Removal efficiency of emerging micropollutants in biofilter wastewater treatment plants in tropical areas.

We studied the removal of 61 emerging micropollutants, including illicit drugs, in a biofilter wastewater treatment plant located in the French Indies (Martinique). Raw wastewater concentrations were the highest for paracetamol followed by caffeine, naproxen, ibuprofen, its metabolite 2-hydroxyibuprofen, atenolol, ketoprofen, furosemide, methylparaben, cocaine, benzoylecgonine, and 11-nor-delta-9-carboxytetrahydrocannabinol (THC-COOH). The calculated removals were better than those reported in the literature, while the cumulative removal efficacy (i.e., removal of the total mass load) was estimated to be 92 ± 4%. However, this good performance may be partly explained by the removal of paracetamol (also named acetaminophen) and caffeine, which represented 86.4% of the total mass load. Our results point to the adsorption of some molecules on sludge, thus raising the question about local soil pollution from sludge spreading.

Substances detected in used syringes of injecting drug users across 7 cities in Europe in 2017 and 2018: The European Syringe Collection and Analysis Project Enterprise (ESCAPE).

BACKGROUND AND AIMS: Injecting drug use is a matter of public health concern, associated with risks of overdoses, addiction and increased risk of bloodborne viral transmissions. Self-reported data on substances injected can be inaccurate or subject to bias or drug users might be oblivious to their injected substances or adulterations. Gathering of robust analytical information on the actual composition of substances injected might provide better information about the drugs that are being used. Therefore, this study aimed to analyse the residual content of discarded syringes collected across 7 European cities, collectively called the European Syringe Collection and Analysis Project Enterprise (ESCAPE). METHODS: Used syringes were collected at street automatic injection kit dispensers or at harm-reduction services in Amsterdam, Budapest, Cologne, Glasgow, Helsinki, Lausanne and Paris. Two sampling periods were executed thus far, in 2017 and 2018. Qualitative chemical analysis of the content of used syringes was performed combining gas chromatographic (GC) and ultra(high)performance liquid chromatographic ((U)HPLC) analytical techniques with detection by mass spectrometry (MS). RESULTS: Substances detected most frequently across both campaigns were cocaine, heroin, buprenorphine, amphetamines and synthetic cathinones. In Amsterdam, Cologne, Lausanne and Glasgow heroin and cocaine were the psychoactive substances most often detected, often in conjunction with each other. Helsinki showed a high presence of buprenorphine and amphetamines. In Budapest and Paris, synthetic cathinones were frequently detected. Less synthetic cathinones and cocaine was detected in 2018, whereas buprenorphine was detected almost twice as much. Inner-city variations were found, probably reflecting the types of people who inject drugs (PWID) in different areas of the city. CONCLUSION: Overall, laboratory-confirmed local data on injected substances showed resemblance to national surveys done among PWID. However, the ESCAPE data also showed some interesting differences, showing it can be used for local interventions and complementing existing monitoring data.

Impact of peri-urban landscape on the organic and mineral contamination of pond waters and related risk assessment.

Ponds are important for their ecological value and for the ecosystem services they provide to human societies, but they are strongly affected by human activities. Peri-urban development, currently one of the most pervasive processes of land use change in Europe, exposes ponds to both urban and agricultural contaminants, causing a potential combination of adverse effects. This study, focused on 12 ponds located in a peri-urban area, has two main objectives: (1) to link the physico-chemical characteristics of the waters and the nature of their contaminants, either organic or mineral, with the human activities around ponds, and (2) to estimate the environmental risk caused by these contaminants. The ponds were sampled during two consecutive years in both spring and in autumn. Although the ponds were distributed over a limited geographical area, their contamination profiles were different and more correlated with the agricultural than the urban land use. In terms of aptitude for biology, half of the ponds were classified in degraded states due to their physico-chemical parameters, but without correlation with the endocrine disrupting activities and the levels of organic pollutants as indicators. The main quantified organic pollutants, however, were pesticides with sufficiently high levels in certain cases to induce an environmental risk exceeding the classical thresholds of risk quotient.

Wastewater-based epidemiology approach to assess population exposure to pesticides: a review of a pesticide pharmacokinetic dataset.

