12-5+Recommendation+Ponceau+4R

To minimize the effect of adding food dye can be identified by analyzing the sample with paper chromatography method, but this is only can be conducted by analyst (actually people can conduct this analysis but they must have the basic).


 * How to analyze it ? **
 * 1) Weigh 2 grams of sample and put into 100 ml beaker glass and add fat-free worsted.
 * 2) Add glacial acetic acid and distilled water.
 * 3) Heat and stir the solution until all of the color attach into worsted (clear solution).
 * 4) Worsted that has been attached by dye is inserted back into the beaker glass.
 * 5) Add diluted ammonia solution.
 * 6) Heat again until the color faded from the worsted.
 * 7) Discard the worsted.
 * 8) Spot the concentrated solution of sample and standard on the chromatography paper.
 * 9) Sample that have been spoted on chromatography paper will emulsified in chamber that has saturated by n-butanol : glacial acetate acid : water, until showing the separation of colors.
 * 10) Compare the Rf of sample with Rf of standard.


 * = [[image:resultforseparationdyeswiththinlayerchromatography1_m.gif caption="chromatography paper in chamber"]] ||= [[image:resultforseparationdyeswiththinlayerchromatography_m.gif caption="The result of color separation"]] ||

[|SEPARATION OF FOOD DYES BY TLC (THIN LAYER CHROMATOGRAPHY)]
__**Material: **__
 * Test solution: a mixture of 7 dyes dissolved in water: Erythrosin, Brillant Black BN, Fast Red E, Naphthol Red S, Yellow Orange S, Ponceau 4R, Tartrazine.
 * Reference solutions: Yellow Orange S and Brillant Black, each dissolved in water.
 * Freshly prepared developing solvent: 2.5 % sodium citrate solution, ammonia 25 %, 2-propanol (20 : 5 : 3).

__**Apparatus: **__
 * Developing chamber (jam glass bottle with a screw cover: h = 11 cm, d = 5 cm)
 * Fertigfolie POLYGRAM® CEL 300 plate (Macherey Nagel)
 * Glass capillaries (1 µL)

__**Experimental: **__  Using a soft pencil, a line is drawn approximately 1,5 cm from the bottom of the plate. The spotting points are numbered (1,2,3). At the spotting points 1 and 3 the reference solutions are applied onto the plate, at the spotting point 2 the dye mixture. Using capillaries approx 0.25 µL of the dye solutions are applied to the TLC plate. The capillaries fill themselves quickly when dipped into organic sample solutions. Before emptying the submerged end of the capillary is rolled horizontally on filter paper. The clean upper end of the capillary is placed on the layer vertically and carefully, vertically so that the capillary empties itself and carefully to avoid damage to the layer. Easy application of samples is allowed with a spotting guide. When the solvent is completely evaporated (approx. 10 min) from the plate, the loaded TLC plate is carefully placed in the TLC chamber with the sample line toward the bottom. The plate whose top is leaned against the jar wall should sit on the bottom of the chamber and be in contact with the solvent (solvent surface must be below the extract line). The TLC chamber is covered. The TLC plate is allowed to remain undisturbed. When the solvent front has reached three quarters of the length of the plate, the plate is removed from the developing chamber and the position of the solvent front is immediately marked. The solvent on the plate is allowed to evaporate.

The dye mixture is separated into individual components. The progress of their separation may be followed by observing the movement of colored spots. The use of referenve solutions allows to identify Yellow Orange S and Brillant Black in the mixture.
 * __Result: __**


 * For people who can't conduct the analysis, don't worry! Because you still can identify it visually or using your senses such as: **


 * In the manufacture product we can see the ingredients. What we have to do with the ingredients? We can check it, and make sure there is no Ponceau 4R.
 * A product that contains dangerous food dye will feel bitter or less bitter when consumed and leave the color on our tongue.
 * In long time, dangerous food dye can cause broken the brain nerve and disturb the balance of body or can cause hyperactivity for children.

