목차

[표제지 등]=0,1,2

연구개발 최종보고서=1,3,1

요약문=2,4,11

목차=13,15,4

표차례=17,19,1

그림차례=18,20,1

제1장 서론=19,21,1

제1절 연구개발의 목적 및 필요성=19,21,3

제2절 연구개발의 범위=21,23,1

제2장 국내ㆍ외 기술개발 현황=22,24,1

제3장 연구개발수행 내용 및 결과=23,25,1

제1절 연구개발 수행 내용=23,25,1

1. 실험재료=23,25,1

가. 민속주=23,25,1

나. 민속주 제조에 첨가되는 식물성 부재료=23,25,1

다. 시약=23,25,7

2. 시료의 소재=30,32,1

가. 표준시약=30,32,1

나. 민속주에 함유된 asarone, coumarin, thujone의 추출 및 분리=30,32,1

다. 식물성 부재료로부터 asarone, coumarin, thujone의 추출 및 분리=30,32,1

3. 실험방법=30,32,1

가. Asarone, coumarin, thujone의 분석을 위한 최적 기기분석조건 수립=30,32,1

(1) Gas chromatograph(GC-FID)=30,32,2

(2) Gas chromatograph/mass spectrometry(GC/MS)=31,33,1

(3) Retention index(RI)=31,33,1

(가) 머무름 지수(RI)의 수립=31,33,2

(나) 머무름 지수의 수립을 위한 n-alkane의 retention time=32,34,2

(다) 머무름 지수(RI) 수립을 위한 basic program=34,36,6

나. 전통 민속주에 함유된 독성 관련 향기성분의 추출 방법 수립=40,42,1

(1) Liquid liquid extraction & perforation(LLEP) 방법=40,42,1

(가) LLEP 방법에 의한 독성 관련 향기성분의 추출=40,42,1

(나) 추출물에서의 acid fraction 분리=40,42,1

(다) 분리된 성분의 농축=40,42,1

(2) Simultaneous steam distillation and extraction(SDE) 방법=40,42,1

(가) SDE 방법에 의한 독성 관련 향기성분의 추출=40,42,2

(나) 추출물에서의 acid fraction 분리 및 농축=41,43,1

(3) Solid phase extration(SPE) 방법=41,43,1

(가) SPE 방법에 의한 독성 관련 향기성분의 추출=41,43,1

(나) 추출물의 농축=41,43,3

다. Solid phase extraction(SPE) 방법에 의한 독성 관련 향기성분의 최적 추출 조건 수립=44,46,1

(1) 추출 및 분리를 위한 최적 SPE 컬럼의 선택=44,46,1

(2) 추출 및 분리를 위한 용리 용매의 선택=44,46,1

(3) 염의 첨가에 의한 회수율 비교=44,46,1

(4) Asarone, coumarin, thujone의 농도에 따른 회수율 비교=45,47,1

(5) Asarone, coumarin, thujone의 GC에 의한 분석=45,47,1

(6) Asarone, coumarin, thujone의 GC/MS에 의한 확인=45,47,2

(7) Selected ion monitoring(SIM) 방법에 asarone, coumarin, thujone의 정량적 분석=46,48,1

라. Solid phase microextraction(SPME) 방법에 의한 독성 관련 향기성분의 최적 추출 조건=46,48,1

(1) Asarone, coumarin, thujone의 흡착을 위한 최적 SPME fiber의 선택=46,48,1

(2) Asarone, coumarin, thujone의 흡착을 위한 최적 조건 설정=46,48,2

제2절 연구결과=48,50,1

1. Asarone, coumarin, thujone 분석을 위한 최적 기기분석 조건=48,50,1

가. Gas chromatograph(GC-FID)=48,50,1

나. Gas chromatograph/mass spectrometry(GC/MS)=49,51,3

2. 추출방법에 따른 asarone, coumarin, thujone의 회수율 비교=51,53,1

가. 분리된 향기성분의 농축=51,53,1

나. 추출방법에 따른 회수율 비교=51,53,2

3. SPE 방법에 의한 asarone, coumarin, thujone의 분석=53,55,1

가. Asarone, coumarin, thujone의 GC에 의한 분석=53,55,1

나. Asarone, coumarin, thujone의 GC/MS에 의한 확인=54,56,1

다. Asarone, coumarin, thujone의 최적 추출을 위한 고정상의 선택=55,57,3

라. 최적 용리용매의 선택=58,60,2

마. 염의 첨가가 회수율에 미치는 영향=60,62,2

바. Asarone, coumarin, thujone 농도가 회수율에 미치는 영향=62,64,2

사. GC/MS의 SIM 방법에 의한 asarone, coumarin, thujone의 검량선 작성=64,66,4

4. SPME 방법에 의한 asarone, coumarin, thujone의 분석=68,70,1

