Drug-facilitated Sexual Assault Research Articles

Detection of nitrated benzodiazepines by indirect laser-induced fluorescence detection on a microfluidic device

Recently there has been concern regarding the use of flunitrazepam and other low-dose benzodiazepines in drug-facilitated sexual assault. These compounds are placed in drinks of unsuspecting victims and produce a sedative effect with anterorgrade amnesia. Chip-based microfluidic systems can provide a quick and disposable procedure for the detection of flunitrazepam and other nitrated benzodiazepines used in these crimes. This paper describes the application of indirect quenching of cyanine dye (Cy5) for detection of nitrated benzodiazepines. The separation is performed on a microfluidic device with a separation channel 8 cm long and 50 μm wide and utilizes indirect fluorescence detection with 635 nm laser excitation. The optimization of the separation using micellar electrokinetic chromatography with organic modifiers is described. A borate buffer containing 2.6 μM Cy5 dye, 15 mM sodium dodecyl sulphate (SDS) and 20% methanol is used. Complete separation of four target drugs occurs in under 2 min with limits of detection in the low μg/ml range. Overall the method provides a rapid and simple analysis for the presence of nitrated benzodiazepines in beverages and other similar preparations.

A Simple and Rapid Method for the Identification of Zolpidem Carboxylic Acid in Urine

Zolpidem is a non-benzodiazepine hypnotic that has been implicated in both drug-facilitated sexual assault and drink spiking. Detection of the drug in urine is extremely difficult because of its extensive metabolism. A method is presented for the detection and quantitation of zolpidem carboxylic acid (ZCA), the major urinary metabolite of zolpidem. The metabolite was extracted from urine at pH 4.5-5.0 with chloroform/isopropanol alkylated with ethyl iodide and identified by gas chromatography-mass spectrometry in the selected ion-monitoring mode. Following a single ingestion of 10 mg zopidem, ZCA is detected for up to 72 h. The limit of detection is 2 ng/mL, with an overall recovery of 80%. Using this procedure, zolpidem was identified in two cases of alleged drug-facilitated sexual assault.

Drug facilitated sexual assault using an over-the-counter ocular solution containing tetrahydrozoline (Visine ®)

Drug facilitated sexual assault (DFSA) has been defined as sexual activity occurring where consent is invalid or absent due to the effects of drugs and or alcohol. We report the use of a commonly available over-the-counter drug to induce an obtunded compliant victim with no memory of the period during the sexual assault. An adult male repeatedly used tetrahydrozoline to induce a comatose state in an adult female and four female children for the purposes of sexual assault. It may be warranted to include imidazoline derivatives in the testing for cases of DFSA.

Toxicological investigation of drug-facilitated sexual assaults

Drug-facilitated sexual assaults have drawn a lot of attention during the last years. The role of the non-forensic doctors in the early recognition and management of such cases is crucial. Diagnosis assisted by samplings must be ascertained without delay, in victims initially brought to the emergency room for a sleepy state, a suspected abuse of drug or a pseudo-alcoholic clinical state. Further, where amnesia is present, it may be prudent to assume that sexual activity has occurred and offer pregnancy and sexually transmitted disease prophylaxis. Forensic doctors should entertain the possibility of poisoning in any presentation involving amnesia. Forensic toxicological laboratories should develop sensitive and specific methods to assure a low threshold of detection of the drugs. The awareness of a delinquent or criminal use of alcohol and/or drugs could permit a better medical recognition, so that proper and timely forensic examination is achieved, and adequate evidence is collected for the establishment of an effective investigation and prosecution.

