BIOACTIVE COMPOUNDS IN BENGKOANG (Pachyrhizus erosus) AS ANTIOXIDANT AND TYROSINASE INHIBITING AGENTS

In Indonesia, the roots of bengkoang (Phacyrhizus erosus) have been used as the excipient for sun screening and skin whitening paste. Since the active compounds exhibiting skin whitening or sun screening effect have not previously been studied, the aim of this study was to identify compounds with antioxidant and tyrosinase inhibitor activities. Soxhlet extraction was used as the method of isolation with petroleum ether as the solvent and it was followed by fractionation using ethyl acetate to obtain three isoflavonoids (i.e. daidzein (2); daidzein-7-O-ß-glucopyranose (3); 5-hydroxy-daidzein-7-O-ß-glucopyranose (4)), and a new pterocarpan (i. e. 8,9-furanyl-pterocarpan-3-ol (1)) which antioxidant activities (SC50% values) of 2.11; 11.86; 0.69 and 7.86 respectively. All compounds showed tyrosinase inhibiting activities with IC50 values of 4.38; 5.35; 7.49 and 22.20 mM, respectively for compound 4, 2, 1 and 3. These compounds can be used as antioxidant and skin whitening materials

A New Compound (8,9) -Furanyl-Pterocarpan-3-Ol Used for Standardization of Bengkuang (Pachyrhizus erosus) Extract as Sunscreen and Skin Whitening Agent

Bengkuang (Pachyrhizus erosus) has been traditionally used as sun screening and skin whitening. The active compounds in bengkuang extract already published included their activities in antioxidant and skin whitening. However, standardization of bengkuang extract has not been studied. This research aims to find out the analysis procedure by High Performance Liquid Chromatography to make standardization bengkuang extract.The first step of this research was collecting bengkuang from Prembun, Central Java, Indonesia in dry season. After cleaning and peeling, bengkuang root was sliced, dried and ground to make powder. Then followed by extraction using Soxhlet in petroleum ether and subsequently in methanol. Methanol extract was evaporated and then partitioned with ethyl acetate-water. Ethyl acetate fraction was evaporated and then separated in open column chromatography using silica gel as stationary phase and a gradient mixture of chloroform-ethyl acetate-methanol as mobile phase. Bio guided fraction method was used for separation and purification to get isolated compounds. The isolated compounds obtained from this fractionation were then elucidated and analyzed their activities.A new compound (8,9-furanyl-pterocarpan-ol) has been selected as a biomarker for extract standardization. The optimum of HPLC condition for standardization consisted of a column (Zorbax SB-C18; i.d. 0.46 cm; 5 μm particle size), mobile phase (gradient elution of MeOH-water) with flow rate of 1 ml/min and detector (UV-detector at 293 nm). The obtained LOD value was 0.51 ± 0.02 µg. The potentials of this compound to absorb UV ray, antioxidant and anti-tyrosinase were 4.018 mAU*S/mml; 2.113±0.001mM (SC50); 7.19±0.11 mM (IC50), respectively.Keywords : bengkuang (Pachyrhizus erosus) extract, (8,9)-furanyl-pterocarpan-3-ol, standardization, sunscreen, skin whitenin

Bioaccumulation of poly-aromatic hydrocarbons in plankton, algae and fish in south sea waters in Jogjakarta

As  a  pollutant,  the  presence  of Poly  Aromatic  Hydrocarbons  (PAH)  in  the environment must be always monitored, because of their ability to spread widely through  the  food  chain  and  also  their  carsinogenic  properties.  The  solubility  of PAH  in  water  is  very  low,  therefore  it  is  difficult  to  analyze  their  presence  in water environment by using water as a sample. By utilizing the phenomenon of the  accumulation  of  PAH  in  biolipids,  the  analysis  method  has  been  developedusing  bioindicators.  In  this  research,  the  concentration  of  four  kinds  of  PAH (pyrene,  benzo(a)anthracene,  benzo(k)fluoranthene  and  perylene)  in  several samples  (water,  plankton,  algae  and  fish)  that  collected  from  the  south  sea Jogjakarta  has  been  determined. The bioaccumulation  factor  (BAF)  of  PAH  in each  sample  has  been  calculated  and  the  results  were  from  4498  to  432754; 2552  to  49265;  14156  to  730991,  respectively  for  plankton,  algae  and  fish. Based on the BAF values, plankton can be used as a bioindicator for short term PAH monitoring, while the Upeneus moluccensisfish primarily gills organ can be used as a bioindicator for medium term (months) PAH monitoring.Key  words  :  poly  aromatic  hydrocarbons,  bioaccumulation  factor,  bioindicator,  Upeneus moluccensi

