SOCIALIZATION OF ANALYSIS OF TOTAL TERPENOIDS FROM MANILTOA GRANDIFLORA (A. GRAY) SCHEFF LEAVES USING TLC METHODS

Saputangan leaves contained terpenoids which appeared reddish brown rings when tested using Liebermann Burchard, Salkowski and cerric sulfate. Saputangan leaves were macerated, then partitioned to obtain total terpenoids using a separatory funnel while checking filtrate partition with cerric sulfate. Extracts dissolved with methanol were partitioned with n-Hexane and then partitioned between aquadest and ethyl acetate in a ratio of 1: 1 to obtain 50 g of total terpenoids. TLC analyses was performed on total terpenoids using n-hexane: acetone (80:20 v/v) solvent. The total terpenoids were proved that terpenoids have 11 terpenoids as red stain based on their Rf. Based on the results of tests conducted on participants involved in socialization, it was stated that around 98% of participants had understood terpenoid compounds.

Characterization and Correlation of Aliphatic Hydrocarbons in Oil Polluted Waters of Bonny Local Government Area, Rivers State

Forensic chemical analyses was used to characterize, determine the source and correlate aliphatic hydrocarbons (AHCs) in water samples collected from Bonny L.G.A, Rivers State. AHCs were extracted from the water samples with n-hexane in a separatory funnel, clean-up achieved by column chromatography using n-hexane and analysis by Gas Chromatography Flame Ionization Detector (GC-FID). The GC chromatogram showed well resolved AHC peaks which distributed from nC9 –nC40. The GC chromatograms of AHCs, with a wide range of low molecular weight hydrocarbons, indicate minimal biodegradation of the oil hydrocarbons in the samples. CPI values from 0.68 to 0.78 for BY-A1, BY-A3, BY-A4 and 0.97 for BY-A2 suggest two sources of oil hydrocarbons. Calculated Pr/Ph ratios showed BY-A1 (2.08), BY-A3 (2.00) and BY-A4 (2.20) were derived from mixed marine and terrestrial source deposited in suboxic environment, while BY-A2 (3.70) was mainly terrigenous source in an oxic environment. A cross-plot of Pr/n-C17 and Ph/n-C18 ratios also showed a tight cluster for samples BY-A1, BY-A3 and BY-A4 which were degraded with sample BY-A2 separate and slightly degraded. Results from this study revealed that the spilled oil was derived from mixed organic sources deposited in an oxic environment with different oil input from external sources showing more terrestrial contribution.

Pengaruh Perbedaan Metode Ekstraksi Metabolit Sekunder Streptomyces sp. GMR22 terhadap Toksisitas pada Sel BHK-21

Streptomyces sp. GMR22 is local isolate from Wanagama 1 Forest in Yogyakarta. They have the potential to be developed to produce active compounds because have PKS and NRPS genes.The active compounds from isolation are strongly influenced by various factors, one of them is extraction techniques. Effect difference of extraction technique will be affected by the quality of secondary metabolites produced.The purpose of this study was to compare the cytotoxicity effects of secondary metabolites of Streptomyces sp. GMR22 which have extracted with different stages from previous studies. The extraction technique was carried out by multilevel separatory funnel extraction methods, which was first extracted using non-polar solvent (n-hexane) and then extracted using semi-polar solvent (ethyl acetate). This research is important because in previous studies (separatory funnel only extracted using ethyl acetate) with the use of the lowest concentration in the dengue virus antiviral test (further test) caused 100% of deaths in BHK-21 cells.This study indicate that multilevel extraction result in lower CC50 value than previous studies. There are 49.160 µl/ml (n-hexane extract) and 284.56 µl/ml (ethyl acetate extract) while water extract is 464,38 µl/ml. FTIR compound analysis show that the three extracts produced have different spectrum patterns, especially in the n-hexane and ethyl acetate extract. Value of CC50 is not too high, it is expected that the secondary metabolites contained in the extracts can be used for further analysis such as antiviral testing because it is safe for normal host cells such as BHK-21 cells

NEW UNIVERSAL METHOD FOR DETERMINATION OF ANTICOAGULANTS IN RODENTICIDES

Solid paraffin briquettes are one of the most modern types of rodenticide baits, because they protect it from the environmental exposure and do not reduce palatability of rodents. This paper proposes a simple universal method for the determination of brodifacoum, bromadiolone and difenacoum in solid briquettes, which are a mixture of paraffin, poison and filler, which can be used as grain, flour and the like. The difficulty in the analysis of baits containing paraffin, is to get rid of it. For this, we propose to use hexane-acetonitrile extraction in the system. Paraffin dissolves in hexane, while brodifacoum, bromadiolone and difenacoum do not. The resulting two-phase system was separated using a separatory funnel and the bottom fraction was collected. If any fillers were present in the sample, they were filtered until the mixture was separated, the residue was washed with acetonitrile. Before introducing the sample into the chromatograph, it was filtered twice with PTFE filters with a porosity of 0.45 µm. The best separation was achieved using a Thermo Acclaim® Surfactant column using acetonitrile and 0.1 M aqueous ammonium acetate solution (pH 5.4) as the mobile phase in a gradient elution mode. All three poisons were detected at a wavelength of 264 nm. This method has a sensitivity of 0.028 mg. The linearity range is from 0.00067 to 0.01%. The recovery rate for bromadiolone is 94%, for brodifacoum, 98%, and for difenacoum, 90%.

