ANALYSIS OF SECONDARY METABOLITES OF CALLUS OF RAMBUTAN Nephelium lappaceum L

Rambutan plant (Nephelium lappaceum L.) is a member of the Sapindaceae family. The rambutan plant is one of the natural ingredients that can be developed as traditional medicine. Rambutan peel has the potential for good antioxidant and anticancer activity. Rambutan fruit does not grow every time it needs efforts to produce the active substance in rambutan, using plant tissue culture techniques. The use of the correct variety of mediums and hormones at the right concentration is the key to thriving tissue culture. Explants derived from rambutan leaves were planted precisely on solid media Murashige and Skoog (MS) and WoddyPlant Medium (WPM) containing Indole-3-Butyric Acid (IBA) and Kinetin. After seven days, the callus was subcultured, then after 35 days, the subculture callus was collected and dried. Dry callus and rambutan leaves (Wild type) were macerated with n-hexane, ethyl acetate, and ethanol. The concentrated extract was then applied to a GF 254 silica gel plate with the mobile phase Toluene-Acetone (7: 3) and n-hexane-EthylAsetate (3: 7). The results showed that the concentration of IBA 2 ppm and kinetin three ppm was the best combination because it produced callus. TLC results of rambutan leave with plant tissue culture containing flavonoids and triterpenoids. This study provides new information regarding the induction of rambutan callus and can become the basis for producing active metabolites in rambutan with cell suspension culture development.  Tanaman rambutan (Nephelium lappaceum L.) Merupakan salah satu anggota famili Sapindaceae. Tanaman rambutan merupakan salah satu bahan alami yang berpotensi untuk dikembangkan sebagai obat tradisional. Kulit rambutan mempunyai potensi aktivitas antioksidan dan antikanker yang baik. Buah rambutan tidak tumbuh setiap saat maka perlu upaya untuk memproduksi zat aktif dalam rambutan, salah satunya dengan menggunakan teknik kultur jaringan tanaman. Penggunaan variasi medium dan hormone yang tepat pada konsentrasi yang tepat merupakan kunci sukses kultur jaringan. Eksplan yang berasal dari daun rambutan ditanam secara tepat pada media padat Murashige dan Skoog (MS) dan Woddy Plant Medium (WPM) yang mengandung Indole-3-Butric Acid (IBA) dan Kinetin. Setelah 7 hari kalus disubkultur, kemudian setelah 35 hari kalus subkultur dikumpulkan dan dikeringkan. Kalus kering dan daun rambutan (Wlid type) dimaserasi dengan n-heksan, etilasetat dan etanol, kemudian ekstrak pekat diaplikasikan pada plate silika gel GF 254 dengan fasa gerak Toluen-Aseton (7: 3) dan n-heksan-EtilAsetat (3: 7). Hasil penelitian menunjukkan bahwa konsentrasi IBA 2 ppm dan kinetin 3 ppm merupakan kombinasi terbaik karena menghasilkan kalus. Hasil Kromatografi lapis tipis (KLT) daun rambutan dengan kultur jaringan tanaman mengandung flavonoid dan triterpenoid. Hasil penelitian ini memberikan informasi baru mengenai induksi kalus rambutan dan bisa menjadi dasar produksi metabolit aktif dalam rambutan dengan pengembangan ke arah kultur suspensi sel

THE GROWTH OF PIMPINELLA ALPINA HOST CALLUS AT VARIOUS TREATMENTS OF PLANT GROWTH REGULATOR CONCENTRATIONS OF NAA. 2,4 D AND ITS COMBINATION WITH BAP

