다중 표적 프로파일 기법을 이용한 인삼 및 홍삼 가공식품 중 유사원료 혼입 판별 연구 = Determination of Adulteration in Ginseng(Panax ginseng) Products by Multi-target Profiling Method

다국어 초록 (Multilingual Abstract)

The consumption of medicinal food to maintain good health has increased, resulting to extend the markets for functional foods. Especially, ginseng(Panax ginseng, ginseng radix) and red ginseng; a processed ginseng product in South Korea, are commonly used as foods in general and healthy functional food in particular from thousands of years ago. Due to high market demands, inexpensive morphologically similar materials may be intentionally mixed into ginseng or red ginseng products to derive benefits. It is very important to know whether the ginseng or red ginseng products are adulterated or not. The genetic analysis cannot be applied to processed red ginseng products, as they involve high temperature and pressure processing and genes may be destroyed. Therefore, this study was designed to find out some physicochemical indicators that would not be changed in various processing processes. Several possible substances such as Codonopsis lanceolata, Platycodon grandiflorum, and Pueraria lobata were tested as adulterants and investigated in the study.
To approached with multi-taret profiling method in subject samples and report a suitable method, the inorganic components were analyzed by using inductively coupled plasma/mass spectrometry(ICP/MS) and inductively coupled plasma/optical emission spectrometry(ICP/OES), volatile organic components were analyzed by using gas chromatography-mass spectrometry(GC-MS), phytochemical screening via qualitative and quantitative analyses were done using standard reference methods and analysis of nonvolatile organic components were determined using high performance liquid chromatography(HPLC) and LC-mass spectrometry/mass spectrometry(MS/MS).
Six macro elements(Na, Mg, Ca, K, Fe, and Al) in individual dried and steamed dried samples were analyzed through ICP-OES and 19 trace elements(Ba, Cr, Cu, Mn, Ni, Rb, Sr, Zn, Ga, Se, Tl, Be, Co, V, Li, Cs, Bi, Pb, Cd) in the samples were analyzed using ICP-MS. No significant difference due to processing was shown between raw samples and steamed samples and it was difficult to discriminate among the samples based on the analyzed contents of inorganic components. Among the statistical analyses, the Linear Discriminant Analysis (LDA) of the inorganic profile enabled discrimination among the individual samples. The raw samples and steamed samples were clearly distinguished from each other on the graph. Thus discrimination among the samples using inorganic elements was possible by LDA statistics.
The volatile organic components were extracted using the Solid phase micro extraction(SPME) method, and analyzed using GC-MS. From the results, (E)-2-hexen-1-ol and barbatene were identified in the raw samples of C. lanceolata and P. gradiflorum, but not in the raw samples of the P. ginseng and P. lobata. Furaneol was detected only from the steamed sample of P. gradiflorum. There were difficulties in the analysis of volatile organic components because the reproducibility was poor and the expected quantities of indicators were very small. Therefore, it was judged that applying volatile organic components as a method to identify the similar materials mixed into processed ginseng or red ginseng products would be difficult.
In phytochemical screening tests, terpenoids, phytosterols, phenolic compounds, coumarins, flavonoids, and alkaloids were screened and changes in colors were observed. Since high reactions of flavonoid and phenolic components were identified in the methanol extract of P. lobata, it was mixed into red ginseng extract at ratios of 10, 20, 30, 40 and 50% and screening tests were conducted. The intensity of the colors changed according to the mixing ratio, so it was easy to check whether or not the P. lobata were mixed.
Analyses were conducted using HPLC, to explore indicators that do not exist in ginseng but are detected from C. lanceolata, P. grandiflorum and P. lobata. In the results, peak 1, which was detected in C. lanceolate and P. grandiflorum in the same time zone, and peak 2, which was detected only in P. lobata but not in other samples, were identified. Expected indicators were verified by measuring their accurate masses through LC-MS/MS analyses, and as components not detected in ginseng, the same indicator of lobetyolin was identified in C. lanceolate, P. grandiflorum and an indicator ononin was identified in P. lobata. Therefore, it was concluded that the HPLC method developed in this study can identify whether or not C. lanceolate, P. grandiflorum, or P. lobata has been mixed into ginseng or red ginseng.
A total of 61 general and health functional foods made of ginseng or red ginseng distributed in the market were monitored and there were no C. lanceolate, P. grandiflorum or P. lobata mixed into the foods. When samples of processed products of C. lanceolate and P. grandiflorum, which are similar materials, were analyzed respectively, the common indicator lobetyolin was identified. Also when samples of processed P. lobata products were analyzed, ononin was detected. The method developed from this research can be used not only for ginseng processing food but also for the quality examination of bonnet, balloon flower roots, and arrochards.
In nutshell the indicator of similar materials, basic physicochemical components, macro inorganic and trace inorganic elements, and volatile organic components, were examined. Although it was difficult to set them as a method for clear discrimination, the examination was meaningful for the collection of basic data. Through this study, a method to identify whether or not similar materials were mixed into ginseng or red ginseng products using the HPLC analysis method was established. The identification method using HPLC was developed in this study and can be applied for development of methods to identify other adulterated foods.

