Incubations

After dissolving the lyophilisate in the added volume, incubation is an essential step to obtain the radiolabeled medicinal product. In this phase, the chemical reactions take place, resulting in Tc labeling. If incubation is inadequate, the labeling reaction may not be completed, and the radiopharmaceutical may not be suitable for administration. 

Each kit requires specific incubation conditions, but in general, this process is carried out at room temperature in a clean area. In certain cases, the incubation must be performed in a boiling water bath in such cases it is necessary to operate carefiilly. 

Incubation is normally performed with occasional agitation of the shielded reaction vial. 

Quality is directly related to the labeling yield, which is measured by the amount of unbound Tc activity. Limits of radiochemical impurities are stated in the official monographs (Council of Europe 2005). 

The determination of the radiochemical purity lies in the responsibility of the user (Theobald 1994). To assure safety and efficacy of a Tc radiopharmaceutical, the product should be tested regularly, in certain cases before application to the patient. 

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Figure A3.13.2 illustrates the origin of these quantities. Refer to [47] for a detailed mathematical discussion as well as the treatment of radiative laser excitation, in which incubation phenomena are unportant. Also refer to [11] for some classical examples in thennal systems.

As a rule, in diemial unimolecular reaction systems at modest temperatures, is well separated from the other eigenvalues, and thus the time scales for incubation and relaxation are well separated from the steady-... 

Note that in the low pressure limit of iinimolecular reactions (chapter A3,4). the unimolecular rate constant /fu is entirely dominated by energy transfer processes, even though the relaxation and incubation rates... 

Figure A3.13.3. Dissociation incubation ( iiK.-) and relaxation rate constants for the...

Figure Bl.14.8. Time course study of the arrival and accumulation of labelled sucrose in the stem of a castor bean seedling. The labelled tracer was chemically, selectively edited using CYCLCROP (cyclic cross polarization). The first image in the upper left comer was taken before the incubation of the seedlmg with enriched hexoses. The time given in each image represents the time elapsed between tire start of the incubation and the acquisition. The spectmm in the lower right comer of each image shows the total intensity...

Figure C 1.5.6. Single Ag nanoparticles imaged with evanescent-wave excitation. (A) Unfiltered photograph showing scattered laser light (514.5 nm) from Ag particles immobilized on a polylysine-coated surface. (B) Bandpass filtered (540-580 nm) photograph taken from a blank Ag colloid sample incubated witli 1 mM NaCl and...

To the remainder of the casein solution add 0 5 to o 8 g. of finely powdered commercial trypsin, shake to dissolve, and place in a thermostat (or in an incubator) at 40 . After 15 minutes, remove 25 ml. and add a few drops of phenolphthalein it will now be found that the solution remains colourless. Run in carefully Mj 10 NaOH solution until the colour of the solution is just pink, add 5 ml, of neutralised formalin and then titrate against Mj 10 NaOH solution until the pink colour is just restored note the amount required. Remove fiirther quantities (rf 25 ml. at intervals which must be determined by the speed of the reaction. The following will probably make a suitable series i, 2, 3,... 

To the remainder of the gelatin solution, add 0 5 to o 8 g. of finely powdered commercial trypsin and incubate at 40 . Carry out the formaldehyde titration on 25 ml. samples at intervals as above. 

Miscellaneous Alkaloids. Stukimic acid (57) is a precursor of anthranihc acid (28) and, in yeasts and Escherichia coli (a bacterium), anthranHic acid (o-aminobenzoic acid) is known to serve as a precursor of tryptophan (26). A similar but yet unknown path is presumed to operate in higher plants. Nonetheless, anthranHic acid itself is recognized as a precursor to a number of alkaloids. Thus damascenine [483-64-7] (134), C qH NO, from the seed coats of JSHgella damascena has been shown (95) to incorporate labeled anthranHic acid when unripe seeds of the plant are incubated with labeled precursor. 

Pish protein concentrate and soy protein concentrate have been used to prepare a low phenylalanine, high tyrosine peptide for use with phenylketonuria patients (150). The process includes pepsin hydrolysis at pH 1.5 ptonase hydrolysis at pH 6.5 to Hberate aromatic amino acids gel filtration on Sephadex G-15 to remove aromatic amino acids incubation with papain and ethyl esters of L-tyrosine and L-tryptophan, ie, plastein synthesis and ultrafiltration (qv). The plastein has a bland taste and odor and does not contain free amino acids. Yields of 69.3 and 60.9% from PPG and soy protein concentrate, respectively, have been attained. 

Pyridine herbicides are not strongly sorbed to soils and ate readily leached. The mobiUty of flutoxypyt [69377-81-7] has been found to decrease with increasing incubation time (399) this is attributed to entrapment of the herbicide within the soil organic matter. 

