Malaysian Journal of Analytical Sciences Vol 26 No 2 (2022): 415 - 428

 

 

 

 

OPTIMISATION OF THE EXTRACTION METHOD OF RED Christia vespertilionis LEAVES TO YIELD BIOACTIVE PHTYOCHEMICALS AS MONITORED BY GAS CHROMATOGRAPHY-MASS SPCTROMETRY

 

(Pengoptimuman Kaedah Pengekstrakan Pada Daun Merah Christia vespertilionis untuk Menentukan Fotokimia Bioaktif Melalui Kromatografi Gas-Spektrometri Jisim)

 

Izzah Farhah Zambari¹, Sitti Rahma Abdul Hafid², Nur Airina Muhamad¹*

 

¹Institute of Biological Sciences, Faculty of Science,

Universiti Malaya, 50603 Kuala Lumpur, Malaysia.

²Nutrition Unit, Product Development and Advisory Services Division,

Malaysian Palm Oil Board, 43000 Kajang, Selangor, Malaysia.

 

*Corresponding author:  nurairina@um.edu.my

 

 

Received:  31 August 2021; Accepted:  30 December 2021; Published:  28 April 2022

 

 

Abstract

Christia vespertilionis (L. f.) Bakh. f. is well-known for treating various contagious diseases. This plant has been recognised among researchers and locals to have anti-inflammatory properties and has thus become popular for treating cancer. Two types of C. vespertilionis are acknowledged which are green and red. The green C. vespertilionis has been widely studied by many researchers, however just a few have studied the red type. This study was carried out to optimise the extraction method of red C. vespertilionis leaves by different extraction techniques (maceration and Soxhlet extraction) and solvents (methanol and ethanol) to yield bioactive phytochemicals using gas chromatography-mass spectrometry (GC-MS). The components were identified through GC-MS via comparisons as guided by the National Institute of Standards and Technology Mass Spectral Library 2011 (NIST 11, version 2.0g). According to the four (4) samples of red C. vespertilionis leaves using maceration of methanol (RMM), maceration of ethanol (RME), Soxhlet extraction of methanol (RSM), and Soxhlet extraction of ethanol (RSE), seventy-one (71) bioactive phytochemicals were identified. Eleven (11) major bioactive phytochemicals (abundance of > 4%) were identified namely, acetic acid, butyl ester; 1-butanol, 3-methyl-, acetate; heptanoic acid, propyl ester; hexanoic acid, 3-oxo-, ethyl ester; phenol, 3,5-bis(1,1-dimethylethyl)-; 1-octadecene; 4-O-methylmannose; .alpha.-d-mannofuranoside, methyl; 2-undecene, 9-methyl-, (E)-; n-hexadecanoic acid; and 1-octadecanol. Only seven (7) out of eleven (11) compounds were reported to have biological activities. Among those samples, RSM was the most effective using correlation coefficient between abundance (%) and retention time (minute(s)) with a significant difference at P < 0.05.

 

Keywords: Christia vespertilionis, gas chromatography-mass spectrometry, phytochemicals, maceration, Soxhlet extraction

 

