Indones. J. Chem.

, 2016, 16 (1), 59 - 64 59

Potassium Permanganate-Catalyzed Alpha-Pinene Oxidation:

Formation of Coordination Compound with Zinc(II) and Copper(II), and Growth Inhibition
Activity on Staphylococcus aureus and Escherichia coli

Masruri*, Rekfa Wika Amini, and Mohammad Farid Rahman

Department of Chemistry, University of Brawijaya
Jl. Veteran Malang 65145, East Java Indonesia

Received January 21, 2015; Accepted September 4, 2015

ABSTRACT

Catalytic oxidation of alpha-pinene was investigated using potassium permanganate as an oxidant. The
reaction consumed catalyst following stoichiometric amount instead of the catalytic one. The keto-carboxylate
compound 2 was afforded as the oxidation product. Further study of its complex compound with copper(II) and
zinc(II) was also reported including their activity for inhibiting the growth of Staphylococcus aureus and Escherichia
coli. In overall, the complex compound shows important result by inhibiting the bacterial growth.

Keywords: alpha-pinene; catalytic oxidation; antibacterial; natural product

ABSTRAK

Telah dilakukan reaksi oksidasi alfa-pinena menggunakan kalium permanganate. Reaksi ini bekerja dengan
menggunakan katalis dalam jumlah yang stoikiometris. Produk reaksi yang diperoleh adalah senyawa keto-
karboksilat 2. Studi lebih lanjut terhadap pembentukan senyawa komplek produk 2 dengan tembaga(II) dan seng(II)
juga telah dilakukan, termasuk pengujian aktivitas penghambatan pertumbuhan pada bakteri Staphylococcus aureus
dan Escherichia coli. Secara umum, senyawa-senyawa kompleks yang dihasilkan menunjukkan aktivitas
penghambatan pertumbuhan kedua bakteri.

Kata Kunci: alfa-pinena; katalitik oksidasi; antibakteri; bahan alam

INTRODUCTION up with an issue related activity below the antibiotic

standard, volatility and solubility in water.
The alpha-pinene is a monoterpenoid compound, a The oxidation catalyzed by potassium
member group of secondary metabolite natural product. permanganate, osmium tetroxide and chromic acid of
It contains C10 with a single C=C double bond and the alpha-pinene double bond can proceed to provide
bicyclic [2.2.1]. This compound composes Indonesian two carbonyl groups or a carbonyl and one carboxyl
turpuntine oil about 60-70% [1-3]. It has also been groups [9] by the cleavage of double bond. Some
reported in some literatures contains in essential oil report has been devoted using different catalyst such
extracted from Cosmos caudatus, Eupatorium odoratum, as ozone [10-12], complex [Mn(TPP-PEO)Cl] catalyst
Hyptis species [4], Cananga odorata [5], and Thymus [13], ruthenium [14-17], rhodium [18], palladium [19],
pectinatus [6]. Purification under a reduced pressure of complex of metal-oxalate [20], and silica-supported
fractional distillation of Indonesian turpuntine oils permanganate [21]. The oxidized of alpha-pinene, then,
provided almost 99% purity alpha-pinene. It was also can coordinate with some metal ions such as Zn, Cu,
reported active in the reducing inflammation in cell level Pt, and Mo. Moreover, many reports show the
[7], inhibiting of bacterial growth but it was lesser important antibacterial activity of diketone complex
effective than the antibiotic control [2]. Further compound with Cu and Zn [22], aromatic O-N-O ligand
transformation of its functional group by catalytic with Zn, Cu, Pt, and Co [23], oxime complex compound
hydroxylation and acetyloxylation in acidic condition with Cu [24-25], amino acid complex compound with
provided a hydroxylated and acetyloxylated of alpha- Co [26], and imidazol-Co [27]. This paper discloses the
pinene derivatives. These new structures slightly recent investigation on permanganate catalyzed alpha-
increase the solubility in water, and also improve in the pinene oxidation and its complex compounds with
activity on S. aureus and E. coli [8]. However, still come zinc(II) and copper(II). The activity in inhibiting bacterial
growth is reported as well.

