Application of FTIR fingerprints coupled with chemometric for comparison of stingless bee propolis from different extraction methods


  • Nur Basyirah Md Zin Universiti Sultan Zainal Abidin
  • Azierah Azemin Universiti Sultan Zainal Abidin
  • Muhammad Muslim Mohd Rodi Universiti Sultan Zainal Abidin
  • Zalilawati Mat Rashid Universiti Sultan Zainal Abidin
  • Khamsah Suryati Mohd Universiti Sultan Zainal Abidin



Propolis, stingless bee, FTIR, PCA, HCA


Propolis is a mixture of resin collected by the bees to build their hive. These mixtures contain natural remedy and used to treat variety of health-related problems. Unlike honeybee propolis, the study of stingless bee propolis still lacking. The important part of propolis study is the optimization of extraction procedures. The aim for this study is to employ chemometrics on FTIR data in order to discriminate the chemical fingerprinting of Malaysian stingless bee, Heterotrigona itama propolis yielded from different extraction methods, which were maceration, sonication and soxhlet. The chemical fingerprinting was obtained through Fourier Transform Infrared (FTIR). Principle component analysis (PCA) and hierarchical cluster analysis (HCA) were applied as pattern recognition methods for FTIR spectra. PCA of FTIR data for different extraction methods of stingless bee’s propolis revealed that variability of PC1 and PC2 is 84.76%. PCA’s variation in propolis by different extraction methods was classified based on specific functional groups arisen from the peaks. The FTIR fingerprinting of HCA of stingless bee’s propolis were distributed into three clusters based on percentages of ethanol, intensity of peaks and different fingerprint region. FTIR coupled with chemometric analysis showed classification of different extraction methods of propolis from PCA and HCA based on vibration of functional groups presence in the samples. Taken together, these results showed that different extraction methods play important role in detaining of chemical in propolis of stingless bee.

Author Biographies

Nur Basyirah Md Zin, Universiti Sultan Zainal Abidin

School of Agriculture Science and Biotechnology, Faculty of Bioresources and Food Industry

Azierah Azemin, Universiti Sultan Zainal Abidin

School of Agriculture Science and Biotechnology, Faculty of Bioresources and Food Industry

Muhammad Muslim Mohd Rodi, Universiti Sultan Zainal Abidin

Centre for Farm Management

Zalilawati Mat Rashid, Universiti Sultan Zainal Abidin

School of Food Industry, Faculty of Bioresources and Food Industry

Khamsah Suryati Mohd, Universiti Sultan Zainal Abidin

School of Agriculture Science and Biotechnology, Faculty of Bioresources and Food Industry


Ami, D., Mereghetti, P., Doglia, S., M. 2012. Multivariate analysis for fourier transform infrared spectra of complex biological systems and processes. Intech Open, 10, 190-220.

Araujo, K. S. D. S., Joaquim, D. S. J., Marcello, O. S., Fernanda, D. B., Ilsamar, M. S., Robson, B., Tarso, D. S. A., Sergio, D. A., Sandra, M. B. M. 2016. Physicochemical properties and antioxidant capacity of propolis of stingless bees (Meliponinae) and Apis from two Regions of Tocantins, Brazil. Acta Amazonica. 46(1), 61-68.

Azemin, A., Md-Zin, N. B., Mohd-Rodi, M. M., Kim- Chee, A. S., Zakaria, A. J., Mohd, K. S. 2017. Application of metabolite profilling and antioxidant activity in assesing the quality of the processed and unprocessed stingless bee’s propolis. Journal of Fundamental and Applied Science. 9(2S), 637-660.

Azwanida, N. N. 2015. A review on the extraction methods use in medicinal plants, principle, strength and limitation. Medicinal and Aromatic Plants, 4(196), 2167-0412.

Bankova, V., Popova, M., Trusheva, B. 2014. Propolis volatile compounds: chemical diversity and biological activity: a review. Chemistry Central Journal, 8(28), 1-8.

Chalmers, J. M. 2006. Mid‐infrared spectroscopy: Anomalies, artifacts and common errors. Handbook of Vibrational Spectroscopy. Wiley Online Library, 2327-2347.

