A new method for rapid and quantitative detection of the Bacillus cereus emetic toxin cereulide in food products by liquid chromatography-tandem mass spectrometry analysis.

The Bacillus cereus emetic toxin cereulide causes foodborne intoxication, which may occasionally result in severe disease, and even death. To differentially diagnose the emetic-type of foodborne disease caused by B. cereus and assess the safety of commercial food, we developed a rapid method to quantitate cereulide. This method was combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis for the extraction of cereulide from food using a normal-phase silica gel cartridge. The limits of detection and quantification were 0.1 and 0.5 ng of cereulide ml(-1), respectively. Spiked cereulide was reproducibly recovered with over 67% efficiency from nine diverse foods implicated in cereulide food poisoning. The recovery rate, reproducibility, and intermediate precision for this single laboratory validation using boiled rice were 87.1%, 4.4%, and 7.0%, respectively. Further, we detected a wide range of cereulide concentrations in leftover food and vomitus samples from two emetic foodborne outbreaks. LC-MS/MS analysis correlated closely with those acquired using the HEp-2 cell assay, and quantitated cereulide from 10 food samples at least five times faster than the bioassay. This new method will provide clinicians with an improved tool for more rapidly and quantitatively determining the presence of cereulide in food and diagnosing food poisoning caused by cereulide.

Analysis of chlorothalonil by liquid chromatography/mass spectrometry using negative-ion atmospheric pressure photoionization.

A highly sensitive and simple method for the analysis of chlorothalonil was presented using a liquid chromatograph/mass spectrometer equipped with an atmospheric pressure photoionization (APPI) source. Chlorothalonil is one of the most extensively used fungicides. The major degraded product of chlorothalonil, 4-hydroxy-2,5,6-trichloroisonaphthonitrile (4OH-TPN), was also quantified with sensitivity similar to that of chlorothalonil. The method was applied to the determination of chlorothalonil in aqueous environment and food samples. The method detection limits (MDLs) of chlorothalonil for aqueous samples and cucumber were determined to be 0.18 and 3.2 ng g(-1), respectively. At several estuarial locations, chlorothalonil was detected with a maximum of 1.1 ng L(-1). On the other hand, 4OH-TPN was detected not from estuaries but from rivers with a maximum of 14 ng L(-1).

Rapid analysis of 56 pesticide residues in natural medicines by GC/MS with negative chemical ionization.

Many methods for determining pesticide residues in food have been reported. Although natural medicines should be confirmed to be as safe as food, few methods for determining pesticide residues in natural medicines have been reported. In this study, 56 pesticides were detected in natural medicines with a simple, rapid sample preparation method. This study indicates that the proposed method is useful for analyzing pesticides in natural medicines.

[Determination of polysorbates in foods by solid-phase extraction and high-performance liquid chromatography].

A simple and rapid method using refractive index high-performance liquid chromatography (RI-HPLC) was developed for the determination of polysorbates (PS) in processed foods. PS were extracted with ethyl acetate containing 5% methanol. The extract was cleaned up on a multimode cartridge (300 mg) and an Alumina-N cartridge (500 mg) to remove fats and food color. HPLC separation was performed on a C18 column (4.6 i.d. x 150 mm) with methanol as the mobile phase. The recoveries of PS80 from nine kinds of foods fortified at the levels 1-5 g/kg were 80-99%. The limit of quantitation for PS80 in foods was 0.10 g/kg. The proposed method was applied to Worcestershire sauce that was PS-positive by TLC, and PS was confirmed to be present as PS80 at the concentration of 0.13 g/kg.

Simultaneous analysis of 17 organochlorine pesticides in natural medicines by GC/MS with negative chemical ionization.

Many methods for the determination of pesticide residues in food have been reported. Although natural medicines should be confirmed to be as safe as food, few methods for the determination of pesticide residues in natural medicines have been reported. In this study, 17 organochlorine pesticides were detected in natural medicines using GC/MS with negative chemical ionization (NCI). GC/MS with NCI can detect halogenated pesticides selectively and thus is suitable for the detection of organochlorine pesticides. This study indicates that GC/MS with NCI is useful for analyzing organochlorine pesticides in natural medicines.

