Name: 저자 스포트라이트: Rhodiola crenulata 에 대한 식별 방법 개발 및 자원 분포 및 약리학적 효과 조사
Uploaded: 2023-10-27T13:20:00
Description: Watch this Scientific Journal Video about Field Collection and Laboratory Routine Identification of Rhodiola crenulata at JoVE.com

이 연구는 중국 국립 자연 과학 재단(81973569, 82274207 및 82104533), 청두 중의학 대학의 Xinglin Scholar Research Promotion Project(XKTD2022013) 및 Ningxia의 주요 연구 개발 프로그램(2023BEG02012)의 지원을 받았습니다.

홍경 천은 중국 쓰촨성 간쯔 티베트족 자치현 간쯔현 주오다 설산(N 31.44570°, E 99.96086°, 4892m)에서 채취됩니다. 이 식물은 청두 전통 중국 의학 대학 민족 의학 학교의 Yi Zhang 교수에 의해 진품으로 인증되었습니다.
1. 홍경천 채취
<ol><li>Rhodiola crenulata의 서식지 지도를 촬영하십시오.</li>
	<li>Rhodiola crenulata의 전체 식물, 잎, 꽃받침 및 뿌리...

Rhodiola crenulate의 현미경 특성 분석 및 TLC 기반 식별

<ol>
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K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Demystifying+traditional+herbal+medicine+with+modern+approach.">Demystifying traditional herbal medicine with modern approach.</a> Nature Plants. 3, 17109 (2017).</li><li>da Fonseca, L. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Herbal+medicinal+products+from+Passiflora+for+anxiety:+An+unexploited+potential.">Herbal medicinal products from Passiflora for anxiety: An unexploited potential.</a> The Scientific World Journal. 2020, 6598434 (2020).</li><li>Aziato, L., Antwi, H. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Facilitators+and+barriers+of+herbal+medicine+use+in+Accra,+Ghana:+an+inductive+exploratory+study.">Facilitators and barriers of herbal medicine use in Accra, Ghana: an inductive exploratory study.</a> BMC Complementary and Alternative Medicine. 16, 142 (2016).</li><li>Tao, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rhodiola+species:+A+comprehensive+review+of+traditional+use,+phytochemistry,+pharmacology,+toxicity,+and+clinical+study.">Rhodiola species: A comprehensive review of traditional use, phytochemistry, pharmacology, toxicity, and clinical study.</a> Medicinal Research Reviews. 39 (5), 1779-1850 (2019).</li><li>Wang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+dietary+supplement+Rhodiola+crenulata+extract+alleviates+dextran+sulfate+sodium-induced+colitis+in+mice+through+anti-inflammation,+mediating+gut+barrier+integrity+and+reshaping+the+gut+microbiome.">The dietary supplement Rhodiola crenulata extract alleviates dextran sulfate sodium-induced colitis in mice through anti-inflammation, mediating gut barrier integrity and reshaping the gut microbiome.</a> Food &amp; Function. 12 (7), 3142-3158 (2021).</li><li>Xie, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rhodiola+crenulate+alleviates+hypobaric+hypoxia-induced+brain+injury+via+adjusting+NF-&#954;B/NLRP3-mediated+inflammation.">Rhodiola crenulate alleviates hypobaric hypoxia-induced brain injury via adjusting NF-&#954;B/NLRP3-mediated inflammation.</a> Phytomedicine. 103, 154240 (2022).</li><li>Wang, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Salidroside,+a+phenyl+ethanol+glycoside+from+Rhodiola+crenulata,+orchestrates+hypoxic+mitochondrial+dynamics+homeostasis+by+stimulating+Sirt1/p53/Drp1+signaling.">Salidroside, a phenyl ethanol glycoside from Rhodiola crenulata, orchestrates hypoxic mitochondrial dynamics homeostasis by stimulating Sirt1/p53/Drp1 signaling.</a> Journal of Ethnopharmacology. 293, 115278 (2022).</li><li>Ma, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+characteristics+of+Rhodiola+Crenulata+and+its+mechanism+in+acute+mountain+sickness+using+UHPLC-Q-TOF-MS/MS+combined+with+network+pharmacology+analysis.">Chemical characteristics of Rhodiola Crenulata and its mechanism in acute mountain sickness using UHPLC-Q-TOF-MS/MS combined with network pharmacology analysis.</a> Journal of Ethnopharmacology. 294, 115345 (2022).</li><li>Wang, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rhodiola+crenulata+attenuates+apoptosis+and+mitochondrial+energy+metabolism+disorder+in+rats+with+hypobaric+hypoxia-induced+brain+injury+by+regulating+the+HIF-1&#945;/microRNA+210/ISCU1/2(COX10)+signaling+pathway.">Rhodiola crenulata attenuates apoptosis and mitochondrial energy metabolism disorder in rats with hypobaric hypoxia-induced brain injury by regulating the HIF-1&#945;/microRNA 210/ISCU1/2(COX10) signaling pathway.</a> Journal of Ethnopharmacology. 241, 111801 (2019).</li><li>d'Avigdor, E., Wohlmuth, H., Asfaw, Z., Awas, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+current+status+of+knowledge+of+herbal+medicine+and+medicinal+plants+in+Fiche,+Ethiopia.">The current status of knowledge of herbal medicine and medicinal plants in Fiche, Ethiopia.</a> Journal of Ethnobiology and Ethnomedicine. 10, 38 (2014).</li><li>S&#225;nchez, M., Gonz&#225;lez-Burgos, E., Divakar, P. 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B. 6 (6), 477-482 (2005).</li><li>Dong, X., Guo, Y., Xiong, C., Sun, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+two+major+Rhodiola+species+and+the+systemic+changing+characteristics+of+metabolites+of+Rhodiola+crenulata+in+different+altitudes+by+chemical+methods+combined+with+UPLC-QqQ-MS-based+metabolomics.">Evaluation of two major Rhodiola species and the systemic changing characteristics of metabolites of Rhodiola crenulata in different altitudes by chemical methods combined with UPLC-QqQ-MS-based metabolomics.</a> Molecules. 25 (18), 4062 (2020).</li><li>Zhao, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HPLC+fingerprint+differentiation+between+natural+and+ex+situ+populations+of+Rhodiola+sachalinensis+from+Changbai+Mountain,+China.">HPLC fingerprint differentiation between natural and ex situ populations of Rhodiola sachalinensis from Changbai Mountain, China.</a> PloS One. 9 (11), 112869 (2014).</li><li>Yu, Y. 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세계에는 90 종 이상의 홍경천 식물이 있으며 모든 종의 60 % 이상이 중국에서 발견되며 홍경천, 홍경천, 홍경천 achalinensis 및 홍경천 17을 포함한 일반적인 식물이 있습니다. 중국 약전(2020)의 첫 번째 부분에 기록된 홍경천은 높은 고도에서 재배되는 티베트 전통 약재입니다. 홍경천에 대한 시장 수요는 매년 증가하고 있으므로 공급원의 올바른 ?...

