Inorganic arsenic alters the development of dopaminergic neurons but not serotonergic neurons and induces motor neuron development via Sonic hedgehog pathway in zebrafish.

The mechanism of inorganic arsenic-induced neurotoxicity at the cellular level is not known. In zebrafish, teratological effects of inorganic arsenic have been shown at various concentrations. Here, we used similar concentrations of inorganic arsenic to evaluate the effects on specific neuron types. Exposure of zebrafish embryos at 5 h post fertilization (hpf) to sodium arsenite induced developmental toxicity (reduced body length) in 72 hpf larvae, beginning at a concentration of 300 mg/L concentration. Mortality or overt morphological deformity was detected at 500 mg/L sodium arsenite. While 200 mg/L sodium arsenite induced development of tyrosine hydroxylase-positive (dopaminergic) neurons, there was no significant effect on the development of 5-hydroxytryptamine (serotonergic) neurons. Sodium arsenite reduced acetylcholinesterase activity. In the hb9-GFP transgenic larvae, both 200 and 400 mg/L sodium arsenite produced supernumerary motor neurons in the spinal cord. Inhibition of the Sonic hedgehog (Shh) pathway that is essential for motor neuron development, by Gant61, prevented sodium arsenite-induced supernumerary motor neuron development. Inductively coupled plasma mass spectrometry (ICP-MS) revealed that with 200 mg/L and 400 mg/L sodium arsenite treatment, each larva had an average of 387.8 pg and 847.5 pg arsenic, respectively. The data show for the first time that inorganic arsenic alters the development of dopaminergic and motor neurons in the zebrafish larvae and the latter occurs through the Shh pathway. These results may help understand why arsenic-exposed populations suffer from psychiatric disorders and motor neuron disease and Shh may, potentially, serve as a plasma biomarker of arsenic toxicity.

Occurrence of 3-monochloropropane-1,2-diol (3-MCPD) esters and glycidyl esters in infant formulas from Germany.

Fatty acid esters of 3-monochloropropane-1,2-diol (3-MCPD), 2-monochloropropanediol-1,3-diol (2-MCPD), and glycidol are heat-induced processing contaminants formed during the deodorisation step of edible oil refining. Because these compounds are potentially carcinogenic and/or genotoxic, their presence in refined oils and fats and foods containing these oils/fats poses possible health concerns. In particular, formula-fed infants may be exposed to these compounds due to their presence in commercial infant formulas. For this reason, in 2018, the European Union established maximum limits for glycidyl esters in oils/fats and infant formulas and recently, as of January 2021, implemented regulations for the sum of free 3-MCPD and 3-MCPD fatty acid esters as well. This work involved the analysis of 3-MCPD and glycidyl ester contents in 45 infant formula products purchased from German supermarkets in 2019. The data produced in this study showed that average contaminant concentrations across all manufacturers were exceptionally low, with 100% of the products analysed falling below the current EU regulation for glycidyl esters in powdered formulas and 96% of the products analysed meeting the recently implemented regulation for the sum of free and bound 3-MCPD. In addition, a comparison of the 2019 data set to occurrence data collected for German formulas purchased in 2015 showed that average bound 3-MCPD and glycidol concentrations have decreased (from 0.094 to 0.054 µg g-1 and from 0.010 to 0.006 µg g-1, respectively), likely indicating that additional/improved mitigation measures for reducing contaminant concentrations in infant formulas have been implemented over the 4-year period. The data collected in this occurrence study are suitable for estimating levels of exposure to these compounds for German infants.Abbreviations: 2-MCPD: 2-monochloropropane-1,3-diol; 3-MCPD: 3-monochloropropane-1,2-diol; ARA: arachidonic acid; CE: collision energy; CXP: collision cell exit potential; DHA: docosahexaenoic acid; DP: declustering potential; EP: entrance potential; EPA: eicosapentaenoic acid; EtOAc: ethyl acetate; IPA: isopropanol; LC-MS/MS: liquid chromatography-tandem mass spectrometry; MeOH: methanol; MTBE: methyl tert-butyl ether; SPE: solid-phase extraction.

Changing the Landscape: An Introduction to the Agricultural and Food Chemistry Technical Program at the 258th American Chemical Society National Meeting in San Diego.

