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)

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

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

Radiation-Induced Degradation of Pirimiphos Methyl in Aerated Solution

Degradation of pirimiphos methyl (1) as an active ingredient in Minawet insecticide 250 EC formulation in aqueous solution was studied. The absorbance, pH, COD (Chemical Oxygen Demand) in aerated solution, and the analyses of degradation products at various irradiation doses with dose rate of 5 kGy/h were observed. The absorbance decreased rapidly at low doses (£ 10 kGy), while at high doses (> 10 kGy) decreased slowly. The optimum irradiation dose for pirimiphos methyl degradation in aerated solution was found to be 15 kGy at pH 3.6. At that condition, more than 99% of pirimiphos methyl has been degraded and the COD of solution decreased about 82%. The analysis of irradiated samples by GC-MS and HPLC showed that 2-diethylamino-6-methyl-4-oxo-3,4-dihydropyrimidine (3) and oxalic acid were clarified as degraded product

Pengaruh Iradiasi Gamma Pada Aktivitas Sitotoksik Daging Buah Mahkota Dewa (Phaleria Macrocarpa (Scheff) Boerl.)

Iradiasi gamma telah digunakanoleh industri obat herbal untuk pengawetan simplisia tanaman obat, tetapi pengaruh iradiasiterhadap khasiatnya belum diteliti. Tujuan penelitian adalah memperoleh dosis iradiasi optimaluntuk pengawetan simplisia daging buah mahkota dewa tanpa merusak khasiatnya. Telahdilakukan iradiasi gamma terhadap simplisia daging buah kering mahkota dewa pada variasidosis 0; 5; 7,5 ; 10; 15; 20 kGy. Cemaran mikroba diuji dengan metode yang mengacu padaSNI, yang menunjukkan bahwa dosis 5 kGy telah dapat membunuh seluruh mikroba. Masingmasingsampel dimaserasi dengan etanol, lalu ekstrak yang diperoleh difraksinasi dengankromatografi kolom, diperoleh 8 fraksi. Uji sitotoksisitas fraksi-fraksi terhadap sel leukemiaL1210 menunjukkan bahwa Fr.3 merupakan fraksi paling sitotoksik. Untuk menentukan dosisiradiasi optimal dalam menghambat pertumbuhan serta membunuh semua bakteri dankapang/khamir pada simplisia daging buah mahkota dewa tanpa menurunkan aktivitassitotoksik, dilakukan analisis kromatografi lapis tipis (KLT) dan kromatografi cair kinerja tinggi(KCKT) terhadap Fr.3. Hasil penelitian menunjukkan bahwa iradiasi dengan dosis > 5 kGy padasimplisia daging buah mahkota dewa dapat menghambat pertumbuhan dan membunuh semuabakteri serta kapang khamir yang ada tanpa menurunkan aktivitas sitotoksik ekstrak etanolsecara nyata terhadap sel leukemia L1210. Penurunan aktivitas sitotoksik ekstrak etanolterhadap sel leukemia L1210 secara nyata terjadi setelah iradiasi pada dosis > 10 kGy. Padadosis 10 kGy, aktivitas sitotoksik sudah terlihat menurun meskipun belum melampaui batassuatu fraksi dinyatakan tidak aktif dan hasil analisis profil kromatogram KLT menunjukkanbahwa Fr. 3 sedikitnya mengandung 10 komponen. Iradiasi sampai dengan dosis 20 kGymengakibatkan intensitas salah satu puncak mayor menurun, dan penurunannya sebandingdengan besarnya dosis. Dosis 5 sampai 10 kGy merupakan dosis optimum untuk tujuanpengawetan tanpa merusak aktivitas sitotoksiknya

PENGARUH IRADIASI GAMMA PADA AKTIVITAS SITOTOKSIK DAGING BUAH MAHKOTA DEWA (Phaleria macrocarpa (Scheff) Boerl.)

Iradiasi gamma telah digunakanoleh industri obat herbal untuk pengawetan simplisia tanaman obat, tetapi pengaruh iradiasiterhadap khasiatnya belum diteliti. Tujuan penelitian adalah memperoleh dosis iradiasi optimaluntuk pengawetan simplisia daging buah mahkota dewa tanpa merusak khasiatnya. Telahdilakukan iradiasi gamma terhadap simplisia daging buah kering mahkota dewa pada variasidosis 0; 5; 7,5 ; 10; 15; 20 kGy. Cemaran mikroba diuji dengan metode yang mengacu padaSNI, yang menunjukkan bahwa dosis 5 kGy telah dapat membunuh seluruh mikroba. Masingmasingsampel dimaserasi dengan etanol, lalu ekstrak yang diperoleh difraksinasi dengankromatografi kolom, diperoleh 8 fraksi. Uji sitotoksisitas fraksi-fraksi terhadap sel leukemiaL1210 menunjukkan bahwa Fr.3 merupakan fraksi paling sitotoksik. Untuk menentukan dosisiradiasi optimal dalam menghambat pertumbuhan serta membunuh semua bakteri dankapang/khamir pada simplisia daging buah mahkota dewa tanpa menurunkan aktivitassitotoksik, dilakukan analisis kromatografi lapis tipis (KLT) dan kromatografi cair kinerja tinggi(KCKT) terhadap Fr.3. Hasil penelitian menunjukkan bahwa iradiasi dengan dosis > 5 kGy padasimplisia daging buah mahkota dewa dapat menghambat pertumbuhan dan membunuh semuabakteri serta kapang khamir yang ada tanpa menurunkan aktivitas sitotoksik ekstrak etanolsecara nyata terhadap sel leukemia L1210. Penurunan aktivitas sitotoksik ekstrak etanolterhadap sel leukemia L1210 secara nyata terjadi setelah iradiasi pada dosis > 10 kGy. Padadosis 10 kGy, aktivitas sitotoksik sudah terlihat menurun meskipun belum melampaui batassuatu fraksi dinyatakan tidak aktif dan hasil analisis profil kromatogram KLT menunjukkanbahwa Fr. 3 sedikitnya mengandung 10 komponen. Iradiasi sampai dengan dosis 20 kGymengakibatkan intensitas salah satu puncak mayor menurun, dan penurunannya sebandingdengan besarnya dosis. Dosis 5 sampai 10 kGy merupakan dosis optimum untuk tujuanpengawetan tanpa merusak aktivitas sitotoksiknya

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