Naoki Fukui

Simultaneous measurement of diazolidinyl urea, urea, and allantoin in cosmetic samples by hydrophilic interaction chromatography

A new HPLC method for simultaneous measurement of diazolidinyl urea (DU), urea, and allantoin by hydrophilic interaction chromatography using a column packed with triazol-bonded silica particles is described. The calibration curves of DU, urea, and allantoin were linear over the ranges 2.5-125.0, 30-1250, and 0.25-18.75mg/L, respectively. The recoveries of DU, urea, and allantoin from homemade cosmetic samples ranged from 92.84% to 101.69%, 98.19% to 103.22%, and 95.75% to 102.09%, respectively. The peak relative standard deviation (RSD) values for the recovery tests of 3 concentrations/5 replicates were 3.03% for all compounds. In day-to-day measurement of recovery tests from homemade lotions over 3 consecutive days, the RSDs were less than 2.5% in all cases. This method was well validated and would be helpful for quickly analyzing these compounds in cosmetic samples.

[Validation Study on a Multi-Residue Method for Determination of Pesticide Residues in Vegetables and Fruits by using General Matrix Standard Solutions].

Quantitative methods using the matrix-matched standard solutions approach are widely used for multi-residue pesticide determination by GC-MS/MS to deal with the issue of matrix effects. However, preparing matrix-matched standard solutions in analyses of many kinds of samples is very time-consuming. In order to solve this problem, a method that employs general matrix standard solutions has been developed using polyethylene glycol (PEG), extract of vegetables-fruit juice (VFJm) and triphenyl phosphate (named the PEG-VFJm method). Here, a validation study for 168 pesticides was performed on three kinds of samples [potato, spinach and apple] at concentrations of 0.010 and 0.050 μg/g. In these three commodities, 144 to 158 pesticides satisfied the required criteria using the matrix-matched method and 129 to 149 pesticides satisfied the same criteria using the PEG-VFJm method. Our results suggest that application of general matrix standard solutions would enable rapid and effective analyses of pesticides.

Determination of irradiation histories of raw beef livers using liquid chromatography-tandem mass spectrometry of 5,6-dihydrothymidine.

A method for detecting irradiation histories of raw beef livers was developed by measuring 5,6-dihydrothymidine (DHdThd) using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Liver DNA was extracted using phenol-chloroform extraction followed by precipitation in 50% ethanol. DNA was then enzymatically digested and nucleosides were purified using an OASIS MCX column. DHdThd and thymidine (dThd) contents of resulting test solutions were analyzed using LC-MS/MS. DHdThd was detected specifically after γ-irradiation. Concentration ratios of DHdThd to dThd in the test solutions increased dose-dependently after irradiation at 1.0-11.3kGy, which included the practical dose for sterilization of 2-7kGy. Dose-response curves from beef livers of individual animals almost overlapped. Thus, this method is a candidate for the detection of irradiation histories of foods from which DNA can be extracted.

Enhancement of Pesticide Peak Response in GC-MS in the Presence of Multiple Co-Existing Reference Pesticides

We carried out a collaborative study in six laboratories to confirm the universality of the enhancing effect of co-existing reference pesticides on the GC-MS peak response to a target pesticide (malathion, procymidone, or flucythrinate). First, we confirmed the response enhancement of the target pesticides with increasing numbers of co-existing reference pesticides in solution. Then, using diluted green soybean matrix, we analyzed the target pesticides with two types of matrix-matched calibration, containing the target pesticides or 166 other pesticides. In both cases, the response-enhancing effect of co-existing pesticides was confirmed in all laboratories. The enhancement was reduced by addition of green soybean matrix to the sample and calibration solutions. Our results show that it is necessary to estimate the peak response-enhancing effect of co-existing pesticides in the calibration solution to obtain accurate results with GC-MS determination. The enhancing effect could be reduced by addition of food matrix to the sample and calibration solutions.

Problems and solutions of polyethylene glycol co-injection method in multiresidue pesticide analysis by gas chromatography-mass spectrometry: evaluation of instability phenomenon in type II pyrethroids and its suppression by novel analyte protectants

AbstractPolyethylene glycol 300 is commonly used as a base material for “analyte protection” in multiresidue pesticide analysis via gas chromatography-mass spectrometry. However, the disadvantage of the co-injection method using polyethylene glycol 300 is that it causes peak instability in α-cyano pyrethroids (type II pyrethroids) such as fluvalinate. In this study, we confirmed the instability phenomenon in type II pyrethroids and developed novel analyte protectants for acetone/n-hexane mixture solution to suppress the phenomenon. Our findings revealed that among the examined additive compounds, three lipophilic ascorbic acid derivatives, 3-O-ethyl-L-ascorbic acid, 6-O-palmitoyl-L-ascorbic acid, and 6-O-stearoyl-L-ascorbic acid, could effectively stabilize the type II pyrethroids in the presence of polyethylene glycol 300. A mixture of the three ascorbic acid derivatives and polyethylene glycol 300 proved to be an effective analyte protectant for multiresidue pesticide analysis. Further, we designed and evaluated a new combination of analyte protectant compounds without using polyethylene glycol or the troublesome hydrophilic compounds. Consequently, we obtained a set of 10 medium- and long-chain saturated fatty acids as an effective analyte protectant suitable for acetone/n-hexane solution that did not cause peak instability in type II pyrethroids. These analyte protectants will be useful in multiresidue pesticide analysis by gas chromatography-mass spectrometry in terms of ruggedness and reliable quantitativeness. Graphical abstractComparison of effectiveness of the addition of lipophilic derivatives of ascorbic acid in controlling the instability phenomenon of fluvalinate with polyethylene glycol 300

