Oral Fluid Collection Device Research Articles

Evaluation of an Oral Fluid Collection Device and a Solid-Phase Extraction Method for the Determination of Coca Leaf Alkaloids by Gas Chromatography-Mass Spectrometry.

Some South American countries have ancient traditions that may pose legal problems, such as the consumption of coca leaves, as this can provide positive results for cocaine use after the analysis of biological samples. For this reason, it is necessary to find specific markers that help differentiate legal from illegal consumption, such as tropacocaine, cinnamoylcocaine, and especially hygrine and cuscohygrine. In this work, two techniques for collecting biological samples are compared: the Quantisal® Oral Fluid collection device and passive drooling. Once the samples were collected, they were subjected to solid-phase extraction for subsequent injection into GC-MS. Different validation parameters included in international guides have been studied to evaluate whether the proposed method is valid for the defined purpose, placing special emphasis on the study of the matrix effect and little value on GC-MS analyses. With respect to this parameter, an increase in the signal was found for CUS and t-CIN, but it was not significant for the rest of the substances studied. The recoveries have varied significantly depending on the way of working, being higher when working with standardized areas. After carrying out work with the oral fluid samples collected from laboratory volunteers, the method was applied to two real samples. The results obtained support the need for further research to overcome certain limitations presented by the device.

The analysis of drugs in neonates using an ad hoc oral fluid collection device to develop a prediction model for neonatal abstinence syndrome and provide maternal support

To use oral fluid specimens collected in an ad hoc device to test for opioid exposure and to help assess the risk of withdrawal in potential neonatal abstinence syndrome (NAS) cases. Testing for Neonatal drug exposure typical uses meconium, urine, blood, hair, or umbilical cord tissue/blood specimens. Due to the invasiveness, challenges, and limitations of collection, and/or analytical difficulties of these matrices, oral fluid may be a more desirable specimen when testing for opioid exposure and assessing the risk of neonatal abstinence syndrome (NAS). Traditional oral fluid collection devices are not viable options for neonatal specimen collection due to their large size and potential chemicals additives that induce salivation. Unstimulated and stimulated infant oral fluid samples have been used for therapeutic drug monitoring as an alternative matrix to blood. Oral fluid samples were collected under Virginia Commonwealth University's Institutional Review Board from ten neonates born to mothers in methadone or buprenorphine treatment programs. Oral fluid samples were collected by rolling three foam-tipped swabs along the mouth, inner cheeks, and tongue until saturated. The three swabs containing approximately 300 μL oral fluid were placed in 2.0 mL of phosphate-buffered saline, centrifuged, and analyzed by ultra-high-pressure liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for 26 drugs and/or metabolites after solid phase extraction (SPE) with OFXQ columns. Results were compared to maternal and the neonatal medical history. The following analytes were detected in the neonatal oral fluid: 2-Ethyl-5-methyl-3,3-diphenyl-1-pyrroline (EMDP), 2-Ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), 6-acetylmorphine, amphetamine, benzoylecgonine, cocaine, cotinine, ethyl sulfate, methadone, methamphetamine, morphine, nicotine, and norfentanyl. Maternal history and infant medications were available for 9 of 10 neonates. The detection of norfentanyl was consistent with epidurals given during birth. The following analytes detected were inconsistent with patient histories; 6-acetylmorphine, amphetamines, benzoylecgonine, ethyl sulfate, and nicotine. Three of the ten neonates were treated for withdrawal symptoms by an oral suspension of methadone given twice daily. These neonates were released from the hospital on day of life 18, 20, or 24. Prior to treatment, four to eleven drugs and/or metabolites were identified in the three neonates, including 6-acetlymorphine, amphetamine, benzoylecgonine, cocaine, cotinine, EDDP, EMDP, ethyl sulfate, methadone, morphine, nicotine, and/or norfentanyl. In two neonates, methadone oral fluid concentrations increased in the first few days of life then significantly decreased in concentration prior to treatment. No methadone was detected in the third neonate, but a variety of other drugs were identified. None of the other neonates presented symptoms of withdrawal and were released from the hospital on day 3 or 4 of life, except for one neonate born pre-term (35 weeks gestation) was not released until day of life 9. An easy-to-use collection device for neonatal oral fluid must be developed. Identifying trends between neonates who demonstrate withdrawal symptoms and those who do not could be vital to developing a predictive model for the determination of NAS. When analyzing neonatal samples, complete clinical histories of mother and neonate are necessary to prevent the misinterpretation of data, which could lead to unwarranted criminal or legal complications.

