Test suspension

Stainless steel disks are contaminated with a bacterial test suspension and dried. The disinfectant is applied on the dried film on the disk and kept at a specified temperature for a defined time. The disk is than transferred to a previously validated neutralization medium to stop the action of the disinfectant. The cfii of surviving bacteria recovered from the surface is determined quantitatively. 

The iotai proteolytic activity of pancreas powder is determined by comparing the quantity of peptides nonprecipitabie by a 556 m/V solution of trichloroacetic acid R released per minute from a substrate of casein solution with the quantity of such peptides released by pancreas powder (protease) UK from the same substrate in the same conditions. For the test suspension and the reference suspen-sion, prepare the suspension and carry out tiie dilution at (W-°C. 

Determine the activity of the substance to be examined using the corrected absorbance for the test suspension (T-Tb) and the callbraticm curve, taking into account the dilution factors. 

In a small mortar cooled to OM C, triturate carefully a quantity of the substance to be examined equivalent to about 2500 Ell. Eur, U. of lipolytic activity with 1 mL of cooled maleate buffer solution pH 7.0 R (lipase advent) until a very fine suspension is obtained. Dilute the suspension with cold maleate buffer solution pH 7.0 R. transfer quantitatively to a volumetric flask, and dilute to 100-0 mL with the cold buffer solution. Keep the flask containing the test suspension in iced water during the titration. 

Ctirry out three determinations in the same manner for the test suspension (Tl, Tj, and T3), If the quantity of 0.1 N sodium hydroxide used is outside the limits 0,08-0.16 mLAuin, the assay should be started again widt a quantity of test suspension that is more suitable but situated between 0.4 and 0.6 mL. Otherwise the quantity of the substance to be examined should be adjusted to comply with the conditions of the test... 

Iodoform can be recognized by its odor and yellow color and, more securely, from the melting point (119-123°C). The substance can be isolated by suction filtration of the test suspension or by adding 0.5 mL of dichloromethane, shaking the stoppered test tube to extract the iodoform into the small lower layer, withdrawing the clear part of this layer with a Pasteur pipette, and evaporating it in a small tube on the steam bath. The crude solid is crystallized from methanol-water. 

Bacteria were grown in nutrient broth (Difco) at 37 for 18 hours. Test suspensions were diluted to approximately 5 X 10 cells per ml. in O.IM phosphate buffer... 

Test suspension cell lines directly from a culture at approx 5 x 10s cells/mL. 

Sun=100mg/Test Suspension, Sun and also Sunless Tanning Lotion (2 times a day), 3cc/Synthol into Biceps and Calf Muscles (3cc each / 12cc total)... 

Q4--The table in the article shows that slin and test suspension are taken EOD each is this correct ... 

Prepare a suspension of pancreas powder (protease) BRP as described for the test suspension but without addition of enterokinase so as to obtain a known final concentration of about 0.065 Ph. Eur. U. per milliliter calculated on the basis of the stated activity. 

Carry out the titrations immediately after preparation of the test suspension and the reference suspension. Place 29.5 mL of olive oil emulsion in the reaction vessel equilibrated at 37 0.5°C. Fit the vessel with the electrodes, a stirrer, and the burette (the tip being immersed in the olive oil emulsion). 

Nti - Nt2 = average volume of 0.02 N sodium hydroxide used per minute during the titration of the test suspension Nri - Nr2 = average volume of 0.2 N sodium hydroxide used per minute during the titration of the reference suspension mt = mass in milligrams of the substance to be examined mr = mass in milligrams of the reference preparation... 

Incubation and Drugs. The RA-233 employed in this study was obtained from Wilbur Benson, Boehringer-Ingelheim, of Stamford, Connecticut. PGEi was provided by Joel Moake of the University of Texas School of Medicine at Houston. Cell suspensions were maintained at room temperature until addition of antiplatelet agents, at which time both drug-exposed suspensions and control suspensions were incubated at 37°C. Test samples contained 1 X 10-6Af RA-233 and 3 x 10 6M PGEi, while control samples received equal volumes of sterile saline. Both control and test suspensions were incubated for 10 min in a water bath at 37°C, then were sheared for 10 min in the Rice University ROM-8 viscometer which was maintained at 37 1°C with warmed forced air. 

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