__** The table of food dyes that is not allowed to consume in Indonesia : **__



As we know, Ponceau 4R is a synthetic food dye that gives red color to many foods such as : dessert topping, jelly, seafood dressing, tinned strawberry, packeted cake mix, cheesecake, and soup. These are the specification of Ponceau 4R :


 * Colour Index No. || 16255 ||
 * Food Colour No. || Red 7 ||
 * CAS NO. || 2611-82-7 ||
 * E.E.C. No. || E-124 ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">I.S. No. || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">2558 ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Total Dye Content Corrected for Sample (Min.) || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">82.00% ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Volatile Matter (chloride & sulphate) (Max) || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">18.00% ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Solubility In Water (20oc) || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">120 gram / litre ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Combined Ether Extract. (Max) || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">0.20% ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Water Insoluble (Max) || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">0.20% ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Class || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Monoazo ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Light Stability || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">4 ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Heat Stability (100oc) || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">5 ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Alkali Stability || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">3 ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Subsidiary Dyes (MAX) || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">1.00% ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Dyes Intermediate (Max) || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">0.50% ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Lead || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">< 10 ppm ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Arsenic || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">< 3 ppm ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Mercury || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">< 1 ppm ||
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Heavy Metals || <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">< 20 ppm ||



<span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">media type="youtube" key="YaNjdidIreQ?hl=en" height="347" width="405"
 * <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Core of the video above are : **


 * 1) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;"> A dangerous dye is a kind of dyes that could be digested by our bodies and the substrates is a dangerous substrate that will be transferred by our blood to our brain.
 * 2) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">There are positive and negative effects of the dyes, but if we consumed it temporary and excess it will be a negative to our body. And usually a synthetic dye made from tar of coal.
 * 3) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Tar of coal is a product of dry distillation of coal. And it will become quokkas and melted to become a tar. And almost all of our dye that used everyday except the food grade dyes is come from tar of coal. But in reality, some of the snack seller adds this substance to their snack, and the dyes are the product of the dry distillation of coal. Almost all of the color components are on the tar.
 * 4) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Our body can tolerate until the limit of or body itself, if we consume out of the limit, it will be a toxic and be a carcinogenic that caused cancer, but there is rarely fast reaction of the dyes. No one immediately die after consuming the product that contains synthetic dyes, because people just add it a few so it doesn’t matter.
 * 5) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">First this one, the double bond of the Nitrogen atom and second is the sulfides bonds at the sodium atom that can caused hyperactivity.
 * 6) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">If children too much consume it, they will very hard to fast asleep and are not obey to their parent.
 * 7) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">By using chromatography we can analyze qualitatively the presence of Ponceau 4R. By know the solubility of this substance it could be able to analyze by using paper chromatography.
 * 8) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">If we want to analyze Ponceau 4R, firstly the sample must be extracted by using wool. Because all of the dyes could be absorb by the protein in the wool in the acid condition. Approximately at pH 4, the wool from sheep will absorb the azo compounds and after that will be diluted again by using base (ammonia). You can dilute it by using test tube or beaker glass that filled with ammonia 1:1 and soak the wool into the ammonia after that shake it, all of the color will be rinsed out. After that throw out the wool that turn to be white again from the solution. So you will get the solution of the dyes in the ammonia. Evaporate it until the solution left about 0,1ml. Take that solution by using small capillary pipe to be analyzed by chromatography method. You can use Paper Chromatography or Thin Layer Chromatography. Drop it on the layer and drop the pure Ponceau 4R beside it and rinse it with the eluen in the chamber until perfectly separated. The composition of the eluen is very specific, so you must read it from literature. To identify the containing of the dyes, we should compare the RF of each spot that formed during the chromatography process.
 * 9) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">As long as sources know, all of the dyes could be absorbed with wool. And synthetic dye is made from Hydro-Carbon reaction (Tar). The reagent that usually used to identify are Conc. Sulfuric Acid, Conc. Hydrochloric Acid, Ammonia10% and Sodium Hydroxide 12%. Afterthat, there is a table of dyes reaction. Dye A will react with Conc. Sulfuric acid to form another color, etc. The wool is used to do this analysis not the solution. You cut the wool into 4 parts. By porcelain glasses you react the wool with each reagent and stir it. Compare the color changing by the table. Years ago, this practicum is did in this laboratory.
 * 10) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">The quantitative analysis of Ponceau 4R better use GC (Gas Chromatography) and the qualitative one better use Visible Spectrophotometer by knowing the wavelength of Ponceau 4R it self.
 * 11) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Ponceau 4R is including Aromatic salt class.
 * 12) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">This dyes is a carcinogenic substance if consumed temporary.
 * 13) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">The negative effect of all of the dyes will not appear immediately, except allergic.
 * 14) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">This substance will composed in our body to produce other dangerous substances (Histamine).
 * 15) <span style="font-family: Tahoma,Geneva,sans-serif; font-size: 150%;">Some of substance that go into our digestion process will react with our body, but some will not react and will throw away by urine or feces.

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