Seonghee Ahn

Accurate determination of ochratoxin A in Korean fermented soybean paste by isotope dilution-liquid chromatography tandem mass spectrometry.

Ochratoxin A (OTA), a naturally occurring mycotoxin, has been frequently detected in doenjang, a traditional fermented soybean paste, when it is fermented under improper conditions. Reliable screening of OTA in traditional fermented soybean paste (doenjang) is a special food-safety issue in Korea. Our laboratory, the National Metrology Institute of Korea, established an isotope dilution-liquid chromatography tandem mass spectrometry (ID-LC/MS/MS) method as a higher-order reference method to be used for SI-traceable value-assignment of OTA in certified reference materials (CRMs). (13)C20-OTA was used as an internal standard. Sample preparation conditions and LC/MS measurement parameters were optimised for this purpose. The analytical method was validated by measuring samples fortified with OTA at various levels. Repeatability and reproducibility studies showed that the ID-LC/MS/MS method is reliable and reproducible within 2% relative standard deviation. The analytical method was applied to determine OTA in various commercial doenjang products and home-made doenjang products.

Final report on key comparison CCQM-K55.b (aldrin): An international comparison of mass fraction purity assignment of aldrin

Under the auspices of the Organic Analysis Working Group (OAWG) of the Comit? Consultatif pour la Quantit? de Mati?re (CCQM) a key comparison, CCQM K55.b, was coordinated by the Bureau International des Poids et Mesures (BIPM) in 2010/2011. Nineteen national measurement institutes and the BIPM participated. Participants were required to assign the mass fraction of aldrin present as the main component in the comparison sample for CCQM-K55.b which consisted of technical grade aldrin obtained from the National Measurement Institute Australia that had been subject to serial recrystallization and drying prior to sub-division into the units supplied for the comparison. Aldrin was selected to be representative of the performance of a laboratorys measurement capability for the purity assignment of organic compounds of medium structural complexity [molar mass range 300 Da to 500 Da] and low polarity (pKOW < ?2) for which related structure impurities can be quantified by capillary gas phase chromatography (GC). The key comparison reference value (KCRV) for the aldrin content of the material was 950.8 mg/g with a combined standard uncertainty of 0.85 mg/g. The KCRV was assigned by combination of KCRVs assigned by consensus from participant results for each orthogonal impurity class. The relative expanded uncertainties reported by laboratories having results consistent with the KCRV ranged from 0.3% to 0.6% using a mass balance approach and 0.5% to 1% using a qNMR method. The major analytical challenge posed by the material proved to be the detection and quantification of a significant amount of oligomeric organic material within the sample and most participants relying on a mass balance approach displayed a positive bias relative to the KCRV (overestimation of aldrin content) in excess of 10 mg/g due to not having adequate procedures in place to detect and quantify the non-volatile content?specifically the non-volatile organics content?of the comparison sample. There was in general excellent agreement between participants in the identification and the quantification of the total and individual related structure impurities, water content and the residual solvent content of the sample. The comparison demonstrated the utility of 1H NMR as an independent method for quantitative analysis of high purity compounds. In discussion of the participant results it was noted that while several had access to qNMR estimates for the aldrin content that were inconsistent with their mass balance determination they decided to accept the mass balance result and assumed a hidden bias in their NMR data. By contrast, laboratories that placed greater confidence in their qNMR result were able to resolve the discrepancy through additional studies that provided evidence of the presence of non-volatile organic impurity at the requisite level to bring their mass balance and qNMR estimates into agreement. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

Characterization of Recombinant Human Erythropoietin Isoforms by Capillary Isoelectric Focusing with Hydroxypropyl Cellulose-Coated Capillaries