STUDI PEMISAHAN SENYAWA IDDROKARBON

ABSTRAK: Beberapa jenis senyawa hidrokarbon poliaromatik yang mempunyai struktur Mania dengan jumlah cincin aromatilc yang sama ternyata menvliki sifat-sifat yang hampir sama, sehingga sulit untuk dipisahkan Dalam penelitian ini akan dipelajari pemisahan campuran 8 jenis PAH yaitu fluorantena, pirena, benzo(a)antrasena, krisena, benzo(k)fluorantena, benzo(a)-pirena, perilena dan dibenzo(a,h)antrasena Dan kedelapan senyawa tersebut yang mempunyai jumlah cincin sama adalah pirena/krisena dengan empat cincin, perilena / benzo (a) pirena / dibenzo (a,h) antrasena dengan lima cincin. Sampai saat ini, pemisahan senyawa semi volati/ yang terbaik diperoleh apabila cligunakan kromatografi gas kolom kapiler. Agar diperoleh pemisahan yang optimal perlu dilakukan studi pemisahan senyawa PAH tersebut dalam sistem kromatografi gas kolom kapiler. Hasil penelitian menunjukkan bahwa kromatografi gas kolotn kapiler yang dilengkapi detektor FID dan kolom CP-sil 8 CB (25m X 0,23mm) dengan kondisi sebagai berikut : komposiSi gas : nitrogen 28 ml/min, oksigen 230 ml/min, hidrogen 33 ml/min, make-up gas pada 5 skalapemanasan kolom terprogramtemperatur injektor dan detektor 300°C: volume injeksi maksimal 2 aL: temyata memberikan efisiensi pemisahan sebesar 22747 - 167119, sedangkan kualitas pemisahan yang dinyatakan sebagai parameter resolusi harganya berkisar antara 2,782 - 18,418. Batas kepekaan dari kedelapan senyawa yang diuji sekitar 7,07 - 44,00ng. Apabila diaplikasikan pada penetapan PAH dalam contoh sedimen diperoleh harga recovery sebesar 77.38% - 88.92% dan harga koefisien variansi (CV) antara 8.12% - 12.08%. Analisis standard reference material 1647C rnemberikan harga recovery sebesar 95,16% - 102,10%. Dengan demikian metode ini dapat diaplikasikan untuk analisis kandungan PAH dalam contoh lingkungan yang banyak mengandung bahan pembawa seperti contoh sedimen dan tanah. Kata kunci : hidrokarbon poliaromatilc, GC-kapiler, efisiensi pemisaha

Pengaruh Suhu Terhadap Stabilitas Propagermanium dengan Metode High Performance Liquid Chromatography

Propagermanium merupakan senyawa organo germanium sintetik yang memiliki efek farmakologi terutama sebagai obat anti kanker, dan belum diketahui secara pasti profil kecepatan degradasi. Penelitian ini bertujuan untuk mengetahui pengaruh suhu terhadap stabilitas senyawa propagermanium dengan analisis menggunakan metode HPLC. Reaksi degradasi propagermanium dalam pH dapar fosfat 3,6 dilakukan pada suhu 60˚, 70˚, dan 80˚C. Hasil penelitian dengan menggunakan metode grafik diperoleh kinetika reaksi orde pertama. Peningkatan suhu menyebabkan harga tetapan kecepatan degradasi propagermanium relatif meningkat dan harga waktu paruh propagermanium relatif menurun. Suhu berpengaruh pada stabilitas propagermanium. Harga tetapan kecepatan degradasi (kobs) 0,0266 jam-1, t1/2 26,05 jam, t90 3,95 jam, Ea 5896 kal/mol pada suhu 25˚C. Kata kunci : propagermanium, stabilitas, suhu, HPL

