Potensi Steroid Alami Tumbuhan Pasak Bumi (Eurycoma Longifolia Jack) Untuk Maskulinisasi Ikan Nila (Oreochromis Niloticus)

Yusuf, Noor Syarifuddin (2019) Potensi Steroid Alami Tumbuhan Pasak Bumi (Eurycoma Longifolia Jack) Untuk Maskulinisasi Ikan Nila (Oreochromis Niloticus). Doktor thesis, Universitas Brawijaya.

Abstract

Laporan hasil disertasi dengan judul “Potensi Steroid Alami Tumbuhan Pasak Bumi (Eurycoma longifolia Jack) Untuk Maskulinisasi Ikan Nila (Oreochromis niloticus)“ bertujuan menganalisis kandungan senyawa bioaktif steroid dari bagian tumbuhan Pasak Bumi untuk mendapatkan pelarut, metode ekstraksi yang menghasilkan steroid tertinggi, mengisolasi stigmasterol dari bagian tumbuhan pasak bumi yang memiliki kandungan steroid tertinggi dan menganalisis performa ekstrak steroid dan ekstrak kolom stigmasterol tumbuhan Pasak Bumi pada maskulinisasi ikan Nila. Penelitian ini dilaksanakan dalam tiga tahapan dengan perincian sebagai berikut: Tahapan Pertama. Melakukan analisis dan karakterisasi senyawa aktif tumbuhan Pasak Bumi yang diawali dengan penyiapan bahan baku bagian tumbuhan Pasak Bumi berupa akar, batang, kulit, daun dan ranting berbentuk serbuk halus atau tepung (Powder). Pengujian proximat untuk mengetahui persentase kandungan protein, lemak, air , karbohidrat dan kadar abu. Pengujian fitokimia bagian tumbuhan Pasak Bumi secara kualitatif dengan menganalisis golongan senyawa alkaloid, flavanoid, saponin, terpenoid, polifenol dan steroid pada bagian akar, batang, kulit, daun dan ranting. Analisis kandungan senyawa bioaktif steroid dari bagian tumbuhan Pasak Bumi memperlihatkan efektivitas metode pengekstraksi pada bagian tumbuhan Pasak Bumi adalah akar yang direflux dengan pelarut metanol (rendemen 9,22%), ranting yang disoklet dengan pelarut metanol (rendemen 6,09%), batang yang direflux dengan pelarut metanol (rendemen 4,30%), daun yang direflux dengan pelarut metanol (rendemen 3,87%) dan kulit yang disoklet dengan pelarut metanol (rendemen 3,37%). Berdasarkan analisis Kromatografi Lapis Tipis (KLT) ekstrak metanol, kloroform, aseton dari daun, ranting, kulit, batang dan akar tumbuhan Pasak Bumi yang dibandingkan terhadap kedua senyawa standar (stigmasterol dan testosteron) dan analisis data ekstrak yang dihasilkan dengan variasi metode ekstraksi (maserasi, sokletasi dan reflux) dengan variasi pelarut (metanol, aseton, kloroform, kloroformmetanol (1:1)) dapat dijelaskan bahwa dari kedua senyawa target stigmasterol dan testosteron, hanya stigmasterol yang teridentifikasi pada bagian daun, ranting, kulit, batang dan akar sedangkan testosteron tidak teridentifikasi pada kelima bagian tumbuhan Pasak Bumi tersebut. Tahapan Kedua. Mengisolasi stigmasterol dari bagian tumbuhan Pasak Bumi yang memiliki rendemen ekstrak tertinggi dan kandungan steroid tertinggi yaitu bagian akar. Pemisahan ekstrak metanol akar tumbuhan Pasak Bumi dilakukan dengan metode kromatografi kolom untuk mendapatkan kristal stigmasterol, dari 1.095 gram total sampel tepung