정공등의 기원식물 정공등(丁公藤) Erycibe obtusifolia Bentham을 추출하여, 극성에 따라 chloroform, ethyl acetate, butanol, water로 분획하여, 이 중 butanol와 water분획을 사용하여 성분분리를 실시하였다. Butanol와 water분획을 sephadex LH-20과 MCI를 이용한 gel chromatography를 통해 총 4개의 화합물을 얻었다. 얻어진 화합물은 물리화학적 성상과 문헌조사, 또는 각종 기기분석(1H-NMR, 13C-NMR, Mass)을 통하여 구조동정을 실시하였고 그 결과 compoundⅠ 은 (1R,3S,4R,5S)-4-((E)-3-(3,4-dihydroxyphenyl)acryloyloxy)-1,3,5-trihydroxycyclohexanecarboxylic acid 인 crypto-chlorogenic acid (C16H18O9)로, compound Ⅱ는 7-(beta-D-glucopyranosoyloxy)-6-methoxy-2H-1-benzopyran-2-one 인 scopolin (C16H18O9)으로, compound Ⅲ은 (1R,3S,4R,5S)-5-((E)-3-(3,4-dihydroxyphenyl)acryloyloxy)-1,3,4-trihydroxycyclohexanecarboxylic acid 인 neochlorogenic acid(C16H18O9)으로, compound Ⅳ는 3,4-di-O-caffeoylquinic acid 로 확인 동정 하였다. 분리된 Compound Ⅱ, Ⅲ, Ⅳ와 표준품 scopoletin(Ⅴ) 총 5개의 화합물을 지표물질로 하여 정공등의 HPLC를 통한 분석법 개발 및 함량 평가를 실시하였다. HPLC분석을 위하여 AKZO NOBEL Kromasil 100-5 C18 (250×4.6 mm, 5 μm) column을 사용하였고 이동상으로는 0.5% acetic acid와 메탄올을 85 : 15비율에서 30분간 50 : 50으로 변화시겨 사용하였으며, UV 330 nm파장에서 확인하였다. HPLC/UV를 이용하여 국내 및 중국에서 유통되고 있는 정공등 20종에 대한 함량평가를 실시한 결과 전체 시료에서 scopolin 0.25 ± 0.062%, scopoletin 0.2 ± 0.068 %, neochlorogenic acid 0.102 ± 0.062%, 3,4-di-O-caffeoylquinic acid 0.073 ± 0.042 %를 함유하고 있다는 것을 확인할 수 있었다. 또한 HPLC를 이용한 함량평가 결과를 이용한 다변량분석 (PCA)을 통하여 종간 유사성 실험을 실시하였으며 그 결과 지역별로 시판되고 있는 정공등 유통품은 함유된 성분이 비슷하지만 각 성분함량에서 큰 차이를 보였으며, 이는 검체에 성분함량 차이에서 나타나는 것으로 확인되었으며, 또한 재배 지역별 차이를 나타내는 것으로 볼 수 있었다.
Erycibe obtusifolia Benth. (Convolvulaceae) was distributed in the southern China. The genus Erycibe included 11species in China. Its roots, stems and twigs were used in Chinese folk medicine to relieve the symptoms of rheumatoid arthritis. Pharmacological study indicated that Erycibe obtusifolia showed a strong anti-inflammatory activity and the effect of muscarinic agonist. The dried roots of Erycibe obtusifolia Benth. were exhaustively extracted with methanol and concentrated under reduced pressure. Then the residue was suspended in H2O and extracted successively with chloroform, ethyl acetate and butanol. Four compounds were isolated from the butanol and water portions. Their structures were elucidated by spectroscopic methods including UV, 1H-NMR, 13C-NMR, FAB-Mass and ESI-Mass as (1R,3S,4R,5S)-4-((E)-3-(3,4-dihydroxyphenyl)acryloyloxy)-1,3,5-trihydroxycyclohexanecarboxylic acid(1, crypto-chlorogenic acid), 7-(beta-D-glucopyranosoyloxy)-6-methoxy-2H-1-benzopyran-2-one(2, scopolin),(1R,3S,4R,5S)-5-((E)-3-(3,4-dihydroxyphenyl)acryloyloxy)-1,3,4-trihydroxycyclohexanecarboxylic acid(3, neochlorogenic acid) and 3,4-di-O-caffeoylquinic acid(4).CompoundⅡ, Ⅲ, Ⅳ and scopoletin were decided as representative components of Erycibe obtusifolia Benth. We established HPLC analytical method by using the representative components and 20 commercial samples which were collected considering to various cultivated area. The HPLC fingerprinting was successfully achieved with an AKZO NOBEL Kromasil 100-5C18 (250×4.6 mm, 5 μm) column. The mobile phase consisted of 0.5% acetic acid in water(A) and methanol(B) using gradient method of 85(A) to 50(A) for 35min. The fingerprints of chromatograms were recorded at an optimized wavelength of 330 nm. This developed analytical method was validated with specificity, selectivity, accuracy and precision. And it is suggested that scopolin, scopoletin, neochlorogenic acid, 3,4-di-O-caffeoylquinic acid were more than 0.162%, 0.133 %, 0.057%, 0.044 %, respectively. In addition, principal component analysis (PCA) was performed on the analytical data of 20 different Erycibe obtusifolia samples in order to classify samples collected from different regions. We hope that this assay can be readily utilized as quality control method for Erycibe obtusifolia.