* Corresponding author. Tel/Fax : +62-341-575838/554403

Email address : masruri@ub.ac.id

Masruri et al.
60 Indones. J. Chem., 2016, 16 (1), 59 - 64

EXPERIMENTAL SECTION afford the product (540 mg, 92.14%) as pale yellow
solid. Further analysis was conducted using FTIR and
Materials UV-Vis spectrophotometers. Similar procedure was
undertaken using copper chloride.
Chemicals used for the research included
potassium permanganate (Merck), ethyl acetate (Smart Evaluation of bacterial growth inhibition
Lab), n-hexane (Smart Lab), magnesium sulfate The antibacterial evaluation was conducted using
anhydrate (Merck), sodium sulfate anhydrate (Merck), disk-diffusion method. The paper disk was loaded in
silica gel 60 for column chromatography (Merck), pre- various concentrations of chemicals tested, and
coated TLC Silica F254 (Merck). Alpha-pinene was aseptically placed on the bacteria. Staphylococcus
prepared from turpuntine oil provided by PT. Perhutani aureus and Escherichia coli was taken as a bacterial
Anugerah Kimia, Indonesia and was purified using tested, and the activity was determined by measuring
fractional distillation under reduced pressure following the inhibition diameter (mm) of the bacterial growth
procedure Masruri et al. [2]. each paper disk. Beside that, tube dilution method was
also performed following procedure from Masruri et al.
Instrumentation [2].

Instrument is applied for analysis included gas RESULT AND DISCUSSION

chromatography-mass spectrometer (Shimadzu GCMS-
QP2010S), infrared spectrophotometer (Shimadzu FTIR- Catalytic Oxidation of Alpha-Pinene
8400S), UV-Vis Spectrophotometer (UV-Vis Shimadzu
1601), refractive index, analytical balance (Mettler The alpha-pinene as starting material was
Toledo). The chemical drawing was performed using isolated following procedure Rekfa et al. [1] from
ChemBioDraw Ultra 12.0 while 3D structure with turpuntine oils of local producer. Degree of purity was
ChemBio3D Ultra 12.0 and was optimized using MM2 determined from GCMS chromatogram. It was a clear
method, minimum RMS gradient 0.10, and 10000 oil with 98% purity. This was further applied for
iteration. oxidation study. A mixture of alpha-pinene and
potassium permanganate was added with solution of
Procedure sodium hydroxide. Stirring under reflux condition
transforms the color of reaction mixture from violet to
Oxidation of alpha-pinene deep black. A few aliquot of the reaction sample was
Alpha-pinene (10 mL, d 0.864 g/mL, 63.42 mmol), taken with syringe and subjected for thin layer
acetone (5.0 mL) and potassium permanganate (10.02 g, chromatography to monitor the reaction progress. The
63.42 mmol) were placed in a 50 mL-round-bottom flask disappearing of alpha-pinene spot on the TLC plate as
and solution sodium hydroxide (10 mL, 0.5 M) was an indication the reaction was accomplished. After
added. This reaction mixture was refluxed until all alpha- completion, reaction mixture was neutralized, and the
pinene was consumed (monitored on TLC). The product product was filtered off. Separation and purification of
was neutralized with solution of hydrochloric acid 0.2 M, the product was further undertaken under flash
filtered, and extracted with ethyl acetate (3 × 10 mL). chromatography using ethyl acetate-hexane provided
The ethyl acetate layers was combined, dried over pure compound 2.
sodium sulfate anhydrate and concentrated under Theoretically oxidation of olefins using potassium
vacuum. The product was purified with flash permanganate in a base condition affords two type
chromatography using ethyl acetate/n-hexane as solvent products, keto-aldehyde product 3 and keto-
and isolated as yellow-brown oil (3.10 g, 26.53% yield). carboxylate product 2 (Fig. 1). However, separation
+
Mass Spectrum (EI): m/z 184 (M , 1%), 111 (20), using flash chromatography did not observed the
108 (15), 99 (88), 83 (16), 82 (17), 81 (20), 79 (22), presence of product 3. The infrared spectra revealed
-1
71 (100), 70 (24), 69 (34), 67 (12), 55 (28), 53 (20), the presence of hydroxyl (band at 3419 cm ), carbonyl
-1
43 (98), 41 (49), 40 (5). Calculated for C10H16O3 was group (band at 1714 cm ), and other supporting band
-1
184.1099, found 184.11; FTIR (thin film, cm ) 3419 (O- such as methyl and methylene groups between 3000
-1
Hstretching), 2952-2873 (C-Hstretching), 1714 (C=Ostretching). and 2800 cm . No double bond band was observed as
indication that all alkene was converted to carboxylate
Synthesis coordination compounds and carbonyl group. And also, no C-H band for
Oxidation product of alpha-pinene (250 mg, aldehyde was recorded as well. These results confirm
1.36 mmol) in 5.0 mL ethanol was added zinc chloride the existence of the keto-carboxylate product 2 instead
(1.0 equiv.). This reaction mixture was refluxed for 2 h to of keto-aldehyde product 3 (Fig. 2). Mass spectra from

