N-acetyl[3H]leukotriene D4: a stable internal standard for automated BIO-FAST HPLC extraction and separation of LTE4 from human urine.
The BIO-FAST (Fully Automated Sample Treatment) HPLC can be used for the isolation and separation of leukotriene E4 (LTE4) from the urine of asthmatic patients. A chemically related leukotriene, N-acetyl[14,15-3H]leukotriene D4 (NAc[3H]LTD4), has been evaluated as an internal standard to allow full automation of the BIO-FAST method. NAcLTD4 is not a human metabolite, does not co-elute with endogenously produced LTs and is stable in native urine at 37 degrees C for at least 18 h. Recovery and stability studies were conducted by adding NAc[3H]LTD4 and [3H]LTE4 to the baseline urine of four asthmatic patients. Automated extraction of these four samples over 22 hours, using the BIO-FAST system, yielded recoveries of 80.5% (6.6 %CV, n = 12) and 72.4% (10.0 %CV, n = 12) for the NAc[3H]LTD4 and [3H]LTE4, respectively. The ratio of NAc[3H]LTD4 to [3H]LTE4 was 1.12 (6.3 %CV, n = 12) demonstrating the consistent relative extraction of these two leukotrienes.
Nicoll-Griffith
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Department of Medicinal Chemistry, Merck Frosst Centre for Therapeutic Research, Pointe Claire-Dorval, Quebec, Canada.
Zamboni
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D001331:Automation; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation; D004355:Drug Stability; D006801:Humans; D017999:Leukotriene E4; D012015:Reference Standards; D013189:SRS-A / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000652:urine; D014316:Tritium
D013189:SRS-A; D014316:Tritium; D017999:Leukotriene E4; C077266:N-acetylleukotriene D4