Analysis of anabolic steroids in hair: Time courses in guinea pigs

Sensitive, specific, and reproducible methods for the quantitative determination of eight anabolic steroids in guinea pig hair have been developed using LC/MS/MS and GC/MS/MS. Methyltestosterone, stanozolol, methandienone, nandrolone, trenbolone, boldenone, methenolone and DHEA were administered intraperitoneally in guinea pigs. After the first injection, black hair segments were collected on shaved areas of skin. The analysis of these segments revealed the distribution of anabolic steroids in the guinea pig hair. The major components in hair are the parent anabolic steroids. The time courses of the concentrations of the steroids in hair (except methenolone, which does not deposit in hair) demonstrated that the peak concentrations were reached on days 2-4, except stanozolol, which peaked on day 10 after administration. The concentrations in hair appeared to be related to the physicochemical properties of the drug compound and to the dosage. These studies on the distribution of drugs in the hair shaft and on the time course of their concentration changes provide information relevant to the optimal time and method of collecting hair samples. Such studies also provide basic data that will be useful in the application of hair analysis in the control of doping and in the interpretation of results.     

Simultaneous determination of dimethylamphetamine and its metabolites in rat hair by gas chromatography–mass spectrometry

In order to study the disposition of dimethylamphetamine (DMAP) and its metabolites, DMAP N-oxide, methamphetamine (MA) and amphetamine (AP), from plasma to hair in rats, a simultaneous determination method for these compounds in biological samples using gas chromatography–mass spectrometry with selected ion monitoring (GC–MS-SIM) was developed. As DMAP N-oxide partially degrades to DMAP and MA during GC–MS analysis, it was necessary to avoid conditions which co-extract the N-oxide in the sample preparation so as to assure no contribution of artifactual products from DMAP N-oxide in the detection of the other compounds. For confirmation of the satisfactory separation of DMAP N-oxide from the others, the internal standards used for quantification were labeled with different numbers of deuterium atoms. Determination of unchanged DMAP was performed without any derivatization, that of DMAP N-oxide was carried out after conversion into trifluoroacetyl-MA by reaction with trifluoroacetic anhydride, and MA and AP were quantified after trifluoroacetyl-derivatization.After intraperitoneal administration of DMAP HCl to pigmented hairy rats (5 mg kg⁻¹ day⁻¹, 10 days, n=3), concentrations of DMAP and its metabolites in urine, plasma and hair were measured by GC–MS-SIM. The area under the concentration versus time curves (AUCs) of DMAP, DMAP N-oxide, MA and AP in the plasma were 397.2±97.5, 279.7±68.3, 18.4±1.2 and 15.9±2.2 μg min ml⁻¹, while their concentrations in the hair newly grown for 4 weeks after administration were 4.82±0.67. 0.45±0.09, 3.25±0.36 and 0.89±0.05 ng mg⁻¹, respectively. This fact suggested that the incorporation tendency of DMAP N-oxide from plasma into hair was distinctly low in comparison with the other compounds.     

Hair analysis for drugs of abuse XXI. Effect of para-substituents on benzene ring of methamphetamine on drug incorporation into rat hair

In order to study the effect of para-substituents on the benzene ring of methamphetamine on drug incorporation into hair from blood, the plasma AUCs and hair concentrations of 7 methamphetamines [methamphetamine(MA), p-hydroxymethamphetamine(OHMA), p-bromomethamphetamine (BMA), p-aminomethamphetamine (AMA), p-nitromethamphetamine (NMA), p-methoxymethamphetamine (MOMA) and 3,4-methylenedioxymethamphetamine (MDMA)] plus propylhexedrine(PHX) in DA rats was determined after intraperitoneal injection at 5 mg/kg, with single dose for the plasma AUC and 10 doses for the hair concentration. Drug incorporation rates into hair (ICRs) were calculated by dividing each hair concentration by each plasma AUC. Comparing the highest value (NMA) to the lowest one (OHMA), the ICR of NMA was 31.7 times larger than that of OHMA. Using the ICR of MA which has no substitute on the benzene ring as a base point, nitro, bromo, methylenedioxy, methoxy and amino groups raised the drug incorporation into rat hair in this order. On the other hand, hydroxy substitution showed a negative effect on the ICR. In comparison between the ICRs of MA and PHX, it was found that the benzene ring shows higher affinity to melanin and less lipophilicity than the cyclohexyl ring. Our results showed that there is a relatively strong effect of the functional groups on drug incorporation into hair. The combination of melanin affinity and lipophilicity are clearly correlated with their ICR.     