Simultaneous analysis of 10 pyrethroid pesticides in natural medicines by GC/MS with negative chemical ionization.

Many methods for the determination of pesticides residues in food have been reported. Although natural medicines should be confirmed as safe as food, only a few methods for the determination of pesticide residues in natural medicines have been reported. In this study, 10 pyrethroid pesticides were detected in natural medicines by GC/MS with negative chemical ionization (NCI). GC/MS with NCI can detect halogenated pesticides selectively, and thus is suitable for the detection of pyrethroid pesticides. This study indicates that GC/MS with NCI is useful for analyzing pyrethroid pesticides in natural medicines.

CMV DNA detection in dried blood spots for diagnosing congenital CMV infection in Japan.

Human cytomegalovirus (CMV) is a leading congenital infectious agent in developed countries. In the past, the incidence of congenital infection has been rather low in Japan because a high seroprevalence of CMV present in young women. However, this seroprevalence has been decreasing in recent years, so that the incidence of congenital CMV infection in Japanese neonates may increase and approach the level seen in other developed countries. The method was used for detecting CMV DNA reported by Barbi et al. [Barbi et al. (1996): Clin Diagn Virol 6:27-32] using a dried blood spot on filter paper, to diagnose congenital CMV infection in Japanese neonates. This method is effective and less laborious than virus isolation both for epidemiological studies and for identifying asymptomatic infected babies. Japanese neonates (1,176) were examined; two of who were asymptomatic were found to be infected.

Surveillance of polychlorinated biphenyl congeneric patterns in human breast milk from 1973 to 2000 in Osaka, Japan.

OBJECTIVE: Because the analysis of polychlorinated biphenyls (PCBs) in 1970's was performed using a packed column gas chromatograph, with an electron capture detector to determine total-PCB concentration, analytical data obtained by this analysis do not meet the requirement for the risk assessment of PCB congeners. In this context, the present study was carried out to reevaluate the congeneric analysis data by analyzing breast milk fat specimens that have been kept frozen. METHODS: PCB congeners in human breast milk were analyzed by high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS) using a capillary column for the selected ion monitoring (SIM) of PCBs. RESULTS: Twelve major PCB congeners were detected in breast milk. The concentrations of all congeners of PCBs in breast milk were found to decrease annually, and their changes over time differed greatly for each congener. Between 1973 and 2000, almost no changes in the proportions of the 12 congeners of heptachlorinated biphenyls (HpCBs) and hexachlorinated biphenyls (HxCBs) were found whereas a marked and a slight decrease in the proportion of pentachlorinated biphenyls (PeCBs) and tetrachlorinated biphenyls (TeCBs), respectively, were observed. CONCLUSIONS: The PCB contamination of the Japanese population is estimated to be largely caused by seafood intake, but the proportions of PCB congeners in fish commodities were different from those in breast milk. The absorption, metabolism and therefore the accumulation of PCBs in the human body differ greatly depending on the congener.

Time-trend (1973-2000) of polybrominated diphenyl ethers in Japanese mother's milk.

The time-trend and recent concentrations of polybrominated diphenyl ethers (PBDEs) in Japanese mother's milk were investigated. The time-trend of 16 PBDEs (BDE-28, 37, 47, 66, 71, 75, 77, 85, 99, 100, 119, 153, 154, 138, 183, and 190) in pooled milk samples from mothers living in Osaka between 1973 and 2000 was analyzed. Additionally, PBDE concentrations in individual milk samples collected from 13 mothers living in Kanagawa and Okayama in 1999 were measured. The total concentration of all PBDE congeners (SigmaPBDEs) measured in the pooled samples increased during the period between 1973 (<0.01 ng/g lipid) and 1988 (1.64 ng/g lipid), and remained low afterwards while showing remarkable changes in PBDE congener profiles. The SigmaPBDEs in the 1999 individual milk samples were also low (0.56-3.97 ng/g lipid), except for a single sample (291 ng/g lipid). The source of this exposure could not be identified.