High-performance liquid chromatographic evaluation of the effect of heat treatment on trimethoprim and sulfamethoxazole stability in serum.
Heat treatment of patient samples is utilized as a method to decrease the risk of accidental transmission of human immunodeficiency virus (HIV). Heat treatment has been reported to affect the outcome of drug analysis. In this study, the effects of heat treatment (56 degrees C for 5 h) and storage for 2 months at -20 degrees C on the stability of trimethoprim (TMP) and sulfamethoxazole (SMX) at three different concentrations in serum (10/100, 1/20, and 0.1/5 microgram/mL) each were evaluated. Simultaneous determination of TMP, SMX, and sulfamethazine (SMeth), the internal standard, in serum was performed by a reversed-phase high-performance liquid chromatographic (HPLC) procedure with isocratic elution and ultraviolet detection. The peak/height ratios (PHRs) for each sample from untreated and heat-treated groups were compared. No statistically significant differences were found between untreated and heat-treated groups for TMP. Heat treatment decreased the PHR for SMX at 100 micrograms/ml concentration (p = 0.042) and increased the PHR for SMX at 20 micrograms/ml concentration (p = 0.049). These marginal differences are unlikely to be significant. Storage of samples for 2 months at -20 degrees C had no statistically significant effect on sample PHRs. Thus, heat treatment of serum does not alter clinical interpretation of TMP and SMX at clinically relevant concentrations and may protect laboratory workers from accidental HIV exposure.
D000890:Anti-Infective Agents / Q000097:blood / Q000627:therapeutic use; D000892:Anti-Infective Agents, Urinary / Q000097:blood; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D005615:Freezing; D006358:Hot Temperature; D006801:Humans; D013420:Sulfamethoxazole / Q000097:blood* / Q000627:therapeutic use; D014295:Trimethoprim / Q000097:blood* / Q000627:therapeutic use
D000890:Anti-Infective Agents; D000892:Anti-Infective Agents, Urinary; D014295:Trimethoprim; D013420:Sulfamethoxazole