A study of Irish secondary school student's views on mental health supports in school.

OBJECTIVES: An increasing number of young people are experiencing mental health difficulties and schools have been identified as environments that can support them. However, it is unclear how students feel about the current supports in school and whether they are used. The aim of this study is to explore the perceptions of young people in Irish post-primary schools regarding mental health and well-being supports in schools. METHODS: An online survey was conducted with (n=109) young people to determine the perceptions of mental health and well-being supports in post-primary Irish schools. Using a convenience sampling method, an online survey was distributed via gatekeepers in local youth and sporting groups. Data collection was completed using the Barriers to Seeking Help-brief version (BASH-B) and additional tailored questions. RESULTS: Qualities like being 'trustworthy' and 'a good listener' were reported as key for adults to be considered a good support in schools. Current mental health and well-being supports were not considered adequate with 65.1% of the participants feeling 'somewhat supported' and 22.9% feeling 'not at all supported'. CONCLUSIONS: Better advertising of mental health support services should be implemented in schools to promote awareness. This study can inform the development of such services which are urgently needed.

In vivo chronic nicotine exposure differentially and reversibly affects upregulation and stoichiometry of α4ß2 nicotinic receptors in cortex and thalamus.

Studies with heterologous expression systems have shown that the α4ß2 nicotinic acetylcholine receptor (nAChR) subtype can exist in two stoichiometries (with two [(α4)2(ß2)3] or three [(α4)3(ß2)2] copies of the α subunit in the receptor pentamer) which have different pharmacological and functional properties and are differently regulated by chronic nicotine treatment. However, the effects of nicotine treatment in vivo on native α4ß2 nAChR stoichiometry are not well known. We investigated in C57BL/6 mice the in vivo effect of 14-day chronic nicotine treatment and subsequent withdrawal, on the subunit expression and ß2/α4 subunit ratio of (3)H-epibatidine labeled α4ß2*-nAChR in total homogenates of cortex and thalamus. We found that in basal conditions the ratio of the ß2/α4 subunit in the cortex and thalamus is different indicating a higher proportion in receptors with (α4)2(ß2)3 subunit stoichiometry in the thalamus. For cortex exposure to chronic nicotine elicited an increase in receptor density measured by (3)H-epibatidine binding, an increase in the α4 and ß2 protein levels, and an increase in ß2/α4 subunit ratio, that indicates an increased proportion of receptors with the (α4)2(ß2)3 stoichiometry. For thalamus we did not find a significant increase in receptor density, α4 and ß2 protein levels, or changes in ß2/α4 subunit ratio. All the changes elicited by chronic nicotine in cortex were transient and returned to basal levels with an average half-life of 2.8 days following nicotine withdrawal. These data suggest that chronic nicotine exposure in vivo favors increased assembly of α4ß2 nAChR containing three ß2 subunits. A greater change in stoichiometry was observed for cortex (which has relatively low basal expression of (α4)2(ß2)3 nAChR) than in thalamus (which has a relatively high basal expression of (α4)2(ß2)3 nAChR).

Rare human nicotinic acetylcholine receptor α4 subunit (CHRNA4) variants affect expression and function of high-affinity nicotinic acetylcholine receptors.

Nicotine, the primary psychoactive component in tobacco smoke, produces its behavioral effects through interactions with neuronal nicotinic acetylcholine receptors (nAChRs). α4ß2 nAChRs are the most abundant in mammalian brain, and converging evidence shows that this subtype mediates the rewarding and reinforcing effects of nicotine. A number of rare variants in the CHRNA4 gene that encode the α4 nAChR subunit have been identified in human subjects and appear to be underrepresented in a cohort of smokers. We compared three of these variants (α4R336C, α4P451L, and α4R487Q) to the common variant to determine their effects on α4ß2 nAChR pharmacology. We examined [(3)H]epibatidine binding, interacting proteins, and phosphorylation of the α4 nAChR subunit with liquid chromatography and tandem mass spectrometry (LC-MS/MS) in HEK 293 cells and voltage-clamp electrophysiology in Xenopus laevis oocytes. We observed significant effects of the α4 variants on nAChR expression, subcellular distribution, and sensitivity to nicotine-induced receptor upregulation. Proteomic analysis of immunopurified α4ß2 nAChRs incorporating the rare variants identified considerable differences in the intracellular interactomes due to these single amino acid substitutions. Electrophysiological characterization in X. laevis oocytes revealed alterations in the functional parameters of activation by nAChR agonists conferred by these α4 rare variants, as well as shifts in receptor function after incubation with nicotine. Taken together, these experiments suggest that genetic variation at CHRNA4 alters the assembly and expression of human α4ß2 nAChRs, resulting in receptors that are more sensitive to nicotine exposure than those assembled with the common α4 variant. The changes in nAChR pharmacology could contribute to differences in responses to smoked nicotine in individuals harboring these rare variants.