akar pasak bumi dihasilkan ekstrak metanol sebanyak 54,98 gram dengan persentase massa ekstrak rata-rata sebesar 4,93 % tepung akar pasak bumi. Stigmasterol sebanyak 590,1 mg dengan persentase stigmasterol total dalam ekstrak metanol sebesar 5,40 % dan rata-rata persentase stigmasterol dalam akar tumbuhan pasak bumi adalah 1,35 %. Penentuan struktur kimia menggunakan spektrometer FTIR menunjukkan bahwa isolat merupakan senyawa steroid dengan gugus fungsi OH, C=C nonkonjugasi, C-O, dan C-H serta memiliki massa molekul m/z 396. Data interpretasi menunjukkan kemiripan dengan senyawa sterol. xii GC-MS Ekstrak metanol akar tumbuhan Pasak Bumi ditemukan 31 jenis senyawa. Persentase kandungan senyawa berkisar antara 0,93% sampai dengan 10,39% dan terdapat 5 jenis senyawa yang tidak diketahui namanya. Kandungan senyawa stigmasterol dalam ekstrak metanol yang di uji sebanyak 5,97%. Jenis steroid lainnya berupa sitosterol sebanyak 3,22%; senyawa spinasteron sebanyak 4,62% dan sebanyak 1,12% senyawa estratriennol sedangkan pengujian melalui GC-MS ekstrak kolom stigmasterol akar tumbuhan Pasak Bumi memperlihatkan ekstrak kolom yang dihasilkan masih belum sepenuhnya senyawa stigmasterol. Sebanyak 15 jenis senyawa yang teridentifikasi. Persentase senyawa Stigmasterol hanya sebesar 32,82%. Senyawa lainya yang tergolong tinggi dari jenis steroid adalah Beta sitosterol 23,41% dan Kampesterol sebanyak 13,21%. Jenis steroid lainnya yang teridentifikasi adalah Spinastenon sebesar 2,1%; Sitostenon sebesar 1,27%;Stimastenon sebanyak 0,96% dan terdapat satu senyawa yang tidak teridentifikasi sebesar 0,9%. Pengujian LC-MS/MS dari 22 jenis asam amino yang di analisis hanya teridentifikasi sebanyak 19 jenis asam amino pada ekstrak metanol akar tumbuhan Pasak Bumi. Jumlah Arginin sebesar 2.360 mg/kg, Asam glutamat sebesar 1.230 mg/kg, Prolin sebesar 802 mg/kg, Asam aspartat 633 mg/kg dan lainnya sementara pada ekstrak kolom Stigmasterol hanya teridentifikasi 2 jenis asam amino yaitu Serin sebesar 0,42 mg/kg dan Fenil alanin sebesar 0,27 mg/kg. Tahapan Ketiga. Persentase Jantan. Maskulinisasi ikan Nila menggunakan ekstrak metanol (EPB) dan ekstrak kolom stigmasterol (SPB) akar tumbuhan pasak bumi dengan analisis anova menunjukkan pengaruh pada persentase Jantan (P<0,05). EPB melalui pakan menghasilkan persentase jantan (80,36% - 82,10%) lebih tinggi dari EPB melalui perendaman (72,66% - 76,36%) sedangkan persentase jantan SPB melalui peredaman (65,82% - 67,14%) memperlihatkan hasil yang tidak jauh berbeda dengan SPB melalui pakan (63,92% - 66,97%). Persentase jantan menggunakan 17α-Metil testosteron (MT) baik melalui perendaman (83,99%) dan melalui pakan (88,53%) masih lebih lebih tinggi dibandingkan pemberian EPB dan SPB. Persentase Interseks. Analisis Anova menunjukkan pengaruh pada interseks Ikan Nila (p<0,05). Ikan nila yang yang tidak diberikan hormon (kontrol negatip) tidak tejadi fenomena interseks (0%) tetapi EPB melalui perendaman interseks yang terjadi (7,83% - 9,99%), EPB