Cocaethylene presence in oral fluids from cocaine users

Simultaneous consumption of cocaine and alcohol is very common amongst cocaine users and many addiction centres now warn of the dangers of combined consumption of cocaine and alcohol. Research by the National Treatment Agency highlighted that 61% of powder cocaine users always use alcohol at the same time. Another study published in Italy reported 5.6% of drivers under the influence had consumed both alcohol and cocaine (Snengi R., Alcohol and alcoholism, 2018, 53(6):735–741). It is well documented that the use of alcohol with cocaine has serious additive effects such as increased cardiotoxicity, cardiovascular effects and prolonged euphoria. Simultaneous consumption of cocaine and alcohol results in the formation of cocaethylene, the ethyl ester of benzoylecgonine. Cocaethylene is pharmacologically active, and its pharmacological activity is similar to that of cocaine. In this study we evaluated the commercially available screening tests to understand their ability to cross-react with cocaethylene. We also analysed data from over 14,000 oral fluid samples submitted for cocaine analysis from drug treatment and probation sites in the UK. Available commercial screening product inserts were reviewed to assess reported cross-reactivity of the cocaine assay with cocaethylene. Confirmation data from oral fluid samples received at Abbott Toxicology, UK between January 2019 and April 2022 was reviewed. All samples had been collected using the Certus Oral Fluid Collection device and analysed using LC- MS/MS for cocaine, benzoylecgonine, norcocaine, cocaethylene as well as anhydroecgonine methyl ester. Cross-reactivity data from commercial products showed that there was a large variation in cross reactivity to cocaethylene between different vendors ranging between 4%–80%. The assay target was either benzoylecgonine or cocaine. Several product inserts did not list cocaethylene under the assays cross reactivity information. In total, 14,842 samples were received in the laboratory, of which 75% ( n = 11,194) were positive for either cocaine or one of its metabolites. Cocaine was present in 98% ( n = 11,019), benzoylecgonine in 89% ( n = 10,017) and cocaethylene in 6.6% ( n = 740) of the positive samples. The longer lasting and increased effects of cocaethylene may cause prolonged impairment. The data presented highlights that the cross reactivity to either cocaethylene or benzoylecgonine should not be a forgotten while developing cocaine testing strategy.

Poor correlation between alcohol concentration in oral fluid and breath in subjects consuming beverages immediately before testing.

IntroductionIn previous research projects and clinical settings, alcohol analysis in oral fluid (saliva) has been used as an alternative to breath or blood alcohol testing. In this study we examined whether it is possible to obtain clinically relevant data regarding alcohol consumption in individuals who recently consumed alcohol by analysing oral fluid samples when the recommended rinsing of the mouth is impossible before sample collection.Materials and methodsWe conducted a study of 89 nightclub patrons in Norway. Before collecting oral fluid samples and performing breath alcohol testing, participants were required to drink a glass of water to remove residual alcohol from the mouth. Oral fluid samples were collected with the Quantisal oral fluid collection device and analysed using an enzymatic method for alcohol. The alcohol concentration in the neat (undiluted) oral fluid was then calculated. Breath alcohol testing was performed using Lion Alcolmeter 500 instruments.ResultsNo false-negative or false-positive results for alcohol were detected in the oral fluid when compared with those in the breath. The Intraclass Correlation Coefficient of 0.40 indicated a poor correlation between alcohol concentrations in the two sample types.ConclusionsThe procedure for collecting oral fluid was suitable for the qualitative determination of alcohol intake but not for quantitative assessment. We recommend that oral fluid samples should not be used for estimating blood or breath alcohol concentrations in people who have recently consumed alcohol or non-alcoholic beverages, as recommended in the instructions for use.

Impact of Quantisal® Oral Fluid Collection Device on Drug Stability

The stability of drugs can affect drug tests and interpretations. A comprehensive study to verify drug stability in Quantisal® oral fluid (OF) collection device was undertaken in accordance with Australian standard, AS/NZS 4760:2019 (SAI-Global, 2019). The evaluation was performed for the following drugs: (±) amphetamine, (±) methylamphetamine, (±) 3,4-methylenedioxymethylamphetamine (MDMA), (−)Δ9-tetrahydrocannabinol (THC), cocaine, benzoylecgonine, morphine, codeine, and oxycodone. Stability was assessed at four different storage temperatures over seven time points at ±50% cut-off concentrations (Appendix A, Para A4-4.1, AS/NZS 4760:2019) (SAI-Global, 2019). All drugs were found to be significantly more stable at 4 and –20°C, with stability spanning at least 14 days with percentage change within ±20% from the cut-off concentrations (SAI-Global, 2019). In addition, we report a variation trend with cocaine and benzoylecgonine at elevated temperatures, suggesting hydrolytic decomposition of cocaine and a concomitant increase in benzoylecgonine quantitative values. We confirm the cross-talk by showing that the percentage change in the profile of average cocaine-benzoylecgonine measurement is within the acceptance concentration range of ±20%. This finding highlights the importance of precaution during storage and careful considerations during subsequent interpretation of liquid chromatography-mass spectrometry (LCMS) measurements.