가. Asarone, coumarin, thujone의 GC에 의한 분석=68,70,1

나. Asarone, coumarin, thujone의 GC/MS에 의한 확인=68,70,1

다. Asarone, coumarin, thujone의 분석을 위한 최적 SPME fiber의 선택=69,71,2

라. 온도에 따른 asarone, coumarin, thujone 흡착율의 변화=70,72,1

마. SPME fiber의 노출시간=71,73,2

바. 시료내의 염의 농도가 SPME fiber의 흡착에 미치는 영향=73,75,1

사. 시료의 양에 따른 흡착율의 변화=73,75,2

아. SPME 방법에 의한 asarone, coumarin, thujone의 검량선 작성=75,77,1

(1) SIM 방법에 의한 asarone, coumarin, thujone의 검량선 작성=75,77,3

5. Asarone, coumarin, thujone의 정량적 분석=78,80,1

가. 전통 민속주에서 asarone, coumarin, thujone의 정량적 분석=78,80,5

나. SPE 방법에 의한 민속주 부재료의 20% ethanol 추출물에서 asarone, coumarin, thujone의 분석=83,85,1

(1) 민속주 부재료 20% ethanol 추출물중의 asarone 분석=83,85,1

(2) 민속주 부재료 20% ethanol 추출물중의 coumarin 분석=83,85,2

(3) 민속주 부재료 20% ethanol 추출물중의 thujone 분석=84,86,5

다. 국외에서의 asarone, coumarin, thujone에 대한 규제 및 고찰=89,91,2

6. 독성관련 향기성분이 함유된 민속주의 재분석=90,92,1

가. Sample A에서 thujone의 함량 분석=90,92,2

(1) SPE 방법에 의한 Sample A 부재료의 20% ethanol 추출물에서 thujone의 분석=91,93,1

(2) 부재료 양을 달리한 Sample A 제조=91,93,2

나. Sample B에서 thujone의 함량 분석=93,95,1

(1) SPE 방법에 의한 Sample B 부재료의 20% ethanol 추출물에서 thujone의 분석=93,95,1

(2) Sample B 제조=93,95,2

제4장 연구개발목표 달성도 및 대외기여도=95,97,2

제5장 연구개발결과의 활용성과 및 계획=97,99,1

제6장 참고문헌=98,100,6

최종보고서 요약(초록)=104,106,2

Table 1. List of traditional folk wine of Korea=24,26,3

Table 2. List of medicinal plant material used for traditional folk wine of Korea=27,29,3

Table 3. Retention time of n-alkane standards for gas chromatographic retention index=33,35,1

Table 4. Sample condition used for 100μm PDMS(polydimethylsiloxane) SPME fiber=47,49,1

Table 5. GC condition for analysis of toxic relevant flavor components in traditional folk wine=48,50,1

Table 6. GC/MS condition for identification of toxic relevant flavor components in traditional folk wine=49,51,1

Table 7. Analytic data of asarone, coumarin and thujone identified by GC-MS=51,53,1

Table 8. Recovery rate of asarone, coumarin and thujone by extraction methods=52,54,1

Table 9. GC condition for analysis of asarone, coumarin and thujone by SPE method=53,55,1

Table 10. GC/MS condition for identification of asarone, coumarin and thujone by SPE method=54,56,1

Table 11. Recovery rate of asarone, coumarin and thujone on adsorbent type of SPE=55,57,1

Table 12. Recovery rate of asarone, coumarin and thujone on varying proportions of mixture for elution solvent=58,60,1

Table 13. Effect of salt addition to sample solution on recovery rate of asarone, coumarin and thujone=60,62,1

Table 14. Recovery rate under concentration of asarone, coumarin and thujone=62,64,1

