Determination of residual 4'-aminomethyl-4,5',8-trimethylpsoralen and mutagenicity testing following psoralen plus UVA treatment of platelet suspensions.
Psoralens and UVA light have been used in the laboratory to study the inactivation of viruses that may be infrequently present in platelet concentrates that are prepared for transfusion. In order to evaluate safety aspects of the treatment of platelet suspensions with 4'-aminomethyl-4,5',8-trimethylpsoralen (AMT), we have investigated the residual levels and mutagenic potential of AMT after UVA phototreatment. 4'-aminomethyl-4,5',8-trimethylpsoralen, at a final concentration of 40 micrograms/mL, was added to platelet suspensions which contained 16% plasma and a synthetic medium. Platelet suspensions containing AMT were irradiated with up to 7.2 J/cm2 UVA light under normal oxygen levels. Residual levels of AMT were determined by HPLC and a bioassay based on bacteriophage phi 6 inactivation. The photodestruction of AMT or its activity by UVA was characterized by a D37 value of 0.6 and 0.3 J/cm2 with HPLC or bioassay, respectively. At 2.4 J/cm2 UVA, which results in approximately 5 log10 inactivation of vesicular stomatitis virus (VSV) and retention of platelet in vitro properties, 12% (HPLC) to 9% (bioassay) AMT remained. Like other psoralens, AMT was found to bind to serum proteins as shown by ultrafiltration. Results are consistent with approximately 36% of the initial drug load binding primarily to serum albumin. It was determined using 3H-AMT that 9 to 18% of radioactivity was bound to platelets in the absence of irradiation. Similar fractions (13 to 18%) of AMT were bound to platelets after 3.6 J/cm2 UVA irradiation, and 8 to 10% of total AMT was associated with saline-washed irradiated platelets and is presumably tightly bound.(ABSTRACT TRUNCATED AT 250 WORDS)
Wagner
S J
SJ
American Red Cross Blood Services, Jerome H. Holland Laboratory for the Biomedical Sciences, Rockville, MD 29855.
White
R
R
Wolf
L
L
Chapman
J
J
Robinette
D
D
Lawlor
T E
TE
Dodd
R Y
RY
D001435:Bacteriophages / Q000187:drug effects; D001769:Blood; D001792:Blood Platelets / Q000187:drug effects* / Q000528:radiation effects; D006801:Humans; D009152:Mutagenicity Tests / Q000379:methods; D009153:Mutagens / Q000633:toxicity*; D017319:Photosensitizing Agents / Q000633:toxicity*; D012486:Salmonella typhimurium / Q000187:drug effects; D012709:Serum Albumin / Q000494:pharmacology; D014307:Trioxsalen / Q000031:analogs & derivatives* / Q000633:toxicity; D014466:Ultraviolet Rays; D010948:Viral Plaque Assay
D009153:Mutagens; D017319:Photosensitizing Agents; D012709:Serum Albumin; C018822:aminomethyltrioxsalen; D014307:Trioxsalen