Malaysian Journal of Analytical Sciences, Vol 28 No 5 (2024): 995 - 1002

BIOASSAY-GUIDED FRACTIONATION OF POTENTIAL ANTIOXIDANT CONSTITUENTS FROM Geniotrigona thoracica PROPOLIS AND IDENTIFICATION OF ITS FLAVONOID COMPOUNDS VIA Q-TOF LCMS

(Pengasingan Sebatian Antioksidan Berpotensi Berpandukan Pencerakinan Biologi daripada Propolis Geniotrigona thorasica dan Pengenalpastian Sebatian Flavonoid

Menggunakan Q-TOF LCMS)

Ainur Awanis Mohd Badiazaman^1*, Khamsah Suryati Mohd^2*, and Wan Fitriyyah Hanani Wan Kamarul A’rifin²

¹Faculty of Applied Sciences, Universiti Teknologi MARA, 26400 Bandar Tun Abdul Razak, Jengka, Pahang, Malaysia

²Faculty of Bioresources and Food Industry, Universiti Sultan Zainal Abidin, 22200 Besut, Terengganuu, Malaysia

^*Corresponding author: khamsahsuryati@unisza.edu.my, ainurawanis@uitm.edu.my

Received: 5 May 2024; Accepted: 9 July 2024; Published: 27 October 2024

Abstract

Recently, stingless bee propolis has gained attention in the food market, mainly due to its pharmacological properties. Previous studies have reported the higher antioxidant properties of Geniotrigona thoracica propolis extract. In this paper, we report on the bioassay-guided fractionation of the active n-hexane fraction of G. thoracica propolis and its flavonoid compounds by Quadrupole-Time of Flight Liquid Chromatography Mass Spectrometry (Q-TOF LCMS) analysis. Chromatographic separation of n-hexane fraction was carried out using column chromatography. Antioxidant activities were evaluated using 2,2-diphenyl-1-picrylhydrazyl hydrate (DPPH) and 2,2’-azinobis (3- ethylbenzthiazolin-6-sulfonic acid) (ABTS) free radical scavenging assays. The identification of flavonoid compounds was carried out using Q-TOF LCMS analysis. Eleven sub-fractions resulting from the chromatographic separation of n-hexane fraction showed DPPH and ABTS radical scavenging activity with IC₅₀ values ranging from 468.60 to 5.83 µg/mL and 248.30 to 4.00 µg/mL, respectively. A total of seven flavonoid, phenolic and anthocyanin compounds were identified such as naringenin (1), acacetin (2), eriodictyol (3), myricetin (4), chlorogenic acid (5), eriodictyol-7-O-glucoside (6) and cyanidin chloride (7). Most of these compounds have been reported to possess antioxidant activity in previous studies. This indicate that G. thoracica propolis has potential source of antioxidants for the development of food and nutraceutical products.

Keywords: Geniotrigona thoracica, stingless bee propolis, bioassay-guided fractionation, flavonoid, Q-TOF LCMS

Abstrak

Kebelakangan ini, propolis lebah kelulut telah menarik perhatian di dalam pasaran makanan, terutamanya disebabkan oleh sifat famarkologi yang dimilikinya. Kajian terdahulu melaporkan sifat antioksidan yang tinggi untuk estrak propolis Geniotrigona thorasica. Dalam kajian ini, kami melaporkan pengasingan sebatian fraksi aktif n-heksana propolis G. thorasica dan sebatian flavonoidnya melalui analisis Spektrometri Jisim Kromatografi Cecair Penerbangan- Empat Masa (Q-TOF LCMS). Pemisahan kromatografi fraksi n-heksana dilakukan menggunakan kromatografi lajur. Aktiviti antioksidan dinilai menggunakan ujian pembebasan radikal bebas 1,1-difenil-2-pikrilhidrazil (DPPH) dan 2,2’-azino-bis (3-etilbenzotiazolin-6-sulfonik asid) (ABTS)). Pengenalpastian sebatian flavonoid dilakukan menggunakan analisis Q-TOF LCMS. Sebelas sub-fraksi dihasilkan daripada pemisahan kromatografi di dalam fraksi n-heksana telah menunjukkan aktiviti pembebasan radikal bebas DPPH dan ABTS dengan nilai IC₅₀ masing-masing berjulat antara 468.60 sehingga 5.83 µg/mL dan 248.30 sehingga 4.00 µg/mL. Sebanyak tujuh sebatian flavonoid, fenolik dan antosianin telah dikenalpasti seperti naringenin (1), asetin (2), eriodiktiol (3), mirisetin (4), asid klorogenik (5), eriodiktiol-7-O-glukosida (6) dan klorida sianidin (7). Kebanyakan sebatian ini telah dilaporkan memiliki aktiviti antioksidan di dalam kajian terdahulu. Ini menunjukkan bahawa propolis G. thorasica merupakan sumber potensi antioksidan untuk pembangunan produk makanan dan nutraseutikal.

Kata kunci: Geniotrigona thorasica, propolis lebah kelulut, pencerakinan berpandukan biologi, flavonoid, Q-TOF LCMS

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