Drug-facilitated sexual assaults

Drug-facilitated sexual assaults

LC-MS-(TOF) Analysis Method for Benzodiazepines in Urine Samples from Alleged Drug-Facilitated Sexual Assault Victims

In recent years there has been an increase in the number of reports in the U.S. of the use of drugs to commit sexual assault. In 1994, a nationwide urine testing program was developed to assess the incidence of the use of drugs to facilitate sexual assault and provide information for use in the investigation of these crimes. Urine samples were collected from victims of suspected drug-induced sexual assault by law enforcement agencies, emergency rooms, and rape crisis centers. The most implicated drug class was benzodiazepines, either alone or in combination with alcohol. In this report, a procedure was developed for the screening of 22 benzodiazepines in human urine by liquid chromatography-time of flight-mass spectrometry [LC-MS-(TOF)]. The limit of quantitation for all benzodiazepines ranged from 2 to 10 ng/mL, and the limit of detection was 0.5 to 3.0 ng/mL. These results suggest that the method sensitivity is suitable to screen for all 22 benzodiazepines in human urine at low levels. The method was used to analyze samples previously reported to have screened positive for benzodiazepines by immunoassay at 50 ng/mL cut off but failed to confirm by a gas chromatography-MS method. The results of reanalysis of these samples using this LC-MS method are reported.

The ability of two commercially available quick test kits to detect drug‐facilitated sexual assault drugs in beverages

Assessment of the sensitivity and specificity of two commercially available 'drug-facilitated sexual assault' drug detector kits, Drink Guard and Drink Detective. Experimental. Laboratory. Gamma hydroxybutyrate (GHB) sodium salt, ketamine hydrochloride, temazepam, flunitrazepam and diazepam were dissolved (Tween added to benzodiazepine solutions) as separate stock solutions and added to 330 ml samples of cola (Pepsi Max), beer (Stella Artois), 'alcopop' (Bacardi Breezer) and placebo (distilled water). The doses used are reported to be common in cases of intoxication. Each kit was tested 10 times for each drink/drug combination. Two blind, independent observers scored each test (presence/absence of drug) in accordance with kit instructions; chi 2 was used to compare the proportion of times raters scored tests correctly and incorrectly. Sensitivity and specificity were calculated overall, for each drink, and sensitivity was calculated for each drug. Inter-observer agreement was evaluated using the kappa statistic. While both raters were able to score significantly more tests correctly than incorrectly using Drink Detective, and one rater scored similarly using Drink Guard, the overall sensitivity of Drink Detective and Drink Guard was 69.0% (95% CI 64.2-73.5%) and 37.5% (95% CI 30.1-45.5%), respectively. Sensitivity was drink-dependent. Drink Detective was unable to detect our dose of GHB in water, with all tests scored negatively by both raters for this drink/drug combination (n = 20 negative scores). Overall, specificity was 76.6% (95% CI 71.5-81.0%) and 87.9% (95% CI 83.0-91.6%) for Drink Guard and Drink Detective, respectively, but was affected by the beverage. Inter-rater agreement was poor for Drink Guard (kappa = 0.278 +/- 0.069) but excellent for Drink Detective (kappa = 0.894 +/- 0.245). Inter-observer agreement was drug-dependent. Use of drug detector kits by the public in the night-time environment needs further investigation and may create a false sense of security (false negatives) and undue concern (false positives) among kit users.