Jenis Pelarut Metanol Dan N-Heksana Terhadap Aktivitas Antioksidan Ekstrak Rumput Laut Gelidium sp. Dari Pantai Drini Gunungkidul – Yogyakarta

Type of Solvent Methanol And N-Hexane Against Antioxidant Activity Seaweed Extract Gelidium sp. From Drini Beach Gunungkidul – Yogyakarta Gelidium sp. the one of red seaweed which has potential as naturalantioxidant. The researh was to know the activity of methanol extract, n-hexane Gelidium sp., and determined totalphenolic compound of pigments (chlorophyll a and carotenoid). Decriptiveexplorative method was used in this research and it was taken from Drini Beach, Gunungkidul, Yogyakarta. Maseration to this sample was done by methanol as solvent, evaporated by rotary evaporator and partitied by n-hexane solvent with separatoryfunnel. Antioxidant activity was determined by transfer electron method using DPPH (1,1-diphenyl-2-picrylhidrazyl) as free radicals. Totalphenolic compound being tested using Folin-Ciocalteu solution with gallic acid as standard and measured at a wavelength of 725 nm, while the chlorophylls were spectrophotometry method and measured at a wavelength of 663 nm and 646 nm as well as carotenoidswere measured at a wavelength of 470 nm. The results showed that IC50 value of methanol extract was 340,10 ppm and n-hexane was 66,25 ppm. IC50 value of methanol extract was categorized as very weak in antioxidant activity, while n-heksan extract was categorized as strong. Total phenolic content in each extract were 46,55 and 135,62 (mg GAE/g extract), chlorophyll a 8,47 and 10,88 (mg/g extract sample) and carotenoids 50,38 and 84,27 (μmol/g extract sample). Keywords: Gelidium sp., Antioxidant, DPPH Abstrak Gelidium sp. merupakan salah satu rumput laut merah yang berpotensi sebagai sumber antioksidan alami. Penelitian ini bertujuan untuk mengetahui aktivitas antioksidan ekstrak metanol dan n-heksan Gelidium sp. (segar), menentukan kadar total fenolat dan pigmen (klorofil a dan karotenoid). Metode deskriptif eksploratif digunakan dalam penelitian ini dan diperoleh diambil dari Pantai Drini, Gunungkidul, Yogyakarta dibersihkan dengan air tawar, dimaserasi dengan pelarut metanol, diuapkan dengan rotary evaporator dan dipartisi dengan pelarut n-heksan menggunakan corong pemisah (separatory funnel). Aktivitas antioksidan ditentukan dengan metode transfer elektron menggunakan DPPH (1,1-diphenyl-2-picrylhidrazyl) sebagai radikal bebas. Kadar total fenolat diuji dengan metode Folin-Ciocalteu dengan asam galat sebagai larutan standar dan diukur pada panjang gelombang 725 nm. Kadar klorofil a diukur dengan metode spektrofotometri pada panjang gelombang 663 nm dan 646 nm sedangkan kadar karotenoid diukur pada panjang gelombang 470 nm. Hasil penelitian menunjukkan nilai IC50 ekstrak metanol sebesar 340,10 ppm dan ekstrak n-heksan 66,25 ppm. Nilai IC50ekstrak metanol termasuk kategori aktivitas antioksidan sangat lemah sedangkan n-heksana termasuk kategori kuat. Kadar total fenolat pada masing-masing ekstrak 46,55 dan 135,62 (mg GAE/g ekstrak), kadar klorofil a sebesar 8,47 dan 10,88 (mg/g sampel) dan kadar karotenoid sebesar 50,38 dan 84,27 (µ mol/g sampel).Kata kunci: Gelidium sp., Antioksidan, DPPH

Effect of Salinity on the Effectiveness of Solidifiers for Crude Oil Spill Remediation