Abstrak

Christia vespertilionis (L. f.) Bakh. f. dikenali secara meluas dalam merawat pelbagai penyakit berjangkit. Tumbuhan ini diakui di kalangan penyelidik dan penduduk tempatan tentang kelebihannya sebagai salah satu ubat tradisional yang mempunyai sifat anti-radang serta digunakan secara meluas dalam merawat barah. Terdapat dua jenis C. vespertilionis iaitu jenis hijau dan merah. C. vespertilionis hijau telah banyak dikaji oleh penyelidik dan hanya sedikit yang dikaji untuk jenis merah. Kajian ini dijalankan untuk mengoptimumkan kaedah pengekstrakan daun C. vespertilionis merah dengan teknik pengekstrakan yang berbeza (pengekstrakan maserasi dan Soxhlet) dan pelarut (metanol dan etanol) untuk menentukan fitokimia bioaktif melalui kromatografi gas-spektrometri jisim (GC-MS). Komponen tersebut dikenal pasti melalui GC-MS dengan membuat perbandingan dari data rujukan Institut Piawaian dan Teknologi Nasional 2011 (NIST11-MS, version 2.0g). Berdasarkan 4 sampel daun merah C. vespertilionis melalui maserasi metanol (RMM), maserasi etanol (RME), pengekstrakan Soxhlet methanol (RSM) dan pengekstrakan Soxhlet etanol (RSE), tujuh puluh satu (71) sebatian fitokimia dikenal pasti. Sebelas (11) sebatian fitokimia utama (> 4 % kawasan puncak) juga dikenalpasti seperti asid asetik, butil ester; 1-butanol, 3-metil-, asetat; asid heptanoik, propil ester; asid heksanoik, 3-okso-, etil ester; fenol, 3,5-bis (1,1-dimetiletil)-; 1-oktadekena; 4-O-metilmanos; .alpha.-d-manofurosida, metil; 2-undekena, 9-metil-, (E)-; asid n-heksadekanoik dan 1-oktadekanol. Hanya tujuh (7) daripada sebelas (11) sebatian yang dilaporkan mempunyai aktiviti biologi. Di antara sampel tersebut, RSM adalah yang paling berkesan dengan menggunakan pekali korelasi antara kebanyakan (%) dan masa pengekalan (min) dengan perbezaan yang signifikan pada P < 0.05.

 

Kata kunci:  Christia vespertilionis, kromatografi gas-spektrometri jisim, fitokimia, maserasi, pengekstrakan Soxhlet

 

 

 

Graphical Abstract

 

References

1.      Nguyen-Pouplin, J., Tran, H., Phan, T. A., Dolecek, C., Farrar, J., Tran, T. H., Caron, P., Bodo, B. and Grellier, P. (2007). Antimalarial and cytotoxic activities of ethnopharmacologically selected medicinal plants from South Vietnam. Journal Ethnopharmacology, 109: 417-427.

2.      Hofer, D., Schwach, G., Tabrizi-Wizsy, N.G., Sadjak, A., Sturm, S., Stuppner, H. and Pfragner, R. (2013). Christia vespertilionis plant extracts as novel antiproliferative agent against human neuroendocrine tumor cells. Oncology Reports, 6(29): 2219-2226.

3.      Osman, M. S., Ghani, Z. A., Ismail, N. F., Razak, N. A., Jaapar, J. and Ariff, M. A. (2017). Qualitative comparison of active compounds between red and green Mariposa Christia vespertillonis leaves extracts. AIP Conference Proceedings, 1885(020282): 1-6.

4.   Sharma, A. and Cannoo, D.S. (2016). Effect of extraction solvents/techniques on polyphenolic contents and antioxidant potential of the aerial parts of Nepeta leucophylla and the analysis of their phytoconstituents using RP-HPLC-DAD and GC-MS. RSC Advance, 6: 78151-78160.

5.      Bunawan, H., Bunawan, S. N. and Baharum, S. N. (2015). The red butterfly wing (Christia vespertilionis): A promising cancer cure in Malaysia. International Journal of Pharmacy and Pharmaceutical Sciences 7(8): 1.

6.      Mohd Yasin, Z. N., Zakaria, M. A., Nik Mat Zin, N. N. I., Ibrahim, N., Mohamad, F. S., Mat Desa, W. N. S., Sul’ain, M. D. and Abu Bakar, N. (2020). Biological activities and GCMS analysis of the methanolic extract of Christia vespertilionis (L.F.) Bakh. F. leaves. Asian Journal of Medicine and Biomedicine 4(1): 1-11.

7.      Haida, Z. and Hakiman, M. (2019). A review of therapeutics potentials of Clinacanthus nutans as source for alternative medicines. Sains Malaysiana, 48(12): 2683-2691.