This special issue of the Journal of Agricultural and Food Chemistry (JAFC) is a highlight of the Agricultural and Food Chemistry Division (AGFD) technical program at the 258th National Meeting of the American Chemical Society (ACS) in San Diego, CA, U.S.A., on August 25-29, 2019. At the conference, AGFD had 44 oral sessions at 19 symposia and 100 poster presentations with more than 400 abstract submissions. The technical program covered a broad range of current research and development topics in agricultural and food chemistry, including bioactive food components, diet and human nutrition, utilization of agricultural materials in food systems, food packaging, nanotechnology, and food safety, as well as several special award symposia. This is the first JAFC special issue that highlights an ACS national meeting program with joint efforts from AGFD.

Assessment of intestinal absorption/metabolism of 3-chloro-1,2-propanediol (3-MCPD) and three 3-MCPD monoesters by Caco-2 cells.

3-chloro-1,2-propanediol (3-MCPD) and 3-MCPD esters are contaminants present in a variety of processed foods, including infant formulas. Toxicological data are unavailable in humans, but rodent studies have demonstrated renal and testicular toxicity from 3-MCPD and 3-MCPD esters. There is evidence that 3-MCPD esters are hydrolyzed in the digestive system, releasing 3-MCPD that would be absorbed and induce damage. We assessed absorption and metabolism of 3-MCPD and three 3-MCPD monoesters, 1-oleoyl (1-Ol), 1-linoleoyl (1-Li) and 1-palmitoyl (1-Pa) commonly found in U.S. infant formula using differentiated Caco-2 cells. After 1-hour incubation, all three monoesters released free 3-MCPD and free fatty acids (FFA) into Caco-2 cell supernatants. Free 3-MCPD had a high apparent permeability (Papp = 30.36 ± 1.31 cm/s × 10-6) suggesting that it is freely diffusible and highly absorbed by intestinal epithelium. 1-Li released 3-4-fold more 3-MCPD than 1-Ol and 1-Pa over 1 h, suggesting that this variable release rates might contribute to the overall in vivo exposure to 3-MCPD. None of the monoesters or FFA were detected in basolateral supernatants, suggesting that these compounds do not cross the intestinal wall without further transformation. In summary, this study provides relevant data to advance knowledge of in vivo intestinal absorption and metabolism of 3-MCPD monoesters.

Updated occurrence of 3-monochloropropane-1,2-diol esters (3-MCPD) and glycidyl esters in infant formulas purchased in the United States between 2017 and 2019.

Fatty acid esters of 3-monochloropropane-1,2-diol (3-MCPD) and glycidol are potentially carcinogenic and/or genotoxic processing contaminants that are formed during the process of edible oil refining. Because of their toxicological properties, the presence of these compounds in refined oils and foods containing these oils, particularly infant formula, poses a potential food safety concern. For this reason, recent research efforts have focussed on the development of methods for the analysis of MCPD and glycidyl esters in infant formula in order to estimate levels of exposure. This work presents occurrence data for 3-MCPD and glycidyl esters in 222 infant formulas purchased in the United States between December 2017 and January 2019. The results of this study show a wide range of contaminant concentrations across four different manufacturers, with average bound 3-MCPD concentrations ranging from 0.035 µg g-1 to 0.63 µg g-1 and average bound glycidol concentrations ranging from 0.019 µg g-1 to 0.22 µg g-1. The data suggest that manufacturers B and C source palm oil produced with mitigation measures, leading to reduced amounts of 3-MCPD and glycidyl esters in their infant formulas. Additionally, comparison with a previously published study in our laboratory of the occurrence of 3-MCPD and glycidyl esters in infant formula purchased in the U.S. between 2013 and 2016 revealed that, since 2016, contaminant concentrations have decreased in products produced by manufacturers A, B, and C, while contaminant amounts in formulas from manufacturer D have slightly increased.

The impact of infant formula production on the concentrations of 3-MCPD and glycidyl esters.

Fatty acid esters of 3-monochloropropanediol (3-MCPD), 2-monochlorpropanediol (2-MCPD), and glycidol are process-induced chemical contaminants found in refined vegetable oils. Due to their toxicological properties, there is potential concern regarding exposure to these compounds, particularly for formula-fed infants where refined edible oils are the primary fat source in commercial infant formulas. In order to assess exposure, 55 commercial oil samples, specifically intended for use in infant formula, were collected in 2015 from various infant formula manufacturers in the United States and analysed using a LC-MS/MS direct detection method. At the time of collection, there were no validated methods for the analysis of MCPD and glycidyl esters in infant formula. Therefore, analysis of these commercial oil samples served as an alternative for confirming the presence of these ester contaminants in infant formula. Bound 3-MCPD and glycidol concentrations in these oils ranged from below the limit of quantitation (

Estimated US infant exposures to 3-MCPD esters and glycidyl esters from consumption of infant formula.