Determination of oxcarbazepine and its metabolites in postmortem blood and hair by means of liquid chromatography with mass detection (HPLC/APCI/MS)

A typical use of hair analysis in forensic toxicology is the documentation of previous drug administration. This is illustrated in a suicidal death of a 58-year-old epileptic patient who was treated with oxcarbazepine and probably with levomepromazine. The toxicological analysis carried out by HPLC/APCI/MS included also the hair (6 cm length) besides postmortem blood. The method was validated for levomepromazine, oxcarbazepine (OXCBZ) and its two metabolites, 10-hydroxycarbazepine (CBZ-10OH) and trans-diol-carbazepine (CBZ-diOH) in various biological matrices. The analysis of the postmortem blood indicated oxcarbazepine and its two main metabolites were present at therapeutic concentrations; levomepromazine was detected at a fatal concentration. In three 2-cm segments of hair, oxcarbazepine and its two metabolites were detected; however, levomepromazine was not detected in this specimen. As a result of complex chemical-toxicological investigation it was confirmed the information that the decedent. was an epileptic patient and was treated with oxcarbazepine for at least 6 months before death. In addition, he took a toxic dose of levomepromazine in order to commit suicide. The analysis revealed differences between the concentration levels of oxcarbazepine and its active metabolite CBZ-10OH in postmortem specimens and hair, suggesting different mechanisms of penetration of metabolites and their precursors into this matrix.     

Effect of N-methyl-N′-Nitro-N-nitrosoguanidine on amino acid incorporation into mucosal protein of rat stomach

Administration of the carcinogenic N-nitroso compound, N-methyl-N′-nitro-N-nitrosoguanidine in drinking water (0.5 mg/mL) to male Wistar rats for 1 week caused impairment of in vivo and in vitro incorporation of [¹⁴C]leucine into stomach mucosal protein. This impairment gradually returned to normal after 4 weeks. Uptake of [¹⁴C]leucine into mucosal protein was significantly inhibited after in vitro treatment of stomach mucosa with the carcinogen. Addition of the N-nitroso compound in a cell-free system using postmitochondrial supernatant prepared from stomach mucosa also showed inhibition of amino acid incorporation. Using a more defined system consisting of purified polyribosome from stomach mucosa and pH 5 enzyme fraction derived from liver it was further demonstrated that the carcinogen purturbed protein synthesizing ability of polyribosome, under both in vivo and in vitro treatment conditions. In these respects this carcinogen has similar action on the target tissue of stomach as in the liver, although the in vivo effect may be related more to toxicity than carcinogenicity.     

Study of the age and sex dependence of trace elements in hair by correspondence analysis

The aim of the study was to examine the potential of multidimensional analysis, and in particular of correspondence analysis (CA), in bringing to light the influence of sex and age on trace element (TE) concentrations in hair from an unselected French population. Sixteen elements (S, Hg, Se, Zn, Pb, Cd, Ni, Co, Mn, Fe, Cr, Mg, Al, Ca, Cu, Ag) were assayed by inductively coupled argon plasma (ICAP) emission specrroscopy in the scalp hair of 135 men and 346 women. In spite of the high background noise, CA was able to reveal the differing patterns in males and females. For instance, in this population, higher relative levels of the essential elements, Ca, Mg, Zn, and Cu, but also of Ag, characterized women’s hair, whereas higher relative levels of the heavy metals, Fe and Pb, were associated with men’s hair. Al and Ag were unexplainedly high in the hair of the youngest members of the population. The Cu and Co of youth seemed to give way to a predominance of Zn in maturity. The hair of individuals in their forties tended to be richest in Ca and Mg, but these elements decreased with advancing age. Heavy metals (Hg, Pb, Fe) accumulated with age, whereas Se, Mn, and Cr seemed independent of age. CA is manifestly a very useful tool for revealing underlying dimensions in complex dynamic systems and unsuspected relationships among variables. Clearly, the significance of the high Al and Ag contents in the hair of certain members of the population, especially of the very young, needs to be investigated from both physiological and toxicological aspects.     