Wastewater-based epidemiology is an innovative approach to estimate a population's intentional and unintentional consumption of chemicals based on biomarker assays found in wastewater. This method can provide real-time objective information on the xenobiotics to which a population is directly or indirectly exposed. This approach has already been used to assess the population exposure to four classes of pesticides: organochlorines (chlordecone), triazines, organophosphates, and pyrethroids. This review aims to obtain the data (excretion rates) and characteristics (pesticide and metabolites stability, including in-sewer one) for other pesticides to broaden the scope of this new method. Excretion rates and stability descriptions for 14 pesticides, namely 2,4-D, aldrin, carbaryl, chlorobenzilate, dieldrin, diquat, ethion, glufosinate, glyphosate, folpet, malathion, parathion, penconazole, and tebuconazole, will be discussed in a practical framework.

Reprint of: The effect of the Music Day event on community drug use.

Illicit drugs consumption can be back-calculated based on the analysis of drug residues in wastewater using the wastewater-based epidemiology method. The Music Day, held on June 21 in France since 1982, has grown to global proportions and is now celebrated as World Music Day. This large outdoor event takes place in many cities with people allowed to play music in the streets. As psychotropic drugs are often associated with music events, the goal of this study is to investigate the use of illicit drugs on this day in Bordeaux, the fifth largest urban area in France. Daily sampling campaigns of composite wastewater were carried out for seven days in two wastewater treatment plants in Bordeaux in 2017 (Music Day) and 2018. World Music Day in Bordeaux has no observable effect on illicit drug consumption even if this event has massive public participation: this is the first report of the absence of an illicit drug consumption increase in a festival of such magnitude, corroborating the effect of others' views and opinions, because this event takes place publicly in the street and not among peers. Different hypothesizes are put forward to explain this fact: inappropriate type of event for drug consumption, effect of other festivals, and influence of the event's timing on a weekday.

The effect of the Music Day event on community drug use.

Illicit drugs consumption can be back-calculated based on the analysis of drug residues in wastewater using the wastewater-based epidemiology method. The Music Day, held on June 21 in France since 1982, has grown to global proportions and is now celebrated as World Music Day. This large outdoor event takes place in many cities with people allowed to play music in the streets. As psychotropic drugs are often associated with music events, the goal of this study is to investigate the use of illicit drugs on this day in Bordeaux, the fifth largest urban area in France. Daily sampling campaigns of composite wastewater were carried out for seven days in two wastewater treatment plants in Bordeaux in 2017 (Music Day) and 2018. World Music Day in Bordeaux has no observable effect on illicit drug consumption even if this event has massive public participation: this is the first report of the absence of an illicit drug consumption increase in a festival of such magnitude, corroborating the effect of others' views and opinions, because this event takes place publicly in the street and not among peers. Different hypothesizes are put forward to explain this fact: inappropriate type of event for drug consumption, effect of other festivals, and influence of the event's timing on a weekday.

Spatio-temporal assessment of illicit drug use at large scale: evidence from 7 years of international wastewater monitoring.

BACKGROUND AND AIMS: Wastewater-based epidemiology is an additional indicator of drug use that is gaining reliability to complement the current established panel of indicators. The aims of this study were to: (i) assess spatial and temporal trends of population-normalized mass loads of benzoylecgonine, amphetamine, methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA) in raw wastewater over 7 years (2011-17); (ii) address overall drug use by estimating the average number of combined doses consumed per day in each city; and (iii) compare these with existing prevalence and seizure data. DESIGN: Analysis of daily raw wastewater composite samples collected over 1 week per year from 2011 to 2017. SETTING AND PARTICIPANTS: Catchment areas of 143 wastewater treatment plants in 120 cities in 37 countries. MEASUREMENTS: Parent substances (amphetamine, methamphetamine and MDMA) and the metabolites of cocaine (benzoylecgonine) and of Δ9 -tetrahydrocannabinol (11-nor-9-carboxy-Δ9 -tetrahydrocannabinol) were measured in wastewater using liquid chromatography-tandem mass spectrometry. Daily mass loads (mg/day) were normalized to catchment population (mg/1000 people/day) and converted to the number of combined doses consumed per day. Spatial differences were assessed world-wide, and temporal trends were discerned at European level by comparing 2011-13 drug loads versus 2014-17 loads. FINDINGS: Benzoylecgonine was the stimulant metabolite detected at higher loads in southern and western Europe, and amphetamine, MDMA and methamphetamine in East and North-Central Europe. In other continents, methamphetamine showed the highest levels in the United States and Australia and benzoylecgonine in South America. During the reporting period, benzoylecgonine loads increased in general across Europe, amphetamine and methamphetamine levels fluctuated and MDMA underwent an intermittent upsurge. CONCLUSIONS: The analysis of wastewater to quantify drug loads provides near real-time drug use estimates that globally correspond to prevalence and seizure data.

