Determination of opiates in human fingernail--comparison to hair.

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (2014-08-03)
Min Shen, Hang Chen, Ping Xiang
ABSTRACT

6-Monoacetylmorphine in keratinized matrices can be used to discriminate between heroin users and individuals exposed to other sources of morphine alkaloids. Frozen pulverization is effective in preventing 6-monoacetylmorphine hydrolysis. The main aim of this study was to develop an LC-MS/MS method for the determination of five opiates in human fingernails using a frozen pulverization preparation method and to investigate the correlation between the concentration of opiates in nail and hair samples from subjects whose urine specimens were positive for morphine. Borate buffer (500 μL; pH 9.2) was added to 20mg of pulverized fingernail, followed by ultrasonication and liquid-liquid extraction. Analytes were analyzed on an Allure PFP propyl column by gradient elution. The mass spectrometer was operated in the positive electrospray ionization mode and multiple reactions monitoring mode. A total of 12 of 18 fingernail samples contained detectable 6-monoacetylmorphine (mean=0.43 ng/mg, range=0.10-1.37 ng/mg), morphine (mean=1.74 ng/mg, range=0.58-3.16 ng/mg) and codeine (range from <limit of quantification to 0.27 ng/mg). Similarly, 12 of 18 hair samples obtained from the same subjects were positive at the revised Society of Hair Testing cutoff level of 0.2 ng/mg. The concentrations of 6-monoacetylmorphine, acetylcodeine and codeine in hair were significantly higher than those in nails. However, the concentration of MOR in nails was significantly higher than that in hair, except for one sample. All of the ratios of 6-MAM/MOR were below 0.57. It is proposed that nails may be an alternative to hair for documenting heroin abuse.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
2-Propanol, BioReagent, for molecular biology, ≥99.5%
Sigma-Aldrich
2-Propanol, suitable for HPLC, 99.9%
Sigma-Aldrich
2-Propanol, ACS reagent, ≥99.5%
Sigma-Aldrich
Chloroform, contains 100-200 ppm amylenes as stabilizer, ≥99.5%
Sigma-Aldrich
Formic acid, reagent grade, ≥95%
Sigma-Aldrich
2-Propanol, anhydrous, 99.5%
Supelco
Ammonium acetate, suitable for mass spectrometry (MS), LiChropur, eluent additive for LC-MS
Sigma-Aldrich
Chloroform, anhydrous, ≥99%, contains 0.5-1.0% ethanol as stabilizer
Sigma-Aldrich
Isopropyl alcohol, ≥99.7%, FCC, FG
Sigma-Aldrich
Ammonium acetate, for molecular biology, ≥98%
Sigma-Aldrich
Chloroform, anhydrous, contains amylenes as stabilizer, ≥99%
Sigma-Aldrich
Formic acid, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥98%
Sigma-Aldrich
Chloroform, suitable for HPLC, ≥99.8%, contains 0.5-1.0% ethanol as stabilizer
Sigma-Aldrich
Chloroform, HPLC Plus, for HPLC, GC, and residue analysis, ≥99.9%, contains amylenes as stabilizer
Sigma-Aldrich
Formic acid, ACS reagent, ≥96%
Sigma-Aldrich
Isopropyl alcohol, meets USP testing specifications
Sigma-Aldrich
Ammonium acetate, reagent grade, ≥98%
Sigma-Aldrich
2-Propanol, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)
Sigma-Aldrich
2-Propanol, HPLC Plus, for HPLC, GC, and residue analysis, 99.9%
Sigma-Aldrich
2-Propanol, BioUltra, for molecular biology, ≥99.5% (GC)
Sigma-Aldrich
Ammonium acetate, ACS reagent, ≥97%
Sigma-Aldrich
Ammonium acetate solution, for molecular biology, 7.5 M
Sigma-Aldrich
Formic acid, puriss., meets analytical specifications of DAC, FCC, 98.0-100%
Sigma-Aldrich
Chloroform, contains ethanol as stabilizer, ACS reagent, ≥99.8%
Sigma-Aldrich
Ammonium acetate, ≥99.99% trace metals basis
Sigma-Aldrich
2-Propanol, Laboratory Reagent, ≥99.5%
Sigma-Aldrich
2-Propanol, puriss. p.a., ACS reagent, ≥99.8% (GC)
Sigma-Aldrich
Chloroform, suitable for HPLC, ≥99.8%, amylene stabilized
Supelco
2-Propanol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Formic acid, ACS reagent, ≥88%