A Case Study Evaluating the Efficacy of an Ad Hoc Hospital Collection Device for Fentanyl in Infant Oral Fluid.

Neonatal drug exposure is currently assessed using meconium, urine, blood, hair, or umbilical cord tissue/blood. Due to the invasiveness, challenges, and limitations of collection, and/or analytical difficulties of these matrices, oral fluid may be a more desirable matrix in diagnosing opioid exposure and risk for opioid withdrawal in neonatal abstinence syndrome. Traditional oral fluid collection devices are not viable options as they are too large for neonates' mouths and may contain chemicals on the collection pad. Unstimulated and stimulated infant oral fluid samples have been used for therapeutic drug monitoring as an alternative matrix to blood. The objective of this study was to assess the viability of a simple oral fluid collection system using a sterile foam-tipped swab rinsed in phosphate-buffered saline. Two infants were administered fentanyl for post-operative pain relief while hospitalized in the Neonatal Intensive Care Units at the Children's Hospital of Richmond of Virginia Commonwealth University. Oral fluid samples were collected at 16h, 2days, and/or 7days following the start of intravenous infusion of fentanyl. Samples were analyzed by ultra-high-pressure liquid chromatography-tandem mass spectrometry for fentanyl and norfentanyl after solid-phase extraction. In one of the three samples tested, fentanyl and norfentanyl were detected at concentrations of 28 and 78ng/mL, respectively. Based on the infusion rate, the theoretical oral fluid fentanyl concentration at steady state was calculated to be 33ng/mL.

User experience and operational feasibility of four point-of-collection oral fluid drug-testing devices according to Brazilian traffic agents

Objective: Traffic fatalities in Brazil still rank among the highest worldwide, with an overall rate of 23.4 deaths/100,000 inhabitants/year. Although alcohol and drug use play an important role in traffic accidents, national data about their relative influence are scarce. Drug screening is not routinely performed by traffic agents because alcohol is the only substance regularly investigated in roadblocks. Therefore, we aimed to describe the initial traffic agent user experience for 4 handheld point-of-collection oral fluid drug testing devices used in routine roadblocks in Brazil, focusing on usage perceptions in hopes of generalizing this approach for other developing countries.Methods: Four different oral fluid collection devices were evaluated: The DDS2, the DOA MultiScreen, the Dräger DrugTest 5000, and the Multi-Drug Multi-Line Twist Screen Device. Fourteen trained traffic agents obtained oral fluid from 164 drivers and performed 37 qualitative evaluations of the devices. Traffic agents filled out a questionnaire focusing on 9 feasibility criteria: Overall simplicity for roadside operation; operational success; saliva sample collection time; sample analysis time; ease of sample preparation and analysis; agreement with observed clinical signs; overall hygiene and safety; sufficient operating instructions; and hygiene of saliva collection. These were weighted based on an expert panel and yielded an overall composite device experience score that ranged from 1 (poor) to 100 (excellent).Results: Ease of use, operational success, and acceptable collection and analysis time were considered the most important criteria by the expert panel. The results ranged from 27.3 to 88.9% for simplicity of use; 45.5 to 100.0% for operational success; 27.3 to 100% for acceptable collection time; and 36.4 to 100.0% for acceptable analysis time. The final device scores, based on the agents’ user experiences, ranked as follows: DOA MultiScreen: 49.3/100; Dräger DrugTest 5000: 82.4/100; Multi-Drug Multi-Line Twist Screen Device: 84.3/100; DDS2: 88.4/100.Conclusion: Based on the selected criteria, 3 of the 4 devices were considered useful by traffic agents in routine roadblock operations. The weighted evaluations suggest that their ease of use (handling, sampling analysis, and reliability), as well as their agreement with findings obtained by other means, defined their utility to traffic agents, although such appraisals must be further analyzed in future studies.

