Cytotoxic Activity Against L1210 Leukemia Cells from the Ethyl Acetate Fraction of Kenikir Leaves (Cosmos. Caudatus) Preserved by Gamma Irradiation

Kenikir leaves (Cosmos caudatus) Has been used as traditional medicine, especially as an anti-cancer, this plant has been in Indonesia both in herbs and capsules. Microbial contamination of herbal medicinal raw materials occurs when the storage process is done. One of the preservation techniques used in the industry is using gamma irradiation techniques to reduce microbial and fungal contamination. The purpose of this research was to study the effect of gamma irradiation for the preservation of kenikir leaves (C. caudatus) as an anti-cancer based on cytotoxic activity using L1210 leukaemia cells. The simplicia was gamma-irradiated by Co-60 source with variation doses of 0 (control); 5; 7.5; 10; and 15 kGy. Then the irradiated and control samples were macerated successively using n-hexane, ethyl acetate, and ethanol. The active extract (ethyl acetate) was further fractionated using column chromatography, obtained seven fractions (F1 - F7). The seven fractions' cytotoxic activity against L1210 leukaemia cells showed that fraction 3 (F3) was the most active fraction with an IC50 of 1.26 µg/mL. Each dose's F3 cytotoxic activity showed that the IC50 7.5 kGy irradiation sample did not change significantly with control (0 kGy) based on ANOVA analysis using SPSS 24 with a 95% confidence level. In comparison, irradiation samples of 10 and 15 kGy showed a change in the IC50 value is significant with the control (0 kGy). These results indicate that gamma irradiation can be used as an alternative for preserving C. caudatus with a maximum dose of 7.5 kGy, so that its anti-cancer properties do not change with those without irradiation

The Effect of Gamma Irradiation on Ethanolic Extract of Temulawak (Curcuma xanthorrhiza Roxb.) Against Human Cancer Cell Lines

The Effect of Gamma Irradiation on Ethanolic Extract of Temulawak (Curcuma xanthorrhiza Roxb.) Against Human Cancer Cell Lines. One of the medicinal plants that are widely used by the people of Indonesia is temulawak (Curcuma xanthorrhiza Roxb.) with the active ingredient is curcuminoid. The curcuminoid is a yellow compound that is believed to have anti-cancer properties, therefore its properties must be maintained in the post-harvest process. This research aimed to study the effect of gamma irradiation on the simplicia of C. xanthorrhiza rhizomes in inhibiting the proliferative activity against human cancer cell lines. The dry powder of C. xanthorrhiza rhizome was irradiated with gamma at a dose of 10 kGy in 2 replications, then extracted using ethanol solvent, evaporated, to obtain an ethanol extract. As a control, an un-irradiated sample was prepared and extracted in the same manner. Each ethanol extract from irradiated and control samples was then tested for their antioxidant activity by DPPH method and antiproliferative activity against human cancer cell lines (HUT78, A549, HeLa, and THP1). The results showed that the ethanol extract of irradiated C. xanthorrhiza rhizome with a dose of 10 kGy still had antioxidant activity and anticancer properties based on the bioassay against four those human cancer cell lines, although the antiproliferative activity decreased between 8-30% compared to the control sample. The highest antiproliferative activity was shown by the ethanol extract against HUT78 cancer cell line with IC50 values of 5.4 and 10.7 µg/ml for control and irradiated samples respectively. Based on the silica gel TLC plate, the ethanolic extracts of C. xanthorrhiza rhizome both unirradiated- and irradiated samples contained curcumin (Rf = 0.58) and demethoxycurcumin (Rf = 0.38)

Gamma Irradiation for Preservation of Suruhan Herbs (Peperomia pellucida L. Kunth.) and its Bioactivity Against L1210 Leukemia Cells

Peperomia pellucida includes the Piperaceae family that has anti-cancer, anti-inflammatory, antioxidant, and anti-microbial activities. Microbes easily contaminate dry herbs during storage, so special handling is needed. One of the preservation techniques is using irradiation technology. This technique can be used to extend the shelf life of herbal medicinal ingredients. This research aimed to study the effects of irradiation on the anti-cancer activity of P. pellucida leaves. P. pellucida leaves' dried powder was irradiated at doses of 5, 7.5, 10, and 15 kGy. The anti-cancer activity test was carried out against L1210 leukaemia cells. The un-irradiated irradiated samples were macerated successively with n-hexane, ethyl acetate, and ethanol, then be concentrated. The ethyl acetate extract was fractionated using silica gel column chromatography, obtained seven fractions. Their IC50 values < 20 mg/mL, with fraction F4 was the most active fraction (IC50 1.9 mg/mL). The result showed that gamma radiation at doses of 5 - 15 kGy reduced their cytotoxic activity significantly. However, the fractions were still in the active category as anti-cancer (IC50 values were < 20 mg/mL)