Development of a rapid screening method to determine primary aromatic amines in kitchen utensils using direct analysis in real time mass spectrometry (DART-MS).

Primary aromatic amines (PAAs) are a group of substances with undesirable health effects, that are used in a variety of commercial products. Several recent studies, using a number of screening and confirmatory methods, have reported the migration of PAAs from some kitchen utensils into acetic acid 3% (w/v). Many of these methods require significant sample preparation, therefore the aim of this work was to determine if direct analysis in real time mass spectrometry (DART-MS) could be utilised as a rapid screening tool for the determination of PAAs in kitchen utensils. DART-MS results from direct analysis of the utensil have been compared with results of PAA migration by ultra high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method. The UPLC-MS/MS method had excellent linearity, appropriate sensitivity (LOD ≤ 1.5 µg L(-1); LOQ ≤ 4.5 µg L(-1)), repeatability from 2.4 to 13.2% and acceptable recoveries. DART-MS results were in good agreement with UPLC-MS/MS data, with 100% of non-compliant (PAA positive) samples successfully identified by DART-MS.

Migration of perfluoroalkyl acids from food packaging to food simulants.

A broad range of fluorochemicals is used to impart oil and water barrier properties to paper and paperboard food packaging. Many of the fluorochemicals are applied to paper and paperboard as complex mixtures containing reaction products and by-products and unreacted starting materials. This work primarily focussed on the determination of seven perfluorocarboxylic acids (PFCAs) in two commercially available food contact papers: a di-perfluoro-alkyloxy-amino-acid and a perfluoroalkyl phosphate surfactant. In addition, the migration of the PFCAs into five food simulants from two commercial packages was evaluated. All seven PFCAs were detected in the range of 700-2220 µg kg⁻¹ of paper, while three perfluoroalkyl sulphonates were under the LOD. Results from migration tests showed that migration depends on paper characteristics, time and food simulant. The percentage of migration after 10 days at 40°C ranged from 4.8% to 100% for the two papers and different food simulants.

Non-visible print set-off of photoinitiators in food packaging: detection by ambient ionisation mass spectrometry.

Direct analysis in real time coupled to time-of-flight mass spectrometry (DART/TOF-MS) was used to detect the non-visible set-off of photoinitiators on the food contact surface of three different packages. The samples were intentionally under-cured to provoke set-off. Twelve commercially available photoinitiators were included in the ink formulations including α-amino-, morpholino, and α-hydroxy benzophenones, thioxanthones, aryl-phosphine oxide and three polymeric versions of these. Major colours of the packages' prints were analysed, as well as the specific areas of the inner surface in contact with them. Larger quantities of photoinitiators were detected on the food contact areas in contact with the darker colours of the images. Speed-cure 7005 and 4-phenylbenzophenone were the compounds most susceptible to set-off in each of the samples by DART response. An identification protocol for unknown set-off compounds was tested, resulting in the set-off detection of diethylene glycol ethers, erucamide and acrylates, and confirmed by solvent extraction GC-MS analysis. Finally, DART/TOF-MS was scanned across transects of the food contact side of packages to map the presence of photoinitiators. Higher photoinitiator signals were observed in patterns corresponding to the printed image, suggesting DART/TOF-MS might "image" print set-off.

Assessing direct analysis in real-time-mass spectrometry (DART-MS) for the rapid identification of additives in food packaging.

The ambient ionization technique direct analysis in real time (DART) was characterized and evaluated for the screening of food packaging for the presence of packaging additives using a benchtop mass spectrometer (MS). Approximate optimum conditions were determined for 13 common food-packaging additives, including plasticizers, anti-oxidants, colorants, grease-proofers, and ultraviolet light stabilizers. Method sensitivity and linearity were evaluated using solutions and characterized polymer samples. Additionally, the response of a model additive (di-ethyl-hexyl-phthalate) was examined across a range of sample positions, DART, and MS conditions (temperature, voltage and helium flow). Under optimal conditions, molecular ion (M+H+) was the major ion for most additives. Additive responses were highly sensitive to sample and DART source orientation, as well as to DART flow rates, temperatures, and MS inlet voltages, respectively. DART-MS response was neither consistently linear nor quantitative in this setting, and sensitivity varied by additive. All additives studied were rapidly identified in multiple food-packaging materials by DART-MS/MS, suggesting this technique can be used to screen food packaging rapidly. However, method sensitivity and quantitation requires further study and improvement.