약초는 중국에서 오랜 역사와 풍부한 응용 경험을 가지고 있으며 Shennong의 약초 고전1에서 최초의 체계적인 기록이었습니다. 말라리아에 적용된 아르테미시닌의 발견은 약초의 새로운 단계1로의 발전을 촉진했습니다. 현대 과학 기술을 사용하여 약초의 정확한 메커니즘을 밝혀냄으로써 약초의 이용률과 수요가 증가하여 약초에 대한 새로운 국제 시장이 열렸습니다 2,3,4. 그러나 이것은 일련의 부정적인 영향으로 이어졌습니다. 비전문가는 한약의 특성에 대해 막연하게 이해하고 있어 한약의 사용이 큰 안전 위험에 직면하게 됩니다5.
홍경천의 식물 중 하나인 홍경천은 주로 티베트, 윈난 북서부, 중국 쓰촨성 서부에 분포한다(그림 1)6,7. 홍경천은 살리드로사이드, 티로졸, 갈산 및 기타 화합물로 구성되어 있으며, "기를 활성화하고 혈액 순환을 촉진하며 맥박을 맑게 하고 천식을 진정시키는 기능"8,9,10,11 기능을 통해 저산소증 관련 질병을 치료합니다. 현장 조사에 따르면 홍경천은 고도 4,000-5,600m의 고산 거골 지대, 협곡 경사면 및 암석 틈새에서 발견 될 수 있습니다. 성장 환경은 춥고 햇빛과 강렬한 방사선으로 가득 차 있으며 고산 초원 생태계에 속합니다. Rhodiola crenulata는 성장 지형에 따라 라멜라 및 점형 개체군에 분포할 수 있으며, 유전자 흐름은 교차 수분을 통해 수행될 수 있습니다.
홍경천 속의 꽃가루 낙태, 불법 발굴 및 퇴화 된 생태 환경은 홍경천 crenulata를 멸종 위기에 처한 종으로 만듭니다 6,12. Rhodiola crenulata의 높은 약용 가치를 고려할 때 위조 제품이 시장에 유입될 것으로 예상됩니다. 이 기사에서는 Rhodiola crenulata의 서식지와 몇 가지 편리한 실험실 식별 방법을 제시합니다. 첫째, 홍경천의 생육 환경과 약효를 관찰하였다. 둘째, 약용 분말의 미세 구조를 현미경으로 관찰하였다. 마지막 단계가 핵심입니다. Rhodiola crenulata의 대표 성분은 특정 물질에서 이들 성분의 상이한 흡착 또는 용해 특성에 따라 분리되고 확인되었습니다. 약용 식물의 DNA 기반 인증 또는 대사체학 분석 방법은 복잡하고 비용이 많이 듭니다13. 이러한 기본적이고 편리하며 경제적인 방법으로 홍경천을 신속하게 식별할 수 있습니다.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.22 &#956;m millipore filter</td>
			<td>Millipore</td>
			<td>SLGP033RB</td>
			<td></td>
		</tr>
		<tr>
			<td>Automatic sampling machine</td>
			<td>CAMAG</td>
			<td>ATS 4</td>
			<td></td>
		</tr>
		<tr>
			<td>Chloral hydrate</td>
			<td>Fuzhou Brunei Technology Co., Ltd</td>
			<td>ST1002</td>
			<td></td>
		</tr>
		<tr>
			<td>Chromatographic&#160;sample&#160;bottles</td>
			<td>Zhejiang ALWSCI Technology Co., Ltd</td>
			<td>C0000008</td>
			<td></td>
		</tr>
		<tr>
			<td>Conical flask</td>
			<td>Sichuan Shubo Co., Ltd</td>
			<td>1121</td>
			<td></td>
		</tr>
		<tr>
			<td>Cover glass</td>
			<td>Citotest Labware Manufacturing Co., Ltd</td>
			<td>10211818c</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting needle</td>
			<td>Shanghai Bingyu Fluid Technology Co., Ltd</td>
			<td>BY-5026</td>
			<td></td>
		</tr>
		<tr>
			<td>Electronic balance</td>
			<td>SHIMADZU</td>
			<td>ATX124</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethyl acetate</td>
			<td>Chengdu Kelong Chemical Co., Ltd</td>
			<td>2022120901</td>
			<td></td>
		</tr>
		<tr>
			<td>Formic acid</td>