목차 (Table of Contents)

• 제 1 장 서 론 1
• 제 1 절 불량식품(Adulterated food) 1
• 1. 불량식품의 정의 1
• 2. 불량식품 관련 주요 사건·사고 3
• 제 2 절 인삼, 홍삼의 정의 및 유사원료 선정 근거 5
• 1. 인삼, 홍삼의 정의 및 기준규격 5
• 2. 인삼, 홍삼의 산업 현황 6
• 가. 건강기능식품의 생산량 및 매출현황 6
• 나. 인삼 및 홍삼제품류의 매출 현황 10
• 3. 유사원료 선정 근거 12
• 제 3 절 인삼, 홍삼 및 유사원료의 연구 동향 14
• 1. 인삼과 홍삼의 기능성 및 지표성분 연구 현황 14
• 2. 유사원료에 대한 연구 현황 15
• 제 4 절 연구 목적 17
• 제 2 장 재료 및 방법 19
• 제 1 절 실험재료 및 기기 19
• 1. 시료 19
• 가. 원시료 및 가공식품의 구입 19
• 나. 시료의 전처리 19
• 2. 시약 및 재료 23
• 3. 기기 24
• 제 2 절 일반성분 분석 25
• 제 3 절 무기성분 분석 26
• 1. 시료의 분해 및 무기성분 분석조건 26
• 가. 시료 분해를 위한 기기조건 및 방법 26
• 나. 무기성분 분석을 위한 기기조건 및 방법 26
• 다. 무기성분 표준용액 제조 27
• 라. 분석법 검증 27
• 제 4 절 휘발성 유기성분 분석 31
• 1. Solid phase micro extraction(SPME)에 의한 휘발성 유기성분 추출 31
• 가. 휘발성 유기성분 포집을 위한 최적 SPME fiber의 선택 31
• 나. SPME에 의한 휘발성 향기성분 포집 방법 31
• 2. 휘발성 유기성분 분석을 위한 기기분석 조건 및 방법 33
• 가. 휘발성 유기성분 분석을 위한 분석기기 조건 33
• 나. 머무름 지수(Retention index)의 수립 방법 35
• 다. 휘발성 유기성분의 확인 및 정량 35
• 제 5 절 비휘발성 유기성분 분석 36
• 1. Phytochemical 성분 분석 36
• 가. 시험용액 제조 36
• 나. Phytochemical screening 실험 방법 36
• 2. HPLC 및 LC-MS/MS를 이용한 비휘발성 유기성분 분석 39
• 가. HPLC를 이용한 유사원료의 지표성분 탐색 39
• 1) LC 분석을 위한 시험용액 조제 39
• 2) 지표성분 탐색을 위한 HPLC screening 분석 조건 40
• 3) 인삼, 홍삼 및 유사원료의 HPLC screening 분석 40
• 나. LC-MS/MS를 이용한 지표성분 분석 방법 42
• 1) LC-MS/MS 분석기기 조건 42
• 2) LC-MS/MS를 이용한 시료 및 표준물질 분석 42
• 3. 유사원료의 지표성분 분석을 통한 혼입여부 판별법 확립 44
• 가. 유사원료의 지표성분 분석을 위한 HPLC 분석 조건 44
• 나. HPLC 분석법의 유효성 검증(Method validation) 46
• 다. 지표물질의 정성 및 정량시험 48
• 제 6 절 인삼, 홍삼 가공식품 중 유사원료 혼입여부 모니터링 49
• 1. 가공식품 시료 49
• 2. 가공식품의 전처리 방법 49
• 가. 분말 형태 제품의 전처리 방법 49
• 나. 액(즙) 형태 제품의 전처리 방법 50
• 다. 농축액 형태 제품의 전처리 방법 50
• 3. 가공식품 모니터링을 위한 HPLC 분석조건 52
• 4. 가공식품 중 유사원료 지표물질의 정성 및 정량방법 52
• 제 7 절 통계분석 53
• 1. 분산분석 (Analysis of variance, ANOVA) 53
• 2. 선형판별 분석 (Linear discriminant analysis, LDA) 53
• 제 3 장 결과 및 고찰 54
• 제 1 절 일반성분 분석 결과 54
• 제 2 절 무기성분 분석 결과 56
• 1. 무기성분 분석법의 유효성 검증 56
• 가. 분석법의 유효성 검증 56
• 나. 인증표준물질을 이용한 분석법의 소급성 확인 58
• 다. 국외 숙련도 시험(FAPAS) 61
• 2. 다량 무기원소 분석 결과 62
• 3. 미량 무기원소 분석 결과 65
• 4. 무기원소 함량을 이용한 선형판별분석(LDA) 결과 68
• 제 3 절 휘발성 유기성분 분석 결과 71
• 1. n-Alkane의 머무름 지수 수립 71
• 2. SPME 방법에 따른 휘발성 유기성분의 분석 결과 73
• 가. 휘발성 유기성분 포집을 위한 최적 SPME 조건 수립 73
• 나. SPME에 의한 휘발성 유기성분 분석 결과 76
• 제 4 절 비휘발성 유기성분 분석 결과 83
• 1. Phytochemical screening 실험 83
• 가. Phytochemical screening 실험결과 83
• 나. 유사원료 혼합비율에 따른 홍삼농축액의 Phytochemical screening 결과 85
• 2. HPLC 및 LC-MS/MS를 이용한 비휘발성 유기성분 분석 88
• 가. HPLC를 이용한 유사원료의 지표물질 탐색 88
• 나. LC-MS/MS를 이용한 지표성분 및 구조 확인 102
• 3. 더덕, 도라지, 칡의 지표성분 분석을 통한 혼입여부 판별법 확립 111
• 가. 유사원료의 지표성분 분석을 위한 HPLC 분석 조건 확립 111
• 나. HPLC 분석법의 유효성 검증(Method validation) 111
• 다. 지표성분의 정성 및 정량 분석 결과 121
• 제 5 절 인삼, 홍삼 가공식품 중 유사원료 혼입여부 모니터링 123
• 제 4 장 요 약 127
• 참고문헌 130