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

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 (C 18 ) silica solid phase and eluent with a ratio of 40% MeOH: water. The separation of compounds was carried out using a C 18 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-Orutinoside 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. AbstrakKuersetin-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 ISSN 1394 -2506 Norliza et al: LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY FOR THE DETECTION AND VALIDATION OF QUERCETIN-3-O-RUTINOSIDE AND MYRICETIN FROM FRACTIONATED Labisia pumila var. alata 818 diperingkatkan menggunakan fasa pepejal oktadesil silika (C 18 ) dengan kadar 40% metanol: air. Pengasingan komponen dicapai menggunakan fasa turus berbalik C 18 (Acquity, 150 mm × 4.6 mm × 1.7 µm). Pengionan negatif telah digunakan untuk mengasingkan pencetus ion m/z 609 untuk kuerse...

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“…Compound 1 gave a precursor ion (M-H) − at m / z 609.197 and a fragmentation ion at m / z 300.028 due to the loss of the rhamnose and glucose sugars was seen as a base peak. Therefore, this compound was identified as quercetin rutinoside [ 44 ]. Compound 2 gave a precursor ion (M-H) − at m / z 505.098 and showed a fragment ion at m / z 445.078 due to the loss of the acetyl moiety (60 amu) and a further loss of the hexosyl moiety (162 amu) resulting in the fragment at m / z 300.…”