			<td>Chengdu Kelong Chemical Co., Ltd</td>
			<td>2021110801</td>
			<td></td>
		</tr>
		<tr>
			<td>Gallic acid</td>
			<td>Chengdu Herbpurify Co., Ltd</td>
			<td>M-017</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycerol</td>
			<td>Sinopharm Chemical Reagent Co., Ltd</td>
			<td>10010618</td>
			<td></td>
		</tr>
		<tr>
			<td>High speed&#160; crusher</td>
			<td>Beijing Zhongxingweiye Instrument Co., Ltd</td>
			<td>FW-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Chengdu Kelong Chemical Co., Ltd</td>
			<td>20230108</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope</td>
			<td>Chongqing Oprec Nistrument Co.,&#160; Ltd</td>
			<td>B203</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope slide</td>
			<td>Citotest Labware Manufacturing Co., Ltd</td>
			<td>7105P-G</td>
			<td></td>
		</tr>
		<tr>
			<td>Oven</td>
			<td>Shanghai Yuejin Medical Equipment Co., Ltd</td>
			<td>DHG-8145</td>
			<td></td>
		</tr>
		<tr>
			<td>Pharmacopoeia sieve</td>
			<td>Hangzhou Xingrun sieve factory</td>
			<td>572423281330</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipette</td>
			<td>Changde BKMAM Biotechnology Co., Ltd</td>
			<td>120302008</td>
			<td></td>
		</tr>
		<tr>
			<td>Salidroside</td>
			<td>Chengdu Herbpurify Co., Ltd</td>
			<td>H-040</td>
			<td></td>
		</tr>
		<tr>
			<td>Saturate tank&#160;</td>
			<td>Yancheng Liegu Technology Co., Ltd</td>
			<td>10*20 P-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Silica gel plate</td>
			<td>Yantai Jiangyou Silica Gel Development&#160; Co., Ltd</td>
			<td>HSG20211227</td>
			<td></td>
		</tr>
		<tr>
			<td>Trichloromethane</td>
			<td>Chengdu Kelong Chemical Co., Ltd</td>
			<td>20221013-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Tweezer&#160;</td>
			<td>Changde BKMAM Biotechnology Co., Ltd</td>
			<td>130302027</td>
			<td></td>
		</tr>
		<tr>
			<td>Tyrosol</td>
			<td>Chengdu Herbpurify Co., Ltd</td>
			<td>L-042</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound equipment</td>
			<td>Ningbo Xinyi Ultrasonic Equipment Co., Ltd</td>
			<td>SB-8200DTS</td>
			<td></td>
		</tr>
		<tr>
			<td>Volumetric pipet</td>
			<td>Changde BKMAM Biotechnology Co., Ltd</td>
			<td>120301006</td>
			<td></td>
		</tr>
	</tbody>
</table>

