Hydrilla verticillata contains some active compounds that potential as an antioxidant, antibacterial, anticancer, antimicrobial and antitumor. One of the active compounds in Hydrilla verticillata is steroids. This research aimed to isolate, to identify and to determine the toxicity and antioxidant activity of steroid compounds in petroleum ether (PE) fraction of Hydrilla verticillata. Hydrilla verticillata biomass powder was extracted by maceration using ethanol solvent. The ethanol extract was hydrolyzed with 2 N of hydrochloric acid and then partitioned with petroleum ether solvent. The steroid compounds from petroleum ether fraction were separated with Preparative Thin Layer Chromatography (TLC) and Column Chromatography. The steroid isolates were identified by UV-Vis and FTIR spectrophotometer. The toxicity level and antioxidant assay of steroid isolates were determined by BSLT and DPPH method, respectively. The result of the study showed that extraction through maceration produced 4.54% yield, whereas the product yield of the partition using petroleum ether was 65.41%. The steroid isolates from TLC and Column Chromatography separation has toxicity and antioxidant properties. The LC50 value of steroid TLC isolate was 1.41 and 12.2 ppm. The LC5o value of steroid Column Chromatography isolates (H2, H4 and H10) were 4.32, 8.24 and 10.35 ppm. The EC50 value of H2 Column Chromatography isolate was 23.00 ppm.
Hydrilla verticillata is one of Allah SWT creations that potential as an antioxidant. This study’s objectives were to determine antioxidant activity and identify steroid isolates of n-hexane fraction of H. verticillata. The steroid compounds were extracted using methanol solvent. The methanol extract was hydrolyzed with HCl 2 N and partitioned using n-hexane. The n-hexane fraction was tested phytochemically and separated using thin-layer chromatography (TLC). The steroids compounds were identified using UV-Vis spectrophotometer, FTIR, and LC-MS/MS. The result showed that maceration extraction produced 5.14% yield, whereas n-hexane fraction produce 47.95% yield. Steroid separation through analytical TLC revealed that n-hexane: ethyl acetate (4:1) as the best eluant with 12 spots, while separation using preparative TLC yielded 19 spots. Steroid isolate from TLC preparative has antioxidant activity with EC50 of 5.109 ppm. Identification of steroid compounds using UV-Vis produced maximum wavelengths at 203.9 and 276 nm, while using FTIR indicated the presence of O-H group, geminal dimethyl, C=O, C=C, secondary C-OH, and =C-H (alkene) which might contain steroid compounds. The result of LC-MS/MS showed the presence of β-sitosterol. Hydrilla verticillata merupakan salah satu ciptaan Allah SWT yang memiliki potensi sebagai antioksidan. Penelitian ini bertujuan mengetahui aktivitas antioksidan dan mengidentifikasi senyawa steroid pada isolat hasil pemisahan fraksi n-heksana H. verticillata. Senyawa steroid diekstraksi menggunakan pelarut metanol. Ekstrak pekat metanol dihidrolisis dengan HCl 2 N dan dipartisi dengan n-heksana. Fraksi n-heksana hasil partisi diuji fitokimia dan dipisahkan senyawanya menggunakan KLT (Kromatografi Lapis Tipis). Identifikasi dilakukan menggunakan UV-Vis, FTIR, dan LC-MS/MS. Hasil penelitian menunjukkan ekstraksi maserasi menghasilkan rendemen 5,14%, sedangkan rendemen partisi n-heksana sebesar 47,95%. Pemisahan steroid menggunakan KLT analitik menunjukkan bahwa variasi eluen terbaik adalah n-heksana: etil asetat (4:1) dengan 12 noda, sedangkan pemisahan KLT preparatif menghasilkan 19 noda. Isolat steroid hasil KLT preparatif memiliki aktivitas antioksidan dengan EC50 5,109 ppm. Identifikasi senyawa steroid menggunakan UV-Vis menunjukkan bahwa panjang gelombang maksimum sebesar 203,9 dan 276 nm, sedangkan untuk identifikasi isolat steroid menggunakan FTIR menunjukkan gugus fungsi O-H, geminal dimetil, C=O, C=C, C-OH sekunder dan =C-H (alkena) yang diduga merupakan senyawa steroid. Hasil LC-MS/MS menunjukkan adanya senyawa steroid β-sitosterol.
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