CLEANING EFFECTIVITY OF SEVERAL SURFACTANS TO PESTISIDES RESIDUES ON FRESH APEL FRUITS

Intensification efforts in farming to increase productivity must consider the pesticide utilization, especially insecticide and herbicide. Several pestisides which are still used include carbofuran and organochlorine, some of them have lipofilic properties and might harm to human health. Therefore, an effort is required to washing off pesticides from farming products is one of the effort which can be performed. Since pesticides has lipofilic properties, therefore cleaning pesticides with water is not sufficient. Surfactant is required to increase washing off ability of water. Wash off ability of several surfactants circulated on the market i. e. SL, ML and A were investigated. The result showed that the wash off ability values of surfactants to DDT residues on fresh apples were 79.18 %, 75.19 % and 67.49 % for SL, A and ML respectively. The wash off effectiveness of surfactant A, SL and ML to -metrine were 85.29 %, 80.48 % and 64.47 % respectively.Key words: pesticide, cleaning efficiency surfactant, DDT, -metrine

ANALYTICAL METHOD VALIDATION OF BENZENE USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY IN BEVERAGE CONTAINING SODIUM BENZOATE AND ASCORBIC ACID

Abstract: Several countries reported discovering benzene in beverages containing benzoic acidand ascorbic acid. Benzoic acid decarboxylation by ascorbic acid will form benzene. AmericanBeverage Association (ABA) recommended the use of accelerated testing to test benzene inbeverages. High Performance Liquid Chromatography (HPLC) is one of several methods toanalyze benzene in a wide variety of samples, but there is no much information providedregarding the validation of analysis method of benzene. Therefore, developing analysis methodof benzene and its validation becomes a current need. The HPLC system consists of Hitachi L-2130 pump, sample injector with 20 L sample loop, and UV detector L-2420 operating at 205nm. The analytical column is a LiChrosorb Phenomenex RP-18 (250 x 4 mm, 10 m, 100 ?), themobile phase is acetonitrile:aquabidest (60:40 v/v) and pumped at a flow rate of 0,8 mL/min.Benzene separated from the matrix and follows the validation requirements. The developedanalytical method showed that resolution was 8.37, r = 0.995 with LOD and LOQ 6.52 ppb and19.75 ppb, with a precise of ?11% and recovery of 80-110%. Accelerated testing indicated thatbenzene levels increased with increasing of the temperature. Beverages containing 400 mg/mL ofascorbic acid and benzoic acid formed benzene which was detected as 699.38 ppb at 25 oC,799.61 ppb at 40 oC, and 808.94 ppb at 60 oC in 48 hours. In conclusion, the method was fullyvalidated and can be utilized to analyze benzene in beverages with the accelerated testing at 60 oC in 48 hours, so that benefits the producers and consumers in the end.Keywords : benzene, HPLC, validation method, ascorbic acid, benzoic aci

THE OPTIMIZATION OF RP-HPLC CONDITION USING RESPONSE SURFACE METHODOLOGY BOX-BEHNKEN DESIGN FOR SIMULTANEOUS DETERMINATION OF METFORMIN HCL AND GLIMEPIRIDE IN SPIKED PLASMA

Objective: Aim of this study was to develop and validate the RP-HPLC method using Box-Behnken Design (BBD) for simultaneous analysis metformin HCl and glimepiride in spiked plasma. Methods: The chromatographic system was comprised of acetonitrile-phosphate buffer 0.0125 M+Sodium Dodecyl Sulphate (SDS) 1 mmol as a mobile phase and Ascentis® Phenyl C18 (250 x 4.6 mm i.d.; 5 µm) column as a stationary phase with UV detector at 210 nm. Three independent variables included phosphate buffer (%), pH and flow rate were optimized using Box-Behnken Design. The observed responses were retention time, peak area and resolution. Results: The predicted optimum condition of the RP-HPLC system consisted of phosphate buffer solution of 72%, pH at 4.3 and flow rate at 0.8 ml/min. By using this condition, the duration of analysis was more than 18 min, so it was necessary to modify the flow rate to be 1.0 ml/min to get shorter analysis duration. This condition was then applied to analyze metformin and glimepiride in spiked plasma and validated according to the EMA guideline. AUC of interfering components at the IS retention time between 588-1092 mV, the linearity of metformin was 0.9993 and glimepiride was 0.9991, accuracy and precision were between-13.33% until 16.08%, dilution integrity and metformin stability studies were between-4.01% until 11.82%, and for glimepiride stability studies were between-37.48% until-4.76%. Conclusion: Box-Behnken Design can help optimize the HPLC system, and the optimum condition was valid to analyze metformin and glimepiride in spiked plasma by considering the storage time of plasma samples