United States Department of Transportation
i
Intensity of the Internal Standard Response as the Basis for Reporting a Test Specimen as Negative or Inconclusive
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2007-08-01
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[PDF-520.21 KB]
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Edition:Final Report
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Abstract:Under normal circumstances, a test specimen is reported as “negative” when the response of the analyte is absent. However, if the intensity of the internal standard (IS) is low, indicating interference factors, the test could be considered “inconclusive.” A quantitative hypothesis, A = (R×I×S)/L, serves as the “cutoff” for the acceptable signal-to-noise (S/N) ratio for the IS in making “negative/inconclusive” decisions, where A: acceptable S/N ratio for internal standard; R: relative response of the IS and the analyte (same concentration); I: concentration of the IS; S: (minimal S/N ratio); and L: limit of detection. The hypothesis was empirically tested using 9-carboxy-11-nor-Delta9-tetrahydrocannabinol (THC-COOH) analyte, THC-COOH-d3 IS, and ibuprofen and hydrogen peroxide (H2O2) as interference factors. Urine specimens containing 0–5 ng/mL of THC-COOH were spiked with various quantities of ibuprofen or H2O2, followed by liquid-liquid extraction, derivatization, and GC-MS analysis under selected-ion-monitoring mode. Among the “adulterated” test specimens evaluated, those with a S/N for the internal standard below the acceptable IS S/N “A,” the quantitative criterion was indeed found to provide a useful guide for making negative/inconclusive decisions. This equation could be programmed into the instrument software to flag results as being inconclusive when they do not meet the criteria described in this paper.
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