Sains Malaysiana 51(6)(2022): 1789-1797

http://doi.org/10.17576/jsm-2022-5106-15

Quantitative HPLC and FTIR-based Metabolomics for Clustering Centella asiatica Cultivation Ages and Evaluation of Their Radical Scavenging Activity

(Kuantitatif HPLC dan Metabolomik berasaskan FTIR untuk Pengelompokan Umur Penanaman dan Penilaian Aktiviti Pemusnahan Radikal Centella asiatica)

MOHAMAD RAFI^1,2,*, ANASTASIA GRANDIVORIANA NOMI¹, DEWI ANGGRAINI SEPTANINGSIH^1,2, RUDI HERYANTO^1,2 & SASTIA PRAMA PUTRI³

¹Department of Chemistry, Faculty of Mathematics and Natural Sciences, IPB University, Jalan Tanjung Kampus IPB Dramaga, Bogor 16680, Indonesia

²Tropical Biopharmaca Research Center-Institute of Research and Community Services, IPB University, Jalan Taman Kencana No. 3 Kampus IPB Taman Kencana, Bogor 16128, Indonesia

³Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan

Received: 30 August 2021/Accepted: 2 November 2021

Abstract

Centella asiatica is a medicinal plant used to treat stomachache, cough, sore throat, blood booster, and skin diseases. The difference in harvesting time is among the factors that can significantly affect the chemical composition of C. asiatica. The purpose of this study was to evaluate the difference of C. asiatica cultivation ages through a metabolomics (targeted and untargeted) analysis using FTIR spectra, HPLC analysis of four main components, and evaluation of its radical scavenging activity. The results showed that differences in cultivation ages affected the chemical composition of C. asiatica. It was shown by the FTIR spectrum indicating the vibration of several functional groups such as OH, C-H, C=O, C=C, C-O, C-N, C-O phenol, and alkyl halides with differences in their intensities. The results of the HPLC showed that C. asiatica harvested after four months post-planting (MPP) had the highest content of madecassoside, asiaticoside, madecassic acid, and asiatic acid, respectively. A principal component analysis (PCA) was carried out to clustering C. asiatica based on its cultivation ages. The PCA results showed that each sample could be grouped with a total variance of the first two principal components (PC) using peak are of the four main components analyzed by HPLC and absorbance at wavenumber 400-4000 cm^-1 from FTIR spectra were 98% and 95%, respectively. The radical scavenging activity demonstrated that the 4 MPP had the highest inhibition of about 53.81±0.92 %. So, C. asiatica at 4 MPP gave optimum level four main metabolite content and radical scavenging activity.

Keywords: Antioxidant; Centella asiatica; FTIR spectra; HPLC; metabolomics

Abstrak

Centella asiatica adalah ubat yang digunakan untuk mengubati sakit perut, batuk, sakit tekak, penggalak darah dan penyakit kulit. Perbezaan masa penuaian adalah antara faktor signifikan yang dapat mempengaruhi komposisi kimia C. asiatica. Tujuan kajian ini adalah untuk menilai perbezaan waktu penanaman C. asiatica melalui analisis metabolomik (disasarkan dan tidak disasarkan) menggunakan spektrum FTIR, analisis HPLC daripada empat komponen utama, dan penilaian aktiviti penghapusan radikalnya. Hasil kajian menunjukkan bahawa perbezaan tempoh penanaman mempengaruhi komposisi kimia C. asiatica. Ia dibuktikan oleh spektrum FTIR dengan getaran beberapa kumpulan berfungsi seperti OH, C-H, C=O, C=C, C-O, C-N, C-O fenol dan alkil halida dengan perbezaankeamatan mereka. Hasil HPLC menunjukkan bahawa C. asiatica yang dituai setelah empat bulan selepas penanaman (MPP) mempunyai kandungan madecassoside, asiaticoside, asid madecasic dan asid asiaticoside tertinggi. Analisis komponen utama (PCA) dilakukan untuk pengelompokan C. asiatica berdasarkan tempoh penanamannya. Hasil PCA menunjukkan bahawa setiap sampel dapat dikelompokkan dengan total varians daripada dua komponen utama (PC) pertama yang menggunakan puncak adalah daripada empat komponen utama yang dianalisis oleh HPLC dan penyerapan pada bilangan gelombang 400-4000 cm^-1 daripada spektrum FTIR masing-masing adalah 98% dan 95%. Aktiviti penghapusan radikal menunjukkan bahawa 4 MPP mempunyai perencatan tertinggi sekitar 53.81±0.92%. Oleh itu, C. asiatica pada 4 MPP memberikan tahap optimum bagi empat kandungan metabolit utama dan aktiviti pembersihan radikal.

Kata kunci: Antioksida; Centella asiatica; HPLC; metabolomik; spektrum FTIR

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^*Corresponding author; email: mra@apps.ipb.ac.id