Cleidiane, S., Anaclara, P., Matheus, A. C., Vanderlei, A. L.,Tatiane, L. C. O. 2018. Determination of total phenolic compounds and antioxidant activity of ethanolic extracts of propolis using ATR–FT-IR spectroscopy and chemometrics. Food Analytical Methods, 11(7). 2013-2021.

Gad, H, A., El-Ahmady, S. H., Abou-Shoer, M. I., Al-Azizi, M. M. 2013. Application of chemometrics in authentication of herbal medicines: A review. Phytochemcal Analytical, 24, 1-24.

Herve, A., Lynne, J. W. 2010. Principal component analysis. John Wiley & Sons. 2, Wiley Online Library, 433-459.

Irene, M., Dewi, F. S., Ellyza, H., Zainal A. M. 2012 . Optimization and comparative study of different extraction methods of biologically active components of Indonesian propolis Trigona spp. Journal of Natural Products, 5, 233-42.

Jolliffe, I. T. 2002. Principal Component Analysis. Springer. New York, 2, 32-37.

Khacha-Ananda, S., Tragoolpua, K., Chantawannakul, P., Tragoolpua, Y. 2013. Antioxidant and anti-cancer cell proliferation activity of propolis extracts from two extraction methods. Asian Pacific Journal of Cancer Prevention, 14(11), 6991-6995.

Kothai S., Jayanthi B. 2015. Environmental impact on stingless bee propolis (Tetragonula iridipennis) reared from two different regions of Tamilnadu - a comparative study. International Journal of ChemTech Research, 7(7), 3081-3088.

Magdalena, W., Izabela, R., Kinga, S., Patrycja, K., Bartłomiej, M. 2016. Propolis and organosilanes in wood protection. Part I: FTIR analysis and biological tests. Annals of Warsaw University of Life Sciences – SGGW Forestry and Wood Technology, 91(2015), 218-224.

Milena, P., Efstathia, G., Krystyna, S. K. G., Jaroslaw, W., Vassya, B., Haralabos, K., Gregory, S., K., Beata, A., Ioanna, C. 2017. Characterization and biological evaluation of propolis from Poland. Molecules, 22, 2-13.

Ming, J., J., Shane, R., S., Cheng, C. G., Yen, L. H., Kun, S. H. 2009. Optimization analysis of the experimental parameters on the extraction process of propolis. Proceedings of the International MultiConference of Engineers and Computer Scientists 2009 Vol II , IMECS 2009, March 18 - 20, Hong Kong.

Mohamad-Asri, M. N., Mat-Desa, W. N. S., Ismail, D. 2018. Combined principal component analysis (PCA) and hierarchical cluster analysis (HCA): An efficient chemometric approach in aged gel inks discrimination. Australian Journal of Forensic Sciences, 1-22.

Niken, P., Ritonga, H., Uslinawaty, Z. 2014. Properties and flavonoids content in propolis of some extraction method of raw propolis. International Journal of Pharmacy and Pharmaceutical Sciences, 6(6), 975-1491.

Prashant, T., Bimlesh, K., Mandeep, K., Gurpreer, K., Harleen, K. 2011. Phytochemical screening and extraction: A review. Internationale Pharmaceutica Sciencia. 1(1), 98-106.

Renata, N. O., Maurício, C. M., Fernando, C. S. O., Thayse, M. P., Brid, Q., Rossana, M. M. T., Garrett, B. M. 2016. FTIR analysis and quantification of phenols and flavonoids of five commercially available plants extracts used in wound healing. Revista Materia, 21(3), 767– 779.

Ricardo, S., Fatima., Cristina, A. A. 2015. Propolis : A complex natural product with a phlethora of biological activities that can be explored for drug development. Evidence-based Complementary and Alternative Medicine, 2015, 1-29.

Rohman, A., Salamah, N. 2018. The employment of spectroscopic techniques coupled with chemometrics for authentication analysis of halal pharmaceuticals. Journal of Applied Pharmaceutical Science, 8(10), 63-68.

Tian, Y., Xu, Z., Zheng, B., Lo, Y. M. 2013. Optimization of ultrasonic-assisted extraction of pomegranate (Punica granatum L.) seed oil. Ultrasonics Sonochemistry, 20, 202–208.






Special Issue on International Conference on Agriculture, Animal Sciences and Food Technology 2018 (Applied Sciences)