Use of Oral Fluid in a Rapid Syphilis Test Assay

Background From 2014 to 2015, the syphilis rate in the United States increased by 19%, reaching its highest rate since 1994. Currently, point-of-care syphilis assays use fingerstick or venipuncture whole blood to identify Treponema pallidum (TP) antibodies by qualitative immunoassay. However, patients and providers prefer oral fluid testing to whole blood testing. In this study, we aimed to determine whether a rapid syphilis test intended for use on whole blood could be used to detect TP antibodies in oral fluid.Methods Oral fluid was collected from 72 participants using the Super•SAL™ Oral Fluid Collection Device (Oasis Diagnostics®, Vancouver, WA). The device uses an absorbent cylindrical pad to collect and filter ~1 mlml of oral fluid. Oral fluid filtrate was tested using the SD Bioline Syphilis 3.0 rapid test (Alere Diagnostics, MA) following manufacturer directions for whole blood. TP particle agglutination (TPPA) and rapid plasma reagin (RPR) results derived from participants’ medical records were used as reference values. We used three different definitions as comparators: 1: TPPA reactive; 2: TPPA and RPR reactive and 3: TPPA reactive and RPR titer >1:4. Those with non-reactive TPPA and RPR results were considered seronegative. We calculated the sensitivity and specificity for definition 1 and sensitivity for definitions 2 and 3. We used the exact binomial method to determine 95% confidence intervals (CI).ResultsWith definitions 1, 2, and 3, respectively, sensitivity was 83.3% (CI: 67.2, 93.6), 86.4% (CI: 65.1, 97.1), and 100% (CI: 71.5, 100). Specificity was 47.2% (CI: 36.5, 75.5).Conclusion The high sensitivity of the SD Bioline Syphilis 3.0 test using oral fluid suggests a strong potential for the development of accurate rapid oral syphilis tests. Sensitivity increased with higher RPR titer. False positive results may be due to the presence of non-venereal treponemal antibodies in oral fluid. Further research and development are needed to optimize specificity.Disclosures All authors: No reported disclosures.

Introduction of sample tubes with sodium azide as a preservative for ethyl glucuronide in urine.

Ethyl glucuronide (EtG) is a direct alcohol marker, which is widely used for clinical and forensic applications, mainly for abstinence control. However, the instability of EtG in urine against bacterial degradation or the post-collectional synthesis of EtG in contaminated samples may cause false interpretation of EtG results in urine samples. This study evaluates the potential of sodium azide in tubes used for urine collection to hinder degradation of ethyl glucuronide by bacterial metabolism taking place during growth of bacterial colonies. The tubes are part of a commercial oral fluid collection device. The sampling system was tested with different gram-positive and gram-negative bacterial species previously observed in urinary tract infections, such as Escherichia coli, Staphylococcus aureus, Enterecoccus faecalis, Staphylococcus epidermidis, Klebsiella pneumoniae, Enterobacter cloacae, and Pseudomonas aeruginosa. Inhibition of bacterial growth by sodium azide, resulting in lower numbers of colony forming units compared to control samples, was observed for all tested bacterial species. To test the prevention of EtG degradation by the predominant pathogen in urinary tract infection, sterile-filtered urine and deficient medium were spiked with EtG, and inoculated with E. coli prior to incubation for 4days at 37°C in tubes with and without sodium azide. Samples were collected every 24hours, during four consecutive days, whereby the colony forming units (CFU) were counted on Columbia blood agar plates, and EtG was analyzed by LC-MS/MS. As expected, EtG degradation was observed when standard polypropylene tubes were used for the storage of contaminated samples. However, urine specimens collected in sodium azide tubes showed no or very limited bacterial growth and no EtG degradation. As a conclusion, sodium azide is useful to reduce bacterial growth of gram-negative and gram-positive bacteria. It inhibits the degradation of EtG by E. coli and can be used for the stabilization of EtG in urine samples.

Ebola exposure, illness experience, and Ebola antibody prevalence in international responders to the West African Ebola epidemic 2014-2016: A cross-sectional study.