An Unusual Case of Drug-Facilitated Sexual Assault Using Aromatic Solvents

This report documents a case of drug-facilitated sexual assault (DFSA) under the influence of solvents. The victim was a 13-year-old female. Upon contact with law enforcement, she was still confused and could hardly explain the facts. She told authorities that she had been kidnapped 4 h previously when two individuals with covered faces put a cloth soaked in a solvent over her mouth. She spent a few hours in a room, during which she lost consciousness. The girl awakened semi-nude in the street with memory loss. No alcohol was present in the subject's body; no odor of alcohol was detected on the subject's breath. No lesions were observed during a gynecological exam. A blood sample was taken with the intent to investigate the use of chloroform or similar anesthetics. Toxicological analysis of the victim's blood revealed the presence of 7.6 mg/L of benzene, 24.8 mg/L of toluene, and 0.6 mg/L of xylene (mixture of isomers). As for other analytical findings, diazepam (0.02 mg/L) was also found. The aromatic solvents involved in this case were detected using gas chromatography with flame-ionization detection (GC-FID) and confirmed using GC-mass spectrometry (MS) in full scan mode after liquid-liquid extraction of the whole blood sample. Quantitation of the aromatic solvents was carried out using GC-FID. Diazepam was detected using GC with nitrogen-phosphorus detection (NPD) and confirmed using GC-MS with full scan mode after solid-phase extraction of the whole blood sample using Bond-Elut Certify columns. Quantitation of diazepam was carried out using GC-NPD. No other drugs, including ethanol, were detected. Recoveries for benzene, toluene, and xylene (mixture of isomers) in whole blood at 5 mg/L were 89.2%, 90.8%, and 93.4%, respectively. Intraday precisions were 5.3%, 5.0%, and 4.9%, respectively, and interday precisions were 12.1%, 11.6%, and 11.5%, respectively. The limits of detection (LOD) and quantitation (LOQ) were 30 and 100 microg/L, respectively. The linearity of the blood calibration curves was excellent with r(2) values of > 0.999 (range 0.1-10 mg/L). Recovery for diazepam in whole blood at 0.5 mg/L was 88.2% with intraday and interday precisions of 2.0% and 10.8%, respectively. The LOD and LOQ were 6 and 20 microg/L, respectively. The linearity of the blood calibration curve was excellent with r(2) values of > 0.999 (range 0.1-2 mg/L). We want to alert other toxicologists about new or unexpected products that should be taken into account when the surreptitious use of substances in DFSA is suspected.

GHB Urine Concentrations After Single-Dose Administration in Humans

Gamma-hydroxybutyric acid (GHB) is used as an illicit drug and is implicated in drug-facilitated sexual assault, but it also has some therapeutic uses. Detection of GHB in urine is important for forensic testing and could be of clinical benefit in overdose management. Urine GHB concentration-time profiles have not been well-characterized or correlated with doses used therapeutically. GHB levels were measured by gas chromatography-mass spectrometry in urine collected over 24 h from 16 adults administered single doses of 50 mg/kg GHB (Xyrem) alone and combined with 0.6 g/kg ethanol. Peak GHB urine concentrations averaged 150-200 mg/L and occurred in the 0-3 h urine collection. Significant variability in GHB urine levels between individuals was observed. Caucasians had lower urine concentrations than other races/ethnicities (p = 0.03). Men had lower GHB levels than women in the first 3 h after dosing (p = 0.038). Coingestion of ethanol did not significantly affect renal clearance of GHB, but urine GHB concentrations were lower in the first 3 h when ethanol and GHB were coingested (p = 0.039). At a proposed cut-off of 10 mg/L to distinguish endogenous versus exogenous GHB levels, 12.5% of the samples collected from 3 to 6 h, 81.3% of samples collected from 6 to 12 h, and 100% of urine specimens collected from 12 to 24 h were below this level. We conclude that the detection time for GHB in urine may be shorter than the previously reported 12-h window in some people taking therapeutic doses of GHB.

Liquid Chromatography–Tandem Mass Spectrometry for Detection of Low Concentrations of 21 Benzodiazepines, Metabolites, and Analogs in Urine: Method with Forensic Applications

Commonly used methods for detecting benzodiazepines (BZPs) and BZP-like substances, such as zolpidem and zopiclone, may not detect low concentrations of these drugs. We developed a liquid chromatographic-tandem mass spectrometric method for identifying these drugs and their relevant metabolites. We extracted BZPs from urine by solid-phase extraction with a mixed-mode phase (OASIS HLB cartridges). Chromatographic separation was performed with a Waters XTerra MS C18 [150 x 2.1 mm (i.d.); bead size, 5 microm] reversed-phase column with deuterated analogs of the analytes as internal standards (IS). Detection was performed with a triple-quadruple mass spectrometer that monitored 2 specific transitions per compound in the electrospray, positive-ion selected-reaction monitoring mode. We tested this technique on urine samples from 12 healthy volunteers and 1 forensic sample obtained in a case of alleged drug-facilitated sexual assault. Chromatographic separation was achieved within 18 min. The linear dynamic ranges extended from 0.02 or 0.1 microg/L (depending on the drug or metabolite) to 50 microg/L. Extraction recovery (range) was 77%-110%. Limits of detection were < or = 0.05 microg/L. No ion suppression was seen except for alprazolam, for which baseline decreased by almost 20%. In the forensic urine sample, the method detected alprazolam (3.5 microg/L) and its characteristic metabolite, alpha-hydroxyalprazolam (0.17 microg/L). This method measured low concentrations of BZPs and BZP-like substances and might be useful for analyses of urine in suspected drug-facilitated sexual assault cases.

