[논문]Validation of an analytical method for cyanide determination in blood, urine, lung, and skin tissues of rats using gas chromatography mass spectrometry (GC-MS)

Shin, Min-Chul; Kwon, Young Sang; Kim, Jong-Hwan; Hwang, Kyunghwa; Seo, Jong-Su

doi:10.5806/ast.2019.32.3.88

Validation of an analytical method for cyanide determination in blood, urine, lung, and skin tissues of rats using gas chromatography mass spectrometry (GC-MS) 원문보기

분석과학 = Analytical science & technology, v.32 no.3, 2019년, pp.88 - 95

Shin, Min-Chul (Environmental Chemistry Research Group, Korea Institute of Toxicology) , Kwon, Young Sang (Environmental Chemistry Research Group, Korea Institute of Toxicology) , Kim, Jong-Hwan (Environmental Chemistry Research Group, Korea Institute of Toxicology) , Hwang, Kyunghwa (Analytical Research Group, Korea Institute of Toxicology) , Seo, Jong-Su (Environmental Chemistry Research Group, Korea Institute of Toxicology)

Abstract ▼ AI-Helper

This study was conducted to establish the analytical method for the determination of cyanide in blood, urine, lung and skin tissues in rats. In order to detect or quantify the sodium cyanide in above biological matrixes, it was derivatized to Pentafluorobenzyl cyanide (PFB-CN) using pentafluorobenzyl bromide (PFB-Br) and then reaction substance was analyzed using gas chromatography mass spectrometer (GC/MS)-SIM (selected ion monitoring) mode. The analytical method for cyanide determination was validated with respect to parameters such as selectivity, system suitability, linearity, accuracy and precision. No interference peak was observed for the determination of cyanide in blank samples, zero samples and lower limit of quantification (LLOQ) samples. The lowest limit detection (LOD) for cyanide was $10{\mu}M$. The linear dynamic range was from 10 to $200{\mu}M$ for cyanide with correlation coefficients higher than 0.99. For quality control samples at four different concentrations including LLOQ that were analyzed in quintuplicate, on six separate occasions, the accuracy and precision range from -14.1 % to 14.5% and 2.7 % to 18.3 %, respectively. The GC/MS-based method of analysis established in this study could be applied to the toxicokinetic study of cyanide on biological matrix substrates such as blood, urine, lung and skin tissues.

주제어

표/그림 (8)

그림 Fig. 1. Chemical structures of before and after derivatization.
표 Table 1. GC/MS operating conditions and parameters
그림 Fig. 2. Representative blank chromatogram of sodium cyanide in rat (A) blood, (B) urine, (C) lung and (D) skin tissue.
그림 Fig. 3. Representative LLOQ chromatogram of sodium cyanide in rat (A) blood, (B) urine, (C) lung and (D) skin tissue.
그림 Fig. 4. Calibration of sodium cyanide in rat (A) blood, (B) urine, (C) lung and (D) skin tissue.
표 Table 2. System suitability of sodium cyanide in blood, urine, lung and skin tissues in rats
표 Table 3. Linearity of calibration curves of sodium cyanide in blood, urine, lung and skin tissues in rats
표 Table 4. Accuracy and precision of sodium cyanide in blood, urine, lung and skin tissues in rats

AI 본문요약
AI-Helper

* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.

제안 방법

Selectivity indicates the ability of an analytical method to isolate and quantify the target substance when other substances are coexistent within a biological sample. Accordingly, selectivity was assessed by analyzing whether the peaks of blank, zero blank, and LLOQ samples of blood, urine, lung, and skin tissues obtained from six different individuals are affected by interference peaks. In addition, system suitability was examined by assessing the precision for peak area on the basis of six repeated injections of LLOQ samples.
Linearity was determined on the basis of calibration curve results obtained using standard samples with at least six different concentrations, including LLOQ and upper limit of quantitation, along with the blank and zero blank. Accuracy and precision were assessed on the basis of six repeated measurements of at least four different concentrations, including LLOQ, with respect to the entire operation of the test method. Accuracy was compared using relative error (%RE), whereas precision was compared using coefficient of variation (%CV).
For validation of the analytical method for determination of cyanide in blood, urine, lung, and skin tissues of rats, the selectivity, system suitability, linearity, accuracy, and precision of the method were tested. Validation of the analytical method was carried out in accordance with the Guideline on Bioanalytical Method Validation provided by the FDA and EMA.
Accordingly, selectivity was assessed by analyzing whether the peaks of blank, zero blank, and LLOQ samples of blood, urine, lung, and skin tissues obtained from six different individuals are affected by interference peaks. In addition, system suitability was examined by assessing the precision for peak area on the basis of six repeated injections of LLOQ samples.
1), which was analyzed by GC-MS. The study also validated an analytical method for determination of cyanide in blood, urine, lung, and skin tissues, which could have been absorbed owing to cyanide exposure, in order to assess hazards associated with cyanide exposure.
To test the linearity, the ratio of PFB-CN to IS concentrations (x axis) and ratio of PFB-CN to IS peak area (y axis) were used to construct a calibration curve with a weight of 1/x² . The correlation coefficient (r²) of each biological sample was within the range of 0.

