[논문]Analysis of Benzoic Acid in Quasi-Drug Drink Using Isotope Dilution Liquid Chromatography Mass Spectrometry

Lee, In-Sun; Ahn, Seong-Hee; Kim, Byung-Joo; Hwang, Eu-Jin; Kim, Yong-Seong

doi:10.5012/bkcs.2008.29.11.2125

Analysis of Benzoic Acid in Quasi-Drug Drink Using Isotope Dilution Liquid Chromatography Mass Spectrometry 원문보기

Bulletin of the Korean chemical society, v.29 no.11, 2008년, pp.2125 - 2128

Lee, In-Sun (Division of Metrology for Quality Life, Korea Research Institute of Standards and Science) , Ahn, Seong-Hee (Division of Metrology for Quality Life, Korea Research Institute of Standards and Science) , Kim, Byung-Joo (Division of Metrology for Quality Life, Korea Research Institute of Standards and Science) , Hwang, Eu-Jin (Division of Metrology for Quality Life, Korea Research Institute of Standards and Science) , Kim, Yong-Seong (Department of Chemistry, KyungNam University)

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문제 정의

The objective of this work is to develop a definitive reference method for the determination of benzoic acid using isotope dilution liquid chromatography mass spectrometry (IDLC/MS). The conditions of separation and electrospray ionization efficiency had to be optimized considering the pKa of benzoic acid (pKa: 4.

제안 방법

We tested self-consistency within and among the standard solutions by inter-comparing the eight isotope ratio standards by LC/MS. Based on the inter-comparing results, repeatability and reproducibility of preparing standard solutions and isotope ratio standards were evaluated and one isotope ratio standard was selected and used for further sample analysis.
0 ppm for SRM mode, respectively. Even though the recovery results of the SIM mode are not much different from those of the SRM mode (data not shown), the SRM mode was operated for the quantitative analysis of IDMS measurement. Because only the filtration step was performed for sample preparation, it is possible for the SIM mode to be interfered by sample matrix.
The sample was then analyzed by LC/MS without further cleanup or extraction procedures. For the validation of our method, we prepared samples with known levels of benzoic acid by fortifying blank samples, which were also obtained from local markets and tested that their benzoic acid levels were below the detection limit of this method, by gravimetrically adding a stock solution of benzoic acid.

대상 데이터

API 2000 mass spectrometer from Applied Biosystems (Forster City, CA, USA) which is interfaced with an 1100 Series LC system from Agilent Technologies (Palo Alto, CA, USA) was used in this study. Samples prepared as described above and the isotope ratio standard were injected onto a Waters Corp.
Milli-Q RG purification system. Ammonium acetate was from Aldrich (Milwaukee, WI, USA) and formic acid was from Fluka (Steinheim, Germany).

이론/모형

The blank drink was confirmed by mass spectrometry that the level of benzoic acid was below the detection limit of the LC/MS method. Samples with various levels of benzoic acid were prepared by fortifying the blank drink by gravimetrically spiking a benzoic acid stock solution and analyzed by the ID-LC/MS method. Table 1 shows that the recovery of fortified benzoic acid was over 99% through the benzoic acid level tested, indicating that matrix in commercial drinks do not seriously affect on the determination of benzoic acid by the ID-LC/MS.
Additionally, sample preparation processes were simplified to a single stage of membrane filtering without further procedures. Using ID-LC/MS method, commercial drinks were analyzed. The accuracy, repeatability, and reproducibility of the method were also evaluated if the method provides adequate metrological quality.

성능/효과

The results are listed in the Table 2. The relative standard deviation of the results obtained from the ID-LC MS method with the SRM mode within a time period ranges between 0.30% and 0.76%, indicating that the method has an excellent repeatability. The measurement values from the two period agree to each other within their uncertainties, indicating that the method provide a good reproducibility.
Repeatability and reproducibility test was also carried out. The test results proved that ID-LC/MS/MS using the negative ion mode for determination of benzoic acid has a metrological quality less than 1.2% with 95% confidence.

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Analysis of Benzoic Acid in Quasi-Drug Drink Using Isotope Dilution Liquid Chromatography Mass Spectrometry 원문보기