Interference in the Limulus amebocyte lysate assay for endotoxin determination in peritoneal dialysis fluids and concentrates for hemodialysis.
The interference of the saline concentration of fluids for peritoneal dialysis and concentrates for hemodialysis on the Limulus amebocyte lysate (LAL) assay for endotoxins was investigated. The experiments were carried out individually with each substance that compose fluids for hemodialysis, to determine the possible inhibition or enhancement effects that they could cause on the LAL assay. The compositions were also assayed to investigate the possibility of synergistic effect. They were assayed by the gel-clot method from two different suppliers, and the samples that showed inhibition effect were also assayed by the chromogenic method. The samples were analysed at successive dilutions, with different LAL sensitivities, to satisfy the endotoxin limits of 5 EU/ml for the concentrate and 0.25 EU/ml for the fluid for dialysis peritoneal. The results showed that the major interference on the gel-clot assay occurs in presence of acetic acid and in concentrates containing acid acetic, even the pH being adjusted between 6.5 and 7.5. However, the test, after an adequate dilution, could be validating for all samples. Chromogenic test can be used for peritoneal dialysis fluids considering a limit of 0.25 EU/ml and sample dilution of eight times, but it cannot be used for concentrates for hemodialysis without further dilution. Considering the results and that the chromogenic is a more time-consuming method, endotoxins in fluids for hemodialysis can be satisfactorily assayed by the gel-clot method.
Bohrer
D
D
Departamento de Química, Universidade Federal de Santa Maria, 97110 900 Santa Maria, RS, Brazil. ndenise@quimica.ufsm.br
Hörner
R
R
do Nascimento
P C
PC
Adaime
M
M
Pereira
M E
ME
Martins
A F
AF
Hartz
S A
SA
D015314:Dialysis Solutions / Q000032:analysis*; D004731:Endotoxins / Q000032:analysis*; D005782:Gels / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D008033:Limulus Test; D010530:Peritoneal Dialysis; D012015:Reference Standards; D006435:Renal Dialysis; D012492:Salts / Q000032:analysis
D015314:Dialysis Solutions; D004731:Endotoxins; D005782:Gels; D007202:Indicators and Reagents; D012492:Salts