GHB Urine Concentrations After Single-Dose Administration in Humans

Gamma-hydroxybutyric acid (GHB) is used as an illicit drug and is implicated in drug-facilitated sexual assault, but it also has some therapeutic uses. Detection of GHB in urine is important for forensic testing and could be of clinical benefit in overdose management. Urine GHB concentration-time profiles have not been well-characterized or correlated with doses used therapeutically. GHB levels were measured by gas chromatography-mass spectrometry in urine collected over 24 h from 16 adults administered single doses of 50 mg/kg GHB (Xyrem) alone and combined with 0.6 g/kg ethanol. Peak GHB urine concentrations averaged 150-200 mg/L and occurred in the 0-3 h urine collection. Significant variability in GHB urine levels between individuals was observed. Caucasians had lower urine concentrations than other races/ethnicities (p = 0.03). Men had lower GHB levels than women in the first 3 h after dosing (p = 0.038). Coingestion of ethanol did not significantly affect renal clearance of GHB, but urine GHB concentrations were lower in the first 3 h when ethanol and GHB were coingested (p = 0.039). At a proposed cut-off of 10 mg/L to distinguish endogenous versus exogenous GHB levels, 12.5% of the samples collected from 3 to 6 h, 81.3% of samples collected from 6 to 12 h, and 100% of urine specimens collected from 12 to 24 h were below this level. We conclude that the detection time for GHB in urine may be shorter than the previously reported 12-h window in some people taking therapeutic doses of GHB.

Extraction and analysis of clonazepam and 7-aminoclonazepam in whole blood using a dual internal standard methodology

In this paper, a simple and robust method for the determination of clonazepam and its primary metabolite (7-aminoclonazepam) in whole blood is described. Clonazepam (klonopin) is a popular prescription drug that has been implicated in the field of drug facilitated sexual assaults (DFSA).Clonazepam, 7-aminoclonazepam and the internal standards (deuterated analogues for GC–MS analysis and nitrazepam for analysis by LC–PDA/GC–MS) were spiked into blood samples. The samples were buffered with a pH 6-phosphate solution (5mL) and extracted from phenyl spe columns. The columns were washed with 5% acetonitrile in pH 6-phosphate buffer (3mL) and eluted with ethyl acetate (2×3mL). The eluents were evaporated for further chromatographic analysis. For GC–MS, the samples were derivatized prior to analysis. When performed with LC–PDA the samples were reconstituted in distilled water.From this method LOQs of 5ng/mL of sample is easily achievable by either chromatographic system. By using GC–MS in EI mode, 1ng/mL of sample can be detected.Data is presented here to show the simplicity and efficiency of the extraction scheme. By employing the properties of GC–MS and LC–PDA, this extraction and analysis procedure further extends the number of tools open to the forensic toxicologist for the analysis of this drug.

A case in south-eastern France: A review of drug facilitated sexual assault in European and English-speaking countries

Drug facilitated sexual assaults (DFSA) have been increasingly reported in the medical literature since the 1980s but their legal recognition is more recent, at least in Europe. From a case treated in south-eastern France, whose judicial consequences were known, it seemed of interest to carry out an international study of jurisprudence concerning this type of rape. While from the medical viewpoint the drugs used are well-known and their presence can be clinically verified, the legal consequences of their use in subsequent criminal prosecution is less clear-cut. Some European countries have no jurisprudence in this area, while others consider the use of drugs as an aggravating circumstance. In France, it was only in 2003 that the first case of DFSA was truly punished by the judicial system, with considerable media attention. By contrast, in English-speaking countries, particularly the United States, the use of drugs to facilitate sexual assault has frequently been recognized in legislation and in criminal prosecutions. Prevention is fundamental and is recognised as demonstrated by campaigns in various countries.