Rapid and Reliable Method for Determining Irradiation Histories of Ground Beef and Prawns by Measuring 5,6-Dihydrothymidine

A rapid and reliable method for determining irradiation histories of ground beef and prawns was developed on the basis of a method for determining the irradiation history of beef liver by liquid chromatography-tandem mass spectrometry (LC-MS/MS) of 5,6-dihydrothymidine (DHdThd). Improvements in the method included the following: (1) 50% ethanol precipitation in the DNA extraction step was conducted before the RNase step, (2) snake venom phosphodiesterase I was used for DNA digestion to boost liberation of DHdThd, and (3) a matrix-matched calibration curve was used for determining DHdThd by LC-MS/MS analysis. This method successfully determined irradiation histories of ground beef and prawns. Furthermore, a close correlation between the formation of DHdThd and 2-alkylcyclobutanones, which are an established index of irradiation histories, was observed in ground beef. DHdThd in DNA could be a promising candidate for a new index of irradiation histories of various foods.

LC-MS/MSを用いた迅速で簡便な飲料中の残留農薬一斉分析法の実用化に向けての検討

A rapid and simple multi-residue method for determination of pesticides has been applied to drinking water and beverages. To a disposable polypropylene tube containing 10.0 g sample, 20 mL acetonitrile was added and the mixture was shaken vigorously for 1 min to extract pesticides. Then, 1 g sodium chloride and 4 g magnesium sulfate anhydrous were added, followed by vigorous shaking for 1 min and centrifugation to obtain the organic phase. The organic phase was processed with a graphite carbon black/PSA solid phase column. After concentration and reconstitution with 25% methanol containing aqueous solution, the test solution was analyzed with LC-MS/MS. Recovery tests of 91 pesticides fortified (0.02 μg/g) in 35 kinds of drinking water and beverages were conducted. The decline of recoveries in alcoholic beverages is considered to be due to the increase of organic phase volume owing to ethanol included in the alcoholic beverages. A simulation study was carried out with simulated alcoholic beverages, which consisted of 50% grape juice, with various amounts of ethanol and water, to examine pesticides recoveries and volume of the organic phase. The results suggested this method would be applicable both to alcoholic beverages containing less than 10% ethanol and to alcoholic beverages containing over 10% ethanol after dilution with water to below 10% ethanol prior to the addition of acetonitrile. A sample could be processed and analyzed by LC-MS/MS within 2 h. Thus, this method should be useful for monitoring and screening pesticide residues in drinking water and various beverages.

Rapid and Easy Multiresidue Method for Determination of Pesticide Residues in Foods Using Gas or Liquid Chromatography–Tandem Mass Spectrometry

To obtain a high yield of food grains, many types of pesticides have been developed, which in turn has facilitated the prosperity of the human race and brought relief to farmers. After WWII, organophosphorus and organochlorine pesticides were extensively used worldwide. These early organophosphorus pesticides occasionally caused poisoning, while some organochlorine pesticides persisted in the environment. These facts served as a warning against an improvident use of pesticides and revealed that residual pesticides in crops should be monitored. Thus, national and local governments should monitor imported and regional foods as a policy. In Japan, all pesticides are regulated under a uniform limit (0.01 μg/g), except for a combination of foods and pesticides set under the maximum residue limits (MRLs) (Notification No. 497-499, November 29, 2005). This regulation does not require analysis of all the pesticides; however, it does necessitate pesticide residue analysis of commodities. Thus far, the chemical industry has developed more than 800 pesticides that belong to many classes such as insecticides, fungicides, nematocides, and herbicides. Each class has a different target point and physical properties and this diversity limits the coverage of a single analytical method. It is also impossible to monitor all the pesticides pertaining to one foodstuff using hundreds of methods. Thus, analytical institutes require fast and efficient multiresidue methods in order to maximize the coverage of their monitoring activities. For this reason, researchers have reported many multiresidue analytical methods (Cook, J., et al., 1999, Fillion, J., et al., 2000, Hirahara, Y., et al., 2006, Luke, M. A., et al., 1975, Ueno, E., et al., 2004). These methods were optimized to monitor multiple residues efficiently and are used routinely in quarantine stations and laboratories. Anastassiades et al. reported a rapid approach for the analysis of pesticide residues in fruits and vegetables, named QuEChERS (quick, easy, cheap, effective, rugged and safe) method in 2003 (Anastassiades, M., et al., 2003). The main focus of this report was to shorten the analytical process during extraction and cleanup without employing expensive instruments. The characteristic points of the method are as follows: (1) shaking extraction with acetonitrile in a disposable tube, (2) one step salting out and removal of water from the