대상 데이터

The standard material used in the experiment, sodium cyanide (NaCN; 95.5 %), was purchased from Sigma-Aldrich (Saint Louis, MO, USA). The internal standard (IS), 2,5-dibromotoluene (2,5-DBT; 99.

성능/효과

The accuracy and precision of each biological sample was within -14.1–14.5 % and 2.7–18.3 %, respectively, which satisfied the accuracy criterion of mean concentration being within 15 % (20 % for LLOQ) of the theoretical concentration and precision criterion of %CV <15 % (20 % for LLOQ).
3 %, respectively, which satisfied the accuracy criterion of mean concentration being within 15 % (20 % for LLOQ) of the theoretical concentration and precision criterion of %CV <15 % (20 % for LLOQ). The results confirmed that the analytical method is suitable for the determination of derivatized cyanide.
The results of accuracy (%RE) and precision (%CV) from LLOQ QC, Low QC, Mid QC, High QC samples were within ±15 % (20 % for LLOQ) of the values specified in the FDA and EMA guidelines.
These values were consistent with the criteria for assessing the calibration curve, that is, correlation coefficient ≥0.99 and concentration within 15 % (20 % for LLOQ) of the theoretical concentration, which were determined to be the levels suitable for cyanide determination.
To test the selectivity of the analytical method using GC-MS for determination of cyanide in blood, urine, lung, and skin tissues, blank and zero blank samples using blood, urine, lung, and skin tissues obtained from six individuals and LLOQ sample treated with NaCN and IS were assessed (Fig. 2, 3). No interference peaks were observed in the interval in which cyanide and IS were eluted.

참고문헌 (18)

G. Liu, J. Liu, K. Hara, Y. Wang, Y. Yu, L. Gao, and L. Li, J. Chromatography B, 877, 3054-3058 (2009).

상세보기
S. M. Lee and J. S. Kim, J. Korean Med. Assoc., 56(12), 1076-1083 (2013).

상세보기
S. W. Sauer and M. E. Keim, Ann. Emerg. Med., 37, 635-641 (2001).

상세보기
R. K. Bhandari, R. P. Oda, I. Petrikovics, D. E. Thompson, M. Brenner, S. B. Mahon, V. S. Bebarta, G. A. Rockwood, and B. A. Logue, J. Anal. Toxicol., 38(4), 218-225 (2014).

상세보기
K. Minakata, H. Nozawa, K. Gonmori, I. Yamagishi, M. Suzuki, K. Hasegawa, K. Watanabe, and O. Suzuki, Anal. Bioanal. Chem., 400(7), 1945-1951 (2011).

상세보기
T. T. Chistison and J. S. Rohrer, J. Chromatography A, 1155, 31-39 (2007).

상세보기
A. Tung, J. Lynch, and W. A. McdDade, J. Moss, Anesth. Analg., 85, 1045 (1997).

상세보기
A. M. Calafat and S. B. Stanfill, J. Chromatography B, 772, 131-137 (2002).

상세보기
S. Tsukamoto, T. Kanegae, Y. Matsumura, S. Uchigasaki, M. Kitazawa, T. Muto, S. Chiba, S. Oshida, T. Nagoya, and M. Shimamura, Jpn. J. Legal. Med., 48, 336-342 (1994).
S. H. Chen, S. M. Wu, H. S. Kou, and H. L. Wu, J. Anal. Toxicol., 18, 81-85 (1994).

상세보기
S. Kage, T. Nagata, and K. Kudo, J. Chromatography B, 675, 27-32 (1996).

상세보기
B. D. Paul and M. L. Smith, J. Anal. Toxicol., 30, 511-515 (2006).

상세보기
R. K. Bhandari, R. P. Oda, S. L. Youso, I. Petrikovics, V. S. Bebarta, G. A. Rockwood, and B. A. Logue, Anal. Bioanal. Chem., 404, 2287-2294 (2012).

상세보기
A. Tracqui, J. S. Raul, A. Geraut, L. Berthelon, and B. Ludes, J. Anal. Toxicol., 26, 144-148 (2002).

상세보기
D. Tsikas, J. Chromatography B, 1043, 187-201 (2017).

상세보기
J. B. Lee, S. K. Shon, S. H. Woo, S. Y. Park, J. H. Hwang, O. S. Kwon, N. Y. Kim, and K. J. Paeng, Anal. Sci. Technol., 29(2), 73-78 (2016).

원문보기 상세보기
Guideline for Industry on Bioanalytical Method Validation, FDA/CDER, May 2018.
Guideline on Bioanalytical Method Validation, EMA, Jul, 2011.

LOADING...

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증