Ecstasy (MDMA), Methamphetamine, and Date Rape (Drug-Facilitated Sexual Assault): A Consideration of the Issues

The term “date rape drug” has traditionally been applied by the media to powerful sedatives, such as gamma hydroxybutyrate (GHB) and flunitrazepam (Rohypnol), which can render a person unconscious and hence unable to resist and/or recall an assault. However, some law enforcement agents and others have recently obtained convictions by arguing that the empathy-generating and sensual effects of MDMA. and an occasional increase in disinhibition and sexual desire linked with methamphetamine use, remove a person's ability to give a reasoned consent, turning the person into “a helpless slave” to their own sexual desires and those of the alleged perpetrator. The argument holds that the victim becomes part of the assault because they may appear to be cooperating and colluding with activity which they would not have consented to without taking these drugs. This interpretation of the term “date rape” has been fed by data that sometimes finds MDMA and amphetamines in samples taken from sexual assault victims, and hence these prosecutions sometimes rely on expert testimony from toxicologists, pathologists and police officers rather than psychologists and psychiatrists who are expert in the human effects of these drugs. Some of those in the latter group have dismissed claims that MDMA is an aphrodisiac or a date rape drug as myths propagated by the media. In this article, these arguments and their respective strengths and weaknesses will be examined to assist professionals and others who may become involved in these cases.

The Clinical Development of &amp;#947;-Hydroxybutyrate (GHB)

The discovery of gamma-hydroxybutyrate (GHB) over 40 years ago led to its immediate use as a general anesthetic agent. Subsequent research demonstrated that GHB is an endogenous compound in the mammalian brain and current research suggests that GHB is a probable neurotransmitter. In the United States, reports of anabolic effects lead to its misuse among body builders during the 1980's while the intoxicating properties of the drug lead to its popularization as a substance of abuse during the 1990's. GHB became associated with reports of drug-facilitated sexual assault and cases of physical dependence and withdrawal. Efforts to ban GHB caused increased use of GHB analogues and pro-drugs. Against this backdrop, GHB was being developed for the treatment of narcolepsy, leading to the approval of Xyrem (sodium oxybate) oral solution in 2002 for the treatment of cataplexy in patients with narcolepsy. A risk management program permits the safe handling and distribution of the approved product, minimizes the risk for diversion, provides professional and patient education about the risks and benefits of sodium oxybate, and includes physician and patient registries. Post-marketing surveillance indicates sodium oxybate has an acceptable safety profile and presents minimal risk for the development of physical dependence.

A study of blood and urine alcohol concentrations in cases of alleged drug-facilitated sexual assault in the United Kingdom over a 3-year period

This paper details the alcohol concentrations found in a selection of 1014 cases of claimed drug-facilitated sexual assault analysed at The Forensic Science Service, London Laboratory between January 2000 and December 2002. Where appropriate, either a whole blood sample and/or a urine sample was analysed for alcohol, common drugs of abuse and potentially stupefying drugs. The samples were collected from a complainant within 12 h of an alleged incident in 391 of the 1014 cases analysed. Of these, the majority (81%) contained alcohol. The presence of alcohol itself was not surprising as most of the alleged incidents were associated with social situations such as at a public house, bar, night-club or party, where it is expected that alcohol would have been consumed. However, 233 (60%) of the 391 cases had a high back-calculated figure, where high is defined as greater than 150 milligrams per 100 millilitres (150 mg%). Some of these samples were also found to contain illicit drugs. This is the first paper to our knowledge which discusses in detail the significance of the alcohol concentrations found in cases of this type.