Capillary isoelectric focusing (cIEF) has become a powerful method for protein analysis and has shown promising perspectives in biopharmaceutical research, because it allows fast and automated analysis of proteins with high resolution. Here, an improved cIEF system was set up for exploring better characterization of recombinant human erythropoietin (EPO) with hydroxypropyl cellulose (HPC)-coated capillaries and an offline fast and convenient desalting treatment for EPO real sample. With the cIEF system, several separation conditions for EPO isomers were optimized and high resolution results were obtained. The use of HPC-coated capillary columns allowed for no additive needed for the elimination of EOF. The cIEF system might be applied to further complementary characterization of EPO by comparison with LC and other modes. And the cIEF system was considered promising for online coupling with MS.Graphical Abstract

Accurate Quantification of Saccharin Using Isotope Dilution Liquid Chromatography Mass Spectrometry (ID-LC/MS)

Saccharin is a commonly used artificial sweetener in foodstuffs. However, for its carcinogenic dispute, it has been regulated by government bodies. In this study, isotope dilution mass spectrometry (ID-MS) was introduced for the accurate quantification of saccharin. To employ ID-LC/MS, we obtained its isotope analogue, -sodium saccharin, by customized synthesis. Samples were spiked with -sodium saccharin and analyzed with LC/MS in negative mode. Chromatographic conditions were optimized for the adequate chromatographic retention and separation of saccharin with a column. MS was operated with electrospray ionization by the selected ion monitoring (SIM) mode of for saccharin (m/z 182) and for its isotope analogue (m/z 183). To validate the ID-LC/MS method for accurate measurement, we prepared a batch of a candidate material by sortifying quasi-tea-drinks with saccharin and analyzed samples gravimetrically fortified in various levels of concentration. The repeatability and reproducibility of this method was tested by analyzing the reference material. Result show that ID-LC/MS is a reliable method for the quantitative analysis of saccharin.

Accurate determination of chloramphenicol in meat by isotope dilution liquid chromatography mass spectrometry (ID-LC/MS)

Chloramphenicol is one of the most effective antibiotics for treatment of food-producing animals for typhoid fever. However, it has been reported that it caused severe side effects such as aplastic anemia in human, therefore the use of chloramphenicol for food-producing animal is prohibited by European Union and other countries. In this study, the analytical method using isotope dilution liquid chromatography-mass spectrometry (ID-LC/MS) was established for accurate determination of chloramphenicol in meat. Chloramphenicol was extracted with ethylacetate from porcine and solid phase extraction cartridge was used for enhancing the recovery. The residue of chloramphenicol in porcine was analyzed using the liquid chromatography mass spectrometer (LC/MS) interfaced with electrospray ionization source. Analysis was performed in negative mode with selected reaction mornitoring mode at m/z 321257 of and m/z 326 262 channels for its isotope. The established method was tested using fortified samples at the level of 0.2 1, 10, /kg and analytical results agreed with the gravimetrically fortified values within their uncertainties. This method was validated by analyzing a certified reference material, BCR 445, from IRMM (Institute for Reference Materials and Measurement). Our measurement values agreed with the certified value within their uncertainties. The uncertainty of our measured value was much lower than that the certified value from IRMM.

Accurate determination of hexabromocyclododecane diastereomers in extruded high-impact polystyrene: Development of an analytical method as a candidate reference method

Herein, an isotope dilution-liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) method was developed as a candidate reference method for the accurate quantitation of hexabromocyclododecane (HBCDD) diastereomers in plastics. The repeatability, reproducibility, and uncertainty results showed that developed ID-LC/MS/MS method is reliable and reproducible. As homogeneous samples, HBCDD-containing extruded high-impact polystyrene (HIPS) pellets were fabricated via an extrusion process. Notably, we detected α-, β-, γ-, δ-, ε-, η-, and θ-HBCDDs in the extruded HIPS pellets, and then determined their exact mass fractions using the ID-LC-MS/MS method. The relative contents (average ± standard deviation) of HBCDD diastereomers in HIPS obtained using the phenyl-hexyl column were 67.08 ± 0.41% (α-HBCDD), 19.73 ± 0.37% (β-HBCDD), 11.59 ± 0.16% (γ-HBCDD), and 1.6 ± 0.07% (sum of δ-, ε-, η-, and θ-HBCDDs). These values differed significantly from the ones determined for the technical HBCDD mixtures (10.42% α-HBCDD, 5.30% β-HBCDD, 82.13% γ-HBCDD, 2.15% minor HBCDDs) used to fabricate the HIPS pellets and thus demonstrating the HBCDD isomerization during the extrusion. The proportion of minor HBCDDs was smaller than the uncertainty of the total HBCDD and that the sum of α-, β-, and γ-HBCDDs was comparable to the total HBCDD in the investigated samples. Notably, a real-life sample (expanded polystyrene board obtained from a local construction site) also showed a similar HBCDD profile, being rich in α-HBCDD.

