KAJIAN IN VITRO KAPASITAS ANTIOKSIDAN DAN INHIBISI LIPASE PANKREAS FORMULASI ANGKAK, DAUN KEMANGI, RIMPANG LENGKUAS, DAN DAUN KELOR

Abstract

Obesitas merupakan kondisi yang ditandai oleh akumulasi jaringan lemak tubuh secara berlebihan sehingga dapat meningkatkan risiko berbagai gangguan kesehatan. Orlistat efektif sebagai agen antiobesitas melalui penghambatan lipase pankreas, namun penggunaannya berpotensi menimbulkan efek samping. Indonesia memiliki biodiversitas tanaman yang berpotensi sebagai antiobesitas, seperti Moringa oleifera (daun kelor), Ocimum basilicum (daun kemangi), dan rimpang Alpinia galanga (lengkuas) yang telah dilaporkan aktif secara in vitro. Angkak, beras hasil fermentasi oleh kapang Monascus, diketahui memiliki aktivitas antioksidan serta berpotensi sebagai agen antiobesitas, akan tetapi kajian ilmiah mengenai potensi formulasi keempat bahan tersebut sebagai agen antioksidan dan antiobesitas hingga saat ini masih belum banyak dilaporkan. Penelitian ini bertujuan mengevaluasi kadar fenolik dan flavonoid total, kapasitas antioksidan (DPPH, FRAP, CUPRAC, dan ABTS), serta aktivitas inhibisi lipase pankreas in vitro beserta nilai IC50 menggunakan substrat teremulsi Triton X-100 dan cairan empedu pada ekstrak kasar dan formulasinya. Ekstraksi dilakukan dengan metode sonikasi menggunakan etanol 70%, sedangkan formulasi disusun menggunakan perangkat lunak Design Expert. Ekstrak kasar daun kelor tunggal menunjukkan kadar fenolik dan flavonoid total tertinggi masing-masing sebesar 64,33 ± 1,25 mg GAE/g ekstrak dan 16,85 ± 0,61 mg QE/g ekstrak, serta kapasitas antioksidan tertinggi pada metode DPPH, FRAP, CUPRAC, dan ABTS dengan nilai berturut-turut 90,50 ± 0,54; 384,71 ± 4,80; 526,87 ± 7,22; dan 746,92 ± 5,13 mg TE/g ekstrak. Inhibisi lipase pankreas tertinggi pada substrat teremulsi Triton X-100 diperoleh pada formulasi 3:0:1:0 sebesar 77,96 ± 0,42%, sedangkan pada substrat teremulsi cairan empedu diperoleh dari ekstrak kasar daun kelor tunggal sebesar 77,91 ± 0,38%. Nilai IC50 terendah juga diperoleh pada ekstrak kasar daun kelor yaitu 258,93 ± 3,96 µg/mL (substrat teremulsi Triton X-100) dan 265,73 ± 3,16 µg/mL (substrat teremulsi cairan empedu), namun keduanya masih lebih tinggi dibandingkan orlistat yaitu 58,37 ± 1,04 µg/mL (Triton X-100) dan 58,70 ± 0,40 µg/mL (cairan empedu). Hasil formulasi menunjukkan interaksi aditif, sinergis, dan antagonis pada berbagai uji. Analisis korelasi Pearson menunjukkan kadar fenolik dan flavonoid total berkorelasi positif kuat dengan kapasitas antioksidan, dengan korelasi tertinggi pada fenolik total–ABTS (r = 0,9086) dan flavonoid total–CUPRAC (r = 0,8609). Kadar flavonoid total juga menunjukkan korelasi positif terkuat dengan inhibisi lipase pankreas, yaitu r = 0,9420 (substrat teremulsi Triton X-100) dan r = 0,9052 (substrat teremulsi cairan empedu). Seluruh kapasitas antioksidan berkorelasi positif dengan inhibisi lipase pankreas, dengan metode CUPRAC menunjukkan korelasi tertinggi, yaitu r = 0,7424 (Triton X-100) dan r = 0,7262 (cairan empedu).

Obesity is a condition characterized by excessive accumulation of body fat, which increases the risk of various health disorders. Orlistat is effective as an anti-obesity agent through inhibition of pancreatic lipase; however, its use may cause adverse effects. Indonesia possesses rich plant biodiversity with potential anti-obesity properties, such as Moringa oleifera (moringa leaves), Ocimum basilicum (basil leaves), and the rhizome of Alpinia galanga, which have been reported to exhibit anti-obesity activity in vitro. Angkak, known as Red yeast rice, a rice product fermented by Monascus spp., has been reported to possess antioxidant activity and potential anti-obesity effects. However, scientific studies investigating the combined potential of these four materials as antioxidant and anti-obesity agents remain limited. This study aimed to evaluate total phenolic and flavonoid contents, antioxidant capacity (DPPH, FRAP, CUPRAC, and ABTS), and in vitro pancreatic lipase inhibitory activity, including IC50 values, using substrates emulsified with Triton X-100 and bile fluid in crude extracts and their combined formulations. Extraction was performed using the sonication method with 70% ethanol, while the formulations were designed using Design Expert software. The crude extract of Moringa oleifera leaves exhibited the highest total phenolic and flavonoid contents, with values of 64.33 ± 1.25 mg GAE/g extract and 16.85 ± 0.61 mg QE/g extract, respectively. The same extract also showed the highest antioxidant capacity in the DPPH, FRAP, CUPRAC, and ABTS assays, with values of 90.50 ± 0.54, 384.71 ± 4.80, 526.87 ± 7.22, and 746.92 ± 5.13 mg TE/g extract, respectively. The highest pancreatic lipase inhibition using Triton X-100–emulsified substrate was observed in the 3:0:1:0 formulation (77.96 ± 0.42%), whereas with bile-emulsified substrate the highest inhibition was shown by the crude Moringa oleifera leaf extract (77.91 ± 0.38%). The lowest IC50 values were also obtained from the crude Moringa oleifera leaf extract, namely 258.93 ± 3.96 µg/mL (Triton X-100–emulsified substrate) and 265.73 ± 3.16 µg/mL (bile-emulsified substrate). However, these values were still higher than those of Orlistat, which showed IC50 values of 58.37 ± 1.04 µg/mL (Triton X-100) and 58.70 ± 0.40 µg/mL (bile). The formulation results indicated additive, synergistic, and antagonistic interactions across the various assays. Pearson correlation analysis showed that total phenolic and flavonoid contents were strongly and positively correlated with antioxidant capacity, with the strongest correlations observed between total phenolics and the ABTS assay (r = 0.9086) and between total flavonoids and the CUPRAC assay (r = 0.8609). Total flavonoid content also exhibited the strongest positive correlation with pancreatic lipase inhibition, with r values of 0.9420 for the Triton X-100–emulsified substrate and 0.9052 for the bile-emulsified substrate. All antioxidant assays showed positive correlations with pancreatic lipase inhibition, with the CUPRAC method displaying the highest correlations, r = 0.7424 (Triton X-100) and r = 0.7262 (bile).