Objectives : The root ofCyanchum wifordii(CW) had been used as herbal medicine, 'Baeksuo', in Korea. However, the root ofCynanchum auriculatum(CA) had also been mis-used as 'Baeksuo' in herbal markets, due to its morphological similarity with CW. This study aimed to compare discriminative features a...
Objectives : The root ofCyanchum wifordii(CW) had been used as herbal medicine, 'Baeksuo', in Korea. However, the root ofCynanchum auriculatum(CA) had also been mis-used as 'Baeksuo' in herbal markets, due to its morphological similarity with CW. This study aimed to compare discriminative features and establish the identification-keys between two herbal medicines using microscopic methods.Methods : Microscopic preparations including fixation, rapid dehydration, paraffin infiltration, paraffin embedding, sectioning, mounting, and staining, were performed and the microscopic features between CW and CA samples were compared.Results : The starch grain of parenchymal cells, the amounts of calcium oxalate crystals, cell sizes, and the shape of phloem were not different between CW and CA samples. However, the clusters of stone cells which were conglomerated at outer cortex were apparently observed in CA sample (4~8 layers of stone cells), but not in CW sample (2~4 layers of stone cells, or not). Xylem of CW consisted of several vessels and fiber bundles which were formed as linear shapes and discontinuous radial lines, while that of CA, in which radial lines was not observed, were shaped as a wedge and scattered near cambium with comparatively few fiber bundles enclosed vessels. Especially, in the xylem of CA, fiber bundles located towards cambium were relatively more developed than those located outward cambium.Conclusions : The identification-keys established in this study would be helpful to discriminate microscopic features between CW and CA.
Objectives : The root ofCyanchum wifordii(CW) had been used as herbal medicine, 'Baeksuo', in Korea. However, the root ofCynanchum auriculatum(CA) had also been mis-used as 'Baeksuo' in herbal markets, due to its morphological similarity with CW. This study aimed to compare discriminative features and establish the identification-keys between two herbal medicines using microscopic methods.Methods : Microscopic preparations including fixation, rapid dehydration, paraffin infiltration, paraffin embedding, sectioning, mounting, and staining, were performed and the microscopic features between CW and CA samples were compared.Results : The starch grain of parenchymal cells, the amounts of calcium oxalate crystals, cell sizes, and the shape of phloem were not different between CW and CA samples. However, the clusters of stone cells which were conglomerated at outer cortex were apparently observed in CA sample (4~8 layers of stone cells), but not in CW sample (2~4 layers of stone cells, or not). Xylem of CW consisted of several vessels and fiber bundles which were formed as linear shapes and discontinuous radial lines, while that of CA, in which radial lines was not observed, were shaped as a wedge and scattered near cambium with comparatively few fiber bundles enclosed vessels. Especially, in the xylem of CA, fiber bundles located towards cambium were relatively more developed than those located outward cambium.Conclusions : The identification-keys established in this study would be helpful to discriminate microscopic features between CW and CA.
이에 본 연구는 관련분야 전문가가 아닌 경우 정확한 감별이 어려운 飮片의 외부형태감별과 시간과 비용의 소모가 비교적 많은 유전자감별 이외에 현장에서 비교적 쉽게 적용할 수 있는 내부형태감별을 위한 기준을 제시하고자 한다.
제안 방법
이상 기술된 고정부터 염색까지의 전 과정은 주 등10)이 기술한 방법으로 진행하였다. 결과에 제시된 도해는 각 시료의 특징을 잘 나타내는 프리파레트를 1개씩 취하여 Adobe photoshop CS6 및 photoscape로 밝기와 명암만 조정한 후 조합하여 편집하였다.
대상 데이터
또한 이 연구가 종료된 후 한국한의학연구원에서 자체 개발한 유전자마커를 통해 검증하였음을 표본관측으로부터 확인받았다. 당 연구에 사용한 시료는 현재 한국한의학연구원과 원광대학교 한의과대학 본초학교실에서 표본으로 보관 중이다(Table 1).
한국한의학연구원 한약표준표본관(이하 표본관)에서 기존의 연구8,9)에 따라 1차 동정을 완료한 白首烏 (CW)와 耳葉牛皮消 (CA) 시료를 분양받아 우석대학교 한의과대학 본초학교실에서 제시한 외부형태감별기준9)으로 검색표 고안자가 직접 2차 동정하였다. 또한 이 연구가 종료된 후 한국한의학연구원에서 자체 개발한 유전자마커를 통해 검증하였음을 표본관측으로부터 확인받았다.
이론/모형
각 시료에서 무작위로 5개의 飮片을 취하여 물에 충분히 불린 후 Formalin-acetic acid-alcohol로 고정하고 Rapid dehydration法으로 탈수한 것을 Paraffin/TBA method에 따라 포매하고 절삭한 후 Safranin O, Haemalum, FastGreen FCF가 포함된 Ju's triple statining method로 염색하였다. 프리파레트를 현미경관찰 후 촬영(Olympus CX31 with Dicovery C15, Japan)하였다.