A study investigating the breakdown of clopytaUd [1702-17-6] reported half-Hves on different soils of approximately 2—7 weeks in a laboratory incubation (400) it was indicated that carryover was likely to occur in field soil. Picloram degrades and does not accumulate in field soil although low residue levels do persist for several years (401). The half-life for triclopyr [55335-06-3] is reported to be two weeks in two Canadian soils (402), and it has been shown to be rapidly degraded by aqueous photolysis (403). 

Aliphatic-Garboxylics. There are only two herbicides present in this class, trichloroacetate [76-03-9] (TCA) and dalapon [75-99-0]. These are used primarily for the selective control of annual and perennial grass weeds in cropland and noncropland (2,299). Dalapon is also used as a selective aquatic herbicide (427). Dalapon and TCA are acidic in nature and are not strongly sorbed by sods. They are reported to be rapidly degraded in both sod and water by microbial processes (2,427). However, the breakdown of TCA occurs very slowly when incubated at 14—15°C in acidic sods (428). Timing not only accelerates this degradation but also increases the numbers of TCA-degrading bacteria. An HA has been issued for dalapon, but not TCA (269). 

The preparation of estranes via 19-hydroxy steroids has also been accompHshed microbially. 19-Hydroxycholesterol, prepared chemically (39), is incubated with JS icardia restrictus to provide estrone (20) direcdy (44). Similarly, 19-hydroxyandrost-4-en-3,17-dione is converted to estrone (20) (45). 

The methylene blue and resazurin reduction methods indirectly measure bacterial densities in milk and cream in terms of the time interval required, after starting incubation, for a dye—milk mixture to change color (methylene blue, from blue to white resazurin, from blue through purple and mauve to... 

The agar [9002-18-0] plate method consists of adding a known quantity of sample, usually 1.0 or 0.1 mL, depending on the concentration of bacteria, to a sterile petti plate and then mixing the sample with a sterile nutrient medium. After the agar medium solidifies, the petti plate is incubated at 32°C for 48 hours after which the bacterial colonies are counted and the number expressed ia terms of a 1 mL or 1 g sample. This procedure measures the number of viable organisms present and able to grow under test conditions, ie, 32°C. 

Yogurt is manufactured by procedures similar to buttermilk. Milk with a fat content of 1—5% and soHds-not-fat (SNF) content of 11—14% is heated to ca 82°C and held for 30 minutes. After homogenization the milk is cooled to 43—46°C and inoculated with 2% culture. The product is incubated at 43°C for three hours in a vat or in the final container. The yogurt is cooled and held at <4.4° C. The cooled product should have a titratable acidity of not less than 0.9% and a pH of 4.3—4.4. The titratable acidity is expressed in terms of percentage of lactic acid [598-82-3] which is deterrnined by the amount of 0.1 AiNaOH/100 mL required to neutralize the substance. Thus 10 mL of 0.1 AiNaOH represents 0.10% acidity. Yogurts with less than 2% fat are popular. Fmit-flavored yogurts are also common in which 30—50 g of fmit are placed in the carton before or with the yogurt. 

The use of oxygen in pediatric incubators is an important factor in increasing the survival rate of premature infants who develop cyanosis. However, the use of oxygen is associated with risk of developing the visual defect known as retrolental fibroplasia (38). A careflil monitoring of arterial blood oxygen partial pressure is important. 

A 2-h incubation with another PGE2 analogue, nocloprost (9P-chloro-DMPG) protects normal human fibroblasts but has no effect on the survival of colon adenocarcinoma cells exposed to 10 Gy (1000 rad) (218). Nocloprost protects against radiation-induced DSBs in normal cells but not in tumor cells. Moreover, incubation using nocloprost for 2 h after irradiation enhances the rate of DSB rejoining in fibroblasts but not in adenocarcinoma cells. These data possibly reflect a different distribution of PG receptors on the plasma membrane of the two cell types. 

It is necessary to estabUsh a criterion for microbial death when considering a sterilization process. With respect to the individual cell, the irreversible cessation of all vital functions such as growth, reproduction, and in the case of vimses, inabiUty to attach and infect, is a most suitable criterion. On a practical level, it is necessary to estabUsh test criteria that permit a conclusion without having to observe individual microbial cells. The failure to reproduce in a suitable medium after incubation at optimum conditions for some acceptable time period is traditionally accepted as satisfactory proof of microbial death and, consequentiy, stetihty. The appHcation of such a testing method is, for practical purposes, however, not considered possible. The cultured article caimot be retrieved for subsequent use and the size of many items totally precludes practical culturing techniques. In order to design acceptable test procedures, the kinetics and thermodynamics of the sterilization process must be understood. 

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