보고서 정보
주관연구기관 |
국립원예특작과학원 National Institute of Horticultural and Herbal Science |
보고서유형 | 최종보고서 |
발행국가 | 대한민국 |
언어 |
한국어
|
발행년월 | 2015-03 |
과제시작연도 |
2014 |
주관부처 |
농촌진흥청 Rural Development Administration(RDA) |
등록번호 |
TRKO201500010258 |
과제고유번호 |
1395035170 |
사업명 |
FTA대응경쟁력향상기술개발 |
DB 구축일자 |
2015-07-11
|
초록
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Ⅳ. 연구개발결과
DNA 감식법을 활용한 국내산과 외국산 홍삼 농축액 판별방법 개발 과제(1세부과제)수행을 통해, 국내산과 외국산(미국삼)의 홍삼농축액 원산지 판별을 위하여 다양한 DNA 마커를 적용한 결과, 미토콘드리아 보존부위에서 새로 제작한 PQ91 프라이머로 고려인삼과 미국삼으로 제조된 홍삼농축액이 구분됨을 확인하였다. 개발된 PQ91 프라이머는 식물체(잎, 뿌리)는 물론 홍삼과 홍삼농축액에도 적용 가능한 특징을 갖고 있다.한편 연구수행을 통하여 고려인삼과 중국 전칠삼을 구분할 수 있는 PN418 프라이머도 개발하였다.
Ⅳ. 연구개발결과
DNA 감식법을 활용한 국내산과 외국산 홍삼 농축액 판별방법 개발 과제(1세부과제)수행을 통해, 국내산과 외국산(미국삼)의 홍삼농축액 원산지 판별을 위하여 다양한 DNA 마커를 적용한 결과, 미토콘드리아 보존부위에서 새로 제작한 PQ91 프라이머로 고려인삼과 미국삼으로 제조된 홍삼농축액이 구분됨을 확인하였다. 개발된 PQ91 프라이머는 식물체(잎, 뿌리)는 물론 홍삼과 홍삼농축액에도 적용 가능한 특징을 갖고 있다.한편 연구수행을 통하여 고려인삼과 중국 전칠삼을 구분할 수 있는 PN418 프라이머도 개발하였다. 본 연구 수행을 통하여 개발된 홍삼 농축액 판별용 DNA 판별기술은 국가 농산물품질관리기관 등에 기술 이전되어, 인삼류의 원산지 판별 검정 업무에 유용하게 활용될 수 있을 것으로 생각된다.분석장비를 활용한 홍삼 농축액의 원산지 판별기술 개발 과제(제1협동과제) 수행을 통해, 국내산과 수입산의 홍삼 농축액의 원산지 판별을 위하여 MS-전자코와 LC/MS/MS를 활용하였다. LC/MS/MS를 사용하여 원산지에 따른 특징적인 진세노사이드를 알아내었다. 이를 기반으로 의심이 되는 시료를 선정하였고 국내산과 수입산이 어떤 비율로 혼합되었는지 알아보기 위해서 국내산과 수입산의 비율을 10:0부터 0:10 까지 희석하여 전자코분석을 해 보았다. 그 결과 희석된 국내산 홍삼 농축액양이 많을수록 향기성분이 많이 생성된 것으로 나타났고 의심시료는 해당 희석배수와 동일한 위치에 있었다. 또한 국내산 홍삼농축액에 더덕 농축액, 도라지 농축액, 과당을 혼합하였을 때도 전자코로 판별이 가능한지 알아보기 위해서 각각 10:0부터 0:10까지 혼합하여 분석하였다. 세 가지 첨가액 모두 국내산 홍삼 농축액이 많을수록 향기성분이 많은 것으로 나타났지만, 전자코 분석만으로 그 종류까지는 알 수 없었다. 추후 연구에서 LC/MS/MS를 병행하여 분석한다면 어떠한 물질이 첨가되었는지도 판별이 가능할 것으로 생각된다.
홍삼 농축액 시중 유통품 시료 수집을 통한 분석장비별 Database 구축 및 원산지 판별법 검증 과제(제2협동과제) 수행을 통해, 시중 유통품 시료 총 232점을 수집하여 분석장비별 Database 구축을 완료하였고, 국내 재배 고려인삼과 중국 재배 고려인삼으로 가공한 홍삼농축액 시료와 시중 유통품을 이용하여 원산지 판별법 검증을 수행한 결과, 근적외선분광광도계(NIRS), X-선형광분석기(XRF), MS-전자코를 사용하여 원산지 판별이 어려운 시판 홍삼 농축액의 원산지를 시료의 추출이나 화학반응 등의 전처리 과정 없이 1~2분내에 작성된 검량식을 이용하여 신속하고 정확하게 홍삼 농축액의 원산지를 판별할 수 있는 뛰어난 효과가 있음을 확인하였다. 시중에 유통중인 홍삼농축액의 올바른 원산지 정보 제공과 부정유통 방지를 위해 매우 유용하게 이용이 가능하다. 또한 본 연구에 의한 원산지 판별법은 원산지 단속업무에 과학적인 근거자료를 제공하는데 그 활용도가 높을 것으로 기대된다.