Purwaceng (Pimpinella alpina   Host) merupakan tumbuhan herbal yang akarnya berkhasiat obat afrodisiak, diuretik dan tonik. Pertumbuhan dan perkembangan Purwaceng sangat terbatas serta eksploitasi yang berlebih dan tidak diimbangi dengan upaya konservasi sehingga tanaman tersebut dimasukkan dalam kategori Apendiks 1 (hampir punah) maka diperlukan konservasi. Salah satu upaya konservasi tanaman ini yaitu dengan teknik kultur jaringan tanaman. Metode ini merupakan salah satu cara untuk menginduksi kalus dan metabolit sekunder. Penilitian ini bertujuan untuk mengetahui pengaruh konsentrasi zat pengatur tumbuh terhadap pertumbuhan kalus. Konsentrasi Zat Pengatur Tumbuh NAA (0,5 mg/L, 1 mg/L, 2 mg/L), 2,4-D (0,4 mg/L, 0,8 mg/L, 1,2 mg/L), kombinasi NAA dengan BAP (0,5 mg/L NAA : 1 mg/L BAP, 1 mg/L NAA : 2 mg/L BAP, mg/L NAA : 3 mg/L BAP) dan kombinasi 2,4-D dengan BAP (1 mg/L 2,4-D : 0,5 mg/L BAP, 1 mg/L 2,4-D : 1 mg/L BAP, 2 mg/L 2,4-D : 1 mg/L BAP) kedalam media Murashige Skoog. Kemudian eksplan daun purwaceng ditanam ke dalam media-media tersebut sampai terlihat pertumbuhan. Hasil penelitian menunjukkan bahwa konsentrasi zat pengatur tumbuh memberikan pengaruh yang berbeda pada pertumbuhan kalus yang ditunjukkan oleh hari munculnya kalus, warna dan tekstur kalus. Hasil penelitian didapatkan pada zat pengatur tumbuh tunggal 2,4-D pada konsentrasi 1,2 mg/L, memiliki pertumbuhan kalus yang baik dan sedangkan untuk zat pengatur tumbuh kombinasi yang optimal adalah dengan perlakuan NAA 0,5 mg/L with BAP 1 mg/L.Purwaceng (Pimpinella alpina) is an herbal plant that has aphrodisiac, diuretic, and tonic properties. The imbalance between the growth and the exploitation of this plant makes it included in Appendix I (endangered), so its conservation will be crucial. One method of the conservation of this plant is tissue culture. This method is considered one way to induct the callus. The research objective is to find the effect of various treatments of plant growth regulator concentrations of NAA. 2,4 D and its combination with BAP on the growth of purwoceng callus. Concentration of plant growth regulator NAA (0.5 mg L-1, 1 mg L-1, 2 mg L-1), 2,4-D (0.4 mg L-1, 0.8 mg L-1, 1.2 mg L-1 ), a combination of NAA with BAP (0.5 mg L-1 NAA: 1 mg L-1 BAP, 1 mg L-1 NAA: 2 mg L-1 BAP, mg L-1 NAA: 3 mg L-1 BAP) and a combination of 2, 4- D with BAP (1 mg LL 2,4-D : 0.5 mg L BAP, 1 mg L-1 2,4-D : 1 mg L-1 BAP, 2 mg L-1 2,4-D : 1 mg L-1 BAP) into Murashige Skoog medium. Then purwaceng leaf explants were planted on the medium until the growth was seen. This research shows that the plant growth regulator concentration has a different influence on the growth of callus that is recognized by the day of callus induction, colour and callus texture. MS media with growth regulator 1.2 mg L-1 2,4-D can grow callus faster (17 days) with friable texture

Identification of Secondary Metabolites Compounds From Nampu (Homalomena rosrata Griff.)

Abstract: Nampu (Homalomena rosrata Griff.) is traditionally used to increase sexual activity for men. However, the lack of scientific information on the phytochemical content of Nampu plants needs further research on the secondary metabolite compounds for isolation and identification purposes. The phytochemical screening of raw material, ethanol extract, and water fraction shows positive results for flavonoid, tannin, polyphenol, quinone, monoterpenoid-sesquiterpenoid, and steroid-triterpenoid groups. The chloroform fractions were monitored by the thin-layer chromatography (TLC) method with the addition of KOH 5%. The results showed a blue spot under UV light at 365 nm after the addition of KOH 5%. The fraction was purified by a preparative thin-layer chromatography method (TLC-P), while the isolates' purity was tested by the two-dimensional TLC method and TLC with three different phases of mobile phases. The results of the analysis with the spectrophotometer UV-Visible showed the wavelength of 278.8 nm and 213.2 nm. In addition, the functional group analysis using the I.R. spectrometer showed O.H. stretch at wave numbers 3419.79 cm-1, aromatic C-H at wave number 2922.16 cm-1, C = O at the wave number 1672.28 cm-1, strain C = C aromatic at the wave number 1585.49 cm-1. Based on TLC and spectrophotometric data, the isolates were coumarin compounds.Abstrak: Nampu (Homalomena rosrata Griff.) secara tradisional digunakan untuk meningkatkan aktivitas seksual pria. Namun, kurangnya informasi ilmiah tentang kandungan fitokimia tanaman Nampu, perlu penelitian lebih lanjut tentang senyawa metabolit sekunder untuk tujuan isolasi dan identifikasi. Penapisan fitokimia bahan baku, ekstrak etanol dan fraksi air menunjukkan hasil positif untuk golongan flavonoid, tanin, polifenol, kuinon, monoterpenoid-seskuiterpenoid, dan steroid-triterpenoid. Fraksi kloroform dipantau dengan metode kromatografi lapis tipis (KLT) dengan penambahan KOH 5%. Hasilnya menunjukkan bercak berwarna biru di bawah lampu U.V. 365 nm. Fraksi dimurnikan dengan metode kromatografi lapis tipis preparatif (KLT-P), sedangkan kemurnian isolat diuji dengan metode KLT dua dimensi dan KLT dengan tiga fase gerak yang berbeda. Hasil analisis dengan spektrofotometer UV-Visible menunjukkan panjang gelombang 278,8 nm dan 213,2 nm. Selain itu, analisis gugus fungsi menggunakan spektrometer infra merah menunjukkan adanya gugus OH, regangan pada bilangan gelombang 3419,79 cm-1, C-H aromatik pada bilangan gelombang 2922,16 cm-1, C = O pada bilangan gelombang 1672,28 cm-1, regangan C = C aromatik pada bilangan gelombang 1585,49 cm-1. Berdasarkan data KLT dan spektrofotometri, diduga isolat merupakan golongan senyawa kumarin.Nampu (Homalomena rosrata Griff.) secara tradisional digunakan untuk meningkatkan aktivitas seksual pria. Namun, kurangnya informasi ilmiah tentang kandungan fitokimia tanaman Nampu, perlu penelitian lebih lanjut tentang senyawa metabolit sekunder untuk tujuan isolasi dan identifikasi. Penapisan fitokimia bahan baku, ekstrak etanol dan fraksi air menunjukkan hasil positif untuk golongan flavonoid, tanin, polifenol, kuinon, monoterpenoid-seskuiterpenoid, dan steroid-triterpenoid. Fraksi kloroform dipantau dengan metode kromatografi lapis tipis (KLT) dengan penambahan KOH 5%. Hasilnya menunjukkan bercak berwarna biru di bawah lampu U.V. 365 nm. Fraksi dimurnikan dengan metode kromatografi lapis tipis preparatif (KLT-P), sedangkan kemurnian isolat diuji dengan metode KLT dua dimensi dan KLT dengan tiga fase gerak yang berbeda. Hasil analisis dengan spektrofotometer UV-Visible menunjukkan panjang gelombang 278,8 nm dan 213,2 nm. Selain itu, analisis gugus fungsi menggunakan spektrometer infra merah menunjukkan adanya gugus OH, regangan pada bilangan gelombang 3419,79 cm-1, C-H aromatik pada bilangan gelombang 2922,16 cm-1, C = O pada bilangan gelombang 1672,28 cm-1, regangan C = C aromatik pada bilangan gelombang 1585,49 cm-1. Berdasarkan data KLT dan spektrofotometri, diduga isolat merupakan golongan senyawa kumarin