Migration of fluorochemical paper additives from food-contact paper into foods and food simulants.

Fluorochemical-treated paper was tested to determine the amount of migration that occurs into foods and food-simulating liquids and the characteristics of the migration. Migration characteristics of fluorochemicals from paper were examined in Miglyol, butter, water, vinegar, water-ethanol solutions, emulsions and pure oil containing small amounts of emulsifiers. Additionally, microwave popcorn and chocolate spread were used to investigate migration. Results indicate that fluorochemicals paper additives do migrate to food during actual package use. For example, we found that microwave popcorn contained 3.2 fluorochemical mg kg(-1) popcorn after popping and butter contained 0.1 mg kg(-1) after 40 days at 4 degrees C. Tests also indicate that common food-simulating liquids for migration testing and package material evaluation might not provide an accurate indication of the amount of fluorochemical that actually migrates to food. Tests show that oil containing small amounts of an emulsifier can significantly enhance migration of a fluorochemical from paper.

Diffusion behaviour of additives in polypropylene in correlation with polymer properties.

The migration behaviour of polymer additives in 17 polypropylene (PP) samples is described. These samples cover the major types of PP used in food packaging. The diffusion coefficients of additives with relatively small molecular masses, M(r) = 136 (limonene), as well as the migration of typical antioxidants used in PP up to M(r) = 1178 (IRGANOX 1010), were measured at different temperatures. In addition, the diffusion data and percentages of xylene-soluble fractions were correlated. This enables a prediction of the migration behaviour of a PP sample by testing its 'isotactic index' with xylene. The results clearly indicate that PP can be subdivided, from the migration point of view, into the monophasic homopolymer (h-PP), the monophasic random copolymer (r-PP), and the heterophasic copolymer (heco-PP). The diffusion coefficients for r-PP are at least one order of magnitude higher than those of h-PP and comparable with the values for heco-PP. Upper limits for the diffusion values can be calculated based on the safety margin required by consumer protection laws.

Determination of 2,6-diisopropylnaphthalene (DIPN) and n-dibutylphthalate (DBP) in food and paper packaging materials from US marketplaces.

A gas chromatography-ion-trap tandem mass spectrometry procedure was developed for the determination of 2,6-diisopropylnaphthalene (DIPN) and n-dibutylphthalate (DBP) in domestic and imported paper packages and food sold in US marketplaces. The procedure involved ultrasonic extraction with dichloromethane, followed by analysis with the gas chromatography-ion-trap tandem mass spectrometry. Calibration curves for DIPN and DBP were achieved with concentrations ranging from 0.01 to 10 microg ml(-1) and the corresponding r(2) values were 0.9976 and 0.9956, respectively. In most of the fortified samples the recoveries were higher than 80% with a relative standard deviation (RSD) <10%. Using this procedure, it was found that less than 20% of the tested domestic packages and more than 60% of the tested imported food packages contained both DIPN and DBP. The concentrations of DIPN and DBP ranged from 0.09 to 20 mg kg(-1) and 0.14 to 55 mg kg(-1), respectively, with most of the DINP and DBP levels lower than 20 mg kg(-1). DIPN was not detected (<0.01 mg kg(-1)) in 41 food samples and DBP was only detected in two domestic and four imported food samples with concentrations ranging from <0.01 to 0.81 mg kg(-1).

Interfacial behavior of common food contact polymer additives.

Irganox 1076 (IN1076) and Irganox 1010 (IN1010), phenol containing species often used as antioxidant additives in food packaging polymers have both hydrophilic and hydrophobic functional groups. Consequently these additives are likely to absorb to surfaces where their free energy is minimized. Experiments described in this work examine the two-dimensional phase behavior and vibrational structure of IN1076 and IN1010 films adsorbed to the air/water interface. Surface pressure isotherms show that repeated compression of these films leads to continued irreversible loss of molecules and that on a per molecule basis, this loss is more pronounced for IN1076 than for IN1010. Differences in the surface properties of these two antioxidant additives are interpreted based on differences in molecular structure. Surface specific vibrational measurements of these organic films show very little conformational order, implying that even when closely packed, both antioxidant species have little affinity for forming highly organized domains. These findings have important ramifications for mechanisms that reduce antioxidant activity in polymers as well as descriptions of antioxidant blooming on polymer surfaces.