Abstract
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This study was carried out to develop convenient and reliable DNA markers for authentication of Korean and foreign ginseng in commercial processed products. We used polymerase chain reaction (PCR) with universal rice primers (URP), random amplified polymorphic DNA (RAPD) primer, expressed sequence t
This study was carried out to develop convenient and reliable DNA markers for authentication of Korean and foreign ginseng in commercial processed products. We used polymerase chain reaction (PCR) with universal rice primers (URP), random amplified polymorphic DNA (RAPD) primer, expressed sequence tag (EST)-derived simple sequence repeat (SSR) primer and consensus set of primers(chloroplast primers and mitochondrial primers) for authentication of ginseng species in commercial products such as powder, shredded slices, pellets, red ginseng extracts.These primers could not generated polymorphisms among the Panax species except for three mitochondrial primers. Three primers (cox1, nad1/2-3 and nad2/1-2) generated co-dominant polymorphic banding patterns discriminating Korean ginseng cultivars from P. quinquefolius and P. notoginseng. Primers PQ91 and PN418 were designed from the consensus sequence of nad1/2-3 region.
Two banding patterns (A or B) were detected in PQ91. Korean ginseng cultivars and P. notoginseng shared the same banding pattern (A type) and P. quinquefolius was identified another banding pattern (B type). In the case of PN418, two banding patterns (A or B)were detected in the Korean ginseng cultivars and two foreign Panax species. Korean ginseng cultivars and P. quinquefolius shared the same banding pattern (A type) and P. notoginseng was identified another banding pattern (B type). The combination banding patterns of three Panax species, Korean ginseng cultivars (Panax ginseng C. A. Mey.), P. quinquefolius and P. notoginseng, was identified as‘AA’, ‘BA’ and ‘AB’, respectively. Consequently, PQ91 and PN418 primer sets can be used to distinguish among Panax species. Both markers were successfully applied to evaluate the original species from processed ginseng products such as red ginseng extract and powder.
The geographical origin of red ginseng extract (RGE) was studied using a mass spectrometry based electronic nose. The treated RGE was analyzed, and discriminant function analysis (DFA) was used for discriminating of geographical origins. The DFA plots indicated a significant separation of imported RGE and domestic RGE.The F-value of discriminant function first score (DF1) was much higher than that of discriminant function second score (DF2), indicating that discrimination was mainly affected by DF1. Based on DF1, the concentration of domestic RGE to imported RGE shifted to the left side of DFA plot, and the mixing ratio highly correlated to DF1 value. Unknown sample (#2) was closely located to the sample of mixed imported : domestic (6:4) RGE. In the bar graph, the DF1 value correlated to the mixing ratio. Unknown samples (#2) were thought to be mixed with the imported RGE.
The geographical origin of commercial RGE was studied using liquid chromatography-tandem mass spectrometry(LC-MS/MS) and electronic nose based on mass spectrometry. LC-MS/MS analysis result, in the case of ginseng ginsenoside detected in Ra1, Rb1, Rg2, the larger difference between the domestic and China, is considered as indicator would be available. Also, not detected in the case of ginseng ginsenoside Rh2, F2 was found only in the China RGE. Responses of ion fragments were obtained by electronic nose. Discriminant function analysis of the pattern obtained three grouped Korea, China, suspected origin RGE separately. Moreover, in order to determine the mixture ratio of the origin unknown specimen, the mixture ratio was predicted by mixing the specimens of entirely Korean or Chinese origin according to the mixture ratio. Analysis of the two devices are used together, each other’s advantage and disadvantage can be complement. The adulteration of RGE was studied using electronic nose based on mass spectrometry.
Domestic red ginseng, bellflower root, and Codonopsis lanceolata concentrates were diluted to 10。Brix. F-value of DF1 in the DFA plots was much higher than that of DF2, indicating that discrimination was mainly depending upon DF1. Mixing ratio of domestic red ginseng and bellflower or Codonopsis lanceolata highly correlated to DF1(r2=0.9788, 0.9925). This technique could be used to efficiently differentiate the geographical origin and adulteration of RGE.Discrimination of geographic origin of agricultural products and food is a important issue in Korea because the price difference between Korean and Chinese food is a key among some reasons. NIRS (Near Infrared Reflectance Spectroscopy) has been applied to identify the geographical origin of red ginseng extracts. Total 232 samples (Korean 132 and Chinese 100) were used to obtain calibration equation 9,405–4,000cm-1 wavenumber.
The math treatment with MIN-MAX normalization and 8cm-1 resolution and the partial least squares(PLS) regression was outstanding for calibration equation. The root mean square error of estimation and determination of coefficient (r2) values in calibration set was 10.9 and 0.95, respectively. And ED-XRF(Energy Dispersive X-ray Fluorescence Spectrometer) has been applied to identify the geographical origin of red ginseng extracts. Mineral content ratios, of a total of 13 species, including P, S, Cl, K, Fe, Zn, Rb, Sr, Ag, Sn, Sb, and Te were measured among 232 samples. Macro element content ratios and trace element content ratios were determined using the standardless fundamental parameters (SLFP) analysis. These element ratios of P, Cl, K, Fe and Zn were significantly different between domestic and imported samples. The result from the canonical discriminant analysis showed that the accuracy of geographical origin discrimination was 97.56%; the correlation coefficient was 0.9201.
It was concluded that this technique could be used as a useful method in discriminating the geographical origins between Korean and Chinese red ginseng extracts. And we applied to identify the geographical origin of red ginseng extracts using a mass spectrometry based electronic nose. Total 232 samples (Korean 132 and Chinese 100) were used to measure and discriminant function analysis (DFA). The DFA plots indicated a significant separation of Korean and Chinese red ginseng extracts. The result from the canonical discriminant analysis showed that the accuracy of geographical origin discrimination was 100%. And it showed that the extra 60 samples for validation equation were estimated their authentication correctly. This study showed that the application of NIRS, ED-XRF, MS-Electronic Nose would be possible for discrimination of geographical origin between Korean and Chinese red ginseng extracts.
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