PHARMACOGNOSY
Year : 2009  |  Volume : 1  |  Issue : 1  |  Page : 46-48 Table of Contents     

GC-MS analysis of propolis of Indian origin


1 Lovely Professional University, Phagwara, Jalandhar, India
2 Dept. of Pharmacognosy, Al-Ameen College of Pharmacy, Hosur road, Bangalore, India
3 IIHR (Institute of Horticulture Research), Hessaraghatta, Bangalore, India
4 St. John's Pharmacy College, Bangalore, India

Correspondence Address:
N Kumar
Lovely Professional University, Phagwara, Jalandhar
India
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DOI: 10.4103/0975-1483.51876

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  Abstract 

More than 300 constituents have been identified in different propolis samples. This study investigated the composition of ethanolic extracts of propolis samples collected from the Gujarat zone. A gas chromatography-mass spectrometry was carried out on a Agilent GC-MS 5975 under electron impact ionization (70 eV). The chromatographic column for the analysis was done by the HP5MS capillary column (30 m x 0.25 mm internal diameter). The carrier gas used was helium at a flow rate of 1 ml/min. The oven temperature was 100C to 280C with a constant increase of 10C. Propolis samples were analyzed with the column held initially at 60C for 2 min and then increased to 230C with a 2C/ min heating ramp and then kept at 230C for 3 min. Finally, the temperature was increased to 280C with a 3C/min heating ramp. The injection was performed in split mode at 220C. Sample volumes of 1 μl were injected and analyzed by GC-MS. The following compounds were identified for the first time in the propolis sample: p-coumeric acid, Benzyl cinnamate, 4-pentanoic acid, and Ferulic acid. The main type of the compound identified was fatty acids derivatives. Similar results were found by Silici, et al . [5] for the turkey propolis.

Keywords: GC-MS analysis, propolis, Honeycomb


How to cite this article:
Kumar N, Mueen Ahmed K K, Dang R, Shivananda T N, Das K. GC-MS analysis of propolis of Indian origin. J Young Pharmacists 2009;1:46-8

How to cite this URL:
Kumar N, Mueen Ahmed K K, Dang R, Shivananda T N, Das K. GC-MS analysis of propolis of Indian origin. J Young Pharmacists [serial online] 2009 [cited 2014 Apr 16];1:46-8. Available from: http://www.jyoungpharm.in/text.asp?2009/1/1/46/51876


  Introduction Top


The term propolis derives from the Greek pro (for "in front of", "at the entrance to") and polis ("community or city") and means a substance in defense of the hive.[1] Propolis is a resinous, sticky gum, with a color that varies from yellow-green to dark brown depending on its source and age. A propolis is used to make the protective shield at the entrance of beehive. [2] It is also used to fill the cracks in the hive, to attach the corners of frames to the grooves in the hive, and to polish the cells of the honeycomb. Propolis was used especially in antiquity in Egypt. The propolis was very well known to the priests who had monopolized medicine, chemistry, and the art of mummifying corpses. A literature survey revealed that flavonoids, aromatic acid, diterpenic acid, and phenolic compounds appear to be the principle components responsible for the biological activities of propolis. This study investigated the composition of ethanolic extracts of propolis (GEEP) samples collected from the Gujarat zone. This extract was selected for GC-MS analysis as it was showing maximum activity in case of the antibacterial, [3] anti-fungal, [3] and anthelmintic [3] activity.


  Materials and Methods Top


Chemicals

Bis-(trimethyl-silyl) trifluoroacetamide (BSTFA) and trimethylchlorosilane (TMCS) (Merck-2333) were used as silylation reagents with spectrophotometric grade pyridine (Merck-7460).

Instrument

A gas chromatography-mass spectrometry was carried out on an Agilent GC-MS 5975 under electron impact ionization (70 eV). The interface temperature was 230C, and the MS scan range was 35-450 atomic mass units (AMU). The chromatographic column for the analysis was done by HP5MS capillary column (30 m x 0.25 mm internal diameter). The carrier gas used was helium at a flow rate of 1 ml/min. The oven temperature was 100C to 280C with a constant increase of 10C. Propolis samples were analyzed with the column held initially at 60C for 2 min and then increased to 230C with a 2C/ min heating ramp and then kept at 230C for 3 min. Finally, the temperature was increased to 280C with a 3C/min heating ramp. The injection was performed in split mode at 220C.