The preventive effect of vitamin D3 on radiation-induced hair toxicity in a rat model

Our aim is to investigate the protective effect of vitamin D3 especially from radiation-induced hair toxicity. A model of skin radiation injury was developed and a single fraction of 20 Gy Gamma irradiation was applied to the right dorsal skin of fourteen rats. All animals were randomly divided into 2 groups: Group I: irradiation alone (n = 7) and Group II: irradiation and 0.2 microg vitamin D3 given IM (n = 7). Fifty days after post-irradiation rats were sacrificed. The outcomes were evaluated on the basis of histopathological findings and immunohistochemical staining for Vitamin D receptor (VDR) in skin and hair follicles. The number of hair follicles in the radiation field for the group of animals irradiated without pretreatment was significantly lower than outside of the irradiated area (p = 0.016) as it is expected. Contrarily the number of hair follicles did not show significant difference in the pretreated group between the irradiated field and outside of the fields (p = 0,14). Skin of the vitamin D3 pretreated group demonstrated stronger immunoreactivity for VDR compared to irradiation alone group. These results indicate that administration of vitamin D3 may protect hair follicles from radiation toxicity. Further clinical trials should be conducted to prove the preventive effect of vitamin D3 as well as dosing and timing of the agent on radiation-induced alopecia.     

电喷雾离子化/质谱法(ESI/MS)及其在生物大分子研究中的应用

电喷雾离子化／质谱法（ESI／MS）在各种有机化合物、多肽、蛋白质（含糖蛋白）、核苷酸、糖、脂类及合成高分子物质等分析领域获得了广泛的应用,本文系统介绍了ESI／MS的基本原理,其联用技术,及其在生物大分子研究,包括肽图谱测定,糖分析和核苷酸分析中的应用。     

桐油脂肪酸组成分析和甘三酯结构判定

采用2-氨基-2-甲基丙醇（2-amino-2-methylpropanol,AMP）衍生化、GC／MS法分析桐油的脂肪酸组成：软脂酸3．41％,硬脂酸3．71％,油酸7．07％,亚油酸7．51％,亚麻酸1．31％,十八碳共轭三烯酸73．19％,未定出成分3．80％;采用RP—HPLC／APCI—MS法分离桐油中的甘三酯组分,并根据特定甘三酯断裂生成的特征甘二酯离子的丰度比初步判定主要甘三酯的结构。     

优良菌种发酵葡萄酒优化条件的探讨

以诱变耐低温果酒酵母菌种YU2.28和产香酵母S15.3为发酵菌株,进行了葡萄酒发酵条件优化的试验研究.探讨了菌种生长温度、通氧量等因素,通过对菌种的生长情况和发酵醪液中总酯含量的变化分析,确定了自选酵母酿制葡萄酒的最佳技术参数,并对优化条件下发酵得到的葡萄酒进行GC/MS分析.结果显示：YU2.28和S15.3以1：3比例的混合发酵,接种量3%,调节醪液pH值为4.0,SO2添加量40 mg/L,发酵温度20℃,主发酵6 d内控制以230r/min的摇床转速进行摇瓶发酵,并进行9 h（每天1.5 h）供氧处理,后发酵30 d,酿造出的葡萄酒品质较佳,具有酒体丰盈,酒液澄清透亮,香气醇和的特征.成品酒香气成分共检测出醇类9种,酯类8种,酸类6种和少量的醛类、酮类等成分.     