Rapid quantitative and qualitative confirmatory method for the determination of monofluoroacetic acid in foods by liquid chromatography-mass spectrometry.

A rapid quantitative method and a qualitative confirmatory method for the determination of monofluoroacetic acid (MFA) in complex food matrices are presented. The quantitative method utilizes a water extraction, solid phase extraction clean-up and liquid chromatography-mass spectrometry (LC-MS) for determination of MFA. This method showed a high degree of specificity, detecting MFA in all of the spiked samples, while none of the unfortified samples tested positive for MFA. Spike recoveries were high in all matrices analyzed, varying from 85 to 110%, and comparable at low (2mg/L) and high (20mg/L) spiking levels. Repeatability tests at the low spiking levels yielded RSDs of less than 5% for all matrices analyzed. The qualitative confirmatory method developed is conceptually different from the quantitative method, ensuring that both methods would not be subject to the same interferences. The method uses the formation of the hydrazide of MFA through derivatization with 2-nitrophenylhydrazine. This derivatization is well established for the determination of carboxylic acids, but this is the first application to the determination of MFA. The derivatization yield was matrix dependent, however the limit of detection (LOD) (0.8microg/L) was sufficient to confirm the presence of MFA in all spiked matrices. Repeatability tests at the low spiking levels yielded RSDs of approximately 7% for all matrices analyzed.

Diffusion of limonene in polyethylene.

Diffusion coefficients of limonene in various linear low-density polyethylene (LLDPE) and low-density polyethylene (LDPE) resins have been determined from sorption data using a thermogravimetric methodology. From these data, one can determine whether polymer synthesis parameters such as the choice of catalytic process or co-monomer result in substantial differences in how much food packaging additives might migrate to food. For example, LLDPE is currently manufactured using either one of two distinct catalytic processes: Ziegler-Natta (ZN) and metallocene, a single-site catalyst. ZN catalysis is a heterogeneous process that has dominated polyolefin synthesis over the last half-century. It involves a transition metal compound containing a metal-carbon bond that can handle repeated insertion of olefin units. In contrast, metallocene catalysis has fewer than 20 years of history, but has generated much interest due to its ability to produce highly stereospecific polymers at a very high yield. In addition to high stereospecificity, metallocene-catalysed polymers are significantly lower in polydispersity than traditional ZN counterparts. Absorption and desorption testing of heat-pressed films made from LLDPE and LDPE resins of varying processing parameters indicates that diffusion coefficients of limonene in these resins do not change substantially.

Perfluorochemicals: potential sources of and migration from food packaging.

Perfluorochemicals are widely used in the manufacturing and processing of a vast array of consumer goods, including electrical wiring, clothing, household and automotive products. Furthermore, relatively small quantities of perfluorochemicals are also used in the manufacturing of food-contact substances that represent potential sources of oral exposure to these chemicals. The most recognizable products to consumers are the uses of perfluorochemicals in non-stick coatings (polytetrafluoroethylene (PTFE)) for cookware and also their use in paper coatings for oil and moisture resistance. Recent epidemiology studies have demonstrated the presence of two particular perfluorochemicals, perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) in human serum at very low part per billion levels. These perfluorochemicals are biopersistent and are the subject of numerous studies investigating the many possible sources of human exposure. Among the various uses of these two chemicals, PFOS is a residual impurity in some paper coatings used for food contact and PFOA is a processing aid in the manufacture of PTFE used for many purposes including non-stick cookware. Little information is available on the types of perfluorochemicals that have the potential to migrate from perfluoro coatings into food. One obstacle to studying migration is the difficulty in measuring perfluorochemicals by routine conventional analytical techniques such as GC/MS or LC-UV. Many perfluorochemicals used in food-contact substances are not detectable by these conventional methods. As liquid chromatography-mass spectrometry (LC/MS) develops into a routine analytical technique, potential migrants from perfluoro coatings can be more easily characterized. In this paper, data will be presented on the types of perfluoro chemicals that are used in food packaging and cookware. Additionally, research will be presented on the migration or potential for migration of these chemicals into foods or food simulating liquids. Results from migration tests show mg kg(-1) amounts of perfluoro paper additives/coatings transfer to food oil. Analysis of PTFE cookware shows residual amounts of PFOA in the low microg kg(-1) range. PFOA is present in microwave popcorn bag paper at amounts as high as 300 microg kg(-1).