BackgroundHealthcare and other front-line workers are at particular risk of infection with Ebola virus (EBOV). Despite the large-scale deployment of international responders, few cases of Ebola virus disease have been diagnosed in this group. Since asymptomatic or pauci-symptomatic infection has been described, it is plausible that infections have occurred in healthcare workers but have escaped being diagnosed. We aimed to assess the prevalence of asymptomatic or pauci-symptomatic infection, and of exposure events, among returned responders to the West African Ebola epidemic 2014–2016.Methods and findingsWe used snowball sampling to identify responders who had returned to the UK or Ireland, and used an online consent and questionnaire to determine their exposure to EBOV and their experience of illness. Oral fluid collection devices were sent and returned by post, and samples were tested using an EBOV IgG capture assay that detects IgG to Ebola glycoprotein. Blood was collected from returnees with reactive samples for further testing. Unexposed UK controls were also recruited.In all, 300 individuals consented, of whom 268 (89.3%) returned an oral fluid sample (OFS). The majority had worked in Sierra Leone in clinical, laboratory, research, and other roles. Fifty-three UK controls consented and provided samples using the same method.Of the returnees, 47 (17.5%) reported that they had had a possible EBOV exposure. Based on their free-text descriptions, using a published risk assessment method, we classified 43 (16%) as having had incidents with risk of Ebola transmission, including five intermediate-risk and one high-risk exposure. Of the returnees, 57 (21%) reported a febrile or diarrhoeal illness in West Africa or within 1 mo of return, of whom 40 (70%) were not tested at the time for EBOV infection.Of the 268 OFSs, 266 were unreactive. Two returnees, who did not experience an illness in West Africa or on return, had OFSs that were reactive on the EBOV IgG capture assay, with similar results on plasma. One individual had no further positive test results; the other had a positive result on a double-antigen bridging assay but not on a competitive assay or on an indirect EBOV IgG ELISA. All 53 controls had non-reactive OFSs. While the participants were not a random sample of returnees, the number participating was high.ConclusionsThis is the first study, to our knowledge, of the prevalence of EBOV infection in international responders. More than 99% had clear negative results. Sera from two individuals had discordant results on the different assays; both were negative on the competitive assay, suggesting that prior infection was unlikely. The finding that a significant proportion experienced “near miss” exposure events, and that most of those who experienced symptoms did not get tested for EBOV at the time, suggests a need to review and standardise protocols for the management of possible exposure to EBOV, and for the management of illness, across organisations that deploy staff to outbreaks.

Detection of Hydrocodone and Morphine as Metabolites in Oral Fluid by LC-MS/MS in Patients Prescribed Codeine.

A retrospective analysis of oral fluid drug testing results using LC-MS/MS was performed to determine the prevalence rates in oral fluid for codeine (COD) and 3 COD metabolites-morphine (MOR), norhydrocodone (NHC), and hydrocodone (HCOD). Oral fluid samples were collected using Quantisal oral fluid collection device (Immunalysis Inc.) and submitted to Millennium Health, LLC for the routine drug analysis by LC-MS/MS. Consistent with previously published literature, COD was the primary analyte detected in oral fluid after the use of COD. In COD-positive samples, HCOD, MOR, and NHC were detected at rates of 68.4%, 18.4%, and 6.3%, respectively. Concentration ranges of these analytes were 1.0 to >2000 ng/mL for COD, 1.0-20.2 ng/mL for MOR, 1.0-740.0 ng/mL for HCOD, and 2.1-47.5 ng/mL for NHC. In contrast to urine, where HCOD is typically detected as a minor metabolite of COD, HCOD was the most commonly detected metabolite in oral fluid in samples testing positive for COD with reported prescriptions for COD. This observation suggests that care should be taken when interpreting HCOD positives in oral fluid results, and that the use of COD should be considered as one possible explanation for HCOD positives.

Acceptability of a Community-Based Outreach HIV-Testing Intervention Using Oral Fluid Collection Devices and Web-Based HIV Test Result Collection Among Sub-Saharan African Migrants: A Mixed-Method Study.