ORTHO-CHLOROANILINE DEGRADATION UNDER THE INFLUENCE OF OZONE IN AQUEOUS SOLUTION

The degradation study on 2-chloroaniline (2-ClA) used as a model of water pollutant by ozonation was performed. The change of spectrum and substrate concentration of treated solutions were measured by spectrophotometer as well as by HPLC method. The change of solutions pH after ozonization was observed. The yields of the products formed (chloride ions, ammonium ions, and formaldehyde) were investigated as a function of ozonation time. The results showed that the ozonation gave to an efficient degradation of 2-ClA.   Keywords: degradation, ortho-chloroaniline, ozon

IN VITRO BIOACTIVITYTEST OF IRRADIATEDMAHKOTA DEWA BARK (Phaleria macrocarpa (Scheff.) Boerl.) AGAINST HUMANCANCER CELL LINES

Gamma i"adiation has been used to preserve an herbal medicine, but it has not been known the effects of gamma irradiation on their bioactivity as an anticancer agent yet. In the previous study, the gamma irradiation on mahkota dewa bark with the optimum dose of 7.5 kGy could be used for decontamination of bacteria and fungus/yeast. In this report, the effect of gamma i"adiation with the dose of 7.5 kGy on the bioactivities of mahkota dewa (Phaleria macrocarpa (Scheff) Boer/.) bark against leukemia L1210 cel/s was studied. The control and i"adiated samples were successively macerated with n-hexane and ethyl acetate. In the previous results, silica gel column chromatography of ethyl acetate extract of non irradiated sample (control) gave 8 fractions. Among these fractions, fraction 6 indicated the most cytotoxic-potentialfraction, so that in this experiment,the ethyl acetate extract of irradiated and non irradiated sample were fractionated with the same manner as previous fractionation. The fraction 6 obtained both from control and i"adiated samples were then assayed their inhibitory activities against 4 kinds of human cancer lines, i.e. HeLa, THP-1, HUT-78 and A-549. The results showed that the fraction 6 from control sample gave IC50values of 3.65, 5.59, 3.55, and 4.06 llg/mL, against HeLa, THP-1, HUT-78 and A-549, respectively, meanwhile fraction 6 from irradiated sample gave IC50values of 8.26, 7.02, 5.03, and 5.59 llg/mL, respectively. Gamma i"adiation dose of 7.5 kGy on mahkota dewa bark could decreased the cytotoxic activity of fraction 6 as the most cytotoxic-potential fraction against HeLa, THP-1, HUT-78 and A-549 cancer cel/lines, but decreasing the cytotoxic activity has not exceeded the limit of an extract and the fraction declared inactive. So that the irradiation dose of 7.5 kGy can be use for decontaminationof bacteria and fungus/yeast without eliminating the cytotoxic activity

Toksisitas Akut Ekstrak Etanol Temulawak (Curcuma xanthorrizha Roxb.) Iradiasi yang Mempunyai Aktivitas Antikanker

Pasteurisasi simplisia dan produk obat herbal telah dilakukan oleh beberapa perusahaan obat herbal, namun informasi tentang keamanan obat herbal yang diradiasi masih sedikit, bahkan pengaruh iradiasi gamma untuk tujuan pasteurisasi terhadap toksisitas simplisia temulawak belum pernah diteliti. Ekstrak etanol temulawak mempunyai aktivitas berpotensi sebagai antikanker. Pada penelitian ini dilakukan uji toksisitas akut ekstrak etanol dari rimpang temulawak yang tidak dan yang diradiasi dengan dosis 5 dan 10 kGy. Uji toksisitas akut ekstrak etanol dilakukan pada mencit dengan mengamati pengaruh ekstrak terhadap perilaku hewan (profil farmakologi) setelah pemberian dosis tunggal bahan uji, perkembangan bobot badan hewan percobaan dan kematian setiap hari selama 14 hari serta pengamatan bobot beberapa organ pada hari ke-14.Hasil uji toksisitas akut setelah pemberian ekstrak pada mencit jantan dan betinamenunjukkan bahwa sampai dosis 7500 mg/kg bobot badan (BB) tidak ada kematian dan efektoksik yang bermakna, maka ekstrak etanol dari rimpang temulawak yang tidak dan yangdiradiasi dengan dosis 5 dan 10 kGy dapat dinyatakan aman. Dengan demikian DL50 dariekstrak etanol dari rimpang temulawak yang tidak dan yang diradiasi (5 dan 10 kGy) padamencit lebih besar dari 7500 mg/kg BB

