Quantitative analysis of l-alpha-acetylmethadol, l-alpha-acetyl-N-normethadol, and l-alpha-acetyl-N,N-dinormethadol in human hair by positive ion chemical ionization mass spectrometry.
Quantitative analysis of l-alpha-acetylmethadol, l-alpha-acetyl-N-normethadol, and l-alpha-acetyl-N,N-dinormethadol in human hair by positive ion chemical ionization mass spectrometry.
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采用正离子化学电离质谱法对人发中的 L-α-乙酰美沙多、L-α-乙酰基-N-去甲美沙多和 L-α-乙酰基-N,N-二去甲美沙多进行定量分析。
DOI：
10.1093/jat/21.6.420
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发表时间：
1997
影响因子：
2.5
通讯作者：
Rollins,DE
中科院分区：
文献类型：
--
作者：
Wilkins,DG;Valdez,AS;Krueger,GG;Rollins,DE
建立了人发中I-α-乙酰美沙醇、I-α-乙酰基-N-去甲氧基美沙醇和I-α-乙酰基-N，N-二甲基美沙姆的灵敏、特异的定量分析方法。在该方法的发展过程中，确定了实验室进行的样品前处理方法对测定的头发中药物和代谢物的浓度有很大影响。在20毫克的头发样品中加入氚内标物，并将样品在第八型蛋白酶的缓冲溶液中消化过夜。通过改进我们实验室以前开发的用于血浆和组织分析的液-液萃取程序来提取消化液。衍生萃取物在Finnigan MAT®4500质谱仪上以正离子化学电离模式进行分析，使用甲烷和氨试剂气体、氦载气和DB-5MS(30 m，0.25-µm膜厚)毛细管柱。三种化合物的线性范围均为50 ng/mg头发(r=0.99)，定量限实验确定LAAM为0.5 ng/mg，NorLAAM和DINOR/AAM为0.3 ng/mg。当头发浓度分别为0.5、5.0和25.0 ng/mg时，三种分析物的批内精密度在1.0%到10.5%之间。批间精密度为4.7%~12.9%。该方法还用于检测和定量测定大鼠(n=6)毛发中LAAM、NorLAAM和dinorLAAM的含量。发色毛发中分别检出1.27 ng/mg(±0.04)、1.28 ng/mg(±0.014)和2.89 ng/mg(±0.014)的LAAM、NorLAAM和DinorLAAM。然后评估了五种实验室洗涤剂对大鼠头发中LAAM和代谢物的测量浓度的影响。使用磷酸盐缓冲液和1%的十二烷基硫酸钠洗发液可以将测得的LAAM、NorLAAM和dinorLAAM浓度降低至少30%，这表明在实验室的洗发过程中会去除(提取)头发中含有的药物。使用甲醇、二氯甲烷或水的洗涤程序将测量的浓度降低了不超过20%。由于头发中测得的LAAM、NorLAAM和dinorLAAM浓度似乎取决于实验室使用的特定洗涤程序，因此必须根据样品前处理条件谨慎地解释定量数据。
A sensitive and specific method was developed for the quantitative analysis of I-α-acetylmethadol (LAAM), I-α-acetyl-N-normethadol (norLAAM), and I-α-acetyl-N,N-dinormethadol (dinorLAAM) in hair. In the development of this method, it was determined that sample pretreatment methods performed by the laboratory greatly affect the measured concentrations of drug and metabolite in hair. Deuterated internal standards were added to 20-mg hair samples and the samples digested overnight in a buffered solution of Protease Type VIII enzyme. Digests were extracted by modification of a liquid-liquid extraction procedure developed previously in our laboratory for the analysis of plasma and tissues. Derivatized extracts were analyzed on a Finnigan MAT®4500 mass spectrometer in positive ion chemical ionization mode using methane and ammonia reagent gases, helium carrier gas, and a DB-5MS (30 m, 0.25-µm film thickness) capillary column. The assay was linear to 50 ng/mg hair (r= 0.99) for all three compounds with a limit of quantitation experimentally determined to be 0.5 ng/mg for LAAM and 0.3 ng/mg for norLAAM and dinor/AAM. Intra-assay precision ranged from 1.0 to 10.5% for the three analytes at concentrations of 0.5, 5.0, and 25.0 ng/mg of hair. Interassay precision ranged from 4.7 to 12.9%. The performance of the method was also evaluated for its utility in detecting and quantitating LAAM, norLAAM, and dinorLAAM in hair from rats (n= 6) that had been administered 3 mg/kg LAAM intraperitoneally once daily for five days. LAAM, norLAAM and dinorLAAM were detectable in pigmented hair at concentrations of 1.27 ng/mg (± 0.04), 1.28 ng/mg (± 0.014), and 2.89 ng/mg (± 0.014), respectively. Five laboratory wash solvents were then evaluated for their effect on the measured concentration of LAAM and metabolites in the rat hair. Phosphate buffer and 1% SDS washes substantially reduced the measured LAAM, norLAAM, and dinorLAAM concentrations by at least 30%, which suggests that drug incorporated into hair is removed (extracted) during the laboratory wash procedures. Wash procedures using methanol, methylene chloride, or water reduced the measured concentrations by no more than 20%. Because measured concentrations of LAAM, norLAAM, and dinorLAAM in hair appear to depend on the specific wash procedures used by a laboratory, quantitative data must be interpreted cautiously based on the sample pretreatment conditions.