PHYTOCHEMICAL ANALYSIS OF CALLUS TWO VARIETIES ORTHOSIPHON ARISTATUS (BLUME) MIQ ON MURASHIGE AND SKOOG MEDIA: A STRATEGIC STEP OF SECONDARY METABOLITE PRODUCTION

Objective: The research aimed to provide new information regarding the secondary metabolites content of purple and white-purple Orthosiphon aristatus (Blume) Miq. callus, which can then be used as a basis for developing towards cell suspension and ultimately producing secondary metabolites using bioreactors. Methods: Callus induction of two varieties of O. aristatus were performed by inoculating sterile leaf explants grown on Murashige and Skoog basal media supplemented with 2,4-dichlorophenoxyacetis acid 0.4 ppm. The secondary metabolites were analysed and quantified using high-performance liquid chromatography with gradient elution. Results: The results showed the growth of callus two varieties of O. aristatus in growth media MS with 2,4-D 0.4 ppm. Rosmarinic acid content in the acetone extract of the purple variety callus was 1.28% w/w, and the white-purple variety was 2.22% w/w. Conclusion: This study could form the basis for the development of rosmarinic acid production by In vitro culture modification

Effect of Media Variation on the Induction and Phytochemical Profile of Callus in Two Varieties of Cat's Whiskers (Orthosiphon aristatus Blume Miq)

The levels of rosmarinic acid and sinensetin in purple and white-purple varieties of Orthosiphon aristatus, cat's whiskers, can be increased using modified in vitro culture. This work focused on callus induction of the purple and white-purple variety of cat's whiskers grown on Gamborg (B5) and CHU (N6) with the addition of growth regulators 2,4-dichlorophenoxyacetis acid. Our observation suggested that the callus could grow within three weeks and produce rosmarinic acid and sinensetin. The level of sinensetin from various extraction methods is relatively low; in contrast, the rosmarinic acid from purple callus was detected at about 5% w/w, while the white-purple variety was around 2% w/w. The results of this study also provided new information on the basic media other than MS that can grow cat's whiskers callus while producing active compounds

FORMULASI SEDIAAN LOSION ANTIOKSIDAN EKSTRAK AIR DAUN TEH HIJAU (Camellia sinensis L.)

