Malaysian Journal of Analytical Sciences Vol 22 No 5 (2018): 817 - 827

DOI: 10.17576/mjas-2018-2205-09

 

 

 

LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY FOR THE DETECTION AND VALIDATION OF QUERCETIN-3-O-RUTINOSIDE AND MYRICETIN FROM FRACTIONATED Labisia pumila var. alata

 

(Kromatografi Cecair Spektrometri Jisim Tandem untuk Pengesanan dan Pengesahsahihan Kuersetin-3-O-Rutinosida dan Mirisetin daripada Pemeringkatan Labisia pumila var. alata)

 

Norliza Abdul Latiff^1,3*, Chua Lee Suan², Mohammad Roji Sarmidi³, Ismail Ware¹, Siti Nor Azlina Abdul Rashid^1,3, Maizatulakmal Yahayu¹

 

¹Institute of Bioproduct Development

²Faculty of Chemical and Energy

Universiti Teknologi Malaysia, 81310 Johor Bahru, Johor, Malaysia

³Innovation Centre in Agritechnology for Advanced Bioprocessing,

Universiti Teknologi Malaysia, 84600 Pagoh, Johor, Malaysia

 

*Corresponding author:  norliza@ibd.utm.my

 

 

Received: 6 August 2018; Accepted: 24 September 2018

 

 

Abstract

Quercetin-3-O-rutinoside and myricetin was reported to be important secondary metabolites in Labisia pumila (L. pumila). The reliability of the instrument, its sensitivity and methodology are the key in the analysis of a specific metabolite and liquid chromatography tandem mass spectrometry (LC-MS/MS) provides better accuracy and faster separation in detecting of such chemical compound. Therefore, this study aims to detect and validate the presence of quercetin-3-O-rutinoside and myricetin from fractionated extract of L. pumila (40% MeOH: water). The plant material was fractionated using octadecyl (C₁₈) silica solid phase and eluent with a ratio of 40% MeOH: water. The separation of compounds was carried out using a C₁₈ reversed phase column (Acquity, 150mm × 4.6mm × 1.7 µm). Negative ionization was utilized to fragment the precursor ion of m/z 609 for quercetin-3-O-rutinoside and m/z 317 for myricetin. Three fragments ions; m/z 447, 463, 301 was recognized as quercetin-3-O-rutinoside product ions within 3.3 minutes. Meanwhile, m/z 271,137 and 151 represented characteristics ions for myricetin were recognized within 3.9 minutes. The concentration of quercetin-3-O-rutinoside and myricetin was 0.007 mg/g and 0.009 mg/g respectively. The linearity was found between 0.991-0.998. Detection based on (limit of detection) LOD and (limit of quantification) LOQ were found in the ranged of 0.03-0.04 µg/mL and 0.11-0.13 µg/mL, respectively. Both compounds showed good recovery of above 87%. Intra-day and inter-day (RSD) study was in the acceptable range (below 10%). Based on the result of this study, the LC-MS/MS was shown to be the best selective, fast and sensitive method to determine quercetin-3-O-rutinoside and myricetin in the plant fraction. These findings could be used as a guideline for the detection of the compounds and suitable for quality controls of L. pumila this and another medicinal herb containing quercetin-3-O-rutinoside and myricetin.

 

Keywords:  Labisia pumila, quercetin-3-O-rutinoside, myricetin, validation

 

Abstrak

Kuersetin-3-O-rutinosida dan mirisetin telah dilaporkan sebagai metabolit sekunder yang penting dalam Labisia pumila (L. pumila). Kebolehpercayaan instrumen, kepekaan dan kaedah pembangunan menjadi aspek penting dalam analisis yang disasarkan kepada metabolit tertentu dan kromatografi cecair spektrometri jisim tandem (KC-SJ/SJ) menyediakan pengasingan lebih cepat dan tepat bagi pengesanan sebatian kimia. Oleh itu, kajian ini adalah bertujuan untuk mengesan dan menentusahkan kehadiran kuersetin-3-O-rutinosida dan mirisetin daripada pemeringkatan L. pumila (40% metanol:air). Bahan tumbuhan ini diperingkatkan menggunakan fasa pepejal oktadesil silika (C₁₈) dengan kadar 40% metanol: air. Pengasingan komponen dicapai menggunakan fasa turus berbalik C₁₈ (Acquity, 150 mm × 4.6 mm × 1.7 µm). Pengionan negatif telah digunakan untuk mengasingkan pencetus ion m/z 609 untuk kuersetin-3-O-rutinosida dan m/z 317 untuk mirisetin. Tiga pecahan ion; m/z 447, 463, 301 dikenal pasti sebagai produk molekul ion dari kuersetin-3-O-rutinosida dalam selang masa 3.3 minit. Manakala m/z 271, 137 and 151 mewakili ciri ion untuk mirisetin telah dikenalpasti dalam selang masa 3.9 minit. Kepekatan kuersetin-3-O-rutinosida dan mirisetin masing-masing ialah 0.007 mg/g dan 0.009 mg/g.  Kelinearan adalah di antara 0.991-0.998. Had pengesanan (LOD) dan had pengkuantitian (LOQ) adalah masing-masing dalam julat 0.03-0.04 µg/mL dan 0.11-0.13 µg/mL. Kedua-dua sebatian menunjukkan penghasilan yang baik, lebih daripada 87%. Kajian dalam-hari dan antara-hari (RSD) adalah dalam julat yang boleh diterima (bawah 10%). Berdasarkan keputusan melalui kajian ini, KC-SJ/SJ ditunjukkan sebagai kaedah terpilih, cepat dan sensitif bagi menentukan kuersetin-3-O-rutinosida dan mirisetin di dalam pemeringkat tumbuhan ini. Penemuan ini boleh digunakan sebagai panduan untuk pengesanan sebatian dan sesuai sebagai kawalan kualiti untuk L. pumila dan herba perubatan yang lain yang mengandungi kuersetin-3-O-rutinosida and mirisetin.

 

Kata kunci:  Labisia pumila, kuersetin-3-O-rutinosida, mirisetin, pengesahsahihan

 

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