Table 15. Analysis of asarone, coumarin and thujone on SPME fiber types=70,72,1

Table 16. Concentration of toxic relevant flavor components in traditional folk wine by SPE method=80,82,3

Table 17. Concentration of toxic relevant flavor components in some medicinal plant liquor by SPE method=85,87,4

Table 18. Amount of medicinal plant material used for Sample A=92,94,1

Table 19. Concentration of toxic relevant flavor components in Sample A by SPE method=92,94,1

Table 20. Amount of medicinal plant material used for Sample B=94,96,1

Table 21. Concentration of toxic relevant flavor components in Sample B by SPE method=94,96,1

Figure 1. GC chromatogram of n-alkane standards(Mixture I)=37,39,1

Figure 2. GC chromatogram of n-alkane standards(Mixture II)=38,40,1

Figure 3. GC chromatogram of n-alkane standards(Mixture III)=39,41,1

Figure 4. Likens & Nickerson type simultaneous steam distillation and extraction apparatus=42,44,1

Figure 5. Mass spectrums of asarone, coumarin and thujone peaks=50,52,1

Figure 6. Comparison of three extraction methods for asarone, coumarin and thujone=52,54,1

Figure 7. Effect of adsorbent types on recovery rate of asarone, coumarin and thujone by SPE=56,58,1

Figure 8. Chromatogram of asarone, coumarin and thujone in thaditional folk wine isolated by SPE=57,59,1

Figure 9. Effect of varying proportions of solvent mixture on recovery of asarone, coumarin and thujone by SPE=59,61,1

Figure 10. Change of recovery rate on salt addition in sample solution=61,63,1

Figure 11. Standard curves of asarone, coumarin and thujone by SPE method=65,67,2

Figure 12. Mass spectrums of asarone, coumarin and thujone standards obtained by SIM mode=67,69,1

Figure 13. Sample temperature versus intensity, 15min SPME, MS response relative to 15℃ signal internsity=72,74,1

Figure 14. Sampling time versus intensity, 40℃, MS response relative to 5min fiber exposure time=72,74,1

Figure 15. Amount of NaCl versus intensity, 30℃, 15min MS response relative to no NaCl added=74,76,1

Figure 16. Influence of sample volume on SPME fiber adsorption=74,76,1

Figure 17. Standard curves of asarone, coumarin and thujone by SPME method=76,78,2

Scheme I. Analysis of toxic relevant substances from traditional folk wine=43,45,1

Scheme II. Analysis of asarone, coumarin and thujone in traditional folk wine=63,65,1

영문목차

[title page etc.]=0,1,9

SUMMARY=8,10,1

CONTENTS=9,11,10

Chapter1. Introduction=19,21,1

Section1. Objectives and requirement of the project=19,21,3

Section2. Scope of the project=21,23,1

Chapter2. Current condition of international technical development=22,24,1

Chapter3. Contents and results=23,25,1

Section1. Contents=23,25,1

1. Material=23,25,1

a. Traditional folk wine=23,25,1

b. Medicinal plant material used for traditional folk wine of Korea=23,25,1

c. Reagents=23,25,7

2. Preparation of material=30,32,1

a. Standards=30,32,1

b. Extraction and separation of asarone, coumarin and thujone in traditional folk wine=30,32,1

c. Extraction and separation of asarone, coumarin and thujone in medicinal plant liquor used for traditional folk wine=30,32,1

3. Methods=30,32,1

a. Establishment of optimal condition of asarone, coumarin and thujone by analytic instrument=30,32,1

(1) Gas chromatograph(GC-FID)=30,32,2

(2) Gas chromatograph/mass spectrometry(GC/MS)=31,33,1

(3) Retention index(RI)=31,33,1

(a) Establishment of RI=31,33,2

(b) Retention time of n-alkane to establish retention index=32,34,2

(c) Basic program to establish RI=34,36,6

b. Extraction of toxic relevant flavor components in traditional folk wine=40,42,1