Penelitian tentang formulasi dosis ekstrak air daun teh hijau dengan konsentrasi 0,0002%, 0,002%, 0,02% dan 8,6% telah dilakukan. Penetapan nilai ECso dilakukan berdasarkan perhitungan persamaan regresi linier antara larutan uji dan persentase peredamanl. Formulasi losion dimulai berdasarkan orientasi basis dengan parameter organoleptik, homogenitas, pH dan viskositas. Diketahui bahwa losion dengan 5,5% glyceryl monostearate mempunyai stabilitas pH, viskositas dan konsistensi yang terbaik. Evaluasi formulasi termasuk evaluasi fisik (organoleptis, homogenitas, viskositas dan stabilitas) dan evaluasi kimia (pH dan stabilitas aktivitas anti oksidanJ. Evaluasi aktivitas antioksidan formula yang mengandung larutan 1,1- diphenyl-2-picrylhidrazyl menggunakan spektrofotometri UV-Vis menunjukkan bahwa formula yang mengandung 0.02% dan 8,6% ektrak air daun teh memberikan aktivitas anti oksidan terbaik dan stabilitas selama penyimpanan. The research about lotion dosage formulation that contain green tea leaf water extract with concentration 0.0002%, 0.002%, 0,02%, and 8.6% had been done. The basic of the ECsovalue.Wascalculated from linierity regression equation between tested solution concentration and scavenging percent The lotion formulation began with lotion base orientation included organoleptic, homogeinity, pH, and viscosity. It showed that lotion base with glyceryl monostearate 5.5% have the best pH stability, viscosity, and consistency. Evaluation of lotion dosage include physical evaluation (organoleptic, homogeinity, viscosity and stability) and chemical evalution (pH and the stability of lotion antioxidant activity. Antioxidant activity stability evaluation of lotion with l,l-diphenyl-2-picrylhidrazyl solution using spectrophotometri UV-Visible showed, that formula which contain 0.02% and 8.6% green tea leaf water extract gave the best radical scavenging activity and stable during storage

FORMULASI SEDIAAN LOSION ANTIOKSIDAN EKSTRAK AIR DAUN TEH HIJAU (Camellia sinensis L.)

The research about lotion dosage formulation that contain green tea leaf water extract with concentration 0.0002%, 0.002%, 0,02%, and 8.6% had been done. The basic of the EC50 value. Which calculated from linierity regression equation between tested solution concentration and scavenging percent. The lotion formulation began with lotion base orientation included organoleptic, homogeinity, pH, and viscosity. It showed that lotion base with glyceryl monostearate 5.5% have the best pH stability, viscosity, and consistency. Evaluation of lotion dosage include physical evaluation (organoleptic, homogeinity, viscosity and stability) and chemical evalution (pH and the stability of lotion antioxidant activity. Antioxidant activity stability evaluation of lotion with 1,1-diphenyl-2-picrylhidrazyl solution using spectrophotometri UV-Visible showed, that formula which contain 0.02% and 8.6% green tea leaf water extract gave the best radical scavenging activity and stable during storage. Penelitian tentang formulasi dosis ekstrak air daun teh hijau dengan konsentrasi 0,0002%, 0,002%, 0,02% dan 8,6% telah dilakukan. Penetapan nilai EC50 dilakukan berdasarkan perhitungan persamaan regresi linier antara larutan uji dan persentase peredamanl. Formulasi losion dimulai berdasarkan orientasi basis dengan parameter organoleptik, homogenitas, pH dan viskositas. Diketahui bahwa losion dengan5,5% glyceryl monostearate mempunyai stabilitas pH, viskositas dan konsistensi yang terbaik. Evaluasi formulasi termasuk evaluasi fisik (organoleptis, homogenitas, viskositas dan stabilitas) dan evaluasi kimia (pH dan stabilitas aktivitas anti oksidan). Evaluasi aktivitas antioksidan formula yang mengandung larutan 1,1-diphenyl-2-picrylhidrazyl menggunakan spektrofotometri UV-Vis menunjukkan bahwa formula yang mengandung 0.02% dan 8,6% ektrak air daun teh memberikan aktivitas anti oksidan terbaik dan stabilitas selama penyimpanan

Formulation of Antioxidant Lotion Containing Water Extract of Green Tea Leaf (Camellia Sinensis L)

The research about lotion dosage formulation that contain green tea leaf water extract with concentration 0.0002%, 0.002%, 0,02%, and 8.6% had been done. The basic of the EC50 value. Which calculated from linierity regression equation between tested solution concentration and scavenging percent. The lotion formulation began with lotion base orientation included organoleptic, homogeinity, pH, and viscosity. It showed that lotion base with glyceryl monostearate 5.5% have the best pH stability, viscosity, and consistency. Evaluation of lotion dosage include physical evaluation (organoleptic, homogeinity, viscosity and stability) and chemical evalution (pH and the stability of lotion antioxidant activity. Antioxidant activity stability evaluation of lotion with 1,1-diphenyl-2-picrylhidrazyl solution using spectrophotometri UV-Visible showed, that formula which contain 0.02% and 8.6% green tea leaf water extract gave the best radical scavenging activity and stable during storage