8.      Yusufzai, S. K., Khan, M. S., Hanry, E. L., Rafatullah, M. and Elison, B. (2019). GC-MS analysis of chemical constituents and in vitro antioxidant activity of the organic extracts from the stem of Bridelia stipularis. Sains Malaysiana, 48(5): 999-1009.

9.      NIST/NIH/EPA Mass Spectral Library, Standard Reference Database 1, NIST 11. (2011). Standard Reference Data Program, National Institute of Standards and Technology. Gaithersburg, MD, USA.

10.   Abd Rahim, E. N. A., Ismail, A., Omar, M. N., Rahmat, U. N. and Wan Ahmad, W. A. N. (2018). GC-MS analysis of phytochemical compounds in Syzygium polyanthum leaves extracted using ultrasound-assisted method. Pharmacognosy Journal, 10(1): 110-119.

11.   Mohd Yasin, Z. N., Zakaria, M. A., Nik Mat Zin, N. N.I., Ibrahim, N., Mohamad, F. S., Mat Desa, W. N. S., Sul’ain, M. D. and Abu Bakar, N. (2020). Biological activities and chemical compositions of the methanol extract of Christia vespertilionis leaves. Asian Journal of Medicine and Biomedicine 4(2): 78-88.

12.   Sidek, M. M., Ismail, N. Z., Zain, N. N. M. and Arsad, H. (2019). Cytotoxicity activity of Christia vespertilionis leaves extracts against Hela cells and the detection of compounds by GC-MS. Journal of Biological & Scientific Opinion, 7(6): 71-78.

13.   Gami, B. and Parabia, F. (2011). Screening of methanol & acetone extract for antimicrobial activity of some medicinal plants species of Indian folklore. International Journal Research Pharmaceutical Sciences, 2(1): 69-75.

14.   Truong, D-H., Nguyen, D-H., Ta, N.T.A., Bui, A.V., Do, T.H. and Nguyen, H.C. (2019). Evaluation of the use of different solvents for phytochemical constituents, antioxidants, and in vitro anti-inflammatory activities of Severinia buxifolia. Journal of Food Quality, 2019: 8178294.

15.   Karunaratne, U. K. P. R. and Karunaratne, M. M. S. C. (2012). Evaluation of methanol, ethanol and acetone extracts of four plant species as repellents against Callosobruchus maculatus (Fab.). Vidyodaya Journal of Science 17: 1-8.

16.   Kalia, K., Sharma, K., Singh, H.P. and Singh, B. (2008). Effects of extraction methods on phenolic contents and antioxidant activity in aerial parts of Potentilla atrosanguinea L. and quantification of its phenolic constituents by RP-HPLC. Journal Agriculture Food Chemistry, 56: 10129-10134.

17.   Aparna, V., Dileep, K. V., Mandal, P. K., Karthe, P., Sadasivan, C. and Haridas, M. (2012). Antiinflammatory property of n-hexadecanoic acid: Structural evidence and kinetic assessment. Chemical Biology Drug Design, 80(3): 434-439.

18.   Singab, A. N., El-Taher, E. M. M., Elgindi, M. R and Kassem, M-S. (2015). Phoenix roebelenii O’Brien DNA profiling, bioactive constituents, antioxidant and hepatoprotective activities. Asian Pacific Journal Tropical Disease, 5(7): 552-558.

19.   Mitchell, A. M., Strobel, G. A., Moore, E., Robison, R. and Sears, J. (2010). Volatile antimicrobials from Muscodor crispans, A novel endophytic fungus. Microbiology, 156: 270-277.

20.   Chew, Y. L., Chan, E. W. L., Tan, P. L., Lim, Y. Y., Stanslas, J. and Goh, J. K. (2011). Assessment of phytochemical content, polyphenolic composition, antioxidant and antibacterial activities of Leguminosae medicinal plants in Peninsular Malaysia. BMC complementary and Alternative Medicine, 11(1): 12.