field collection and laboratory routine identification of rhodiola crenulata

의약품 성분의 식별은 의약품 안전성의 전제이자 보증입니다. 과학 연구자들의 대다수는 약초의 간단하고, 빠르고, 효과적이고, 저렴한 식별 과정을 선호할 수밖에 없습니다. 홍경천은 높은 고도에서 자라는 티베트 전통 약재로 주로 중국의 티베트, 윈난, 쓰촨 지역에 분포합니다. 홍경천은 항염증, 항저산소증, 항산화 등 여러 생체 활성을 가지고 있으며 발전 가능성이 큽니다. 시장 수요가 증가하고 자원 함량이 급격히 감소함에 따라 홍경천 crenulata 의 많은 혼란스러운 제품이 사람들을 괴롭히고 있습니다. 따라서 이 프로토콜은 일상적인 실험실 테스트와 결합된 현장에서 Rhodiola crenulata 를 식별하기 위한 표준 프로세스를 도입합니다. 서식지, 미세한 특징 및 박층 크로마토그래피의 조합은 의심할 여지 없이 홍경 천을 빠르고 효율적이며 경제적으로 식별하여 티베트 의학의 지속적인 발전과 의약 재료의 품질 관리에 기여할 것입니다.

Herbal medicine has a long history and rich application experience in China, and it was the first systematic recording in Shennong&#39;s herbal classic1. The discovery of artemisinin applied to malaria promoted the development of herbal medicine into a new stage1. The use of modern scientific technology to uncover the exact mechanism of herbal medicine increases the utilization rate and demand for herbal medicine, opening a new international market for it2,3,4. However, this has led to a series of negative effects. Non-professionals have a vague understanding of the characteristics of herbal medicine, which makes the use of herbal medicine face a huge safety risk5.
Rhodiola crenulata, one of the plants of the Rhodiola species, is mainly distributed in Tibet, northwest Yunnan, and western Sichuan of China (Figure 1)6,7. Rhodiola crenulata comprises salidroside, tyrosol, gallic acid, and other compounds for the treatment of hypoxic-related diseases through the function of &#34;invigorating qi and promoting blood circulation, clearing pulse and calming asthma&#34;8,9,10,11. Field investigation shows that Rhodiola crenulata can be found in the alpine talus zones, gully slopes, and rock crevices at an altitude of 4,000-5,600 m. Its growing environment is cold, full of sunshine and intense radiation, and it belongs to the alpine meadow ecosystem. Rhodiola crenulata can be distributed in lamellar and point-like populations according to the growth terrain, and gene flow can be carried out through cross-pollination.
The pollen abortion of the genus Rhodiola, illegality excavation, and degenerated ecological environment make Rhodiola crenulata an endangered species6,12. In view of the high medicinal value of Rhodiola crenulata, counterfeit products are expected to flow into the market. This article presents the habitat of Rhodiola crenulata and some convenient laboratory identification methods. Firstly, we observed the growth environment of Rhodiola crenulata and its medicinal properties. Secondly, the microstructure of medicinal powder was observed by microscope. The last step is the key point. The representative components of Rhodiola crenulata were separated and identified according to the different adsorption or dissolution properties of these components in a certain substance. DNA-based authentication or metabolomics analysis methods of medicinal plants are complicated and expensive13. These basic, convenient, and economical methods can quickly identify Rhodiola crenulata.