BackgroundLate human immunodeficiency virus (HIV) diagnosis is common among sub-Saharan African migrants. To address their barriers to HIV testing uptake and improve timely HIV diagnoses and linkage to care, the outreach HIV testing intervention, “swab2know,” was developed. It combined a community-based approach with innovative testing methods: oral fluid self-sampling and the choice between Web-based HIV test result collections using a secured website or post-test counseling at a sexual health clinic. The sessions included an informational speech delivered by a physician of sub-Saharan African origin and testimonies by community members living with HIV.ObjectivesThe objectives of this study were to evaluate the intervention’s acceptability among sub-Saharan African migrants and its potential to reach subgroups at higher risk for HIV infection and to identify facilitators and barriers for HIV testing uptake.MethodsThis mixed-method study combined qualitative (participant observations and informal interviews with testers and nontesters) and quantitative data (paper–pencil survey, laboratory data, and result collection files). Data were analyzed using a content analytical approach for qualitative and univariate analysis for quantitative data.ResultsA total of 10 testing sessions were organized in sub-Saharan African migrant community venues in the city of Antwerp, Belgium, between December 2012 and June 2013. Overall, 18.2% of all people present (N=780) underwent HIV testing; 29.8% of them tested for HIV for the first time, 22.3% did not have a general practitioner, and 21.5% reported 2 or more sexual partners (last 3 months). Overall, 56.3% of participants chose to collect their HIV test results via the protected website. In total, 78.9% collected their results. The qualitative analysis of 137 participant observation field notes showed that personal needs and Internet literacy determined the choice of result collection method. Generally, the oral fluid collection devices were well accepted mainly because sub-Saharan African migrants dislike blood taking. For some participants, the method raised concerns about HIV transmission via saliva. The combination of information sessions, testimonies, and oral fluid collection devices was perceived as effectively reducing thresholds to participation. Acceptability of the intervention differed between individual participants and settings. Acceptance was higher among women, in churches and settings where community leaders were engaged in HIV awareness raising. Higher preventive outcomes were observed in settings with lower acceptance. The presence of the intervention team visualized the magnitude of the HIV epidemic to the public and promoted HIV testing uptake at large, for example, those who declined indicated they would take up testing later.ConclusionsWhen accompanied by tailored provision of information, outreach HIV testing interventions adopting a community-based approach and innovative methods such as Web-based result collection and oral fluid collection devices are acceptable and reduce thresholds for HIV testing uptake. The swab2know intervention was able to reach sub-Saharan African migrants at risk of HIV infection, and with limited access to regular HIV testing. Among nontesters, the intervention contributed to awareness raising and therefore has a place in a multipronged HIV test promotion strategy.

Extended Detection of Amphetamine and Methamphetamine in Oral Fluid.

Amphetamine and methamphetamine are popular drugs of abuse worldwide and are important components of drug monitoring programs. Windows of detection for amphetamine and methamphetamine in oral fluid after high doses have not been investigated. Repeated high-dose ingestions are likely to cause positive samples for extended periods. Common routes of administration of amphetamine/methamphetamine in Norway are oral intake or injection. The aim of this study was to investigate windows of detection for amphetamine and methamphetamine in oral fluid from drug addicts under sustained abstinence during detoxification. Twenty-five patients admitted to a closed detoxification unit were included in this study. Oral fluid samples were collected daily in the morning and evening, and urine every morning for 10 days. A blood sample was drawn during the first 5 days after admission if the patient consented. Oral fluid results were compared with urine results to determine whether a new ingestion occurred. Oral fluid was collected with the Intercept oral fluid collection device. In-house cutoff concentrations for amphetamine and methamphetamine were 6.8 and 7.5 mcg/L, respectively, in oral fluid, and 135 and 149 mcg/L, respectively, in urine. Amphetamines were detected in 11 oral fluid, 5 urine, and 2 blood specimens from 25 patients. Patients self-reported amphetamines intake of up to 0.5-2 g daily. Windows of detection for amphetamine and methamphetamine in oral fluid were up to 8 days, longer than in urine at the applied cutoff values. These data confirm that oral fluid is a viable alternative to urine for monitoring amphetamine abuse, and that these substances might be detected in oral fluid for at least 1 week after ingestion of high doses. Such long detection times were, as far as we are aware, never reported previously for oral fluid amphetamines.

Assessment of the use of oral fluid as a matrix for drug monitoring in patients undergoing treatment for opioid addiction.

Drug testing is an important clinical tool that is available to physicians who are assessing the effectiveness of drug treatment as well as patient compliance to the administered program. While urine has traditionally been the matrix of choice for drug monitoring, oral fluid, a filtrate of the blood, has shown great promise as an alternative matrix for such applications. Oral fluid collection can be accomplished without the need for highly trained medical staff through the use of a simple, noninvasive oral fluid collection device, which obtains an adequate sample in only a few minutes. There has been a significant amount of research performed on the use of oral fluid for forensic toxicology application; however, more studies assessing the use of oral fluid drug testing are required to validate its ability to achieve clinical drug monitoring goals. Testing for various drugs in oral fluid may yield a different result when compared to the same drugs in urine, requiring an assessment of the utility of oral fluid for such practices. The purpose of this study was to examine the application of oral fluid drug testing in patients undergoing buprenorphine treatment for opioid dependence. A retrospective analysis of drug testing results obtained from 6,928 patients (4,560 unobserved urine collections and 2,368 observed oral fluid collections) monitored for heroin metabolite, amphetamine, benzodiazepines, buprenorphine, tetrahydrocannabinol, cocaine, codeine, hydrocodone, hydromorphone, methadone, morphine, oxycodone, and oxymorphone was completed. Results of this statistical exercise indicated that patients undergoing observed oral fluid collection tested positive more frequently than those unobserved urine collections for several illicit drugs and prescription medications targeted. Oral fluid was shown to detect illicit drug use as well as noncompliance in this patient population under the studied conditions more often than the urine specimens.