A dietary exposure assessment was conducted for 3-monochloropropane-1,2-diol (3-MCPD) esters (3-MCPDE) and glycidyl esters (GE) in infant formulas available for consumption in the United States. 3-MCPDE and GE are food contaminants generated during the deodorisation of refined edible oils, which are used in infant formulas and other foods. 3-MCPDE and GE are of potential toxicological concern because these compounds are metabolised to free 3-MCPD and free glycidol in rodents and may have the same metabolic fate in humans. Free 3-MCPD and free glycidol have been found to cause adverse effects in rodents. Dietary exposures to 3-MCPDE and GE from consumption of infant formulas are of particular interest because formulas are the sole or primary food source for some infants. In this analysis, US Food and Drug Administration data on 3-MCPDE and GE concentrations (as 3-MCPD and glycidol equivalents, respectively) in a small convenience sample of infant formulas were used to estimate exposures from consumption of formula by infants 0-6 months of age. 3-MCPDE and GE exposures based on mean concentrations in all formulas were estimated at 7-10 and 2 µg/kg bw/day, respectively. Estimated mean exposures from consumption of formulas produced by individual manufacturers ranged from 1 to 14 µg/kg bw/day for 3-MCPDE and from 1 to 3 µg/kg for GE.

Occurrence of 3-monochloropropanediol esters and glycidyl esters in commercial infant formulas in the United States.

This work presents occurrence data for fatty acid esters of 3-chloro-1,2-propanediol (3-MCPD) and glycidol in 98 infant formula samples purchased in the United States. These contaminants are considered potentially carcinogenic and/or genotoxic, making their presence in refined oils and foods a potential health risk. Recently, attention has focused on methodology to quantify MCPD and glycidyl esters in infant formula for risk-assessment purposes. Occurrence data for 3-MCPD and glycidyl esters were produced using a procedure for extracting fat from infant formula and an LC-MS/MS method for analysing fat extracts for intact esters. Infant formulas were produced by seven manufacturers, five of which use palm oil and/or palm olein in their formulations. In formulas containing palm/palm olein, concentrations for bound 3-MCPD and glycidol ranged from 0.021 to 0.92 mg kg-1 (ppm) and from < LOQ to 0.40 mg kg-1 (ppm), respectively. Formulas not containing palm/palm olein, bound 3-MCPD and glycidol concentrations ranged from 0.072 to 0.16 mg kg-1 (ppm) and from 0.005 to 0.15 mg kg-1 (ppm), respectively. Although formulas from manufacturers A and G did not contain palm/palm olein, formulas from manufacturer E (containing palm olein) had the lowest concentrations of bound 3-MCPD and glycidol, demonstrating the effectiveness of industrial mitigation strategies.

Extraction and Liquid Chromatography-Tandem Mass Spectrometry Detection of 3-Monochloropropanediol Esters and Glycidyl Esters in Infant Formula.

A method was developed for the extraction of fatty acid esters of 3-chloro-1,2-propanediol (3-MCPD) and glycidol from infant formula, followed by quantitative analysis of the extracts using liquid chromatography-tandem mass spectrometry (LC-MS/MS). These process-induced chemical contaminants are found in refined vegetable oils, and studies have shown that they are potentially carcinogenic and/or genotoxic, making their presence in edible oils (and processed foods containing these oils) a potential health risk. The extraction procedure involves a liquid-liquid extraction, where powdered infant formula is dissolved in water and extracted with ethyl acetate. Following shaking, centrifugation, and drying of the organic phase, the resulting fat extract is cleaned-up using solid-phase extraction and analyzed by LC-MS/MS. Method performance was confirmed by verifying the percent recovery of each 3-MCPD and glycidyl ester in a homemade powdered infant formula reference material. Average ester recoveries in the reference material ranged from 84.9 to 109.0% (0.6-9.5% RSD). The method was also validated by fortifying three varieties of commercial infant formulas with a 3-MCPD and glycidyl ester solution. Average recoveries of the esters across all concentrations and varieties of infant formula ranged from 88.7 to 107.5% (1.0-9.5% RSD). Based on the validation results, this method is suitable for producing 3-MCPD and glycidyl ester occurrence data in all commercially available varieties of infant formula.