저자 스포트라이트: Rhodiola crenulata 에 대한 식별 방법 개발 및 자원 분포 및 약리학적 효과 조사

Rhodiola crenulata의 현장 수집 및 실험실 일상적인 식별

The scope of our research is centered on conducting a resource survey of ethnic medicine. Specifically, this study aims to develop a rapid and convenient methods for the identification of Rhodiola crenulata. The protocol we offer presents a cost-effective, straightforward, and rapid method for identifying various medicinal herbal materials.Our laboratory will concentrate its future research endeavors on investigation two primary areas, the distribution of research associated with Rhodiola crenulata and the pharmacological impacts of this species.

이 실험 프로토콜은 현장에서 Rhodiola crenulate 를 식별하고 수집하는 방법을 설명합니다. Rhodiola crenulate 는 고산 거골 지역, 협곡 경사면 및 높은 고도의 암석 틈새에 서식하는 경향이 있습니다. Rhodiola crenulate 의 서식지, 전체 식물, 꽃 및 잎은 그림 2에 나와 있습니다. Rhodiola crenulate 는 적갈색의 뿌리줄기를 가지고 있습니다(그림 3A</str...

Watch this Scientific Journal Video about Field Collection and Laboratory Routine Identification of Rhodiola crenulata at JoVE.com

여기에서는 서식지, 식물 형태, 의약 특성, 미세한 특징 및 박층 크로마토그래피에서 Rhodiola Crenulata 의 식별에 대해 설명합니다.

The identification of medicinal materials is the premise and guarantee of drug safety. The majority of scientific researchers are bound to favor the ...

본 연구의 범위는 민족 의학에 대한 자원 조사를 수행하는 데 중점을 두고 있습니다. 특히, 본 연구는 홍경천의 식별을 위한 빠르고 편리한 방법을 개발하는 것을 목표로 한다. 우리가 제공하는 프로토콜은 다양한 약초 재료를 식별하기 위한 비용 효율적이고 간단하며 신속한 방법을 제시합니다.우리 실험실은 두 가지 주요 영역, 즉 홍경천과 관련된 연구의 분포 및 이 종의 약리학적 영향을 조사하는 데 향후 연구 노력을 집중할 것입니다.

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Research

JoVE Journal

Medicine

Field Collection and Laboratory Routine Identification of Rhodiola crenulata

The identification of medicinal materials is the premise and guarantee of drug safety. The majority of scientific researchers are bound to favor the simple, fast, effective, and inexpensive identification process of herbals. Rhodiola crenulata is a traditional Tibetan medicine grown at high altitudes, mainly distributed in Tibet, Yunnan, and Sichuan regions of China. Rhodiola crenulate possesses multiple bioactivities, such as anti-inflammatory, anti-hypoxia, and antioxidant properties, and has great potential for development. With the increasing market demand and a rapid decrease in resource content, a large number of confused products of Rhodiola crenulata have been troubling people. Therefore, this protocol introduces a standard process for the identification of Rhodiola crenulata in the field combined with routine laboratory testing. The combination of habitat, microscopic features, and thin-layer chromatography will undoubtedly identify Rhodiola crenulata quickly, efficiently, and economically, contributing to the continuous development of Tibetan medicine and the quality control of medicinal materials.

