15-Tritiated

Tajima and co-workers [108] determined the surface excess of sodium dode-cyl sulfate by means of the radioactivity method, using tritiated surfactant of specific activity 9.16 Ci/mol. The area of solution exposed to the detector was 37.50 cm. In a particular experiment, it was found that with 1.0 x 10" Af surfactant the surface count rate was 17.0 x 10 counts per minute. Separate calibration showed that of this count was 14.5 X 10 came from underlying solution, the rest being surface excess. It was also determined that the counting efficiency for surface material was 1.1%. Calculate F for this solution. 

Tritium differs from the noble gases. Tritium is very reactive and readily combines to form water. The voloxidation process, which controls the point at which the oxidation occurs and recovers the resulting tritiated water, has been developed. As of this writing it has not yet been used. 

The small synthetic scale used for production of many labeled compounds creates special challenges for product purification. Eirst, because of the need for use of micro or semimicro synthetic procedures, the yield of many labeled products such as high specific activity tritiated compounds is often low. In addition, under such conditions, side reactions can generate the buildup of impurities, many of which have chemical and physical properties similar to the product of interest. Also, losses are often encountered in simply handling the small amounts of materials in a synthetic mixture. As a consequence of these considerations, along with the variety of tracer chemicals of interest, numerous separation techniques are used in purifying labeled compounds. 

Decay products of the principal radionuclides used in tracer technology (see Table 1) are not themselves radioactive. Therefore, the primary decomposition events of isotopes in molecules labeled with only one radionuclide / molecule result in unlabeled impurities at a rate proportional to the half-life of the isotope. Eor and H, impurities arising from the decay process are in relatively small amounts. Eor the shorter half-life isotopes the relative amounts of these impurities caused by primary decomposition are larger, but usually not problematic because they are not radioactive and do not interfere with the application of the tracer compounds. Eor multilabeled tritiated compounds the rate of accumulation of labeled impurities owing to tritium decay can be significant. This increases with the number of radioactive atoms per molecule. 

Since ivermectin (= 22,23-dihydroavermectin B ) is obtained by catalytic reduction of avermectin B, the same procedure using tritium gas convenientiy affords tritiated ivermectin (22,23- [JT]-22,23-dihydroavermectin B ). The preparation of a tritiated derivative containing a 22,23-double bond starts with the readily available 5-ketone, which is reduced with [JT]-sodium borohydride stereospecificaHy to a 5- [JT]-derivative (40). Carbon-14 labeled avermectins can be obtained by a biosynthetic process using sodium (l- C)propionate as labeled precursor (48). 

Properties of T2O. Some important physical properties of T2O are Hsted in Table 2. Tritium oxide [14940-65-9] can be prepared by catalytic oxidation of T2 or by reduction of copper oxide using tritium gas. T2O, even of low (2—19% T) isotopic abundance, undergoes radiation decomposition to form HT and O2. Decomposition continues, even at 77 K, when the water is fro2en. Pure tritiated water irradiates itself at the rate of 10 MGy/d (10 rad/d). A stationary concentration of tritium peroxide, T2O2, is always present (9). AH of these factors must be taken into account in evaluating the physical constants of a particular sample of T2O. 

Hot atom reactions have also been used to label organic compounds with T. Irradiation of helium-3 with neutrons according to the nuclear reaction produces very energetic tritium atoms that can displace ordinary hydrogen in organic compounds. This procedure is not very selective, and the labeling pattern must be determined to enable the tritiated product to be used effectively as a tracer (34). 

Isotopic Concentration. A number of techniques have been reported for concentrating tritium from naturally occurring sources. For example, separation factors (H/T) of 6.6 to 29 were observed (49) for the concentration of tritium by electrolysis of tritiated water. Tritium is concentrated in the undecomposed water. 

It is generally assumed that ingested tritiated water is rapidly absorbed and uniformly distributed in the body fluids, with the result that the entire... 

The radiological hazard of tritium to operating personnel and the general population is controlled by limiting the rates of exposure and release of material. Maximum permissible concentrations (MPC) of radionucHdes were specified in 1959 by the International Commission on Radiological Protection (79). For purposes of control all tritium is assumed to be tritiated water, the most readily assimilated form. The MPC of tritium ia breathing air (continuous exposure for 40 h/wk) is specified as 185 kBq/mL (5 p.Ci/mL) and the MPC for tritium in drinking water is set at 3.7 GBq/mL (0.1 Ci/mL) (79). The maximum permitted body burden is 37 MBq (one millicurie). Whenever bioassay indicates this value has been exceeded, the individual is withdrawn from further work with tritium until the level of tritium is reduced. 

In another experiment tritiated adamantane diazirine fixed to the hydrocarbon core of a membrane gave rise to carbene insertion into the catalytic subunit of ATP-ase. After protolytic degradation adjacent areas of the original structure became evident (80JBC(255)860). 

Deuterium oxide [7789-20-0] M 20, f 3.8°/760mm, b 101.4°/760mm, d 1.105. Distd from alkaline KMnOa [de Giovanni and Zamenhof Biochem 7 92 79 I963. NOTE that D2O invariably contains tritiated water and will therefore be RADIOACTIVE always check the radioactivity of D2O in a scintillation counter before using. 

