Hair analysis for drugs of abuse. XIX. Determination of ephedrine and its homologs in rat hair and human hair.
A sensitive GC-MS method was developed for the quantitative analysis of ephedrine (EP), phenylpropanolamine (PPA) and methylephedrine (ME) in animal and human hair. After washing with 0.1% sodium dodecyl sulfate, hair samples (10 mg) were added with deuterated internal standards, extracted by 1-h sonication and over night soaking in 2 ml of 5 M HCl-methanol (1:20) at room temperature. Following evaporation of the liquid phase, the residue was dissolved in phosphate buffer solution (pH 6.0) and purified using a solid-phase extraction procedure with Bond Elut Certify columns. Two types of derivatization were compared - using trifluoroacetic anhydride (TFAA) and pentafluoropropionic anhydride (PFPA) - for discrimination of EP and methamphetamine (MA). Derivatized extracts were analyzed by GC-MS in the EI mode using a capillary column (OV-1 equivalent). From the results comparing three GC-MS conditions, PFP-derivatives separated with a temperature gradient of 20 degrees C/min from 60 degrees C to 280 degrees C gave the best resolution between EP and MA. ME was analyzed as a trimethylsilyl derivative using N,O-bis-trimethylsilyl acetamide at the above GC condition. The assay was linear from 0.5 to 50 ng/mg (r=0.998) and capable of detecting less than 50 pg of derivatized EP, PPA and ME on-column. Intra-assay precision was characterized by C.V. values from 5 to 16% in the concentration range of 1-10 ng/mg hair. The method was used for the quantitative determination of EP, PPA and ME in the hair obtained from three rats with dark brown hair after ten intraperitoneal injections (5 mg/kg/day) of the three drugs and from three male and one female volunteers with black hair after an oral dose of 50 mg/day of EP-HCl for three days. Hair samples were collected by shaving from the back of rats and cutting from the scalp of humans 28 days after the first dose. The incorporation rates of EP, PPA and ME into hair (the ratios of [hair concentration] to [AUC]) obtained from the animal experiment were 0.10, 0.07 and 0.03, respectively, which are a little lower than those (0.14, 0.10 and 0.04) of their desoxy-compounds, MA, amphetamine and dimethylamphetamine. EP was detected at an average of 2.25 ng/mg (n=4) in human scalp hair and at a range of 1-29 ng/mg (n=3) in human beard hair until day 14, but its metabolite (PPA) was at a trace level in the hair of the four subjects. The method was successfully used for detection of ME and EP in the hair of a neonate and its mother who was abusing Bron syrup containing ME during the pregnancy.
Nakahara
Y
Y
National Institute of Health Sciences, Tokyo, Japan. nakahara@nihs.go.jp
Kikura
R
R
D000662:Amphetamines / Q000032:analysis; D000818:Animals; D000697:Central Nervous System Stimulants / Q000032:analysis* / Q000097:blood / Q000652:urine; D004809:Ephedrine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000097:blood / Q000652:urine; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry; D006197:Hair / Q000737:chemistry*; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D010665:Phenylpropanolamine / Q000032:analysis* / Q000097:blood / Q000652:urine; D011247:Pregnancy; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D015813:Substance Abuse Detection / Q000379:methods*
D000662:Amphetamines; D000697:Central Nervous System Stimulants; D010665:Phenylpropanolamine; C041707:N-methylephedrine; D004809:Ephedrine