EFFECT OF GAMMA IRRADIATION ON PHYTOCHEMICAL CONTENT AND ANTICANCER ACTIVITIES OF ROSELLE (Hibiscus sabdariffa Linn)

Gamma irradiation is widely used in herbal medicine industries as an efficient preservative method in reducing microorganism contaminants. The aim of this study was to evaluate the effect of gamma irradiation at the doses of 5; 7.5; and 10 kGy on H. sabdariffa ethanolic extract (HS-EE). The Co-60 was used for irradiation the samples. The phytochemical content of HS-EE was carried out by total microorganism analysis using dilution method, TPC by Follin-Cicalteu method, TFC by aluminium chloride colorimetric method, antioxidant activity using DPPH method, TLC profiling on silica gel F254, in vitro anticancer activity using A-549, HUT-78, and MCF-7 cancer cell lines. The irradiation at 10 kGy caused the total bacteria decreased, while dose of 5 kGy could eliminate the total mold. Irradiation at 5 kGy caused TPC, TFC, and antioxidant activity decreased by 5-11%. TLC chromatograms analysis confirmed that one of the compounds contained in HS-EE was quercetin. The HS-EE has the strongest anticancer property against HUT-78 (IC50 10.51 µg/mL) followed by against MCF-7 (IC50 13.39 µg/mL), and A-549 (IC50 14.19 µg/mL). It can be concluded that irradiation at a dose of 10 kGy could remove total bacteria and molds, decreasing the phytochemical content and anticancer activities of HS-EE. It is recommended to increase the active ingredient level in the formulation

IRRADIATION EFFECTS ON INSECTICIDES AS A POLLUTANT MODEL IN AQUEOUS SOLUTION

Degradation of some insecticides i.e.: fenitrothion, prothiofos, and cypermetrin as a model pollutant was carried out using gamma irradiation of 60Co. Irradiation-induced of fenitrothion in solution acid medium gave the significant effect on the decrease of its concentration. The optimum condition for degradation of fenitrothion (55.5 mg/L) was irradiation at 6 kGy with aeration in initial pH 5.6. At this condition, the concentration of fenitrothion decreased up to 97%. Determination of organic acids in irradiated product by HPLC method showed that oxalic acid and formic acid were clarified as organic acid-degraded products of fenitrothion with the concentration of 23.0 mg/L and 2.5 mg/L respectively. The irradiation of prothiofos in aqueous solution (50 mg/L), showed that irradiation at a dose of 8 kGy and initial pH 7 gave the optimum degradation. At this condition, the concentration of prothiofos decreased up to 98%. Determination of irradiated product by HPLC method showed that oxalic acid (18 mg/L) was clarified as organic acid-degraded product of prothiofos. In the case of cypermetrin, the optimum condition for its degradation was irradiation with aeration of cypermetrin (40 mg/L) at a dose of 20 kGy, initial pH 11, and adding the catalyst 0.05% of FeCl3. At that condition, cypermetrin decreased up to 87% and COD of solution decreased up to 78%. The organic acid detected at optimum condition as degradation products were oxalic acid (1.1 g/L), maleic acid (9.0 g/L), formic acid (127.0 g/L), and acetic acid (286.0 g/L). From these results, it could be concluded that ionizing radiation can be used as a tool to degrade insecticides as organic pollutants

Aktivitas Sitotoksik Ekstrak Etil Asetat Daun Jati Belanda (Guazuma ulmifolia Lamk.) Terhadap Sel Kanker Leukemia L1210

