L. Pötsch

On cosmetically treated hair — aspects and pitfalls of interpretation

Popular hair cosmetic treatments like bleaching or permanent waving were found to affect the stability of incorporated drugs and to cause alterations of the fibers at an ultrastructural level. This may result in a partial or complete loss of drug substances, depending on the particular drug molecule and on its concentration prior to the cosmetic treatment. Moreover, from literature, there is some evidence that drug molecules are not only incorporated into the growing fiber by passive diffusion from blood into the matrix cells and melanocytes, but that the substances enter the hair also via perspiration such as sweat and sebum. Since permed and bleached hair shows an enhanced sorption capacity, the risk of false positives or an unusually high drug concentration in cosmetically treated hair was under investigation. Virgin, permed, mildly as well as severely bleached tresses were exposed to artificial sweat or sebum containing cocaine, benzoylecgonine, 6-acetylmorphine, morphine and codeine (500 ng/g). Except codeine, the concentrations measured by GC/MS were very small and quite close to the detection limit indicating a minor importance of drug uptake into hair fiber from the endogenous-exogenous shunt via sebum or sweat. From the results it is concluded that an increased risk of false positive results in hair analysis on bleached and permanent waved hair fibers does exist, but is not particularly severe.

Biochemical approach on the conservation of drug molecules during hair fiber formation

A biochemical concept for the endogenous incorporation of drug molecules into growing hair is presented. It is based on the principles of transport across biomembranes, on the principles of biotransformation and drug melanin affinity. The approach gives explanations for current observations in hair analysis, which up to date have not been understood sufficiently. Phenomena such as the ratio of parent drug to metabolite in hair, the dependence of incorporation on the physico-chemical properties of the drug, the independence of drug incorporation on active melanogenesis (incorporation into non-pigmented hair) as well as the dependence of drug content on hair pigmentation are elucidated.

Influence of pigmentation on the codeine content of hair fibers in guinea pigs.

Tortoise shell guinea pigs (n = 7) were administered codeine (1 mg/mL codeine-base) in their drinking water for 3 weeks. Black, reddish-brown and white hair was collected separately from each animal before and after treatment. The hair samples were analyzed by GC/MS. The experiment showed positive results for all hair fibers with large individual variability of drug incorporation. Low drug intake resulted in small differences of the drug content in hair fibers different in color, whereas in cases of high drug intake a strong influence of hair pigmentation on the analytical results was observed. The highest drug content was always found in black hair samples, non-pigmented hair showed the lowest drug concentrations and the drug content in reddish-brown fibers was less than in black hair samples from the same animal. From the results it was concluded, that eumelanins rather than phenomelanins are the decisive factor for codeine-melanin binding in hair and the amount of drug intake was suggested to determine the relevance of hair pigmentation on the analytical results.

Stability of opiates in hair fibers after exposure to cosmetic treatment

The stability of opiates in clipped natural human hair was investigated. Hair fibers were incubated with defined solutions of morphine, codeine and dihydrocodeine (pH 7.4) until saturated. Original opiate-positive hair samples collected from drug addicts also were examined. Commercially available bleaching as well as perming formulas (Poly Blonde Ultra, Poly Lock; Henkel, Düsseldorf, Germany) were applied in vitro to the hair strands of both groups under investigation. After these treatments, the drug concentration had decreased for both bleaching and permanent waving. In the spiked hair, only 2-18% of the starting solution could be found after bleaching. About 20-30% of the drug substances could still be detected after perming. In the authentic hair samples, the drug levels of the formerly opiate positive hair fibers had also been reduced but distinct tendencies could not be observed.

Partition coefficient, blood to plasma ratio, protein binding and short-term stability of 11-nor-Δ9-carboxy tetrahydrocannabinol glucuronide

11-Nor-Delta(9)-carboxy tetrahydrocannabinol glucuronide (THCCOOglu) is a major metabolite of tetrahydrocannabinol in blood. Despite its mass spectrometric identification already in 1980, further physicochemical data of THCCOOglu have not been established. Therefore, the octanol/buffer partition coefficient P and the blood to plasma ratio b/p for THCCOOglu concentrations of 100 and 500ng/ml were investigated. Protein binding of the glucuronide was established from spiked albumin solutions at a level of 250ng/ml as well as from authentic samples. The data were compared to those of 11-nor-Delta(9)-carboxy tetrahydrocannabinol (THCCOOH). In addition, the short-term stability of THCCOOglu in plasma at different storage temperatures was studied. Analysis was performed by LC/MS/MS. The glucuronide partition coefficient P (mean: 17.4 and 18.0 for 100 and 500ng/ml, respectively) was unexpectedly lipophilic at pH 7.4. Its blood to plasma ratios averaged 0.62 and 0.68 at 100 and 500ng/ml, respectively. THCCOOglu was highly reversibly bound to albumin (mean: 97%), and the mean fraction bound did not differ from that determined from authentic samples. THCCOOglu degraded even at a storage temperature of 4 degrees C and THCCOOH was identified as a major decomposition product.