Identification of unknown compounds from polyester cans coatings that may potentially migrate into food or food simulants.

Cross-linked polyester resins are being introduced into the market as alternatives to epoxy resins as coatings for metal food cans. Identification of potential migrants, from these coatings into food, is a significant analytical challenge due to the diversity of substances employed in the manufacture of the coatings. However, such identification is required to assess migration from the can coating into the food and quantify dietary exposure. Polyester can coatings were extracted with acetonitrile at 40°C for 24h and the extracts were analyzed by a variety of analytical techniques, including GC-MS, HPLC-DAD/MS, HPLC-DAD/CAD and UHPL C-HRMS. Twenty nine non-volatile oligomers were tentatively identified using retention times, UV spectra, and accurate mass measurements. Identified oligomers suggest the coating in use for food cans is a polyester coating and is mainly based on the monomers isophthalic acid, terephthalic acid and nadic acid. To give confidence in the identification, one of the tentatively identified oligomer was synthetized and analyzed by (13)C and (1)H NMR and UHPL C-HRMS. The NMR and HRMS results, confirmed the presence of this compound in the can extracts. Finally, to determine if rapid, direct detection of the oligomers was practical, the coatings were analyzed by DART-HRMS. Twenty three out of the 29 oligomers were identified in the coating by direct measurement with DART-HRMS in few minutes.

Method development and survey of Sudan I-IV in palm oil and chilli spices in the Washington, DC, area.

Sudan I, II, III and IV dyes are banned for use as food colorants in the United States and European Union because they are toxic and carcinogenic. These dyes have been illegally used as food additives in products such as chilli spices and palm oil to enhance their red colour. From 2003 to 2005, the European Union made a series of decisions requiring chilli spices and palm oil imported to the European Union to contain analytical reports declaring them free of Sudan I-IV. In order for the USFDA to investigate the adulteration of palm oil and chilli spices with unapproved colour additives in the United States, a method was developed for the extraction and analysis of Sudan dyes in palm oil, and previous methods were validated for Sudan dyes in chilli spices. Both LC-DAD and LC-MS/MS methods were examined for their limitations and effectiveness in identifying adulterated samples. Method validation was performed for both chilli spices and palm oil by spiking samples known to be free of Sudan dyes at concentrations close to the limit of detection. Reproducibility, matrix effects, and selectivity of the method were also investigated. Additionally, for the first time a survey of palm oil and chilli spices was performed in the United States, specifically in the Washington, DC, area. Illegal dyes, primarily Sudan IV, were detected in palm oil at concentrations from 150 to 24 000 ng ml(-1). Low concentrations (< 21 µg kg(-1)) of Sudan dyes were found in 11 out of 57 spices and are most likely a result of cross-contamination during preparation and storage and not intentional adulteration.

Development of a liquid chromatography-tandem mass spectrometry method for the determination of sulfite in food.

Sulfites are widely used food preservatives that can cause severe reactions in sensitive individuals. As a result, the U.S. FDA requires that sulfites be listed on the label of any food product containing >10 mg/kg (ppm) sulfite (measured as sulfur dioxide). Currently, the optimized Monier-Williams (MW) method (AOAC Official Method 990.28) is the most common approach for determining sulfite concentrations in food samples. However, this method is time-consuming and lacks specificity in certain matrices. An improved rapid, sensitive, and selective method has been developed using electrospray ionization (ESI) high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the determination of sulfite in various food matrices. A total of 12 different types of foods were evaluated. These included dried fruits and vegetables, frozen seafood, sweeteners, and juices. The matrix is extracted with a buffered formaldehyde solution, converting free and reversibly bound sulfite to the stable formaldehyde adduct, hydroxymethylsulfonate (HMS). Extracts are prepared for injection using a C18 SPE cartridge to remove any lipophilic compounds. HMS is then separated from other matrix components using hydrophilic interaction chromatography (HILIC) and detected using multiple reaction monitoring (MRM). The method was validated at 5 concentrations in 12 food matrices. Accuracy data showed spiked recoveries ranging from 84 to 115% in representative foods. Six commercially available sulfited products were analyzed using the LC-MS/MS method, as well as the MW method, to determine if differences exist.