(1) Liquid liquid extraction and perforation(LLEP) method=40,42,1

(a) Extraction of toxic relevant flavor components by LLEP method=40,42,1

(b) Separation of acid fraction from extract=40,42,1

(c) Concentration of separated components=40,42,1

(2) Simultaneous steam distillation and extraction(SDE) method=40,42,1

(a) Extraction of toxic relevant flavor components by SDE method=40,42,2

(b) Separation and concentration of acid fraction from extract=41,43,1

(3) Solid phase extraction(SPE) method=41,43,1

(a) Extraction of toxic relevant flavor components by SPE method=41,43,1

(b) Concentration of extract=41,43,3

c. Optimal extract condition of toxic relevant flavor components by SPE method=44,46,1

(1) Selection of SPE column=44,46,1

(2) Selection of elution solvent=44,46,1

(3) Comparison of recovery rate by salt addition=44,46,1

(4) Comparison of recovery rate by concentrations of asarone, coumarin and thujone=45,47,1

(5) Analysis of asarone, coumarin and thujone by GC=45,47,1

(6) Identification of asarone, coumarin and thujone by GC/MS=45,47,2

(7) Quantitative analysis of asarone, coumarin and thujone by selected ion monitoring(SIM) method=46,48,1

d. Extraction and separation of toxic relevant flavor components by solid phase microextraction(SPME) method=46,48,1

(1) Selection of SPME fiber to adsorb asarone, coumarin and thujone=46,48,1

(2) Optimum condition to adsorb asarone, coumarin and thujone=46,48,2

Section2. Results=48,50,1

1. Optimum analytic condition of asarone, coumarin and thujone=48,50,1

a. Gas chromatograph(GC-FID)=48,50,1

b. Gas chromatograph/mass spectrometry(GC/MS)=49,51,3

2. Comparison of recovery rate by extraction method=51,53,1

a. Concentration of extract=51,53,1

b. Comparison of recovery rate by extraction method=51,53,2

3. Analysis of asarone, coumarin and thujone by solid phase extraction(SPE) method=53,55,1

a. Analysis of asarone, coumarin and thujone by GC=53,55,1

b. Identification of asarone, coumarin and thujone by GC/MS=54,56,1

c. Selection of solid phase for optimum extraction=55,57,3

d. Selection of optimum elution solvent=58,60,2

e. Effect of salt addition on recovery rate=60,62,2

f. Effect of concentration on recovery rate=62,64,2

g. Standard curves of asarone, coumarin and thujone by GC/MS with selected ion monitoring(SIM) method=64,66,4

4. Analysis of asarone, coumarin and thujone by solid phase microextraction(SPME)method=68,70,1

a. Analysis of asarone, coumarin and thujone by GC=68,70,1

b. Identification of asarone, coumarin and thujone by GC/MS=68,70,1

c. Selection of SPME fiber=69,71,2

e. Change of adsorption rate=70,72,1

f. Exposure time of SPME fiber=71,73,2

g. Influence of adsorption rate on salt concentration in sample=73,75,1

h. Change of adsorption rate by amount of sample=73,75,2

i. Standard curves of asarone, coumarin and thujone by solid phase microextraction(SPME) method=75,77,1

(1) Standard curves of asarone, coumarin and thujone by GC/MS with SIM method=75,77,3

5. Quantitative analysis of asarone, coumarin and thujone=78,80,1

a. Quantitative analysis of asarone, coumarin and thujone in traditional folk wine=78,80,5

b. Analysis of asarone, coumarin and thujone in 20% ethanol extract of medicinal plant meterial by SPE method=83,85,1

(1) Analysis of asarone in 20% ethanol extract in medicinal plant material=83,85,1

(2) Analysis of coumarin in 20% ethanol extract in medicinal plant material=83,85,2

(3) Analysis of thujone in 20% ethanol extract in medicinal plant material=84,86,5

c. Regulation and investigation for asarone, coumarin and thujone of foreign states=89,91,2

6. Analysis of toxic relevant flavor components in traditional folk wine=90,92,1

a. Content analysis of thujone in Sample A=90,92,2

(1) Analysis of thujone in 20% ethanol extract of medicinal plant material used for Sample A by SPE method=91,93,1

(2) Preparation of Sample A=91,93,2

b. Content analysis of thujone in Sample B=93,95,1

(1) Analysis of thujone in 20% ethanol extract of medicinal plant material used for Sample B by SPE method=93,95,1

(2) Preparation of Sample B=93,95,2

Chapter4. Achievement of research goals and foreign contribution=95,97,2

Chapter5. Application of the results=97,99,1

Chapter6. References=98,100,8