The effect of temperature,irradiance and animal size on incorporation rates of Simocephalus vetulus

Carbon incorporation rates of Simocephalus vetulus were measured to study the effects of the physical state of the animals, size of the animal, varying temperature and light conditions. Physical state of the animal showed little effect on incorporation rates after the first hour. Incorporation rates increased in proportion to the third power of animal size. Experimental animals collected at temperatures of 12, 20 or 25°C fed maximally at 10, 15 and 25°C respectively, when subjected to a feeding temperature range of 5 to 30°C. We have interpreted this as an indication that S. vetulus is able to acclimate and incorporate maximally at various temperatures after prolonged exposure to that temperature. When fed over an irradiation range of 0 to 4.68 × 10^–3 cal cm^–2 s^–1 incorporation rates were indirectly proportional to irradiance. This suggests a response to decreased irradiance in the weedy, littoral habitat of these animals.     

The effects of methamphetamine on serotonin transporter activity: role of dopamine and hyperthermia

Multiple administrations of methamphetamine (METH) rapidly decreased serotonin (5HT) transporter (SERT) function in rat striatum and hippocampus. The purpose of this study was to identify the mechanisms/ factors contributing to this METH-induced decrease in SERT function. Multiple high-dose METH injections rapidly decreased 5HT uptake without altering binding of the 5HT transporter ligand paroxetine. Hyperthermia contributed to this deficit in transporter function in striatum and hippocampus, as prevention of METH-induced hyperthermia attenuated this decrease. A role for dopamine (DA) was suggested by findings that pretreatment with the tyrosine hydroxylase inhibitor alpha-methyl-p-tyrosine, the D1 antagonist SCH-23390, or the D2 antagonist eticlopride attenuated the METH-induced decrease in striatal, but not hippocampal, SERT activity. These effects were independent of the ability of these DA-antagonizing drugs to prevent METH-induced hyperthermia. These results suggest that DA contributes to the decrease in SERT function caused by multiple METH injections in the striatum, but not hippocampus, and that hyperthermia facilitates these deficits in SERT function in both brain regions. In contrast, the response of SERT to a single administration of METH was DA and hyperthermia independent. These findings suggest that the mechanisms/ factors involved in decreasing SERT activity after a single administration of METH are distinct from that caused by multiple administrations.     

Gibberellin A58 and ent-6α,7α,12α-trihydroxykaur-16-en-19-oic acid from seeds of cucurbita maxima

The isolation of gibberellin A₅₈ and ent-6α,7α,12α-trihydroxykaurenoic acid from a cellulase-hydrolysed extract of endosperm ofCucurbita maxima is described. The two compounds are characterized by their MS,¹H NMR and ¹³C NMR.     

The first report on the in vitro antimicrobial activities of extracts of leaves of Ehretia serrata

The increasing resistance of infectious microbial organisms to the existing arsenal of antibiotic drugs is on the rise. There is a growing demand for the new antibiotics that are cost effective and easily available to the common people. In search of new antimicrobial entities, this report deals with the in vitro antimicrobial activities of the crude extracts of leaves of Ehretia Serrata. The methanolic extract and its sub-fractions namely n-hexane, chloroform, ethyl acetate, n-butanol and residual water fraction were screened against a range of 30 different bacterial strains and their zones of inhibition (ZI) and minimum inhibitory concentration (MIC) were subsequently evaluated. Methanolic extract has shown activity against all the tested microorganisms such as Azospirillum lipoferum, Escherichia coli, Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Enterococcus sp. with ZOI ranged from 10.3 to 29.0 mm. Moreover, the MIC values of methanolic extract and its sub-fractions against the tested bacteria ranged from 0.8 to 5.1 mg/mL. GC–MS analysis of sub-fractions revealed the presence of mono (2-ethylexyl) 1,2-benzenedicarboxylate, diisooctyl-1,2-benzenedicarboxylate, 3,5-dehydro-6-methoxypivalate-cholest-22-ene-21-ol, and 3,5-bis (1,1-dimethylethyl)-4-hydroxybenzene-propanoic acid. This is the first report on the in vitro antimicrobial activities of leaves of E. Serrata.     