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the direct detection of 2-monochloropropanediol (2-MCPD) esters in edible oils.

A new analytical method has been developed and validated for the detection and quantification of 2-monochloropropanediol (2-MCPD) esters in edible oils. The target compounds are potentially carcinogenic contaminants formed during the processing of edible oils. As the 2-MCPD esters that occur most frequently in refined edible oils were not commercially available, standards were synthesized with identity and purity (95+%) confirmed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and (1)H NMR. Target analytes are separated from edible oil matrices using a two-step solid-phase extraction (SPE) procedure. The extracts are then analyzed using LC-MS/MS with electrospray ionization (ESI). The method has been validated for 11 2-MCPD diesters and 3 2-MCPD monoesters in soybean oil, olive oil, and palm oil using an external calibration curve. The ranges of average recoveries and relative standard deviations (RSD) across the three oil matrices at three spiking concentrations are 79-106% (3-13% RSD) for the 2-MCPD diesters and 72-108% (4-17% RSD) for the 2-MCPD monoesters, with limits of quantitation at or below 30 ng/g for the diesters and 90 ng/g for the monoesters.

Application of single immunoaffinity clean-up for simultaneous determination of regulated mycotoxins in cereals and nuts.

A rapid and sensitive analytical strategy for the simultaneous determination of twelve mycotoxins (aflatoxins, fumonisins, zearalenon, deoxynivalenol, ochratoxin A, T-2 and HT-2 toxins) using ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was developed and validated. The method was validated for peanuts, barley and maize-breakfast cereals; selected as they represent the matrices most often contaminated by mycotoxins. The method is designed for fast and reliable analyses of mycotoxins in regulatory, industrial and private laboratories. Multi-target immunoaffinity columns containing antibodies for all mycotoxins studied herein were used for sample clean-up. Method optimization was predominantly focused on the simplification of extraction and clean-up procedure recommended by column producers. This newly developed and simplified procedure decreased both the sample preparation time and the solvent volumes used for their processing. The analysis of all regulated mycotoxins was conducted by a newly developed UHPLC-MS/MS method with a sample run time of only ten minutes. The method trueness was tested with analytical spikes and certified reference materials, with recoveries ranging from 71% to 112% for all of the examined mycotoxins.

Occurrence of 3-MCPD and glycidyl esters in edible oils in the United States.

Fatty acid esters of 3-monochloropropanediol (3-MCPD) and glycidol are processing contaminants found in a wide range of edible oils. While both 3 MCPD and glycidol have toxicological properties that at present has concerns for food safety, the published occurrence data are limited. Occurrence information is presented for the concentrations of 3-MCPD and glycidyl esters in 116 retail and/or industrial edible oils and fats using LC-MS/MS analysis of intact esters. The concentrations for bound 3-MCPD ranged from below the limit of quantitation (

Analysis of processing contaminants in edible oils. Part 2. Liquid chromatography-tandem mass spectrometry method for the direct detection of 3-monochloropropanediol and 2-monochloropropanediol diesters.

A method was developed and validated for the detection of fatty acid diesters of 2-monochloropropanediol (2-MCPD) and 3-monochloropropanediol (3-MCPD) in edible oils. These analytes are potentially carcinogenic chemical contaminants formed during edible oil processing. After separation from oil matrices using a two-step solid-phase extraction (SPE) procedure, the target compounds are quantitated using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with electrospray ionization (ESI). The first chromatographic conditions have been developed that separate intact diesters of 2-MCPD and 3-MCPD, allowing for their individual quantitation. The method has been validated for 28 3-MCPD diesters of lauric, myristic, palmitic, linolenic, linoleic, oleic, and stearic acids in coconut, olive, and palm oils, as well as 3 2-MCPD diesters, using an external calibration curve. The range of average recoveries and relative standard deviations (RSDs) across the three oil matrices at three spiking concentrations are 88-118% (2-16% RSD) with maximum limits of quantitation of 30 ng/g (ppb).

Analysis of processing contaminants in edible oils. Part 1. Liquid chromatography-tandem mass spectrometry method for the direct detection of 3-monochloropropanediol monoesters and glycidyl esters.

