Neutral-form base, analyzing

Ammonium chloride is analyzed by treatment with formaldehyde (neutralized with NaOH) and the product HCl formed is analyzed by titration using an acid-base color indicator such as phenolphthalein. Alternatively, it may be mixed with caustic soda solution and distdled. The distillate may be analyzed for NH3 by titration with H2SO4 or by colorimetric Nesslerization or with an ammonia-selective electrode (APHA, AWWA, WEF. 1995. Standard Methods for the Examination of Water and Wastewater. 19th ed. Washington, DC, American Pubhc Health Association). The presence of ammonia or any other ammonium compound would interfere in the test. The moisture content in NH4CI may be determined by Karl—Fischer method. 

The opposite scenario for retention dependence versus pH is observed for basic compounds (Figure 4-21) [56], In the mixture of basic components shown in Figure 4-21 (pyridinal species), increasing the pH of the aqueous portion of the mobile phase from 1.5 to 9 led to the enhancement of the retention of the basic analytes. At aqueous mobile-phase pH values of 7 and greater, the components exhibited a high retention. However, as the aqueous pH was changed to below 4, the compounds eluted close to the void volume. Generally, it is recommended that very polar bases are analyzed at pHs where the analyte is in its neutral form. 

Example 2. Analyzing a Base in its Neutral Form. In this example we will use the same compound as in the example above [2,4-dimethylpyridine (base) with pKa of 6.7] along with the same eluent conditions (50% MeCN and 50% phosphate buffer). The goal is to calculate pH of the buffer, in order to obtain the basic analyte in its fully neutral form. 

If Product M, a diprotic base, is to be analyzed in its neutral form, the higher of Product M (which is 5.3) needs to be considered because the other of 3.3 is less basic. Let us use to try to determine at what wpH the analyte would be in its neutral form at eluent conditions of 30v/v% MeCN and 70 v/v% acidic buffer. 

Acid-base (neutralization) reactions are only one type of many that are applicable to titrimetric analysis. There are reactions that involve the formation of a precipitate. There are reactions that involve the transfer of electrons. There are reactions, among still others, that involve the formation of a complex ion. This latter type typically involves transition metals and is often used for the qualitative and quantitative colorimetric analysis (Chapters 8 and 9) of transition metal ions, since the complex ion that forms can be analyzed according to the depth of a color that it imparts to a solution. In this section, however, we are concerned with a titrimetric analysis method in which a complex ion-forming reaction is used. 

Elemental composition P 38.73%, H 1.26%, O 60.01%. The compound may be identified by physical properties alone. It may be distinguished from ortho and pyrophosphates by its reaction with a neutral silver nitrate solution. Metaphosphate forms a white crystalline precipitate with AgNOs, while P04 produces a yellow precipitate and P20 yields a white gelatinous precipitate. Alternatively, metaphosphate solution acidified with acetic acid forms a white precipitate when treated with a solution of albumen. The other two phosphate ions do not respond to this test. A cold dilute aqueous solution may be analyzed for HPO3 by ion chromatography using a styrene divinylbenzene-based low-capacity anion-exchange resin. 

Most of the older methods of fluorimetric analysis of pesticides involved hydrolysis to form fluorescent anions. Co-ral (coumaphos) [147] was hydrolyzed in alkali to the hydroxybenzopyran, which was subsequently determined by means of its fluorescence. Guthion (azinphosmethyl) was hydrolyzed to anthranilic acid for fluorimetric analysis [148,149]. A method was developed [150] for Maretin (N-hydroxynaphthalimide diethyl phosphate) in fat and meat which involved hydrolysis in 0.5 M methanolic sodium hydroxide followed by determination of the fluorescence of the liberated naphthalimide moiety. Carbaryl (1-naphthyl N-methylcarbamate) and its metabolites have been determined by a number of workers using base hydrolysis and the fluorescence of the resulting naphtholate anion [151-153]. Nanogram quantities of the naphtholate anion could be detected. Zectran (4-dimethylamino-3,5-xylyl N-methylcarbamate) has been determined by the fluorescence of its hydrolysis product [154]. The fluorescence behaviour of other carbamate insecticides in neutral and basic media has been reported [155]. Gibberellin spray used on cherries has been determined fluorimetrically after treatment with strong acid [156]. Benomyl (methyl N-[l-(butylcarbamoyl)-2-benzimidazolyl]carbamate) has been analyzed by fluorimetry after hydrolysis to 2-aminobenzimidazole [157]. 

An alternative to the terrestrial synthesis of the nucleobases is to invoke interstellar chemistry. Martins has shown, using an analysis of the isotopic abundance of 13C, that a sample of the 4.6 billion year old Murchison meteorite which fell in Australia in 1969 contains traces of uracil and a pyrimidine derivative, xanthine. Samples of soil that surrounded the meteor when it was retrieved were also analyzed. They gave completely different results for uracil, consistent with its expected terrestrial origin, and xanthine was undetectable [48], The isotopic distributions of carbon clearly ruled out terrestrial contamination as a source of the organic compounds present in the meteorite. At 0°C and neutral pH cytosine slowly decomposes to uracil and guanine decomposes to xanthine so both compounds could be the decomposition products of DNA or RNA nucleobases. They must have either travelled with the meteorite from its extraterrestrial origin or been formed from components present in the meteorite and others encountered on its journey to Earth. Either way, delivery of nucleobases to a prebiotic Earth could plausibly have been undertaken by meteors. The conditions that formed the bases need not have been those of an early Earth at all but of a far more hostile environment elsewhere in the Solar System. That environment may have been conducive to the production of individual bases but they may never have been able to form stable DNA or RNA polymers this development may have required the less extreme conditions prevalent on Earth. 

In addition to a wide range of polar and nonpolar hydrocarbons that can be analyzed by RP-HPLC, it is also possible to separate ionic species. Because water is used as part of almost all mobile phases, those species which are acids and bases can be neutralized by control of pH. In cases where neutralization is not possible, then the addition of a counterion into the mobile phase so that the analyte will form a neutral complex can be used to enhance RP retention. The same principle can be applied to inorganic species by forming a neutral complex that results in reversed-phase retention. 

The purification procedure should remove potentially interfering compounds and, moreover, fractionate the entire spectrum of cytokinins into groups. It is usually not possible to analyze nucleotides and O-glucosides together with free bases, ribosides and iV-glucosides. Classical liquid-liquid partition steps [274] have been recently replaced by less labor-intensive, rapid and selective solid-liquid extraction. When applied in neutral aqueous solutions, cytokinin nucleotides are retained on weak anion-exchangers (DEA -Sephadex, DEAE-cellulose), while the other cytokinin forms are retained on reversed phase sorbents. Dobrev and Kaminek [275] used mixed-mode solid-phase extraction for step-wise... 

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