Development and validation of a sensitive and selective method using GC/MS-MS for quantification of 5-fluorouracil in hospital wastewater.

Pollution of the environment by pharmaceuticals is a subject of growing scientific and societal concern. However, few quantitative data have been reported concerning hospital wastewater contamination. Among the different molecules used at hospital, antineoplastic drugs appear to be of special interest, and 5-fluorouracil (5-FU) can be considered as a key compound of this therapeutic class. To monitor this pharmaceutical in hospital wastewater, a highly specific and selective method was developed using gas chromatography tandem mass spectrometry after solid-phase extraction. This sensitive method (limit of quantification = 40 ng L(-1)) was then applied to assess sewage contamination of a middle-size hospital with oncology service located in Paris, France. Native 5-FU was detectable in 12 of the 14 analysed samples. In positive samples, concentration range was measured from 0.09 to 4.0 microg L(-1). Finally, a predicting model for the hospital wastewater concentrations is presented, and results of this model are discussed.

Exposure of an urban population to pesticides assessed by wastewater-based epidemiology in a Caribbean island.

Wastewater-based epidemiology is an innovative approach to estimate the consumption of chemicals and their exposure patterns in a population, on the basis of measurements of biomarkers in wastewater. This method can provide objective real-time information on xenobiotics directly or indirectly ingested by a population. This approach was used to examine the exposure of the Martinique population to the three classes of pesticides: triazines, organophosphates and pyrethroids. Martinique island (French West Indies) is a closed market and has been closely monitored since the early 2000's when contamination with chlordecone, an organochlorine insecticide widely applied between 1972 and 1993 in banana plantations, became a critical political issue. The aim of this study was to identify and quantify the patterns of human exposure and compare the results to those from other countries. Wastewater was collected as 24-h composite samples and analysed for selected urinary pesticide metabolites by liquid chromatography-tandem mass spectrometry. Organophosphate and pyrethroid metabolites were found in all the samples up to 330 ng/L, while triazines were found only at trace levels. Mass loads indicated higher exposure to pyrethroids than in some cities in Europe, but lower exposure to triazines and organophosphates. The estimated human intake for pyrethroids was close to the Acceptable Daily Intake, but importation of these pesticides to Martinique was low. This study illustrates the high human exposure with indoor pesticide use in comparison to its use in agriculture.

Theory versus practice, bacteriological efficiency versus personal habits: A bacteriological and user acceptability evaluation of filtering tools for people who inject drugs.

INTRODUCTION AND AIMS: People who inject drugs (PWID) are exposed to associated viral, bacterial and fungal risks. These risks can be reduced by filtration. Large disparities in the quality of filtration exist between the various available filters. This paper compares both performance and user acceptability of three filters for drug injection (cotton filters, Sterifilt® and wheel filters) by combining epidemiological and bacteriological analyses. DESIGN AND METHODS: A cross-sectional epidemiological study (ANRS-Coquelicot) using time-location sampling combined with the generalised weight sampling method was conducted among 985 PWID in France. Two filtration-based bacteriological studies of 0.20- and 0.45-µm wheel filters, Sterifilt filters and cotton filters were also conducted. RESULTS: The bacteriological study highlighted the value of using wheel filters with a porosity of less than 0.5 µm, as they limit the risk of bacterial and fungal infection. The results of this study clearly highlight a distinction between the efficiency of Sterifilt and wheel filters, the latter being more effective. Our epidemiological study highlighted that the use of cotton filters is widespread and routine, but is the subject of much criticism among PWID. Sterifilt is not widely used, and its adoption is slow. Finally, the wheel filter remains a largely untested tool. DISCUSSION AND CONCLUSIONS: Low product retention and ease of use are the two most important factors for filters for PWID. Bacterial and fungal risk filtration is less important. It is essential to educate PWID about the benefits of wheel filters. [Jauffret-Roustide M, Chollet A, Santos A, Benoit T, Péchiné S, Duplessy C, Bara J-L, Lévi Y, Karolak S, Néfau T. Theory versus practice, bacteriological efficiency versus personal habits: A bacteriological and user acceptability evaluation of filtering tools for people who inject drugs. Drug Alcohol Rev 2018;37:106-115].