A new analytical method has been developed and validated for the detection of glycidyl esters (GEs) and 3-monochloropropanediol (3-MCPD) monoesters in edible oils. The target compounds represent two classes of potentially carcinogenic chemical contaminants formed during the processing of edible oils. Target analytes are separated from edible oil matrices using a two-step solid-phase extraction (SPE) procedure. The extracts are then analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with electrospray ionization (ESI). Chromatographic conditions that separate sn-1 and sn-2 monoesters of 3-MCPD have been developed for the first time. The method has been validated for GEs, sn-1 3-MCPD monoesters of lauric, myristic, linolenic, linoleic, oleic, and stearic acids, and sn-2 3-MCPD monoesters of oleic and palmitic acids in coconut, olive, and palm oils using an external calibration curve. The range of average recoveries and relative standard deviations (RSDs) across the three oil matrices at three spiking concentrations are 84-115% (3-16% RSD) for the GEs, 95-113% (1-10% RSD) for the sn-1 3-MCPD monoesters, and 76.8-103% (5.1-11.2% RSD) for the sn-2 3-MCPD monoesters, with limits of quantitation at or below 30 ng/g for the GEs, 60 ng/g for sn-1 3-MCPD monoesters, and 180 ng/g for sn-2 3-MCPD monoesters.

A liquid chromatography-tandem mass spectrometry method for the detection of economically motivated adulteration in protein-containing foods.

A new analytical method was developed to determine the presence of six (6) compounds with the potential to be used in economic adulteration to enhance the nitrogen content in milk products and bulk proteins. Residues were extracted from the matrix with 2% formic acid, after which acetonitrile (ACN) was added to induce precipitation of the proteins. Extracts were analyzed by liquid chromatography using a ZIC-HILIC column with tandem mass spectrometry (LC-MS/MS) using electrospray ionization (ESI). Single-laboratory method validation data was collected in six matrices fortified at concentrations down to 1.0 µg/g (ppm). Average recoveries and average relative standard deviations (RSD) using spiked matrix calibration standard curves were the following: cyromazine (CY) 95.9% (7.5% RSD), dicyandiamide (DC) 98.1% (5.6% RSD), urea 102.5% (8.6% RSD), biuret (BU) 97.2% (6.6% RSD), triuret (TU) 97.7% (5.7% RSD), and amidinourea (AU) 93.4% (7.4% RSD). This method provides a rapid and effective approach to proactively combat economically motivated adulteration in protein-containing products.

Collaborative validation of the QuEChERS procedure for the determination of pesticides in food by LC-MS/MS.

Seven FDA pesticide laboratories collaborated to develop and validate an LC-MS/MS method to determine 173 pesticides in <20 min. The average determination coefficient (r²) was >0.99 for all but two compounds tested. The limits of detection were <20 ng/mL for all compounds and <10 ng/mL for 363 of the 368 transitions reported. The method was used to determine pesticides in two AOAC sponsored proficiency samples. The LC-MS/MS determination was used for the analysis of oranges, carrots and spinach using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, Safe) method. Each matrix was fortified at 20, 100, 400, and 1000 ng/g. No false positive responses were detected in controls of the three matrices. 165 pesticides had recoveries between 70 and 130%, and 161 had minimum detection levels less than 10 ng/g. Recoveries of 169 compounds for the 1000 ng/g spikes were within 50-150%. A matrix effect study indicated all three matrices caused a small net suppressing effect, the most pronounced attributable to the citrus matrix. The procedure proved to be accurate, precise, linear, sensitive and rugged, and adds 100 pesticides to the scope of the FDA pesticide program.

A new fullerene complexation ligand: N-pyridylfulleropyrrolidine.

The subject of this paper is a new fullerene building block design with the potential for defined geometry and good electronic communication. The synthesis and characterization of a new pyridinofullerene ligand capable of forming axially symmetric complexes with metalloporphyrins is reported. X-ray structural and molecular modeling studies, (1)H NMR, UV-vis spectroscopy, electrochemistry studies, and fluorescence quenching data support the formation of a strong complex between the new ligand and the metal center of ZnTPP. On the basis of computational studies, the highest occupied molecular orbital (HOMO) of this ligand is significantly different from a model compound with insulating carbons between the pyridine and the fullerene. The N-pyridinium fulleropyrrolidine salts of the new ligand and model compound were also prepared and their spectral and electrochemical properties are reported.