Masruri et al.
 Indones. J. Chem., 2016, 16 (1), 59 - 64 61

O
(i)
O

[1] OH
[2]
(i) O
O
Condition:
H
(i) KMnO4 , NaOH, 0 to 50 oC [3]
Fig 1. Catalytic oxidation of alpha-pinene with potassium
permanganate

Fig 3. Proposed structure of coordination compound

the oxidized product [2] with copper(II) and zinc(II). 3D
Fig 2. FTIR spectra of the oxidation product from alpha- Molecule model was drawn with ChemBio3D Ultra 12.0
pinene, 2 from Cambridgesoft and optimized using MM2 method

GCMS data also proved product 2. Radical cation of The complex compound from product 2 with
molecule 2 with m/z 184 was detected beside the base copper(II) shifts the carbonyl group absorption to a
peak at m/z 71. lesser value than that in 2 (Fig 4A). It was found in
-1 -1
1650 cm while compound 2 was 1714 cm . On the
Complex Compound of 2 with Copper(II) and Zinc(II) other hand, the complex compound with zinc(II)
-1
recorded on 1640 cm . This absorption is slightly lower
Synthesis of complex compound was undertaken than that with copper(II). In addition, the band
by mixing a mol equivalent of keto-carboxylate product 2 movement was also observed for hydroxyl group (Fig.
with zinc and copper chloride using ethanol as solvent 4B). The original hydroxyl absorption was recorded at
-1
under reflux. The complex formation was easily 3419 cm , while in its complex compound with zinc(II),
observed by changing the reaction color and a solid move to a longer wavelength. It was recorded broad in
-1
formation. In Fig. 3, is showed the proposed structure of 3600-3200 cm . On the other hand, hydroxyl group
complex compound of 2 with copper(II) and zinc(II). The absorption in complex compound with copper(II)
-1
complex compounds occur between metal ions with two provided two new peaks in 3350 and 3450 cm ,
molecule 2. Oxygen atom (O13 and O25) of hydroxyl respectively. Other new vibration pattern was showed
group and oxygen atom (O8 and O21) from carbonyl in Fig. 4C. These were recorded for Cu-O and Zn-O
-1
groups on molecule 2 donates the electron to coordinate vibration. It was identified both on 820 and 505 cm for
-1
with Cu27. Meanwhile a complex compound with zinc(II) Cu-O, and band at 820 and 495 cm for Zn-O vibration.
was resulted from interaction between Zn27 with oxygen These were not sharply observed on FTIR spectra of
atom (O25 and O13) from hydroxyl group and oxygen compound 2.
atom (O9 and O14) from the carbonyl group. These Moreover, the ultra violet spectra also provide a
interactions are significantly supported by the shifting of clear signal that formation of complex compound
band absorptions of the main functional groups (hydroxyl between 2 and copper(II) or zinc (II) was occurred (Fig. 5).
and carbonyl groups) the complex compounds in FTIR The right spectra recorded movement of maximum
spectra (Fig. 4). wavelength of 2 to red-shift. The complex compound