ABSTRACT A laboratory investigation was conducted to test the effectiveness of solidifiers with fresh water and artificial seawater using Prudhoe Bay Crude oil. Experiments were designed to study the effects of salinity, solidifier type, solidifier-to-oil mass ratio (SOR), mixing energy and beaker size using five solidifiers. The U.S. Environmental Protection Agency is developing a protocol for testing the effectiveness of solidifiers in a laboratory setting. This involves measuring the amount of free oil remaining in the water after the solidified product is removed using an ultraviolet–visible spectrophotometer. For these experiments, 0.25 mL of oil was added to salinized beaker containing 80 mL of water. Milli-Q water and sterile GP2 seawater were used as the exposure media. The mass of the solidifier was changed depending on the SOR. Each of the solidifier was added to a slick of crude oil on water. After stirring the mixture for 30 minutes, the solidifier was removed. The water with the remaining oil was transferred from the beaker to 250 mL separatory funnel. The solution in the funnel was extracted three times with 20 mL of dichloromethane and the final volume adjusted to 60 mL. The extracted samples were analyzed for oil content with an Agilent 8452 ultraviolet–visible spectrophotometer. All experiments were carried out in triplicate. An analysis of variance (ANOVA) was performed on the data collected, which helped quantify the main and interactive effects of the variables. Salinity of the water was mostly found to be an insignificant factor. Results indicated that SOR and solidifier type are the most important variables affecting removal efficiency.

Complete Validation of a Unique Digestion Assay To Detect Trichinella Larvae in Horse Meat Demonstrates the Reliability of this Assay for Meeting Food Safety and Trade Requirements

A tissue digestion assay using a double separatory funnel procedure for the detection of Trichinella larvae in horse meat was validated for application in food safety programs and trade. The assay consisted of a pepsin-HCl digestion step to release larvae from muscle tissue and two sequential sedimentation steps in separatory funnels to recover and concentrate larvae for detection with a stereomicroscope. With defined critical control points, the assay was conducted within a quality assurance system compliant with International Organization for Standardization–International Electrotechnical Commission (ISO/IEC) 17025 guidelines. Samples used in the validation were obtained from horses experimentally infected with Trichinella spiralis to obtain a range of muscle larvae densities. One-, 5-, and 10-g samples of infected tissue were combined with 99, 95, and 90 g, respectively, of known negative horse tissue to create a 100-g sample for testing. Samples of 5 and 10 g were more likely to be positive than were 1-g samples when larval densities were less than three larvae per gram (lpg). This difference is important because ingested meat with 1 lpg is considered the threshold for clinical disease in humans. Using a 5-g sample size, all samples containing 1.3 to 2 lpg were detected, and 60 to 100% of samples with infected horse meat containing 0.1 to 0.7 lpg were detected. In this study, the double separatory funnel digestion assay was efficient and reliable for its intended use in food safety and trade. This procedure is the only digestion assay for Trichinella in horse meat that has been validated as consistent and effective at critical levels of sensitivity.

USE OF RESIDUAL BIODIESEL FROM RAW GLYCERIN FROM COMMERCIAL BIODIESEL PRODUCTION

The purpose of this work was the removal of byproducts such as free fatty acids and glycerides present in the crude glycerol. The free fatty acids and the glycerides were separated chemically by hydrolysis with phosphoric acid and mechanically with a separatory funnel. The products separated from the crude glycerin were esterified with an alcoholic solution of dimethyl sulfate and subsequently transesterified with a alcoholic solution of potassium hydroxide to obtain the biodiesel (monoalkyl ester). The biodiesel was purified with Montmorilonite clay. It was analyzed by the acid value test and by solubility in methanol. The volume percent of biodiesel varied between 60% and 80% of the original crude glycerol. Finally the biodiesel (4 liters) was tested as a fuel in a pickup truck.

A Validated Trichinella Digestion Assay and an Associated Sampling and Quality Assurance System for Use in Testing Pork and Horse Meat

A revised digestion method, developed for efficiency and quality assurance, was validated for the detection of Trichinella larvae in pork and horse meat to meet requirements for food safety testing and facilitate access to international markets. The method consisted of a tissue homogenization step and a spin bar digestion procedure conducted at 45°C to free larvae from muscle tissue, followed by two sequential separatory funnel steps to concentrate the larvae for detection using a stereomicro-scope. Critical control points were determined for the method and monitored during testing. Under conditions of a defined protocol, test capacity was suitable for industrial applications, since multiples of up to 100 g of tissue could be analyzed at one time. The overall sensitivity of the test system depended on the size and origin of the sample taken from individual infected carcasses. Data from swine indicated that the currently accepted sample size of 1 g from individual carcasses consistently detected larval loads of ≥3 larvae per gram. Larval loads of 1.0 to 1.9 larvae per gram required 3- to 5-g samples of muscle tissue for reliable detection. Five-gram samples were considered optimal, because they consistently detected more tissues than 3-g samples, although the difference was not statistically significant. Tissue localization studies in experimental pigs indicated that the tongue and diaphragm were the tissues of choice for the most sensitive larval recovery. A system of analyst training, laboratory certification based on ISO guide 25, and on-site proficiency panel testing was used to ensure that external laboratories would consistently produce reliable test results. The system developed for pork was successfully modified for the testing of horse meat.