Kanker merupakan penyakit yang menyebabkan tingkat kematian paling tinggi di dunia. Sel kanker merupakan sel yang pertumbuhannya tidak terkontrol dan berkembang di dalam jaringan sel tubuh. Kandungan flavonoid dan sifat antioksidan yang tinggi dari daun jati belanda (Guazuma ulmifolia Lamk) dapat digunakan sebagai pengobatan terapi kanker. Penelitian ini dilakukan untuk menentukan aktivitas sitotoksik  G. ulmifolia Lamk terhadap sel leukemia L1210. Daun G. ulmifolia kering dimaserasi secara bertahap menggunakan pelarut n-heksana, etil asetat, dan etanol. Masing-masing ekstrak yang diperoleh dievaluasi aktivitas sitotoksisitasnya terhadap sel leukemia L1210. Ekstrak paling aktif difraksinasi menggunakan kromatografi kolom dengan berbagai pelarut yang sesuai. Selanjutnya semua subfraksi yang diperoleh diuji aktivitas sitotoksiknya dan dianalisis dengan kromatografi cair kinerja tinggi (KCKT) untuk mengetahui bentuk kromatogranya. Hasil penelitian menunjukkan bahwa ekstrak etil setat memiliki nilai IC50 paling kecil diantara ekstrak yang lain, yaitu sebesar 6,23 µg/mL. Fraksinasi ekstrak etil asetat diperoleh (1-8) fraksi dengan fraksi 4 memiliki aktivitas paling tinggi yang  berpotensi sebagai anti kanker dengan nilai IC50 sebesar 2,67 µg/mL. Analisis fraksi 4 etil asetat G ulmifolia menggunakan KCKT menunjukkan bahwa kandungan utama adalah flavonoid

Toksisitas Akut Ekstrak Etanol dari Simplisia Daun Sirsak (Annona muricata L.) yang Diawetkan dengan Iradiasi Gamma

Daun sirsak (Annona muricata L.) banyak digunakan sebagai obat tradisional dan suplemen oleh masyarakat Indonesia. Daun sirsak mengandung banyak metabolit sekunder yang berkhasiat sebagai antioksidan, antiinflamasi, anti bakteri dan antikanker.  Penanganan simplisia maupun sediaan obat herbal daun sirsak sangat penting agar terhindar dari kontaminasi jamur dan bakteri. Teknik iradiasi gamma dapat digunakan untuk memperpanjang masa simpan simplisia/sediaan obat herbal. Pada penelitian ini dilakukan iradiasi daun sirsak kering pada dosis 7,5 kGy dan pengujian toksisitas akut pada mencit untuk mengetahui efek toksisitas akut sediaan uji ekstrak etanol daun sirsak (Annona muricata L.) yang tidak diradiasi (0 kGy) dan yang diradiasi (7,5 kGy) pada laju dosis 7,0 kGy/jam. Pengujian toksisitas akut oral dilakukan sesuai prosedur menurut WHO dan OECD guideline 423 (Acute toxic class method) menggunakan mencit galur ddY jantan dan betina, umur 2-3 bulan, bobot badan antara 20-30 g. Ekstrak etanol daun sirsak merupakan bahan natural, sehingga starting dosis dimulai pada uji limit dosis 2000 mg/kg BB. Pengamatan dilakukan terhadap gejala-gejala toksik, kematian hewan uji, perubahan kenaikan berat badan (Average Daily Gain: ADG) dan manifestasi efek toksik sampai hari ke-14. Pada akhir uji, mencit dikorbankan dan diambil organ-organ vital dan dievaluasi adanya kelainan secara gross patologi (makroskopi) dan histopatologi. Hasil uji toksisitas akut menunjukkan bahwa pemberian sediaan uji ekstrak etanol daun sirsak yang tidak diradiasi (0 kGy) dan yang diradiasi (7,5 kGy) pada dosis ekstrak 2000 mg/kg BB dan 5000 mg/kg BB tidak menunjukkan gejala-gejala toksik maupun kematian mencit jantan dan betina. Pada pemeriksaan organ-organ vital secara gross patologi dan histopatologis juga tidak ditemukan adanya perubahan yang berarti pada organ organ vital hewan coba. Nilai LD50 sediaan uji ekstrak etanol daun sirsak adalah > 5000 mg/kg BB pada mencit, termasuk kategori 5 (tidak terklasifikasi= unclassified) atau minimal praktis tidak toksik