Gaillard Y, Vayssette F, Pépin G
Laboratoire TOXLAB, Paris, France.
Forensic Sci Int. 2000 Jan 10;107(1-3):361-79. doi: 10.1016/s0379-0738(99)00179-6.
In France during a famous bicycle race, the newspapers documented the degree in which doping seemed to be supervised in some teams by managers and doctors. Use of anabolic steroids and other substances was officially banned in the mid-seventies by sports authorities. This policy has been enforced through urine testing before competition. It is well known, however, that a latency period is all that is necessary to defeat these tests. Nevertheless, hair analysis could be a promising tool when testing for periods that are not accessible to urinalysis any more. We have developed different sensitive methods for testing hair for amphetamines, anabolic steroids and their esters and corticosteroids. For amphetamines, 50 mg of hair were digested with 1 M NaOH, extracted with ethyl acetate, derivatized with TFA and analyzed by gas chromatography positive chemical-ionization mass spectrometry. For corticosteroids, 50 mg of powdered hair were treated with methanol in an ultrasonic bath and subsequently purified using a C18 solid phase extraction column. Analysis was realized by high performance liquid chromatography coupled to electrospray-ionization tandem mass spectrometry. For anabolic steroids and their esters, 100 mg of powdered hair were treated with methanol in an ultrasonic bath for extraction of esters, then alkaline digested with 1 M NaOH for an optimum recovery of other drugs. The two liquid preparations were subsequently extracted with ethyl acetate, pooled, then finally highly purified using a twin solid phase extraction on aminopropyl and silica cartridges. Residue was derivatized with MSTFA prior to injection. Analysis was conducted by gas chromatography coupled to a triple quadrupole mass spectrometer. Thirty cyclists were sampled and tested both in hair and in urine. Amphetamine was detected 10 times in hair (out of 19 analyses) compared to 6 times in urine (out of 30 analyses). Corticosteroids were detected 5 times in hair (methylprednisolone 1 case, triamcinolone acetonide 3 cases and hydrocortisone acetate 1 case) in hair (out of 12 analyses) compared to 12 times (triamcinolone acetonide 10 cases and betamethasone 2 cases) in urine (out of 30 analyses). Anabolic steroids were detected twice (nandrolone 1 case, and testosterone undecanoate 1 case) in hair (out of 25 analyses) compared to none in urine (out of 30 analyses).
在法国一场著名的自行车比赛期间,报纸记录了一些车队中经理和医生对兴奋剂监管的程度。合成代谢类固醇和其他物质的使用在七十年代中期被体育当局正式禁止。这项政策通过比赛前的尿液检测来执行。然而,众所周知,只需一段潜伏期就能通过这些检测。不过,当检测尿液分析无法触及的时间段时,毛发分析可能是一种有前景的工具。我们开发了不同的灵敏方法来检测毛发中的苯丙胺、合成代谢类固醇及其酯类和皮质类固醇。对于苯丙胺,50毫克毛发用1摩尔/升氢氧化钠消化,用乙酸乙酯萃取,用三氟乙酸衍生化,然后通过气相色谱正化学电离质谱法分析。对于皮质类固醇,50毫克毛发粉末在超声浴中用甲醇处理,随后使用C18固相萃取柱进行纯化。通过高效液相色谱-电喷雾电离串联质谱法进行分析。对于合成代谢类固醇及其酯类,100毫克毛发粉末在超声浴中用甲醇处理以萃取酯类,然后用1摩尔/升氢氧化钠进行碱性消化以实现其他药物的最佳回收率。两种液体制剂随后用乙酸乙酯萃取,合并,最后使用氨基丙基和硅胶柱进行双固相萃取进行高度纯化。残留物在进样前用N-甲基-N-(三甲基硅基)三氟乙酰胺衍生化。通过气相色谱-三重四极杆质谱仪进行分析。对30名自行车运动员进行了毛发和尿液采样检测。毛发中检测到苯丙胺10次(共19次分析),而尿液中检测到6次(共30次分析)。毛发中检测到皮质类固醇5次(甲基泼尼松龙1例、曲安奈德3例和醋酸氢化可的松1例)(共12次分析),而尿液中检测到12次(曲安奈德10例和倍他米松2例)(共30次分析)。毛发中检测到合成代谢类固醇2次(诺龙1例和十一酸睾酮1例)(共25次分析),而尿液中未检测到(共30次分析)。