식품첨가물 카민과 코치닐 추출물은 붉은 색을 띄는 색 소로, 주로 코치닐 곤충에서 유래하며 카민산이 주요 성 분으로 사탕, 어묵과 같이 널리 소비되는 식품에 일반적 으로 사용되고 있다. 최근 카민산은 특정 단백질과 관련된 알레르기 반응과 연관이 있는 것으로 보고되고 있다. 본 연 구는 highperformance liquid chromatography with a diode array detector (HPLC-DAD)와 C18 UG120 컬럼을 사용하 여 카민산을 정량시험법을 개발하고 밸리데이션을 수행하 였다. HPLC 최적 분석조건은 이동상 (A)물-트리플루오로 아세트산(trifluoroacetic acid, TFA) 0.1% 및 (B)아세토니트 릴-TFA 0.1%, 유속 1.0 mL/min, 컬럼온도 30oC에서 수행하 였으며, 검량선(0.2-50 mg/L) 범위에서 우수한 결정계수 (r2≥0.9999)를 보였다. 또한, liquid chromatography-tandem mass spectrometry (LC-MS/MS)를 활용한 정성 분석을 통해 limit of detection (LOD) 0.05 mg/k, limit of quantification (LOQ) 0.15 mg/kg을 보였다. 또한, 일내 및 일간 밸리데이 션에서 정확도(87.3-97.1%, 회수율)와 정밀도(0.48-8.90%, 상대표준편차)이었으며, 측정 불확도 또한 추정하였다. 개 발된 분석법은 다양한 식품 유형에 적용 가능하며, 카민과 코치닐 추출물의 모니터링을 통해 카민산과 관련된 잠재적 인 문제가능성을 확인하는 데 활용될 수 있을 것으로 사료 된다.

Carmine and cochineal extract are vibrant red colored pigments derived from cochineal insects, containing carminic acid as their primary component. These colorants are commonly used in widely consumed food products including candies and fish cakes. Carminic acid has recently been associated with allergic reactions linked to specific proteins. This study aimed to develop and authenticate a method for quantifying carminic acid using highperformance liquid chromatography with a diode array detector (HPLC-DAD) and a C18 UG120 column. Conditions included a water-trifluoroacetic acid (TFA) 0.1% (A) and acetonitrile-TFA 0.1% (B) mobile phase, with a flow rate of 1.0 mL/min and a column temperature of 30oC. Calibration curves (0.2-50 mg/L) show good linearity (r 2≥0.9999). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) facilitates qualitative analysis, with a 0.05 mg/kg limit of detection (LOD) and a 0.15 mg/kg limit of quantification (LOQ). Intra-day and inter-day measurements exhibited accuracy (87.3-97.1%, recovery) and precision (0.48-8.90%, relative standard deviation, RSD. Measurement uncertainty was also estimated. The developed method is applicable for the effective monitoring of carmine and cochineal extracts in diverse food types, this is crucial for understanding and mitigating the potential health concerns associated with carminic acid.

ABSTRACT
Materials and Methods
    Chemicals and reagents
    Sample collection
    Preparation of standard solution and samples
    Analytical instrument
    Method validation
    Inter-laboratory validation
    Measurement uncertainty estimation
Results and Discussion
    Method validation
    Inter-laboratory validation
    Measurement uncertainty
    Application
Conclusion
Acknowledgement
국문요약
Conflict of interests
ORCID
References

• Min-A Kim(Department of Food Science and Biotechnology, Seoul National University of Science and Technology, Seoul, Korea)
• Hyo-Jin Yang(Department of Food Science and Biotechnology, Seoul National University of Science and Technology, Seoul, Korea)
• Ku-Won Kim(Department of Food Science and Biotechnology, Seoul National University of Science and Technology, Seoul, Korea)
• Choong-In Yun(Department of Food Science and Biotechnology, Sungkyunkwan University, Suwon, Korea, Research Institute of Food and Biotechnology, Seoul National University of Science and Technology, Seoul, Korea)
• Young-Jun Kim(Department of Food Science and Biotechnology, Seoul National University of Science and Technology, Seoul, Korea, Research Institute of Food and Biotechnology, Seoul National University of Science and Technology, Seoul, Korea) Corresponding author