Development of a Four-way Interface for Online Capillary Isoelectric Focusing-Electrospray-Mass Spectrometry (CIEF-ESI-MS)

A new interface for coupling CIEF and MS using a four-way cross has been developed in a single mechanical system. This new interface could be operated without the electric discontinuity and reinstallation of lines. Additionally, a bare fused sil- ica capillary was facilitated as a spray needle to produce electrospray and to guide catholyte or sheath liquid. Focusing for CIEF was completed in a hanging droplet at the end of spray needle. This capillary spray needle also provided stable spray, enhanced the ionization efficiency and increased sensitivity. Results with carbonic anhydrase I showed that focusing and spraying were well completed with the new interface and the new spray needle.

Development of Gas Chromatography/Mass Spectrometry for the Determination of Essential Fatty Acids in Food Supplemental Oil Products

A gas chromatography/ mass spectrometric (GC/MS) method was developed as a candidate reference method for the accurate determination of essential fatty acids (linoleic acid, α- and γ-linolenic acids) in food supplemental oil products. Samples were spiked with three internal standards (stearic acid-d35, 13 C18-linoleic acid, and 13 C18-α-linolenic acid). Samples were then subject to saponification, derivatization for methylation, and extraction by organic solvent. For GC/MS measurement, an Agilent HP-88 column, designed for the separation of fatty acid methyl esters, was selected after comparing with other columns as it pro- vided better separation for target analytes. Target analytes and internal standards were detected by selected ion monitoring of molecular ions of their methyl ester forms. The GC/MS method was applied for the measurement of three botanical oils in NIST SRM 3274 (borage oil, evening primrose oil, and flax oil), and measurement results agreed with the certified values. Measure- ment results for target analytes which have corresponding isotope-labeled analogues as internal standard were calculated based on isotope dilution mass spectrometry (IDMS) approach, and compared with results calculated by using the other two internal standards. Results from the IDMS approach and the typical internal standard approach were in good agreement within their mea- surement uncertainties. It proves that the developed GC/MS method can provide similar metrological quality with IDMS meth- ods for the measurement of fatty acids in natural oil samples if a proper fatty acid is used as an internal standard. Key word: Fatty acid, GC/MS, IDMS method, Internal standard method, Botanical oils

Analysis of sorbic acid in tea-drink using isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS)

Isotope dilution mass spectrometry (ID-MS) based on liquid chromatography tandem mass spectrometry (LC/MS/MS) was developed for the accurate determination of sorbic acid in tea-drink. An isotope analogue of sorbic acid, 13 C 2 -sorbic acid, was obtained by custom synthesis. MS was operated in the negative mode with selected reaction monitoring (SRM) mode of [M-H]- → [M-CO 2 H]- channel at m/z 111 → 67 for sorbic acid and at m/z 113 → 68 for its isotope analogue. Chromatographic separation was accomplished with a C18 column and an isocratic mobile phase of 55% of 50 mM ammonium acetate (pH 4.5) and 45% of methanol. Homogeneous reference materials were prepared for validation of this method, including repeatability and reproducibility tests, by fortifying tea-drink with sorbic acid in our laboratory. Repeatability and reproducibility studies showed that the ID-LC/MS method is a reliable and reproducible method which provides less than 3.8% of relative standard deviation (RSD) for the analysis of sorbic acid.