21.   Lee, J. J., Yazan, L. S., Kassim, N. K., Che Abdullah, C. A., Esa, N., Lim, P. C. and Tan, D.C. (2020). Cytotoxic activity of Christia vespertilionis root and leaf extracts and fractions against breast cancer cell lines. Molecules, 25(2610):1-18.

22.   Lee, Y. S., Kang, M. H., Cho, S. Y. and Jeong, C. S. (2007). Effects of constituents of Amomum xanthioides on gastritis in rats and on growth of gastric cancer cells. Achieve Pharmaceutical Research, 30(4): 436-443.

23.   Belakhdar, G., Benjouad, A. and Abdennebi, E.H. (2015). Determination of some bioactive chemical constituents from Thesium humile Vahl. Journal Materials Environmental Sciences, 6(10): 2778-2783.

24.   Duke J. (2002). American Agricultural Research Service-Phytochemical & Ethnobotanical Database. Available at: http://www.ars-grin.gov/duke/index.html [accessed on 09 December 2019].

25.   Mutalib, N. A. and Latip, N. A. (2019). Synergistic interactions between Christia vespertilionis leaves extract and chemotherapy drug cyclophosphamide on WRL-68 Cell Line. Asian Journal of Pharmaceutical Research and Development, 7(3): 109-113.

26.   Mishra, P. M. and Sree, A. (2007). Antibacterial activity and GCMS analysis of the extract of leaves of Finlaysonia obovata (a mangrove plant). Asian Journal Plant Sciences, 6(1): 168-172.

27.   Kumar, P. R. Z. A. and Bhaskar, A. (2012). Phytochemical evaluation by GC-MS and in vitro antioxidant activity of Punica granatum fruit rind extract. Journal of Chemical and Pharmaceutical Research, 4(6): 2869-2873.

28.   Suman, T., Chakkaravarthi, K. and Elangomathavan, R. (2013). Phyto-chemical profiling of Cleistanthus collinus leaf extracts using GC-MS analysis. Research Journal Pharmaceutical and Technology, 6(11): 1173-1177.

29.   Igwe, K. K., Okafor, P. N. and Ijeh, I. I. (2015). GC-MS analysis of phytocomponents in Vernonia amygdalina. Del leaves and its contractile potential in mammary tissue in female albino Wistar rats. Journal of Agriculture and Veterinary Science, 8(11): 25-30.

30.   Johannes, E., Litaay, M. and Syahribulan. (2016). The bioactivity of hexadecanoic acid compound isolated from hydroid Aglaophenia cupressina lamoureoux as antibacterial agent against Salmonella typhi. International Journal Biological Medicinal Research, 7(2): 5469-5472.

31.   Gavamukulya, Y., Abou-Elella, F., Wamunyokoli, F. and El-Shemy, H. A. (2015). GC-MS analysis of bioactive phytochemicals present in ethanolic extracts of leaves of Annona muricata: A further evidence for its medicinal diversity. Pharmacognosy Journal, 7(5): 300-304.

32.   Shukri, N. S. S. M., Perumal, V., Balan, T., Murugesu, S., Khatib, A., Saleh, M. S. M, Hin, L.W., Darus, K. A., Zailani, I.A. and Zahidi, F. A. H. M. (2019). The screening of antioxidant activities and FTIR analysis of Christia vespertilionis leaves extracts. International Journal of Research in Pharmaceutical Sciences, 10(1): 1713.

33.   Guo, L., Wu, J., Han, T., Cao, T., Rahman, K. and Qin, L. (2008). Chemical composition, antifungal and antitumor properties of ether extracts of Scapania verrucosa Heeg. and its endophytic fungus Chaetomium fusiforme. Molecules, 13: 2114-2125.

34.   Wu, X. Y., Tang, A. C. and Lu, Q. Y. (2012). Study on antitumor effect of the extract from Christia vespertilionis in vivo. Chinese Journal of Experimental Traditional Medical Formulae, 8: 202-204.