melalui pakan (6,71% - 6,96%), SPB melalui perendaman (7,13% - 10,36%) dan SPB melalui pakan (7,38% - 9,76%). Terlihat kecenderungan fenomena interseks lebih tinggi terjadi melalui perendaman dibandingkan melalui pakan. Kelangsungan hidup. Analisis anova menunjukkan tidak ada pengaruh perlakuan (p>0,05), kecuali pemberian EPB melalui pakan berpengaruh terhadap kelangsungan hidup (p<0,05). Kelangsungan hidup ikan Nila dengan pemberian EPB melalui pakan (86,00% - 88,67%), EPB melalui perendaman (80,67% - 85,33%), SPB melalui pakan (80,67% - 82,67%) dan SPB melaui rendam (82,00% - 84,00%). Kecenderungan data memperlihatkan pemberian EPB baik melalui perendaman maupun lewat pakan menunjukkan kisaran nilai kelansungan hidup ikan nila yang lebih tinggi. Persentase Laju Pertumbuhan Harian (SGP). Analis anova memperlihatkan pemberian EPB berpengaruh pada persentase laju pertumbuhan harian (SGP) (p<0,05) sedang pemberian SPB tidak berpengaruh (p>0,05). SGP pemberian EPB melalui pakan (0,156% - 0,162%), pemberian EPB melalui perendaman (0,134% - 0,150%), pemberian SPB melalui pakan (0,127% - 0,129%) dan pemberian SPB melalui perendaman (0,125% - 0,127%). EPB memperlihatkan kecenderungan SGP lebih tinggi dibandingkankan SPB dan EPB melalui pakan menunjukkan SGP yang lebih tinggi dibandingkan EPB melalui perendaman, SPB melalui pakan dan SPB melalui rendam. SGP melalui pakan cenderung lebih tinggi dibandingkan SGP melalui perendaman. Pertumbuhan Mutlak atau Pertambahan Berat (gram). Analis anova memperlihatkan pemberian EPB berpengaruh pada pertambahan berat (gram) (p<0,05) sedang pemberian SPB tidak berpengaruh (p>0,05). Pertambahan berat pemberian EPB melalui pakan (9,35 gram – 9,65 gram), pemberian EPB melalui perendaman (8,07 gram – 9,02 gram), pemberian SPB melalui pakan (7,67 gram – 7,73 gram) dan pemberian xiii SPB melalui perendaman (7,49 gram – 7,60 gram). EPB memperlihatkan pertambahan berat lebih besar dibandingkankan SPB dan EPB melalui pakan menunjukkan SGP yang lebih tinggi dibandingkan EPB melalui perendaman, SPB melalui pakan dan SPB melalui rendam. Secara umum pertambahan berat melalui pakan lebih tinggi dari pemberian melalui perendaman. Gambaran histologi. Semua perlakuan menunjukkan adanya perkembangan pada diameter tubulus seminiferus gonad ikan Nila. Gambaran histologi perlakuan yang menerima hormon, perkembangan diameter terlihat lebih cepat (besar) dibandingkan dengan tanpa hormon atau perlakuan kontrol negatip. Profil hormon testosteron. Semua ikan nila yang digunakan telah memiliki testosteron alamiah di dalam tubuhnya yang terlihat dari data ikan yang digunakan sebelum perendaman dan sebelum pemberian pakan memperlihatkan keberadaan hormon. Terdapat perbedaan pola antara perendaman dengan yang melalui pakan. Melalui perendaman hormon testosteron melonjak drastis (H-0) kemudian turun secara Fluktuatif hinga (H-60) sedangkan melalui pakan, hormon testosteron dari (H-0) meningkat hingga (H-15) kemudian turun secara fluktuatif hingga (H-60).