Development of a high-speed MALDI-triple quadrupole mass spectrometric method for the determination of 3,4-methylenedioxymethamphetamine (MDMA) in oral fluid.

3,4-Methylenedioxymethamphetamine (MDMA, ecstasy) is still a widely used illicit designer drug and its detection in different matrices is of major importance for forensic purposes (e.g. driving under the influence) as well as for workplace drug testing or abstinence control. Established analytical methods for the determination of MDMA are mainly employing high performance liquid chromatography (HPLC) or gas chromatography (GC) coupled to mass spectrometric detection. Matrix assisted laser desorption/ionization-triple quadrupole-tandem mass spectrometry (MALDI-QqQ-MS/MS) is so far rarely used in forensics and offers an ultrafast high-throughput platform. The Quantisal™ Oral Fluid Collection Device was used for sample collection. After addition of the deuterated internal standard and a carbonate buffer (0.75 M Na2 CO3 ), oral fluid samples were liquid-liquid extracted (ButOAc/EtOAc, 1:1). As little as 1 microlitre of a mixture of this extract and the MALDI matrix (alpha-cyano-4-hydroxycinnamic acid) was spotted onto the MALDI plate and could directly be analyzed. With MALDI omitting chromatographic separation, very short analysis times of about 10 s per sample were possible. The method was developed and validated according to international guidelines including specificity, recovery, matrix effects, accuracy and precision, stabilities and limit of quantification. All validation criteria were fulfilled except for ion suppression/enhancement. Comparison with a routine liquid chromatography-tandem mass spectrometry (LC-MS/MS) method showed good agreement of the results. Applicability of the method was shown by analyzing about 250 oral fluid samples collected after controlled administration of 125 mg MDMA in a pharmacokinetic study. The whole lot of samples could be analyzed in less than 1 h, proving the ultra-high-speed of the method.

Recovery of drugs of abuse from Dräger DCD5000 oral fluid collection device in Australia.

In Australia, it is a requirement of workplace oral fluid (OF) drugs of abuse testing that drug recovery from collection devices be verified by an accredited laboratory. Recovery data are used in conjunction with collection volume imprecision data and uncertainty of measurement to provide an estimation of drug concentration in neat OF. The manufacturer's product information for the DCD5000 collection device indicates that the collection volume of the swab is 380 µL. Recovery data for the swab when used with the isopropanol provided by the manufacturer are not available. A series of experiments using fortified drug-free OF were performed to assess the collection volume imprecision of the Dräger DCD5000 swab and the recovery of drugs from the swab using isopropanol. The fortified OF was collected with the swabs (n = 16), and swabs were discharged into vials of isopropanol as per the manufacturer's instructions. The mean collection volume of the DCD5000 swab was 487 µL with an imprecision of 1.3%. Recovery of drug from the device ranged from 86 to 98% for drugs listed in the Australian OF workplace testing standard. Recovery of methadone, buprenorphine and norbuprenorphine ranged from 93 to 102%. Recovery of 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidene was 45%, suggesting that urine is more suitable sample if methadone therapy is being monitored. Overall, drug recovery from the device using isopropanol was acceptable when the increased collection volume of the swab was taken into account.

Validation of an EMIT® Screening Method to Detect 6-Acetylmorphine in Oral Fluid

An automated assay was modified and validated to qualitatively screen for 6-acetylmorphine (6-AM) in oral fluid using the Siemens EMIT II(®) Plus 6-AM urine assay. The validation utilized an oral fluid calibrator at the currently proposed Substance Abuse and Mental Health Services Administration cutoff concentration of 4 ng/mL, as well as quality control material prepared and validated through liquid chromatography-tandem mass spectrometry. All calibrator, quality control and unknown specimens were analyzed based on the dilution and buffering system of the Quantisal(®) oral fluid collection device. Immunoassay parameters such as the pipetted sample and reagent volumes as well as photometric read times were adjusted as part of the assay modification process. Validation experiments included the determination of intra- and inter-day precision and reproducibility, limits of detection (LODs), assay selectivity, stability studies and a specimen agreement study (n = 132). The 6-AM assay performed well in all validation experiments, over multiple days and under various laboratory conditions. The LOD was determined to be 1.844 ng/mL. The assay sensitivity, specificity and overall misclassification rate were found to be 90, 100 and 6%, respectively.

