한국식품위생안전성학회지 Vol.37 No.2 (p.46-54)

식품에서 당살초 판별을 위한 LC-ESI-MS/MS 분석법과 KASP 마커 개발

Development of Method using LC-ESI-MS/MS and KASP for Identification of Gymnema sylvestre in Food
키워드 :
Gymnema sylvestre,LC-ESI-MS/MS,KASP,Food analysis

목차

ABSTRACT
Materials and Methods
   시약 및 재료
   LC-MS/MS 분석조건 확립
   DNA 추출 및 PCR
   당살초 특이 KASP 마커 제작
   KASP 분석조건 확립
   식품 중 분석법 적용성 검토
Results and Discussion
   당살초 지표 성분 2종에 대한 LC-MS/MS 조건 확립
   당살초에 대한 LC-MS/MS 분석법 검증
   KASP 마커 확인 및 당살초 원물 적용
   식품에 대한 분석법 적용
Acknowledgement
국문요약
Conflict of interests
ORCID
References

초록

Known for its effectiveness in weight loss and diabetes prevention, Gymnema sylvestre products can be found in the US, Japanese, and Indian markets. However, the recommended dosage or safety of these products has not yet been proven. Therefore, development of an analytical method for detecting the content of Gymnema sylvestre in food products is required. Accordingly, this study proposes an analysis method that can examine Gymnema sylvestre in food using LC-ESI-MS/MS and KASP (Kompetitive Allele-Specific PCR) markers. In LC-ESI-MS/MS, a simultaneous analysis method for gymnemic acid and deacylgymnemic acid was optimized using negative ionization mode, and its validation test was completed for solid and liquid samples. In addition, KASP markers were prepared by finding the specific SNP of G. sylvestre in ITS2 and matK through DNA barcodes. The two KASP markers returned positive FAM fluorescence result when combined with G. sylvestre, and this aspect was confirmed in raw G. sylvestre as well. The applicability of the method was tested on 21 different food and healthy functional products containing G. sylvestre purchased on the internet. As a result, although there was a difference in the ratios of gymnemic acid and deacylgymnemic acid in LC-ESI-MS/MS, the index component was detected in all 21 products samples. In the KASP analysis, 9 products returned positive FAM result, and the rest of the products were found to be containing G. sylvestre extract. This study is the first study to use the dual system of LC-ESI-MS/MS and KASP for the analysis of G. sylvestre. The study has confirmed that these two methods are applicable to the examine G. sylvestre content in food products.