Chemical assay of glutaraldehyde incorporation into pericardial tissue.
This paper describes the development of a chemical assay to determine the incorporation of glutaraldehyde into pericardial tissue. The procedure is based upon the hydrolysis of the Schiff's bases formed between the glutaraldehyde and the E-amino groups of lysine, the liberated glutaraldehyde being determined as its 2,4-dinitrophenyl-hydrazone by high performance liquid chromatography (HPLC). The HPLC system uses a reverse phase column eluted with a gradient of aqueous acetonitrile (from 60%-70% over 30 minutes). The glutaraldehyde 2,4-dinitrophenylhydrazone is detected at 356 nm. The results show that the assay has sufficient sensitivity (minimum detectable limit 20 pmol of glutaraldehyde 2,4-dinitrophenyl-hydrazone) and reproducibility (+/- 9% at 100 pmol +/- 4% at 1 nmol) to estimate glutaraldehyde incorporation into pericardial tissue. A series of subsequent experiments have documented that temperature and buffer pH are the main determinants affecting glutaraldehyde incorporation into pericardial tissue. Immersion of pericardial tissue in glutaraldehyde solution of pH 9 and 37 degrees C yields an approximately six times greater incorporation than can be achieved if the reaction is performed at 0 degrees C. Further study is underway to relate the amount of Schiff's base bound to that of total incorporated glutaraldehyde.
Hughes
H
H
Biological & Medical Research Department, King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia.
Lilburn
S
S
Tipton
S
S
Aboul-Enein
H Y
HY
Duran
C M
CM
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D046508:Culture Techniques; D005976:Glutaral / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D010496:Pericardium / Q000378:metabolism*; D012545:Schiff Bases; D012756:Sheep; D013696:Temperature
D012545:Schiff Bases; D005976:Glutaral