A simple and rapid GC/MS method for the simultaneous determination of gaseous metabolites

We modified and tuned a commercial model of a gas chromatography/mass spectrometry (GC/MS) instrument to develop a simple and rapid method for the simultaneous quantification of a variety of gas species. Using the developed method with the newly modified instrument, gas species such as H₂, N₂, O₂, CO, NO, CH₄, CO₂, and N₂O, which are common components of microbial metabolism, were accurately identified based on their retention times and/or mass-to-charge ratios (m/z) in less than 2.5 min. By examining the sensitivities and dynamic ranges for the detection of H₂, N₂, O₂, CH₄, CO₂, and N₂O, it was demonstrated that the method developed in this study was sufficient for accurately monitoring the production and the consumption of these gaseous species during microbial metabolism. The utility of the new method was demonstrated by a denitrification study with Pseudomonas aureofaciens ATCC 13985^T. This method will be suitable for a variety of applications requiring the identification of gaseous metabolites in microorganisms, microbial communities, and natural ecosystems.     

Identification by GC-EAD of the two-component trail-following pheromone of Prorhinotermes simplex (Isoptera, Rhinotermitidae, Prorhinotermitinae)

GC/MS analysis confirmed that neocembrene is the major component of the trail pheromone in the three species of the termite genus Prorhinotermes (P. simplex, P. canalifrons, P. inopinatus). In addition, EAG and GC-EAD experiments with P. simplex strongly suggest that dodecatrienol is a quantitatively minor component but a qualitatively important component of this trail pheromone. Trail-following bioassays confirmed the two-component nature of the trail pheromone. This is the first report of the use of the GC-EAD for the identification of trail pheromone in termites. These original results underline once again the special phylogenetic status of the Prorhinotermitinae among Rhinotermitidae.     

Metabolic deposition of selenium and cadmium into the hair and other tissues of the guinea pig

To better understand the significance of hair trace-element measurements and their relationships with the trace-element levels in body organs and fluids, a series of controlled animal experiments were conducted in which several trace elements were periodically measured during a 90-day chronic exposure to selenium and cadmium. Chronic selenium exposure appeared to be reflected by elevated selenium levels in the hair, kidneys, and liver. Chronic cadmium exposure, although reflected by kidney and liver elevation, appeared not to be reflected by corresponding increases in its concentration in the hair.     

Qualitative and semi-quantitative analysis of gibberellins

A method for the qualitative and semi-quantitative analysis of gibberellins (GAs) was examined, and a systematic method consisting of six steps was established. By this method endogenous GAs in some organs of Pharbitis nil were quantitatively analysed.     

The use of hair morphology in the classification of galagos (Primates, subfamily galagoninae)

The outer epidermal hair structure of bushbabies or galagos varies considerably between genera and species. Comparative morphology can therefore be useful in taxonomic studies, particularly when assessing the true status of newly nominated or misclassified species. Quantitative measurements of outer epidermal hair scales (scale frequency and scale complexity) were made for populations representing 13 taxa within the subfamily Galagoninae. Intraspecific variations in both measures of hair morphology were relatively minor. By contrast, significant interspecific differences were observed, even between closely related, well-established species (e.g. in the greater galagosOtolemur crassicaudatus andO. garnettii and lesser galagosGalago senegalensis andGalago moholi). Assessment of scale frequency resulted in the recognition of all species of the greater and needle-clawed galago genera and added support for nomination to species status of two former subspecies (O. argentatus andE. pallidus). Similar divisions were made in the lesser and dwarf galagos with respect to scale complexity. Possible explanations for the differences in outer epidermal hair scales are discussed. Characteristics of outer epidermal hair scales provide a useful guide to species identity in the subfamily Galagoninae, and the same may prove to be true for other groups of nocturnal mammals.