Sample Preparation

A collection of 10 mg propolis samples were extracted with ethanol at room temperature to obtain the extract. After filtration, the extracts were combined and evaporated to dryness under a vacuum at 50C. A total of 1 mg of dry extract was reacted with 50 μl pyridine + 100 μl bis-(trimethylsilyl) trifluoroacetamide (BSTFA) including 1% trimethylchlorosilane (TMCS) in a sealed glass tube for 30 min at 100C to prepare samples for gas chromatography. Sample volumes of 1 μl were injected and analyzed by GC-MS.

Identification of Compounds

Peaks were identified by Mr. M. Prabhakara Reddy at the Bangalore Test House in Bangalore. Good spectral matches for some compounds could be found in the Wiley and National Bureau of Standards (NBS) mass spectral library.

Chromatographic Condition

Column - HP5MS (30 m x 0.25 i.d.)

Carrier gas - Helium

Oven temperature - 100 - 280C

Inlet temperature - 280C

Detector temperature (MSD) - 230C

Carrier gas flow rate - 1 ml/min

Injection quantity - 1 μl

Scan mode - 50 - 550 mod range


  Results Top


The chemical composition of propolis, which was collected from the Gujarat zone, was investigated by GC-MS after silylation. More than 13 individual compounds were identified [Table 1] and [Figure 1]. For this reason, the chemical composition is presented by means of the main type of compounds identified not as a percentage of individual substances. As expected, the samples displayed the typical pattern of a "poplar" propolis: they contained pinocembrin, chrysin, galangin, caffeic acid, 3- hydroxy-4-methoxycinnamic acid, 3,4-dimethoxycinnamic acid, etc. and the ratio between the main compound classes corresponded to that in P. nigra bud exudates. [4],[5] However, there may be a variation in the components from one region to the other due to the change in topography. The following compounds were identified for the first time in the porpolis sample: p-coumeric acid, Benzyl cinnamate, 4-pentanoic acid, and Ferulic acid. The main type of the compound identified was fatty acids derivatives.


  Discussion Top


Similar results were found by Silici, et al . for the turkey propolis. [6],[8] He reported the presence of 9- octadecanoic acid, hekzadecanoic acid, benzoic acid, 3- hydroxy-4-methoxycinnamic acid, 3,4-dimethoxycinnamic acid, benzyl benzoate, benzene ethanol, β-eudesmol, β-bisabolol, glycerine, 2-nonadecanone, 2-propen-1-one, 4H - 1-benzopyrane-4-on, 2-propenoic acid, heneicosane, and eicosane in all of the samples of the propolis of Turkey. [6],[7] This sample of the Gujarat zone is obviously not of popular origin, its plant source is yet unknown. It will be the aim of future investigations to uncover this plant and study its biological action. This could be of interest because it has been shown that bees have the ability to find and use a propolis source in their environment, which is the best agent to protect their hives against bacterial and fungal infections. Further studies also can be conducted to determine the "type" of propolis, according to its plant source, which will be the first step for quality control of the bees' glue. This will allow us to define the type of compounds that should be quantified as main active propolis constituents.

 
  References Top

1.Burdock GA. Review of the biological properties and toxicity of bee propolis. Food Chem Toxicol 1998;36:347-63.  Back to cited text no. 1  [PUBMED]  [FULLTEXT]
2.Hegazi AG. Propolis an over view. Internation Symposium on Apitherapy. Cairo, 8 th - 9 th March, 1997.  Back to cited text no. 2    
3.Ghisalberti EL. Propolis: A review. Bee World 1979;60:59-84.  Back to cited text no. 3    
4.Heard TA. The role of stingless bees in crop pollination. Annu Rev Entomol 1999;44:183-206.  Back to cited text no. 4  [PUBMED]  [FULLTEXT]
5.Bankova V, Popova M, Bogdanov S, Sabatini AG. Chemical composition of European propolis: Expected and unexpected results. Z Naturforsch [C] 2002;57:530-3.  Back to cited text no. 5  [PUBMED]  
6.Silici S, Kutluca S. Chemical composition and antibacterial activity of propolis collected by three different races of honey bees in the same region. J Ethnopharmacol 2005;99:69-73.  Back to cited text no. 6  [PUBMED]  [FULLTEXT]
7.Yaghobi SMJ, Ghorbani GR, Soleimanian ZS, Satari R. Antimicrobial activity of Iranian propolis and its chemical composition. Apidologie 2007;15:45-8.  Back to cited text no. 7    
8.Oksuz H, Duran N, Tamer C, Cetin M, Silici S. Effect of propolis in the treatment of experimental Straphylococcus aureus Keratitis in rabbits. Ophthal Res 2005;37:328-34.  Back to cited text no. 8    


    Figures

  [Figure 1]
 
 
    Tables

  [Table 1]



 

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