Evaluation of migration models that might be used in support of regulations for food-contact plastics.

Materials and articles intended to come into contact with food must be shown to be safe because they might interact with food during processing, storage and the transportation of foodstuffs. Framework Directive 89/109/EEC and its related specific Directives provide this safety basis for the protection of the consumer against inadmissible chemical contamination from food-contact materials. Recently, the European Commission charged an international group of experts to demonstrate that migration modelling can be regarded as a valid and reliable tool to calculate 'reasonable worst-case' migration rates from the most important food-contact plastics into the European Union official food simulants. The paper summarizes the main steps followed to build up and validate a migration estimation model that can be used, for a series of plastic food-contact materials and migrants, for regulatory purposes. Analytical solutions of the diffusion equation in conjunction with an 'upper limit' equation for the migrant diffusion coefficient, D(P), and the use of 'worst case' partitioning coefficients K(P,F) were used in the migration model. The results obtained were then validated, at a confidence level of 95%, by comparison with the available experimental evidence. The successful accomplishment of the goals of this project is reflected by the fact that in Directive 2002/72/EC, the European Commission included the mathematical modelling as an alternative tool to determine migration rates for compliance purposes.

Migration of a UV stabilizer from polyethylene terephthalate (PET) into food simulants.

The migration characteristics of the UV stabilizer Tinuvin 234 (2-(2H-benzotriazol-2-yl)-4,6-bis (1-methyl-1-phenylethyl)phenol) into food simulants has been measured from polyethylene terephthalate (PET) using HPLC with UV detection. Ethanol/ water, isooctane and a fractionated coconut oil simulant (Miglyol) were used as food simulating solvents. The migration characteristics were measured at temperatures in the range of 40-70 degrees C. Diffusion coefficients were determined to be in the range of 1 x 10(-14) cm2 s(-1) to 1 x 10(-18) cm2 s(-1). At 40 degrees C, the amount of migration into 95% ethanol after 10 days was 2 microg dm(-2). Isooctane is determined to be a good fatty food simulant that provides similar results for PET to those of fatty foods.

Effectiveness of polypropylene film as a barrier to migration from recycled paperboard packaging to fatty and high-moisture food.

The capability of a polypropylene (PP) film barrier to prevent migration of residual contaminants from recycled paperboard into food simulants was studied. Anthracene, benzophenone, methyl stearate and pentachlorophenol were chosen as chemical surrogates to represent classes of contaminants likely to be found in recycled paper/paperboard. Each surrogate was spiked into a test specimen made of seven thin virgin paper layers at concentrations of 1-50 mg kg(-1). Test specimen were dried, stacked and sandwiched with PP films, laminated with PP film and then subjected to migration experiments using a compression cell maintained at 100 degrees C for 2 h. The concentration of the surrogates in the test specimen and in 95% ethanol, isopropanol and 10% ethanol food-simulating solvents was determined by gas chromatography with flame ionization and electron capture detection. The results show that although the concentrations of the surrogates in the food simulants decreased with an increase in PP film thickness, they were still high and generally resulted in dietary concentrations >0.5 microg kg(-1), the level that US Food and Drug Administration would equate with negligible risk for a contaminant migrating from food packaging. Only at the lowest spiking level (1 mg kg(-1) benzophenone) did migration from the paperboard through a 0.127-mm PP film result in a dietary concentration of

Effects of gamma- and electron-beam irradiation on semi-rigid amorphous polyethylene terephthalate copolymers.