English Abstract

The dissertation report entitled "Potential of Natural Steroids from Pasak Bumi (Eurycoma longifolia Jack) for Masculinization of Tilapia (Oreochromis niloticus)" aimed to analyze the content of bioactive steroid compounds from parts of pasak bumi to obtain solvents, extraction methods that produce the highest steroids. In addition, this study aimed to isolate stigmasterol from the parts of pasak bumi that have the most top steroid content. This study also analyzed the performance of steroid extracts and Stigmasterol column extracts of the pasak bumi on the tilapia masculinization. This research was conducted in three stages with the following details: The first stage, analysis and characterization of the active compounds from Pasak Bumi plant. This process begins with the preparation of raw materials from parts of pasak bumi in the form of roots, stems, bark, leaves, and twigs, which have been in the form of fine powder or flour. Proximate assay to determine the percentage of protein, fat, water, carbohydrate, and ash content. Phytochemical assay of the pasak bumi parts qualitatively by analyzing the compounds of alkaloids, flavonoids, saponins, terpenoids, polyphenols, and steroids on the roots, stems, bark, leaves, and twigs. Analysis of bioactive steroid compounds from pasak bumi parts showed the effectiveness of the extraction method on pasak bumi parts was root refluxed with methanol solvent (yield 9.22%), twigs which were coupled with methanol solvent (yield 6.09%), stem that was refluxed with methanol solvent (4.30% yield), leaves refluxed with methanol (3.87% yield) and skins that were supplemented with methanol solvent (3.37% yield). Based on Thin Layer Chromatography (TLC) analysis of methanol, chloroform, acetone extract from leaves, twigs, bark, stems and roots of the Pasak Bumi compared to the two standard compounds (stigmasterol and testosterone) resulted in the analysis of extract data with various extraction methods (maceration, socletation, and reflux). The results of different extraction methods with multiple solvents (methanol, acetone, chloroform, chloroform-methanol (1: 1)) showed that of the two stigmasterol and testosterone target compounds, only stigmasterol was identified in the leaves, twigs, bark, stems, and roots while testosterone not defined in the five parts of the plant Pasak Bumi. Second stage. Isolation of stigmasterol from parts of Pasak Bumi which has the highest extract yield and most top steroid content, namely the root. Separation from the root of pasak bumi methanol extract was carried out by column chromatography method to obtain stigmasterol crystals. 1095 g of pasak bumi root powder produced 54.98 g of methanol extract with an average extract mass percentage of 4.93%. The amount of stigmasterol in methanol extract was 590.1 mg, with a percentage of 5.40%. The average percentage of stigmasterol in the roots of the post-earth plant was 1.35%. Determination of the chemical structure using FTIR spectrometers showed that the isolate was a steroid compound with OH, C = C non-conjugated, C-O, and C-H functional groups and had a molecular mass of 396 m/z. Interpretation data showed similarities to sterol compounds. GC-MS Pasak Bumi root methanol extract found 31 types of compounds. The percentage of compound content ranges from 0.93% to 10.39%, and there are five types of unknown compounds. The content of stigmasterol compounds in methanol extract was 5.97%. Other types of steroids in the form of sitosterol as much as 3.22%; spinasterone as much as 4.62% and estratriennol as much as 1.12%. GC-MS from the Stigmasterol column xv extract of Pasak Bumi root showed that the extract of the column produced was not yet completely stigmasterol compound. A total of 15 types of compounds were identified. The percentage of Stigmasterol compounds is only 32.82%. Other compounds that were classified as high from this type of steroid are Beta sitosterol 23.41% and Kampesterol as much as 13.21%. Different types of steroids identified were Spinastenone at 2.1%; Sitostenon was 1.27%; Stimastenone was 0.96%, and there was one unidentified compound of 0.9%. LC-MS/MS analysis of 22 types of amino acids analyzed only identified 19 types of amino acids in the Pasak Bumi root methanol extract. The study showed the amount of Arginine was 2,360 mg/kg; Glutamic Acid was 1,230 mg/kg, Proline was 802 mg/kg, Aspartic Acid was 633 mg/kg and others. The results of LC-MS/MS analysis on Stigmasterol column extracts identified only two