Applying sewage epidemiology approach to estimate illicit drug consumption in a tropical context: Bias related to sewage temperature and pH.

Illicit drug consumption can be estimated from drug target residue (DTR) in wastewater, with the reliability of results being partly linked to DTR stability in the sewage network. However, wastewater temperature and pH drive the stability of molecules and, in this context, tropical conditions must be studied to specify the impact of residence time in the sewage network on DTR degradation. Warmth enhances biotic and abiotic processes such as degradation, leading to a decrease in oxygen content, and consequently, early diagenesis conditions in wastewater. In this study, we conduct laboratory studies under acidic pH and high temperature (30°C) conditions to determine the degradation half-lives of cocaine (COC), tetrahydrocannabinol, and heroine targets, allowing COC/benzoylecgonine (BZE) ratio variations to be predicted in sewage networks. A rapid COC degradation is observed, as already reported in the literature but without a short-term significant difference between 20°C and 30°C. Acidic pH seems to prevent degradation. Thus, theoretically, the use of COC as DTR is only reliable in acidic conditions, with the decrease in COC concentration being 6% at 8h, but over 40% in other conditions. By contrast, the use of BZE as DTR to estimate COC consumption, which is performed in practice, can be undertaken with the same back-calculation equation as used in temperate countries. However, 11-nor-delta-9-carboxytetrahydrocannabinol stability is more influenced by high temperature: concentration levels after 24h are 20% lower at 30°C than at 20°C, corresponding to a 20% and 40% decrease, respectively. Based on a mean residence time of 8h, underestimated cannabis consumption is close to 15% in tropical contexts, which is double that of temperate areas.

Validation of an ultra-high-performance liquid chromatography-tandem mass spectrometry method to quantify illicit drug and pharmaceutical residues in wastewater using accuracy profile approach.

The analysis of biomarkers in wastewater has become a common approach to assess community behavior. This method is an interesting way to estimate illicit drug consumption in a given population: by using a back calculation method, it is therefore possible to quantify the amount of a specific drug used in a community and to assess the consumption variation at different times and locations. Such a method needs reliable analytical data since the determination of a concentration in the ngL-1 range in a complex matrix is difficult and not easily reproducible. The best analytical method is liquid chromatography - mass spectrometry coupling after solid-phase extraction or on-line pre-concentration. Quality criteria are not specially defined for this kind of determination. In this context, it was decided to develop an UHPLC-MS/MS method to analyze 10 illicit drugs and pharmaceuticals in wastewater treatment plant influent or effluent using a pre-concentration on-line system. A validation process was then carried out using the accuracy profile concept as an innovative tool to estimate the probability of getting prospective results within specified acceptance limits. Influent and effluent samples were spiked with known amounts of the 10 compounds and analyzed three times a day for three days in order to estimate intra-day and inter-day variations. The matrix effect was estimated for each compound. The developed method can provide at least 80% of results within ±25% limits except for compounds that are degraded in influent.

The removal of illicit drugs and morphine in two waste water treatment plants (WWTPs) under tropical conditions.

The consumption of drugs of abuse has been recently investigated in Martinique using the back-calculation approach, also called the "sewage epidemiology" method. Results demonstrated a very high consumption considering the international data. Wastewater treatment plants (WWTPs) are located just behind the Martinique island shoreline, and effluents could impact the vulnerable corals and marine seagrass ecosystem. The present article aims to determine a WWTP's efficiency by comparing the influent and effluent of two WWTPs, with different residence times and biological treatments, located either outdoors or indoors. In parallel, a degradation study is conducted using spiked wastewater exposed to tropical and ambient temperatures. Results demonstrate the consistent efficiency of the two processes, especially for the outdoor WWTP which uses the activated sludge process. The positive effect of the tropical temperature is showed by the increase of cocaine degradation at 31 °C. Thus, low illicit drug residue concentrations in effluent would indicate that wastewater treatment is efficient and even enhanced under tropical context. This fact should be confirmed with others molecules. Furthermore, our results highlight the need for subsequent studies of sludge contamination because of their local recycling as compost.