Repeatability of oral fluid collection methods for THC measurement

Study objectivesTo determine the influence of sample collection for two different collection methods on THC concentrations and to compare THC concentrations collected by both methods. MethodsA total of 136 pairs of oral fluid samples from subjects who had recently smoked Cannabis were obtained by the non-acidic Statsure oral fluid collection device and by ordinary spit tubes. Oral fluid was analyzed for THC by LC–MS/MS. Bland–Altman plots were used for the quantitative analysis of repeatability, whereas Cohen's kappa was used for qualitative analysis to determine the consistency of the results with regard to the Belgian legal limit. ResultsRepeatability of both sampling methods was very low. The Statsure device had a better rate of agreement when compared with the Belgian legal limit than the spitting method. THC concentrations of samples collected by spit tubes were on average a factor 5.9 higher than the corresponding concentrations in samples collected by the Statsure device. ConclusionsThe repeatability of both the Statsure collection method and the ordinary spit tubes was low when applied to subjects who had consumed Cannabis very recently. Furthermore, THC concentrations were higher in samples obtained by spitting than samples collected with Statsure. These results may have implications for confirmation analysis in oral fluid, when applied for legal purposes.

Could oral fluid be used to evaluate anti-hepatitis A virus status in individuals living in difficult-to-access areas?

A strategy adopted by different countries to reduce the number of new cases of hepatitis A is the vaccination. However, the mosaic of the epidemiological profile in developing countries has hampered the establishment of a unified nationwide vaccination program. To determinate national vaccination policies, the results of epidemiological studies need to be carefully considered. For this monitoring, the use of oral fluid is very important due to the painless and non invasive collection characteristics. There are few studies investigating which oral fluid collection device is optimal to detect low antibody levels and its use in selecting individuals for vaccination. So, the present study aimed to evaluate different oral fluid collection devices to detect humoral immune response against hepatitis A virus and its application in epidemiological studies. Therefore, 90 matched serum and oral fluid samples were collected from volunteers with different immune status, under ideal conditions of collection (optimization panel); and 224 matched samples in difficult-to-access areas (epidemiological study). Serum was collected by venipuncture and the oral fluid was obtained using three commercial devices: Salivette®, OraSure® and ChemBio®. Serum and oral fluid were submitted to a commercial immunoblot to detect total anti-HAV antibodies. The optimization panel demonstrated that ChemBio® device had the best performance (100% agreement), followed by OraSure® (95.4%) and Salivette® (90.8%). The optimal collection device (ChemBio®), tested in a difficult-to-access area and evaluated under precarious conditions of collection, showed similar prevalence of total anti-HAV between serum and oral fluid, 80.8% and 79%, respectively. A follow-up was performed to evaluate the stability of oral fluid and it was observed that 210 days after the collection it was possible to detect anti-HAV antibodies. Oral fluid can be used to detect low levels of specific-antibody, being important to select age groups to be vaccinated. Therewith, the choice of proper collection device is essential to evaluate HAV antibodies in the epidemiological scenario.

Development of an electrospray LC‐MS/MS method for quantification of Δ9‐tetrahydrocannabinol and its main metabolite in oral fluid

A fast and sensitive reference method for quantification of Δ(9) -tetrahydrocannabinol (THC) and its main metabolite 11-nor-9-carboxy-Δ(9) -tetrahydrocannabinol (THCCOOH) in oral fluid is described in this study. Samples were collected using an oral specimen collection device, followed by solid-phase extraction and liquid chromatography-tandem mass spectrometry analysis. Chromatographic separation of the analytes was achieved by gradient elution on a reversed-phase column with subsequent detection by electrospray triple quadrupole mass spectrometry in positive ionization multiple reaction monitoring mode. Quantification was performed by means of deuterated analogues of the analytes as internal standards. Total run time of the assay was 12 min. The method allowed sensitive quantification of both analytes at a limit of quantification of 0.2 ng/ml. This sensitivity is essential for analysis of samples collected with the Intercept Oral Fluid Collection device (OraSure) and an assay for simultaneous quantification of THC and THCCOOH in saliva has not yet been described. The calibration curves for THC and THCCOOH were linear in the range between 0.25 and 8 ng/ml (r(2) > 0.99). Ion suppression effects from endogenous or exogenous interferences were investigated using selected model substances (albumin, ascorbic acid, bilirubin, hemoglobin, breath spray, cigarette, chewing gum, chewing tobacco, candy, tooth whitening, and Tums antacid). These substances were chosen because of the high probability of their presence in the collected samples. None of the 11 endogenous model interferences altered the accuracy of analysis, demonstrating good robustness of the method with respect to interferences in common hygiene products, medicine, tobacco and naturally occurring endogenous substances.