On Pathways for Small Molecules Into and Out of Human Hair Fibers

This paper represents an experimental approach of histology of the human hair fiber in dyeing and diffusion phenomena and its contribution to the interpretation of hair analysis results for drug abuse. Rhodamine B was applied to human hair fibers from either aqueous solution or methanol/ethanol solvent. The experiments were performed on natural hair of different ethnic groups as well as on extensively bleached hair strands. The microscopical study of the pathway of diffusion of rhodamine B into the hair fibers indicated that the reagent had entered the unmodified fibers at the scale edges between the cuticle cells. At the beginning of the diffusion process intercellular diffusion was the preferred route predominantly along the nonkeratinous regions of the cell membrane complex (CMC) and intermacrofibrillar cement. Penetration into the high sulfur regions of the fiber occurred as dyeing in aqueous solution proceeded and resulted in evenly stained cross sections. The dye distribution pattern observed in natural hair exposed to nonaqueous solution showed that rhodamine B did not penetrate the cortex cells as easy as from aqueous solution and selectively stained nonkeratinous regions only. The determination of the amount of dye taken up by the fibers by spectrophotometric analysis demonstrated that samples diffusion generally increased by time and temperature. It also depended on the morphology of the hair sample. The penetration of rhodamine B from aqueous solution was much greater than from methanol/ethanol solvent.

In vitro stability of cocaine in whole blood and plasma including ecgonine as a target analyte

The in vitro stability of cocaine (COC) was monitored in fresh whole blood and plasma stabilized with potassium fluoride (0.25%) for as long as 15 days. The samples were stored at 4°C, 20°C and 40°C. Additionally, fresh plasma samples containing either benzoylecgonine (BZE), ecgonine methyl ester (EME) or ecgonine (ECG) were stored at 4°C and 20°C. Data were established using subsequent solid-phase extraction procedures and high-performance liquid chromatography coupled to atmospheric pressure ionization mass spectrometry for isolation and quantitation of COC, BZE, EME, and ECG. COC, BZE, and EME concentrations decreased with increasing storage temperature and time after an apparent first-order reaction kinetic. Only ECG appeared to be stable at storage temperatures as high as 20°C for the entire observation period. At 40°C, the amount of ECG produced from hydrolysis of COC still totalled 80% of the initial COC concentration. Hydrolysis of COC to EME occurred more rapidly in plasma than in blood. The dynamic degradation profiles obtained were dependent on the storage temperature. The conversion of COC to BZE, EME, and ECG appeared to be stoichiometric at all time intervals at storage temperatures of 4°C and 20°C. The presence of any hydrolysis product of COC in blood or plasma constitutes confirmatory evidence of COC incorporation, and determination of ECG seems most promising even in samples stored under unfavorable conditions.

A case report on drug screening of Nail clippings to detect prenatal drug exposure

In a case of a sudden infant death syndrome-related death of a 3-month-old infant, nail clippings were positive for cocaine by gas chromatography-mass spectroscopy analysis that revealed a prenatal exposure to the drug substance. In utero exposure to drugs has been investigated using amniotic fluid, neonatal urine, meconium, and hair samples. Nail analysis offers some advantage over hair analysis because of its continuous growth and persistence after delivery. Nail material is easy to sample in suitable amounts. Currently, the cocaine finding cannot be related to the underlying cause of death. However, this observation indicates that nail analysis may be a new and valuable tool to screen newborns for intrauterine drug exposure. In addition, it can help collect information on the prevalence of possible embryotoxic effects and the link to postnatal manifestations of different dysfunctions in infants who are born by drug abusing mothers.

Perspiration versus saliva – basic aspects concerning their use in roadside drug testing

Abstract Various aspects concerning the practical application and forensic interpretation of data obtained by saliva drug testing and drug monitoring from the skin surface are discussed. Basic information on the composition of saliva and skin secretions and their particular transport mechanisms, as far as known, are given. For drugs of abuse secretion into saliva is suggested to be by passive diffusion and to depend on lipid solubility, pKa, plasma protein binding and on the pH of saliva. Drug molecules from blood are considered to reach the skin surface by various routes such as by sweat and sebum as well as by inter- and/or transcellular diffusion. The role of the stratum corneum as a temporary drug reservoir exceeding positive drug findings in urine is outlined. Current data on opioids, cocaine metabolites, cannabinoids and amphetamines detected in saliva and on the skin surface are reviewed. Aspects of collection, processing and analysis of the samples for implementation in roadside testing are addressed. The requirement of test sensitivity covering the broad concentration ranges and the importance of test specificity bearing in mind that the parent drug is the main analyte present in those specimens is stressed. Theoretical and practical findings on frequently abused drugs are discussed with regard to the possibilities and limitations of drug monitoring from saliva and perspiration to support a suspicion of actual or recent drug administration.

A preliminary study on the stability of benzodiazepines in blood and plasma stored at 4° C

Abstract An approach to determine the stability of benzodiazepines and some of their metabolites (n = 13) by means of a routinely applied gas chromatographic method using electron capture detection was made in this preliminary study. Validation data of the method are given. Spiked blood and plasma samples were stored at 4° C and analysed at selected times up to 240 days. The concentrations of all analytes had decreased to at least 60% of the original levels at the end of the observation period. A clear pattern of breakdown could not be established. The data obtained suggest that results from long-term stored samples should be interpreted cautiously. Further investigations concerning the stability of drugs in blood and plasma samples, additional methods of identification and determination as well as the establishment of optimal storage conditions seem necessary.