Chrysanthemum boreal, C. indicum, and C. indicum var. albescens are well-known wild Chrysanthemum species used for traditional medicine in Korea. In this study, volatile compounds from three wild Chrysanthemums were identified according to four different flowering stages and analyzed using HS-SPME-GC-MS to determine the temporal variation of the volatiles. As a result, 132, 151, and 142 peaks were identified from C. boreale, C. indicum, and C. indicum var. albescens, respectively. Furthermore, 70 out of 132 peaks were identified in C. boreale with a matching ratio of >90% from library search. In addition, 85/151 and 76/142 peaks were identified from C. indicum and C. indicum var. albescens. Forty-nine volatile compounds were found commonly in all three wild Chrysanthemums through all four different flowering stages. However, six, seven, and five unique compounds were detected only in C. boreale, C. indicum, and C. indicum var. albescens, respectively. One hundred volatile compounds were selected for multivariate analysis considering volatile compounds overlapped with each other. The one-way ANOVA (p < 0.05) detected significant differences from 77 out of 100 volatile compounds. In addition, PLS-DA showed the different profiles of volatile compounds according to four different flowering stages in each wild Chrysanthemum. PC1 of each Chrysanthemum accounted for 45.8 56.9, and 11.9% in C. boreale, C. indicum, and C. indicum var. albescens, respectively. PC1 of C. boreale and C. indicum clearly separated the BF stage and the other three stages. Conversely, PC1 of C . indicum var. albescens showed a difference in the composition of volatile compounds between the BF/BO and HO/FO stages. In addition, the different profiles of volatile compounds could be visualized using a heatmap from three wild Chrysanthemums according to four different flowering stages. This study will help improve particular volatile compounds in three wild Chrysanthemums both in quality and quantity.

본 연구는 HS-SPME/GC-MS 방법과 화학계량학 (chemometrics) 분석으로 차의 등급과 품질 판단의 가능성을 실험한 연구이다. HS-SPME/GC-MS 방법으로 녹차, 재가공차 (화차), 홍차 및 등급별 향기를 분석했고, 주성분분석, 부분최 소제곱회귀분석 등 chemometrics와 관능 평가 결과를 결합하여 차의 종류와 등급 예측 판단을 진행하였다. 주성분분석결과는 차의 종류 판별에 양호한 결과를 내었고, 녹차 Y1(X)=- 7.138-4.094X1-7.876X2, 재가공차(화차) Y2(X)=-7.291- 2.059X1+8.157X2, 홍차 Y3(X)=-3.060+4.022X1+0.746X2 판별 함수를 얻었으며 교차검증 및 식별 인자 모두 100%의 정확률 에 도달했다. GC-MS 시그널과 차의 등급에서 부분최소제곱 회귀분석을 진행하고, 관능 평가 결과와 함께 차의 종류 및 등급 예측 판단을 진행하였다. 예측값과 실제값의 상관성은 각각 대불용정 0.9056, 재스민차 0.8855, 운남홍차 0.9527, 기문 홍차 0.9710으로, 재스민차의 값이 약간 낮은 외에 모두 0.9 이상의 의미있는 결과를 보였다. GC-MS와 화학계량학 분석 으로 차의 등급과 품질 판단 연구를 진행하고, 차의 종류 식별 함수와 등급 예측 모형 수립은 차의 종류와 등급 판별에 참고 가치가 있다는 결론을 내었다.

The change in volatile compound composition of three wild chrysanthemum species (Chrysanthemum boreale, C. indicum, and C. indicum var. albescens) was identified and analyzed according to four flowering stages using HS-SPME-GC-MS (headspace solid-phase microextraction coupled to gas chromatography–mass spectrometry). The top five compounds of each flowering stage were selected because those main compounds accounted for 43.25%, 44.14%, and 54.20% of the total relative content of volatile compounds from C. boreale, C. indicum, and C. indicum var. albescens, respectively. Nine compounds (1S-α-pinene, α-thujone, chrysanthenone, umbellulone, thymol, caryophyllene, germacrene D, α-zingiberene, and α-patchoulene) in C. boreale were ranked in the top five compounds through the whole flowering stages. In C. indicum, camphene, eucalyptol, camphor, umbellulone, bornyl acetate, caryophyllene, β-farnesene, germacrene D, and α-zingiberene were ranked in the top five compounds. However, only five compounds (camphor, bornyl acetate, β-farnesene, germacrene D, and α-zingiberene) were ranked in C. indicum var. albescens showing a more stable composition rather than C. boreale and C. indicum. Flowerheads of three wild Chrysanthemums showed a different profile of volatile compounds according to different flowering stages, varying compositions, and relative content in the top five volatile compounds. This study illustrates how main volatile compounds in wild chrysanthemums change dynamically during the flowering regarding compositions and their relative contents, suggesting that it should provide a useful index for harvesting or blending certain target compounds from wild chrysanthemums.

In this study, the calibration characteristics of six new odorant VOCs along with two reference compound were evaluated based on GC-MS (SIM). For the proper evaluation of the GC/MS method, calibration of 7 VOCs were made both by the SPME and direct injection (DI). The minimum detection limit (MDL) of those VOC determined by SPME method fell in the range of 0.31∼0.38 ppb while those of DI from 6.18∼14.3 ppb. The analytical performance of our SPME/GC-MS method was compared against those tested previously by other study groups. Although more efforts are needed to establish the reliability of our SPME/GC-MS method, the method adapted in this study can be recommended as one of the standard methods for the analysis of the target VOCs in this study to settle formal experiment methods.

참나물의 잎과 줄기부위에서 가장 많이 함유하고 있는 무기 성분은 K, P, Ca, Mg 순이었으며 잎 부위가 줄기부위보다 Ca, P, Mg함유량이 약 4배 정도 많았다 전자코를 이용한 휘발성 향기성분 패턴은 신선한 참나물의 경우 제1주성분 값이 +값을, 음건한 건조 참나물은 - 값을 나타내어서 신선한 참나물과 건조한 참나물 사이에는 뚜렷한 차이를 보였고 건조방법에 따른 시료간의 구별이 가능하였다. SDE법에 의해 신선한 참나물은 aldehydes 3종, alcohols 9종, ester 4종, hydrocarbons 5종, terpen hydrocarbons 34종, ketone 1종, 기타 2종의 총 58 종이 확인되었고, 음건한 참나물은 aldehydes 4종, alcohols 7종, hydrocarbon 1종, terpen hydrocarbons 17종, ketone 1종, 기타 1종의 총 31종이 확인되었다. SDE법에 의한 신선, 음건한 참나물 모두 α-selinene(37.89%, 12.59%)가 가장 많이 확인되었는데 신선할 때보다 음건한 경우 휘발성향기성분의 peak수와 peak area%가 적었다. CAR/PDMS fiber HS-SPME법에 의해 34종이 확인되었는데 aldehydes 2종, alcohols 2종, hydrocarbons 7종, terpen hydrocarbons 23종이며 myrcene(15.50%)가 가장 많이 확인되었다. PDMS fiber HS-SPME법에 의해 aldehydes 1종, alcohols 1종, hydrocarbons 2종, terpen hydrocarbons 17종으로 총 21종이 확인되었고 germacrene D(16.84%)가 가장 많았다. SDE법에 의한 경우가 SPME법보다 향기성분의 종류와 양이 많았고 HS-SPME법의 경우 CAR/PDMS fiber가 PDMS fiber 보다 더 많은 종류의 향기가 확인되었다.