The 5a-isomer is obtained from hydrogenation of A -7-keto steroids over platinum or palladium. In the latter case the addition of pyridine to the reaction mixture greatly increases the amount of 5a- product formedd a Attack also takes place on hydrogenation (deuteration or tritiation )... 

The incorporation of a chapter on deuteration in a steroid monograph is quite reasonable since development of a number of the most important deuteration reactions have actually had their impetus through steroid research. The field of steroid chemistry offers possibly the largest variety of deuteration reactions of any area of organic chemistry. Many of these deuteration techniques have also been used for tritium labeling, which is especially pertinent in view of the large demand for tritiated steroids as tracers in biological experiments. 

A large number of examples have been reported in the literature for the deuteration or tritiation of double bonds and for the hydrogenation of enones labeled at an allylic position. The following list of substrates, in addition to those already discussed, provides a representative cross section ... 

Displacement of aromatic halogen in 2,4-diiodo-estradiol with tritiated Raney nickel yields 2,4-ditritiated estradiol. Aromatic halogen can also be replaced by heating the substrate with zinc in acetic acid-OD or by deuteration with palladium-on-charcoal in a mixture of dioxane-deuterium oxide-triethylamine, but examples are lacking for the application of these reactions in the steroid field. Deuteration of the bridge-head position in norbornane is readily accomplished in high isotopic purity by treatment of the... 

The most frequently encountered examples of cyclopropyl ring opening reactions in the steroid field are usually associated with angular or side chain methylation sequences. In fact, isotope labeling of the C-19 angular methyl group is the only reported application of this reaction for deuteration or tritiation pui poses. 

Cleavage of cyclopropyl rings in the presence of an acid or base is illustrated by tlie tritiation of the 19-angular methyl group in 5, 19-cycloandrostane-3, 17-dione [(239) (240) R = T]. This reaction is carried out by heating... 

How many tritium atoms are incorporated into acetate if a molecule of palmitic acid is oxidized in 100% tritiated water ... 

The production of tritium-labelled organic compounds was enormously facilitated by K. E. Wilz-bach s discovery in 1956 that tritium could be introduced merely by storing a compound under tritium gas for a few days or weeks the radiation induces exchange reactions between the hydrogen atoms in the compound and the tritium gas. The excess of gas is recovered for further use and the tritiated compound is purified chro-matographically. Another widely used method of... 

For exchange of non-labile organic hydrogen atoms, acid-base catalysis (or some other catalytic hydrogen-transfer agent such as palladium or platinum) is required. The method routinely gives tritiated products having a specific activity almost 1000 times that obtained by the Wilzbach method shorter times are required (2-12h) and subsequent purification is easier. 

When specifically labelled compounds are required, direct chemical synthesis may be necessary. The standard techniques of preparative chemistry are used, suitably modified for small-scale work with radioactive materials. The starting material is tritium gas which can be obtained at greater than 98% isotopic abundance. Tritiated water can be made either by catalytic oxidation over palladium or by reduction of a metal oxide ... 

Note, however, that pure tritiated water is virtually never used since 1 ml would contain 2650 Ci it is self-luminescent, irradiates itself... 

A distinction between these four possibilities can be made on the basis of the kinetic isotope effect. There is no isotope effect in the arylation of deuterated or tritiated benzenoid compounds with dibenzoyl peroxide, thereby ruling out mechanisms in which a C5— bond is broken in the rate-determining step of the substitution. Paths (ii) and (iii,b) are therefore eliminated. In path (i) the first reaction, Eq. (6), is almost certain to be rate-determining, for the union of tw o radicals, Eq. (7), is a process of very low activation energy, while the abstraction in which a C—H bond is broken would require activation. More significant evidence against this path is that dimers, Arz, should result from it, yet they are never isolated. For instance, no 4,4 -dinitrobiphenyl is formed during the phenylation of... 

In studying two-component polymerization catalysts, beginning with Feldman and Perry (161), a radioactive label was introduced into the growing polymer chain by quenching the polymerization with tritiated alcohols. The use of these quenching agents is based on the concept of the anionic coordination mechanism of olefin polymerization occurring... 

However, in olefin polymerization by two-component catalysts during polymerization not only active transition metal-polymer bonds are formed, but also inactive aluminum-polymer ones, as a result of the transfer process with the participation of a co-catalyst (11, 162-164). The aluminum-polymer bonds are quenched by tritiated alcohol according to the scheme (25), so an additional tagging of the polymer occurs. The use of iodine (165, 166) as a quenching agent also results in decomposing inactive metal-polymer bonds. 

Olefin polymerization by catalysts based on transition metal halogenides is usually designated as coordinated anionic, after Natta (194). It is believed that the active metal-carbon bond in Ziegler-Natta catalysts is polarized following the type M+ - C. The polarization of the active metal-carbon bond should influence the route of its decomposition by some compounds ( polar-type inhibitors), e.g. by alcohols. When studying polymerization by Ziegler-Natta catalysts tritiated alcohols were used in many works to determine the number of metal-polymer bonds. However, as it was noted above (see Section IV), in two-component systems the polarization of the active bond cannot be judged by the results of the treatment of the system by alcohol, as the radioactivity of the polymer thus obtained results mainly from the decomposition of the aluminum-polymer bonds. 

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