types of amino acids, namely Serin at 0.42 mg/kg and Phenylalanine at 0.27 mg/kg. Third Stage. Percentage of Male. Masculinization of tilapia used pasak bumi roots methanol extracts (EPB), and stigmasterol column extracts of pasak bumi root (SPB) with ANOVA analysis showed an effect on the percentage of males (P< 0.05). EPB through feed produced a higher rate of males (80.36% - 82.10%) than EPB through immersion (72.66% - 76.36%). SPB produced a percentage of males through immersion (65.82% - 67.14%) showed results that are not much different from SPB through feed (63.92% - 66.97%). The percentage of males using 17α-Methyl testosterone (MT) through immersion (83.99%) and feed (88.53%) was still higher than that of EPB and SPB. Percentage of Intersex. ANOVA analysis showed the effect on intersex Tilapia (p<0.05). Tilapia that were not given hormones (negative control) did not occur as an intersex phenomenon (0%) but administration of EPB through immersion occurred intersex (7.83% - 9.99%), EPB through feed (6.71% - 6.96 %), SPB through immersion (7.13% - 10.36%) and SPB through feed (7.38% - 9.76%). The results showed a tendency for the intersex phenomenon to be higher through immersion than through feed. Survival rate. ANOVA analysis showed no treatment effect (p>0.05), except that the administration of EPB through feed affected survival (p<0.05). Survival rate of Tilapia by giving EPB through feed (86.00% - 88.67%), EPB through immersion (80.67% - 85.33%), SPB through feed (80.67% - 82.67%) and SPB through immersion (82.00% - 84.00%). The trend of data showed that the administration of EPB either through immersion or feed showed a higher range of values for the survival of tilapia. Percentage of Daily Growth Rate (SGP). ANOVA analysis showed that EPB administration affected the percentage of daily growth rate (SGP) (p<0.05); while giving SPB did not affect (p>0.05). SGP by giving EPB through feed (0.156% - 0.162%), giving EPB through immersion (0.134% - 0.150%), giving SPB through feed (0.127% - 0.129%) and giving SPB through immersion (0.125% - 0.127%). EPB showed a tendency for SGP to be higher than SPB. Giving EPB through feed showed a higher SGP compared to EPB through immersion. Giving SPB through feed showed a higher SGP compared to SPB through immersion. Absolute Growth or Weight gain (g). ANOVA analysis showed that EPB administration affected weight gain (g) (p<0.05), whereas SPB administration did not affect (p>0.05). Weight gain due to EPB through feed (9.35 g - 9.65 g). Weight gain due to EPB administration through immersion (8.07 g - 902 g). Weight gain due to SPB administration through feed (7.67 g - 7.73 g). Weight gain due to SPB administration through immersion (7.49 g - 7.60 g). Giving EPB shows a higher weight gain than SPB. Weight gain due to EPB through the feed is higher than EPB through soaking, SPB was provided through feed and immersion. In general, weight gain due to administration of EPB and SPB through the feed was higher than through immersion. Histology Analysis. All treatments showed development in the diameter of the seminiferous tubules of the Tilapia gonad. The histological figure in the treatment that received hormones showed that the development of gonad diameter was seen to be faster (larger) compared to without hormones or negative control treatments. xvi The profile of Testosterone hormone. All tilapia that were used already have natural testosterone in tilapia body which can be seen from the data of fish used before immersion and before feeding showed the presence of the hormone testosterone. The results showed there were differences in patterns between immersion and feed. The immersion treatment shows that testosterone increases (Day 0) then decreases fluctuatively until (Day 60), whereas through the feed, testosterone from (Day 0) rises to (Day 15) then decreases fluctuatively until (60th day).

Other obstract

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Item Type:	Thesis (Doktor)
Identification Number:	DIS/572.579/YUF/p/2019/061911226
Uncontrolled Keywords:	Steroid Alami, Pasak Bumi, Maskulinisasi Ikan
Subjects:	500 Natural sciences and mathematics > 572 Biochemistry > 572.5 Miscellaneous chemicals > 572.57 Lipids
Divisions:	S2/S3 > Doktor Ilmu Perikanan dan Kelautan, Fakultas Perikanan dan Ilmu Kelautan
Depositing User:	Endang Susworini
Date Deposited:	24 May 2022 08:29
Last Modified:	24 May 2022 08:29
URI:	http://repository.ub.ac.id/id/eprint/190742

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