Drug analysis of residual content of used syringes: a new approach for improving knowledge of injected drugs and drug user practices.

BACKGROUND: Since their inception, harm reduction services, including needle exchange programs, have aimed to improve and update knowledge about illicit drug consumption and injection practices in order to assess and regularly revise the effectiveness of preventive strategies. METHODS: In this paper we describe the development of a scientific approach to obtaining this type of information through analysis of the residual content of used syringes. This was done using a validated liquid chromatography method with mass spectrometry detection to identify different molecules. Used syringes were collected from automatic injection kit dispensers at 17 sites in Paris and the surrounding suburbs each month for one year. RESULTS: In total, 3489 syringes were collected. No compounds were detected in 245 syringes. Heroin was the most commonly observed compound (42%), followed by cocaine (41%), buprenorphine (29%) and 4-methylethylcathinone (23%). These analyses also showed the increased appearance of 4-methylethylcathinone between the summer and winter of 2012. CONCLUSIONS: Despite the bias involved in this approach, the method can provide rapid data on patterns of drug consumption for specific time periods and for well-defined locations. This kind of analysis enables the detection of new substances being injected and thus enables harm reduction services to revise and adapt prevention strategies.

Assessment of river contamination by estrogenic compounds in Paris area (France).

Wastewater treatment plants (WWTPs) receive a large spectrum of endocrine disrupting compounds (EDCs) that are partially eliminated during treatment processes and discharged into rivers. Given the lack of information in France about river contamination by EDCs, we chose to examine estrogenic potential of WWTP influents, effluents and receiving waters in Paris and its suburbs. Water samples were analyzed using gas chromatography coupled with mass spectrometry for quantifying natural and synthetic estrogens combined with an in vitro estrogenicity bioassay associated to a high pressure liquid chromatography fractionation. The four estrogens investigated, Estrone (E1), 17beta-Estradiol (E2), Estriol (E3) and 17alpha-Ethinylestradiol (EE2) were found in all WWTP and river samples at concentrations ranging from 2.7 to 17.6 ng/l and 1.0 to 3.2 ng/l, respectively. The synthetic estrogen EE2 seems more resistant to biodegradation in WWTPs and thus accounted for 35-50% of the estimated estrogenic activity in rivers. However, fractionation of samples and differences between concentrations of E1, E2, E3 and EE2 and the estrogenic activity measured by the in vitro bioassay suggested a complexity of mechanisms underlying the biological response that could not be attributed only to the investigated molecules.

Calibration and field evaluation of Polar Organic Chemical Integrative Sampler (POCIS) for monitoring pharmaceuticals in hospital wastewater.

The Polar Organic Chemical Integrative Sampler (POCIS) is a new tool for the sampling of organic pollutants in water. We tested this device for the monitoring of pharmaceuticals in hospital wastewater. After calibration, a field application was carried out in a French hospital for six pharmaceutical compounds (Atenolol, Prednisolone, Methylprednisolone, Sulfamethoxazole, Ofloxacin, Ketoprofen). POCIS were calibrated in tap water and wastewater in laboratory conditions close to relevant environmental conditions (temperature, flow velocity). Sampling rates (R(s)) were determined and we observed a significant increase with flow velocity and temperature. Whatever the compound, the R(s) value was lower in wastewater and the linear phase of uptake was shorter. POCIS were deployed in a hospital sewage pipe during four days and the estimated water concentrations were close to those obtained with twenty-four hour composite samples.

Presence of illicit drugs and metabolites in influents and effluents of 25 sewage water treatment plants and map of drug consumption in France.

Consumption of illicit drugs is a new concern for water management that must be considered not only because of the social and public health aspects but also in an environmental context in relation with the contamination of surface waters. Indeed, sewage treatment plant (STP) effluents contain drug residues that have not been eliminated since STP treatments are not completely efficient in their removal. We developed and validated an HPLC-MS/MS analytical method to assess the concentrations of 17 illicit drugs and metabolites in raw urban wastewaters: cocaine and its metabolites, amphetamine and amphetamine-likes (methamphetamine, MDMA, MDEA, MDA), opiates and opiate substitutes (methadone and buprenorphine), and THC-COOH cannabis metabolite. This method has been applied to the analysis of influent and effluent samples from 25 STPs located in France all over the country. The results allowed evaluating the drug consumption in the areas connected to the STPs and the efficiency of the treatment technology implied. We selected STPs according to their volume capacity, their treatment technologies (biofilters, activated sludges, MBR) and their geographical location. In influents, the concentrations varied between 6 ng/L for EDDP (main metabolite of methadone) and 3050 ng/L for benzoylecgonine (cocaine metabolite). Consumption maps were drawn for cocaine, MDMA, opiates, cannabis and amphetamine-like compounds. Geographical significant differences were observed and highlighted the fact that drug consumption inside a country is not homogeneous. In parallel, comparisons between STP technology processes showed differences of efficiency. More, some compounds appear very resistant to STP processes leading to the contamination of receiving water.