Masruri et al.
62 Indones. J. Chem., 2016, 16 (1), 59 - 64

Fig 4. FTIR spectra of complex compound resulted from keto-carboxylate product 2 with copper(II) and zinc(II). A is
peak expansion for carbonyl group absorption, B is a peak expansion for hydroxyl group absorption, and C is peaks
expansion for Zn-O and Cu-O group vibration. OP is keto-carboxylate product 2, red line is spectra for complex
compound with zinc, and blue line is spectra from complex compound with copper

Fig 5. UV-Vis spectra of keto-carboxylate product 2 and its coordination with copper(II) (right) and zinc(II) (left)

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 Indones. J. Chem., 2016, 16 (1), 59 - 64 63

Fig 6. Antibacterial activity of the oxidation product of alpha-pinene, 2 and its coordination compound with copper(II)
and zinc(II) using disk diffusion method. Activity of S. aureus (A) and E. coli (B). All measurement is conducted using
5% margin error

Fig 7. Antibacterial activity of the oxidation product of alpha-pinene, 2 and its coordination compound with copper(II)
and zinc(II) evaluated using tube dilution method. Activity on S. aureus (A) and E. coli (B). All measurement using
margin error 5%

with copper(II) shifts the maximum wavelength from 323 In a range concentration provided (0 until 100%
to 305 nm (right), while complex with zinc(II) shifts to v/v), the average activity of the oxidation product from
301 nm. Finally, the proposed structure for the complex alpha-pinene, keto-carboxylate product 2, gives
compound of keto-carboxylate product 2 with metal inhibition diameter less significant in both of bacteria.
copper(II) and zinc(II) is concluded as it in Fig. 3. Similar result was also recorded for sample
coordination compound with copper(II). Sharply
Antibacterial Evaluation different, the prospecting result is indicated for sample
complex compound with zinc(II). The increasing
The antibacterial activity was tested on S. aureus concentration improves the activity in both bacteria S.
and E. coli and determined the inhibition of bacterial aureus and E. coli (Fig. 6). On the other hand, tube
growth using disk diffusion (Fig. 6) and tube dilution dilution method gives an important result in all range of
method (Fig. 7). The antibacterial activity for first method concentration and in all sample tested for both in S.
was counted based on the diameter inhibition (mm), aureus and E. coli (Fig. 7). The increasing
while the second method using optical density (OD) concentration (from 0 to 90%) improves the bacterial
parameter compared to the control sample. The higher growth inhibition activity. For overall sample tested, the
OD values, the higher bacterial concentration growing in optical density value at 0% recorded between 1.326
the tube, and the lesser its activity. and 1.852. This value dropped to between 0.089 and
0.397 when sample tested concentration was 90%.

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64 Indones. J. Chem., 2016, 16 (1), 59 - 64

CONCLUSION 10. Fisher, G.S., Goldblatt, L.A., and Stinson, J.S.,

1951, Converting alpha-pinene to mixtures of
In summary, the catalytic oxidation of alpha-pinene acids, U.S. Patent 2,750,411
using potassium permanganate as catalyst is working in 11. Zhang, X., Chen, Z., Wang, H., He, S., and Huang,
stoichiometric instead of catalytic amount. The reaction D., 2009, Atmos. Environ., 43 (29), 4465–4471.
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from Faculty of Mathematics and Natural Sciences, Ortuño, R.M., 2000, J. Org. Chem., 65 (13), 3934–
University of Brawijaya under contract number 3940.
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