Two semi-rigid amorphous polyethylene terephthalate copolymer materials (in both sheet and powder forms) containing 3% 1,4-cyclohexane dimethanol (CHDM) and 31% CHDM were irradiated at 5, 25 and 50 kGy at ambient temperature with a (60)Co radiator or an electron-beam accelerator. After irradiation, volatiles were determined using static headspace sampling with capillary gas chromatography and mass selective detection or flame ionization detection (HS/GC/MSD or FID). Non-volatiles were extracted with 10% aqueous ethanol and 100% n-heptane food-simulating solvents, maintained at 40 degrees C for up to 10 days. The non-volatiles in the materials and those migrating into the food-simulating solvents were determined by high-performance liquid chromatography (HPLC) with ultraviolet and/or photodiode array detection. The results obtained from the HS/GC/MSD suggest that no new chemicals were detected by either gamma- or e-beam irradiation when compared with non-irradiated specimens. The major volatiles in the copolymers were acetaldehyde and 2-methyl-1,3-dioxolane. The concentrations of acetaldehyde increased from 1.24-1.96 mg kg(-1) to 1.94-3.65, 3.52-7.23 and 5.45-15.37 mg kg(-1) after exposure to 5, 25 and 50 kGy doses, respectively. The concentrations of 2-methyl-1,3-dioxolane decreased from 2.49-5.26 mg kg(-1) to 2.07-3.13, 1.33-2.14 and 0.64-2.24 mg kg(-1) after exposure to 5, 25 and 50 kGy doses, respectively. The results of analysis of the copolymers for non-volatiles show that irradiation did not produce any new detectable non-volatile chemicals. A 5 kGy dose had no detectable effect on either copolymer. The 25 and 50 kGy doses had slightly different effects with respect to gamma- and e-beam irradiation on low MW oligomers. However, these increased doses did not significantly affect migration. The concentration of most low molecular weight oligomers migrating into 10% ethanol and 100% heptane was < or =2 ng g(-1) of each oligomer for both copolymers. The cyclic trimer migrating from the 3% CHDM copolymer was approximately 4 ng g(-1); it was 3 ng g(-1) for the 31% CHDM copolymer. The overall results suggest that irradiation significantly increased levels of acetaldehyde but had no effect on non-volatile compounds migrating into food simulants.

Evaluating the potential for recycling all PET bottles into new food packaging.

To evaluate the feasibility of recycling all PET bottles into food packaging, realistic estimates of the maximum concentration of contaminants that might be expected in the polymer are needed. To estimate the maximum concentration of a contaminant that might be in PET from the storage of non-food substances, sorption experiments into two types of PET were performed. These test materials were 0.8mm thick amorphous PET (a relative sink for contaminants) and commercial PET bottle wall. Using a commercial shampoo containing 1% lindane (C6H6Cl6), the test materials were stored in contact with the shampoo at 20 and 40 degrees C for 231 days. This commercial shampoo also represents an extreme case because it contains 7% acetone, a solvent which swells PET, further enhancing sorption of chemicals. Additional sorption experiments into PET were performed by preparing solutions of 10% toluene in Miglyol (a fractionated coconut oil), 10% benzophenone in Miglyol, 5% 2-butoxyethoxy ethanol (2-BE) in 50/50 water/ethanol, and 10% methyl stearate in heptane. Sorption data from the shampoo into PET illustrate Fickian behaviour. Specifically, the amount of sorption at room temperature is approximately40 times less than that at 40 degrees C. The amount of lindane sorbed into PET from the shampoo after 231 days was 0.1 and 3.7 mgdm(-2) at 20 and 40 degrees C respectively. These values correspond to 28 and 765 mg kg(-1) on a mass/mass basis. All sorptions are within the ranges measured and published by other authors using surrogate contamination testing schemes. Additionally, actual bottles from recycle bins were analysed for the amout of contamination. Results are discussed in terms of potential consumer exposure to non-food contaminants in food containers made of recycled PET and in relation to the surrogate testing methods recommended by the Food and Drug Administration (FDA) for determining the compatibility of a PET recycling process to produce containers suitable for food-contact use.

Mechanistic studies on thiamin phosphate synthase: evidence for a dissociative mechanism.

Thiamin phosphate synthase catalyzes the coupling of 4-methyl-5-(beta-hydroxyethyl)thiazole phosphate (Thz-P) and 4-amino-5-(hydroxymethyl)-2-methylpyrimidine pyrophosphate (HMP-PP) to give thiamin phosphate. In this paper, we demonstrate that 4-amino-5-(hydroxymethyl)-2-(trifluoromethyl)pyrimidine pyrophosphate (CF(3)-HMP-PP) is a very poor substrate [k(cat)(CH(3)) > 7800k(cat)(CF(3))] and that 4-amino-5-(hydroxymethyl)-2-methoxypyrimidine pyrophosphate (CH(3)O-HMP-PP) is a good substrate [k(cat)(OCH(3)) > 2.8k(cat)(CH(3))] for the enzyme. We also demonstrate that the enzyme catalyzes positional isotope exchange. These observations are consistent with a dissociative mechanism (S(N)1 like) for thiamin phosphate synthase in which the pyrimidine pyrophosphate dissociates to give a reactive pyrimidine intermediate which is then trapped by the thiazole moiety.