성능/효과
연구결과 연구대상인 白首烏(CW)와 耳葉牛皮消(CA)의 내부형태는 柔細胞의 澱粉粒이나 草酸鈣晶의 함유, 각 細胞의 크기, 篩部의 형태적 특징 등에서 형태상의 차이는 없는 것으로 사료된다.
후속연구
이상의 내용을 종합하여 白首烏와 耳葉牛皮消의 내부형태 감별 검색표를 작성하면 아래와 같다. 또한, 이 검색표를 기존의 기원식물 및 외부형태 감별기준8,9)과 병용한다면 두 약물의 판별에 더욱 용이하리라 사료된다.
그러나 기존의 감별법은 시간과 비용의 소요가 많아 1차 검증의 수단으로 용이하지 않는 단점이 있다. 이러한 정황을 고려할 때 비교적 단시간 저비용의 내부형태감별법과 같은 1차 검증법이 보급된다면 유통사용상의 혼란을 배제하기에 용이하겠다는 판단으로 본 연구를 수행하였다.
이상의 감별요점은 일선 현장에서 白首烏와 耳葉牛皮消를 판별하는데 활용할 수 있을 것으로 사료되며 추후 戟葉牛皮消 등에 대한 추가연구가 이루어져야 한다고 생각한다.
참고문헌 (15)
Liu H, Ma Z (974, Song period). Gaeboboncho[開寶本草]. In : Tang SW (宋, 1082, Song period). Jungsujeonghwagyeongsajeungnyubigeupboncho[重修政和經史證類備急本草], photographic ed.. Beijing : People's Medical Publishing House. 1996 : 262.
Su S (1062, Beisong period). Dogyeongboncho[圖經本草]. In : Li SZ (1590, Ming period). Bonchogangmok [本草綱目], revised ed.. Beijing : People's Medical Publishing House. 1996 : 1288.
Heo J (1596, Joseon period). Donguibogam[東醫寶鑑]. In : Sinjeungbo Daeyeok Donguibogam[New Enlarged Translation of Donguibogam], translated ed.. Seoul : Bubin publishers. 2012 : 2067-8.
Korea Food & Drug Administration. The Korean Herbal Pharmacopoeia. The KFDA Notification No. 2014-173. 2014 Oct 17th.
Pharmacopoeia Committee of the DPRK. Pharmacopoeia of Democratic People's Republic of Korea. 7th ed. Pyeongyang : Medicine and Science Press. 2011 : 552.
Pharmacopoeia Commission of the People's Republic of China. Pharmacopoeia of the People's Republic of China, 1977 ed.. Beijing : China Medical Science and Technology Press. 1977 : 178-9.
Kim CM et al. Coloured Illustrations for Discrimination of Herbal Medicine : Internal Morphology. Seoul : Academybook. 2014 : 78.
Lee GS. A study on the key of discrimination in Heshouwu Radix kinds. Woosuk University. 2009.
Ju YS. Abstain from use Cynanchi Auriculati Radix as Baiheshouwu. Akomnews. 2012 ; 1872 : 30.
Ju YS et al. Gaejeong UnGok Boncho-hak Silseup -seo [New Practice in Ungok Herbology]. Jeonju : HakYe-sa. 2009 : 33-53. Korean.
Akomnews. KFDA start cracking down fake Heshouwu. Akomnews [serial online] 2010 Mar 8 [cited 2015 June 15].. Available from : URL : http://www.akomnews.com/?p263569
Li Y, Piao D, Zhang H, Woo MH, Lee JH, Moon DC, Lee SH, Chang HW, Son JK. Quality assessment and discrimination of the roots of Cynanchum auriculatum and Cynanchum wilfordii by HPLC-UV analysis. Arch Pharm Res. 2013 ; 36(3) : 335-44.
Moon BC, Choo BK, Cheon MS, Yoon T, Ji Y, Kim BB, Lee AY, Kim HK. Rapid molecular authentication of three medicinal plant species, Cynanchum wilfordii, Cynanchum auriculatum, and Polygonum multiflorum (Fallopia multiflorum), by the development of RAPD-derived SCAR markers and multiplex-PCR. Plant Biotechnol Rep. 2010 ; 4(1) : 1-7.
Moon BC, Kim HK. Authentication of Cultivated and Commercial Herbal Materials of Cynanchi Wiflordii Radix Using Molecular Discrimination Methods. Korean herb Med Inf. 2014 ; 2(1) : 53-59.
Ryuk JA, Lee HW, Ju YS, Ko BS. Monitoring and identification of Cynanchum wilfordii and Cynanchum auriculatum by using molecular markers and real-time polymerase chain reaction. J Korean Soc Appl Biol Chem. 2014 ; 57(2) : 245-51.
AI-Helper
※ AI-Helper는 부적절한 답변을 할 수 있습니다.