Here, we describe the identification of Rhodiola Crenulata from habitat, plant morphology, medicinal properties, microscopic features, and thin-layer chromatography.

연구

의학

Macroscopic and Microscopic Identification of Rhodiola crenulata Traits and Constituents

To begin, observe the appearance traits of Rhodiola Crenulata with the unaided eye and record the observations. Inhale near the plant to identify the fragrance smell. Next, place a small piece of the root in the mouth, then sip and chew for taste identification.Using a brush, remove the soil on the surface of the plant. Place the plant in an oven at 40 degrees Celsius and turn it every 24 hours. Then powder the dried medicinal materials using a powder machine and filter it through a medicated number three sieve.Spread the powder evenly on a clean slide using a dissecting needle covering 1/3 of the slide within a two millimeter area. Using a glass dropper, add a drop of deionized water to the powder and quickly place a cover glass using a tweezer. Next, place the slide onto the microscope platform.Adjust the light source and the coarse focus spiral to view the powder. Switch to a 40X objective lens and observe the starch granules. Again, spread the powder evenly on a slide using a dissecting needle, then add a drop of chloral hydrate to the powder.Hold the slide with a tweezer and heat on the alcohol lamp thrice for one second each. Add a drop of glycerin to the powder and quickly place a cover glass onto the drop. After adjusting the microscope settings as demonstrated previously, select a 40X objective lens to visualize the catheter, cork cells and fibers, along with the wood parenchyma cells and pigment block respectively.Microscopic examination of our Crenulata revealed key features with cork cells appearing brownish yellow, or colorless with polygonal shapes. Starch grains were observed as single or multiples with distinctive umbilical points and closely arranged spiral vessels. Additionally, xylem parenchyma cells were visualized as achromatic and oval, containing calcium oxalate sand crystals, and irregularly shaped brown red pigment blocks were identified.

Rhodiola crenulata 형질 및 성분의 거시적 및 미시적 식별

Preparing and Analyzing Thin Layer Chromatography Samples of Rhodiola crenulata and Reference Compounds

To begin, weigh three grams of Rhodiola crenulata powder and transfer it into a 100-milliliter conical bottle. Using a large bowl pipette, add 25 milliliters of methanol into the bottle. Place the bottle in the ultrasonic instrument.Set the power to 250 watts, frequency to 40 kilohertz, time to 30 minutes, and start the ultrasonication. Next, rinse the outside of the bottle under running water until it reaches room temperature. Filter the solution through a 0.22-micron microporus membrane.Using a one-milliliter syringe, aspirate 800 microliters of liquid from the bottle and collect 400 microliters of the midstream solution into a chromatographic sample bottle. Measure and add one milligram of salidroside, 0.5 milligrams of tyrosol, and 0.5 milligrams of gallic acid into three separate one-milliliter volumetric flasks. Then, fill each flask to the scale marked with methanol.Sonicate the samples in the ultrasonic instrument using the conditions described previously. Following ultrasonication, aspirate 800 microliters of liquid from the flask. Filter the solution through a 0.22-micrometer microporous membrane and collect 400 microliters of filtrate into the bottle.Add trichloromethane, ethyl acetate, methanol, and formic acid on one side of the double tank chromatography cylinder. Shake the cylinder to mix the contents evenly and cover the upper cylinder head. Arrange the gallic acid, salidroside, tyrosol standard solutions, and our crenulata solution on the sample rack in positions A1 to A4.Place a 5x10-centimeter silicone sheet on the sampling table.Turn on the automatic sampling machine and open the air control valve. Press the Continue button for automatic sampling. After sampling, turn off the machine and air valve.Remove the silicone sheet from the machine. Put the silicone sheet into the other side of the double tank chromatography cylinder. Cover the upper cylinder head and pre-saturate for 20 minutes.Next, use a tweezer to hold the upper end of the thin layer plate and quickly put the silicone sheet into the development agent. Once the organic solvent evaporates, spray the silicone sheet with chromogenic solution. The thin layer chromatographic analysis showed that the R.crenulata sample appeared in spots of the same color, corresponding to the gallic acid, salidroside, and tyrosol standards.