chapter two

Color reactions in identications

About half of the identication tests are based on a methodology in which a selective chemical reaction performed on the substance to be examined yields a colored entity or even a colored precipitate that is directly recognizable as positive. These reactions are most commonly referred to as colorimetric reactions, although the name is also incorrectly used for photometric and spectrophotometric determinations. Historically, in a photometric analysis, an unknown sample is compared to a series of standards of known concentrations using a natural light photometer, whereas spectrophotometry uses a UV-visible spectrophotometer with monochromatic light. In colorimetric determinations, on the contrary, samples and standards are compared and judged visually. Since the identication tests are carried out without the use of positive controls or standards, they owe their robustness and reliability to the selectivity of the chemical reaction and the recognizability of the color produced. Especially the selectivity of the chemical reaction will naturally depend entirely on the actual reaction in question, but a few general remarks should be made. In principle, there are several ways of using a color reaction in an identication test. The substance to be examined is changed to a colored compound by the action of a reagent that does not become a part of the new entity formed. One example is test (b) for 3.19. Iodides, where identication is brought about by the violet or violet-red color produced when iodide is oxidized by dichromate to iodine. The substance to be examined changes an uncolored reagent into a colored compound without becoming a part of the new molecule formed. The substance to be examined reacts with a reagent forming a colored compound in which both the reagent and the substance to be examined

2006 by Taylor & Francis Group, LLC

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Pharmaceutical Chemical Analysis: Methods for Identication & Limit Tests are included. These could be covalently bound condensation products or more loosely associated chalets and salts. Examples of these are numerous: the condensation of certain amines and the reagent -napthol in 3.5. Amines, primary aromatic, or the cobalt complex formed in 3.10. Barbiturates non-nitrogen substituted.

In some cases, the actual reaction will include a combination of the above. An ideal chemical derivative color reaction will possess a number of qualities. It should preferably be unidirectional (not reversible), highly selective, and sensitive. The reaction product should be stable and have an appearance that is very different from the appearance of the substance to be examined and the reagent itself. Finally, it should be rugged toward small variations in the determination procedure. When dealing with an identication reaction even performed without comparing the result obtained with a standard, obviously the selectivity is of the greatest importance. Here it is important to remember that color reactions in their nature are at the best selective and not specic. In the case of a condensation between the substance to be examined and a reagent molecule, a functional group of the reagent reacts with a functional group of the substance to be examined. Since no functional group is found in one substance exclusively, the reagent will in principle react with all sample molecules having this particular functional group in its structure. Some degree of selectivity can be achieved, however, if the substance to be examined reacts very easily and under very mild conditions compared to other compounds of its class due to, for example, a steric hindrance aspect. Other reactions, on the contrary, are not even truly selective with respect to functional groups but will under the right conditions give a positive reaction with compounds of other classes. One example is the Legal reaction used for identication of 3.21. Lactates and 3.17. Citrates. The reagent used here, sodium nitroprusside, can react with all ketones and aldehydes having an unsaturated -carbon. However, the reaction is more or less likely to occur depending on the actual compounds ability to shift into its enol resonans form and also depending on steric hindrance in the actual compound. Another example is the test for esters, where the ester bond is hydrolyzed and the carboxylic acid converted into a hydroxamic acid, which gives a distinct colored complex with iron(III). This reaction has been used for the determination of compounds characterized by a number of functional groups including many carboxylic acids, acid anhydrides, acid halides, acid amides, and imides. It is therefore evident that color reactions cannot afford a selective, not to mention specic, identication of a compound. Their value lies in the relative ease with which they can be performed, and in the fact that they are unconstrained by expensive analytical equipment. Usually they are used together with two or more other identications, which combined, provide a reasonable certainty about the identity of the substance to be examined.

2006 by Taylor & Francis Group, LLC