Zaidi et al.,
2012).

Pak. J. Pharm. Sci., Vol.29, No.4, July 2016, pp.197-200 199

 Evaluation of antimicrobial activities of Harmine, Harmaline, Nicotine and their complexes

CONCLUSION virus infection through down regulating cellular NF-κB

and MAPK pathways induced by oxidative stress.
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maximum antimicrobial activity at 100-200ug/100ul is of Mohamed AM El Gendy, Anatoly A Soshilov, Michael S
Hg-Nico and Mercury (II) harmine. Minimum Denison and Ayman OS El-Kadi (2012). Harmaline
antibacterial activity at 100-200ug/100ul is Harmine 100, and harmalol inhibit the carcinogen-activating enzyme
Harmaline 100 and Harmine 200.As compared to nicotine CYP1A1 via transcriptional and posttranslational
alone; the nicotine (II) complexes are able to inhibit mechanisms. Food Chem. Toxicol., 50(2): 353-362.
majority of the studied gram positive and gram-negative Farhan A. Khan, Aneela Maalik, Zafar Iqbal, Imran
organisms at the higher dose level. Therefore, these Malik, Recent pharmacological developments in β-
complexes are broad-spectrum anti-microbial agents, carboline alkaloid “harmaline” (2013). European J.
active against the variety of gram positive and gram Pharmacol., 721(1-3): 391-394.
negative bacteria, with minimal antifungal activity. Rajesh Arora (2010). Plant biotechnology. Medicinal
According to the average value of the zone of inhibition, Plant Biotechnol., 8(2): 1253-1266.
maximum antimicrobial activity at 100µg/100ul was Karolina Malczewska-Jaskóła, Wojciech Jankowski,
showed by Hg-Nico and Cd-Nico. At 200µg/100µl: Cd- Beata Warżajtis, Beata Jasiewicz, Marcin Hoffmann,
Nico and Co-Nico showed maximum activity. Minimum Urszula Rychlewska (2015). Chalcogenated (S)-(-)-
anti-microbial activity at 100-200ug/100ul was showed by nicotine derivatives as chiral linkers for 1D
Nicotine100 and Nicotine-Nico100. coordination polymers. Polyhedron, 100(4): 404-411.
M Xing, F Shen, L Liu, Z Chen, N Guo, X Wang, W
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