Journal
AAPS JOURNAL
Volume 8, Issue 4, Pages E709-E717Publisher
SPRINGER
DOI: 10.1208/aapsj080480
Keywords
methamphetamine; rats; LC-MS/MS; matrix ion effects
Categories
Funding
- NIDA NIH HHS [P01 DA014361, P01DA14361, P01 DA014361-05, P01 DA014361-04, R01DA11560, P01 DA014361-04S1] Funding Source: Medline
- PHS HHS [K2514601] Funding Source: Medline
- NATIONAL INSTITUTE ON DRUG ABUSE [R01DA011560, P01DA014361] Funding Source: NIH RePORTER
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High-throughput liquid chromatography with tandem mass spectrometric detection (LC-MS/MS) methodology for the determination of methamphetamine ( METH), amphetamine (AMP), 4-hydroxymethamphetamine (4-OH-METH), and 4-hydroxyamphetamine (4-OH-AMP) was developed and validated using simple trichloroacetic acid sample treatment. The method was validated in rat serum, brain, and testis. Lower limits-of-quantitation (LOQ) for METH and AMP were 1 ng center dot mL(-1) using positive ion electrospray tandem mass spectrometry (MS/MS). The accuracy of the method was within 25% of the actual values over a wide range of analyte concentrations. The within-assay precision was better than 12% (coefficient of variation). The method was linear over a wide dynamic range (0.3-1000 ng center dot mL(-1)). Quantitation was possible in all 3 matrices using only serum standards because of minimal matrix-associated ion effects or the use of an internal standard. Finally, the LC-MS/MS method was used to determine serum, brain, and testis METH and AMP concentrations during a subcutaneous infusion (5.6 mg kg(-1) day(-1)) of METH in rats. Concentrations of 4-OH-AMP and 4-OH-METH were below the LOQ in experimental samples. The bias introduced by using serum calibrators for the determination of METH and AMP concentrations in testis and brain was less than 8% and insignificant relative to the interanimal variability.
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