Development of immunoaffinity solid phase microextraction probes for analysis of sub ng/mL concentrations of 7-aminoflunitrazepam in urine

We report on the development of solid phase microextraction probes for drug analysis, prepared with antibodies specific for benzodiazepines covalently immobilized to the surface. In the technique, immobilized antibody probes are exposed to a sample containing the drug for 30 min. Extracted drugs are subsequently desorbed from the probes in 500 μL of methanolic desorption solution, which is dried, reconstituted in a small volume of injection solution and analysed by LC-MS/MS. The antibodies were characterized both before and after immobilization, to facilitate the rational selection of antibodies for such analyses. Polyclonal and monoclonal antibodies were compared as was the impact of affinity purification of the polyclonal antibody to isolate the drug-specific fraction. The probes were evaluated for utility in analyzing 7-aminoflunitrazepam at sub ng/mL concentrations in urine, which is expected to be found several days after a single oral dose of 2 mg of flunitrazepam. Such analyses are required in monitoring for abuse of this drug, both in terms of ‘club drug’ use and in cases of drug-facilitated sexual assault. In these cases drug concentrations in blood and urine are much lower than in chronic abuse cases and are difficult to analyse by conventional methods. The method developed has a limit of detection of 0.02 ng/mL, with accuracy ranging from 1% to 27% and precision (% R.S.D.) ranging from 2% to 10% between the lower and upper limits of quantitation for the analysis of 7-aminoflunitrazepam in urine. The dynamic range of the method is from 0.02 ng/mL, which is limited by the instrument sensitivity, to 0.5 ng/mL, which is approaching the capacity of the probes. This would allow for quantitative analysis of samples at concentrations below that measurable by many other methods for general benzodiazepines analysis from urine, and a highly selective screen for samples at higher concentrations. The method has similar limits of detection to the most sensitive literature methods specifically designed for such analysis but with the advantage of significantly simplified sample preparation. This simplification makes the technique more amenable for use by both professionals and non-professionals.

A case of drug-facilitated sexual assault by the use of flunitrazepam

This article presents a case of drug-facilitated sexual assault on a female intoxicated with flunitrazepam. The male assailant added flunitrazepam (1 mg) to the female’s soft drink, and had sexual intercourse with her while her consciousness was impaired. The complainant did not recall the events due to benzodiazepine-induced anterograde amnesia. The use of flunitrazepam was uncovered when its major metabolite, 7-amino flunitrazepam, was detected in a urine specimen collected when the complainant attended hospital approximately one day after consuming the adulterated drink.

A case of drug-facilitated sexual assault leading to death by chloroform poisoning

The purpose of this investigation was to determine the cause of death of a 13-year-old girl, where none was immediately evident. Our analysis showed it to be a very unusual case of a drug-facilitated sexual assault (DFSA), which led to the tragic death of the young rape victim and then to the suicide of the rapist. The incapacitating agent used was chloroform. The post-mortem analysis revealed a blood concentration of 833.9 mg/l for the girl, whereas the quantitation of chloroform in various fluids and viscera of the rapist proved that he had recently been handling the solvent (with concentrations in fat tissues 20 times higher than in his blood). This case draws attention to the need for broad searches for volatile substances in such investigations.

Toxicological findings in cases of alleged drug-facilitated sexual assault in the United Kingdom over a 3-year period

This paper outlines the toxicology results from 1014 cases of claimed drug-facilitated sexual assault (DFSA) analysed at the Forensic Science Service, London Laboratory between January 2000 and December 2002. Where appropriate, either a whole blood sample and/or a urine sample was analysed for alcohol, common drugs of abuse and potentially stupefying drugs. The results were interpreted with respect to the number of drugs detected and an attempt was made to distinguish between voluntary and involuntary ingestion from information supplied. Alcohol (either alone or with an illicit and/or medicinal drug) was detected in 470 of all cases (46%). Illicit drugs were detected in 344 cases (34%), with cannabis being the most commonly detected (26% of cases), followed by cocaine (11%). In 21 cases (2%), a sedative or disinhibiting drug was detected which had not been admitted and could therefore be an instance of deliberate spiking. This included three cases in which complainants were allegedly given Ecstasy (MDMA) without their knowledge. Other drugs detected included gammahydroxybutyrate (GHB) and the benzodiazepine drugs diazepam and temazepam. Another nine cases (1%) involved the complainant being either given or forced to ingest pharmaceutical tablets or an illicit drug.