국내 미지정 색소인 azorubine에 대한 우리나라의 공인분석법을 설립하기 위해 제외국의 공인분석법들(유럽연합의 EFSA, 영국의 FSA, 우리나라의 식품의약품안전평가원)이 비교되었다. 재현된 HPLC 방법들 중 FSA의 분석법이 가장 우수한 분석 결과를 나타내어 azorubine분석법으로 최종 선정되었다. Azorubine 분석을 위한 위 세 기관의 전처리 방법은 모두 낮은 회수율을 나타내었다. 따라서 식품의약품안전평가원의 전처리법을 개선한 새로운 전처리방법이 개발되었다. 최종 선정된 HPLC 분석법과 본 연구를 통해 개발된 새로운 식품 전처리법에 대한 분석법 밸리데이션을 실시하였다. 검증 실험에서 음료류에서 97~103%, 빵 및 과자류에서 95~101%, 기타 식품에서 93~102%의 회수율을 나타내었고, 실험실간 교차검증에서 -1.29~0.26의 z-score를 보여 신뢰할 수 있는 것으로 판단되었다. 이결과로부터 본 연구에서 확립된 전처리법 및 분석법은 미지정 색소 azorubine이 사용된 식품검사에 활용될 수 있는 분석법으로 확인되었다.

Azorubine is a synthetic tar color containing azo-bond in the molecular structure. This food colorant has been allowed to be used for beverages, cheese and dried fruits in the European Union and for some food in Australia. Even though it is applicable as a food color in many countries, this compound has not been permitted in Korea so far as a food additive. Thus, this study was performed to establish an analysis method for azorubine in Korea by comparison of three HPLC analysis methods for azorubine and other azo-compounds which are officially used in the European Food Safety Authority (EFSA, EU), the Food Standard Agency (FSA, England) and the National Institute of Food and Drug Safety Evaluation (NIFDS, Korea). The analysis method of the FSA for azorubine showed the best linearity (r2 = 0.999), limit of detection (LOD, 0.07 μg/mL), limit of quantification (LOQ, 0.20 μg/mL), precision (0~0.5%) and accuracy (98.6~100.7%) among tested HPLC methods using a C-18 column and diode array detector (DAD) with ammonium acetate solution and acetonitrile as an eluent solution. Finally selected method of FSA was further verified by inter-day and intra-day experiments with linearity, LOD, LOQ, precision and accuracy. Recovery test showed the recover ratios of 97~103%, 95~101%, and 93~102% in beverages, breads/snacks and other foods, respectively. Inter-laboratory test represented the absolute value of z-score of less than 2 which means satisfactory levels in this test. Selected method of FSA showed reliable analytical results in application test using food samples collected in commercial markets in Europe.

Materials and Methods
  Food samples
  Chemicals and reagents
  Equipment
  Analysis methods and pretreatment conditions
  Method validation
  Sample analysis
  Analysis by LC-(ESI)-MS/MS
 Results and Discussion
  HPLC and LC-(ESI)-MS/MS analysis
  Validation of HPLC analysis
  Sample analysis
 Conclusion
 국문요약
 References

• Min-ji Kim(Advanced Food Safety Research group, Brain Korea 21 Plus, School of Food Science and Technology, Chung-Ang University)
• Ju-hee Park(Advanced Food Safety Research group, Brain Korea 21 Plus, School of Food Science and Technology, Chung-Ang University)
• Hee-Jae Suh(Department of Food Science, Sun Moon University)
• Chan Lee(Advanced Food Safety Research group, Brain Korea 21 Plus, School of Food Science and Technology, Chung-Ang University) Correspondence to