Comparing illicit drug use in 19 European cities through sewage analysis.

The analysis of sewage for urinary biomarkers of illicit drugs is a promising and complementary approach for estimating the use of these substances in the general population. For the first time, this approach was simultaneously applied in 19 European cities, making it possible to directly compare illicit drug loads in Europe over a 1-week period. An inter-laboratory comparison study was performed to evaluate the analytical performance of the participating laboratories. Raw 24-hour composite sewage samples were collected from 19 European cities during a single week in March 2011 and analyzed for the urinary biomarkers of cocaine, amphetamine, ecstasy, methamphetamine and cannabis using in-house optimized and validated analytical methods. The load of each substance used in each city was back-calculated from the measured concentrations. The data show distinct temporal and spatial patterns in drug use across Europe. Cocaine use was higher in Western and Central Europe and lower in Northern and Eastern Europe. The extrapolated total daily use of cocaine in Europe during the study period was equivalent to 356 kg/day. High per capita ecstasy loads were observed in Dutch cities, as well as in Antwerp and London. In general, cocaine and ecstasy loads were significantly elevated during the weekend compared to weekdays. Per-capita loads of methamphetamine were highest in Helsinki and Turku, Oslo and Budweis, while the per capita loads of cannabis were similar throughout Europe. This study shows that a standardized analysis for illicit drug urinary biomarkers in sewage can be applied to estimate and compare the use of these substances at local and international scales. This approach has the potential to deliver important information on drug markets (supply indicator).

Estimation of illicit drugs consumption by wastewater analysis in Paris area (France).

Illicit drugs consumption is actually an important public health concern that needs to be well defined to be managed. A new method, expressed as sewage epidemiology has been proposed by Daughton and developed by Zuccato. This method involves estimating the consumption from the measurement of drug residues in sewage. Several studies have been carried out, leading to an assessment of drugs consumption in some European countries. This work, carried out in Paris area (France) brings new data to this assessment and allows a comparison of cocaine and MDMA consumptions with European estimations. Four wastewater treatment plants (WWTPs) have been retained for the study, taking into account biological treatment, volume capacity, geographic location and social environment. Cocaine and its major metabolite benzoylecgonine (BZE), amphetamine, 3,4-methylenedioxymethamphetamine (MDMA) and buprenorphine were measured in raw water and WWTP effluent using HPLC-MS/MS after SPE extraction. Amphetamine was rarely detected. Cocaine and BZE were quantified at levels from 5 to 282 ng L(-1) and 15 to 849 ng L(-1), respectively. MDMA and buprenorphine concentrations remained under 20 ng L(-1). Cocaine consumption was estimated from cocaine or BZE concentrations measured in raw water and the results showed significant difference in drug taking during week or weekend. The estimated doses observed in this study are lower than those reported for others countries, especially Spain and Italy. MDMA consumption was estimated at lower levels than cocaine.

Chromatographic separation and identification of a water-soluble dendritic methano[60]fullerene octadecaacid.

The chromatographic separation of a highly water-soluble dendritic monoadduct methano[60]fullerene octadecaacid (dendrofullerene) with octadecylsilica bonded phases has been studied. It has been found that the RP-HPLC behavior of this dendrofullerene obeys the general rules of stationary-phase and mobile-phase selection for controlling the separation of usually acidic compounds. An RP-HPLC-ESI-MS analysis confirms the identity of the dendrofullerene and allows characterization of the molecular weights of the main impurities contained in the sample. The described methods can control the synthesis and efficiently purify this fullerene derivative, which has been previously shown to be active against mutant infectious clones of HIV-1, which are resistant to AZT and 3TC, drugs that are widely used in AIDS therapy.