In 10 g dried sediment sample added 7 ml 0.2 M NH4Cl solution. A mixture of 100 ml hexane: acetone (1:1) was used as a solvent to extract pesticides with overnight shaking for 12 h on reciprocal or wrist action shaker at 180 rpm. The extract was carefully decanted through activated florisil column (2-3 cm), giving twice wash with25 ml hexane: acetone (1:1) to the sediments. The elute was then washed with 200 ml water and then again aqueous layer was extracted with 50 ml hexane. Finally the hexane layer was washed with 100 ml water and then evaporated to dryness with a vacuum rotary evaporator.
In the round-bottom flask (100 mL), we placed p-aminobenzoic acid (1.2 g) and ethanol (12 mL). We swirled the mixture until the solid dissolved completely. We used Pasteur pipet to add concentrated sulfuric acid (1.0 mL) to the flask. We added boiling stone and assembled the reflux. Then, we did reflux for 75 minutes.
4- Set up reflux system using a clean and dry condenser . 5- Place the flask on the hot plate and heat the reaction for 45 minutes - 1 hour . 6- When the reflux is over , remove magnetic stirrer and allow the reaction to cool to room temperature . 7- Add 20 ml of ice water to a separating funnel
Yellow FeCl3 5% solution is added to the nanoparticle solution with 5% ratio of FeCl3 solution: nanoparticle solution (3 drops: 1 ml). The mixture is then stirred slowly for 30 minutes. The colored nanoparticles solution is then observed with the light microscope (Advanced, Indonesia) using OPTILAB application. Total essential oil Total essential oils analysis method is conducted based on Ali  with modification. 0,1 gr of kaffir lime oil nanocapsule is weighed and diluted to 100 ml using aquadest, taken as much as 1 ml (100x dilution) to put in the reaction tube then 1ml of saturated NaCO3 solution is added to the test tube and incubated for 10 min at room temperature.
Apparatus- Chromatography column: C18 (10 microns particle size), with Guard column Flow rate: 1.2ml/min Pressure: 30-40kgf Wavelength: 326nm Mobile phase: methanol : water (95:5 v/v) Internal standard: retinyl acetate Injection volume: 20µl Procedure for Retinol extraction from serum samples- 1) 100 µl of serum sample and 100 µl of Retinyl acetate were added into 12 X 100mm glass test tubes. Vortex-mixed for 30 seconds. Then, kept them at 4 C for 5 mins. 2) 1mL of hexane was added and vortex-mixed intermittently for 60 sec. 3) Centrifuged at 2500 rpm for 12 mins.
Then, the test media is then incubated at 37 ° C, for 18-24 hours. Rinsing reusable instruments The samples were rinsed with 40 ml of pyrogen-free water using a glass beaker that is free from pyrogens. Endotoxin testing using STV A total of 0,2mL from the water obtained from the rinsing was placed in the STV containing LAL reagent and was shaken for 20 to 30 seconds. Then STV was placed in an incubator at 37 ° C for 60 ± 2 minutes. STV was then observed by reversing the reaction tube in one smooth motion.
al. (1998)  preferred DIDA over wildly known amine extractant TOA, due to its possibility of using as extractant without adding a modifier. The parameters of solvent extraction such as aqueous phase acidity, type of diluents, molybdenum concentration, and DIDA concentration were investigated to obtain the suitable condition for effective extraction of molybdenum. The extraction mechanism was an anion-exchange reaction, which formed a polymeric complex, expressed as (R2NH)n MomOp(HSO4)y, where the values of n, m, p and y are depend on the polymerization degrees of molybdenum and amine in organic and aqueous phase. The polymeric complex changes according to the concentration of DIDA and metal, aqueous acidity, diluents, time, and type of extraction process.
The mobile phase and stationary phase in the HPLC will involve in the mechanism. The stationary phase in HPLC normally will be the silica gel. The silica gel will help to separate the components in the liquid sample as its particle size, surface properties and pore structure will lead to good separation results of solvent by minimize the length of diffusion path. The silica gel is also inert to most solvent so it can separate various type of chemical compound with high reproducibility. During the separation, the component in sample will interact with the adsorbent material within the pores of the stationary phase.
Calibrate the spectrophotometer the same exact way so that it does not mess up the calculations gathered. This week using yellow dye #5 mix .20 milligrams of this will 250 ml of distilled water in a volumetric flask. Make five different solutions, distilling the water each time, of new yellow dye solution and place each of the solutions in a blank cuvette to find the max absorbency of each the solutions. Next during week two the absorbency had to be found from the food that was brought in. Using a blank cuvette place the commercial liquid and calculate the concentration and absorbency and which dyes would be in this liquid.
Dissolve the salt in 60 ml of tap water. Add 30 ml 6 M Hcl and stir the mixture with a glass rod. Add 12 g solid Nacl to the solution and stir the mixture for about 2 minutes. Support a 250 ml separatory funnel on a ring, making sure that the stopcock is closed and that a clean beaker is placed beneath the exit tube. Transfer the aqueous solution from the beaker to the separatory funnel.
One sample had 250ng of plasmid A as well but with no enzymes added. All the digestions tubes were incubated at 37℃ for 30 minutes. After incubation, 5μL of loading buffer (30% glycerol, 10 mM Tris-HCl, pH 8, 1 mM EDTA, 0.025% bromophenol blue) was added to each sample. 50 ml of molten agarose (1% agarose boiled and cooled to 55℃ with added SYBRsafe) was poured into the casting tray for gel electrophoresis. Once the gel hardened, .5X TBE (44.5 mM Tris base, 44.5 mM boric acid, and 1.0 mM EDTA) was added just until the gel was covered with the TBE buffer.
32 100 μL of afore-prepared sample solution and the mixed reference standard were diluted 100 times with ethyl acetate. 50 μL of these dilution solutions were separated on the TLC plate coated with SNISG. The plate was developed with petroleum ether: ethyl acetate (4:1) and the movement of solvent was usually controlled at 1 cm from the upper edge. After completion, the plate was dried until no solvent smell remained. It was sprayed with an ethanol solution containing 10% sulfuric acid, and heated at an infra-red drier until obvious color came up, as shown in Fig.2 (B.ab).
15. Add another 25cm3 of Methanol and Ethyl acetate to the solutions. Stir gently for 20 minutes using a stirring rod this is to allow more of the active ingredients to mix with the solution. 16. Take two funnels and place one in two separate clean measuring beakers making sure the bottoms of the funnels don’t touch the bottom of the measuring beakers.