Name: using multiple light scattering to examine the stability of phyllanthus emblica l. extracts obtained with different extraction methods
Uploaded: 2023-04-14T12:30:00
Description: Watch this Scientific Journal Video about Using Multiple Light Scattering to Examine the Stability of Phyllanthus emblica L. Extracts Obtained with Different Extraction Methods at JoVE.com

这项研究得到了中国国家自然科学基金（81973493）;国家中医药跨学科创新团队（ZYYCXTD-D-202209）;三金药业集团成都中医药大学产学研联合实验室项目（2019-YF04-00086-JH）;四川省科技计划资助项目（2021YFN0100）。笔者感谢成都中医药大学中医药创新研究院在质谱工作方面的技术支持。

1. 提取物制备
<ol><li>准确称取适量的PE，并加入10倍（重量）的去离子水进行回流提取。</li>
	<li>称重后，将五个样品用于回流提取0小时（E1），0.5小时（E2），1小时（E3），1.5小时（E4）和2小时（E5）。</li>
	<li>提取后，将样品冷却至室温，并称重以弥补损失的重量，以确保与提取前的重量一致。</li>
	<li>将样品以8,581× g 离心10分钟，以确保从样品溶液中去除不溶性物质和草药?...

<ol>
	<li>Huang, H. -. Z., 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=Exploration+on+the+approaches+of+diverse+sedimentations+in+polyphenol+solutions:+An+integrated+chain+of+evidence+based+on+the+physical+phase,+chemical+profile,+and+sediment+elements.">Exploration on the approaches of diverse sedimentations in polyphenol solutions: An integrated chain of evidence based on the physical phase, chemical profile, and sediment elements.</a> Frontiers in Pharmacology. 10, 1060 (2019).</li><li>Ran, F., 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=High+or+low+temperature+extraction,+which+is+more+conducive+to+Triphala+against+chronic+pharyngitis.">High or low temperature extraction, which is more conducive to Triphala against chronic pharyngitis.</a> Biomedicine and Pharmacotherapy. 140, 111787 (2021).</li><li>Wei, 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=Hepatoprotective+effects+of+different+extracts+from+Triphala+against+CCl(4)-induced+acute+liver+injury+in+mice.">Hepatoprotective effects of different extracts from Triphala against CCl(4)-induced acute liver injury in mice.</a> Frontiers in Pharmacology. 12, 664607 (2021).</li><li>Huang, H. Z., 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=Study+on+the+stability+control+strategy+of+Triphala+solution+based+on+the+balance+of+physical+stability+and+chemical+stabilities.">Study on the stability control strategy of Triphala solution based on the balance of physical stability and chemical stabilities.</a> Journal of Pharmaceutical and Biomedical Analysis. 158, 247-256 (2018).</li><li>Bhattacharya, A., Chatterjee, A., Ghosal, S., Bhattacharya, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antioxidant+activity+of+active+tannoid+principles+of+Emblica+officinalis+(amla).">Antioxidant activity of active tannoid principles of Emblica officinalis (amla).</a> Indian Journal of Experimental Biology. 37 (7), 676-680 (1999).</li><li>Chao, P. C., Hsu, C. C., Yin, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anti-inflammatory+and+anti-coagulatory+activities+of+caffeic+acid+and+ellagic+acid+in+cardiac+tissue+of+diabetic+mice.">Anti-inflammatory and anti-coagulatory activities of caffeic acid and ellagic acid in cardiac tissue of diabetic mice.</a> Nutrition and Metabolism. 6, 33 (2009).</li><li>Tiwari, V., Kuhad, A., Chopra, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emblica+officinalis+corrects+functional,+biochemical+and+molecular+deficits+in+experimental+diabetic+neuropathy+by+targeting+the+oxido-nitrosative+stress+mediated+inflammatory+cascade.">Emblica officinalis corrects functional, biochemical and molecular deficits in experimental diabetic neuropathy by targeting the oxido-nitrosative stress mediated inflammatory cascade.</a> Phytotherapy Research. 25 (10), 1527-1536 (2011).</li><li>Baliga, M. S., Dsouza, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amla+(Emblica+officinalis+Gaertn),+a+wonder+berry+in+the+treatment+and+prevention+of+cancer.">Amla (Emblica officinalis Gaertn), a wonder berry in the treatment and prevention of cancer.</a> European Journal of Cancer Prevention. 20 (3), 225-239 (2011).</li><li>Rehman, H. -. u., 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=Studies+on+the+chemical+constituents+of+Phyllanthus+emblica.">Studies on the chemical constituents of Phyllanthus emblica.</a> Natural Product Research. 21 (9), 775-781 (2007).</li><li>Jang, Y., Koh, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterisation+and+storage+stability+of+aronia+anthocyanins+encapsulated+with+combinations+of+maltodextrin+with+carboxymethyl+cellulose,+gum+Arabic,+and+xanthan+gum.">Characterisation and storage stability of aronia anthocyanins encapsulated with combinations of maltodextrin with carboxymethyl cellulose, gum Arabic, and xanthan gum.</a> Food Chemistry. 405, 135002 (2022).</li><li>Fu, 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=Novel+phenylalanine-modified+magnetic+ferroferric+oxide+nanoparticles+for+ciprofloxacin+removal+from+aqueous+solution).">Novel phenylalanine-modified magnetic ferroferric oxide nanoparticles for ciprofloxacin removal from aqueous solution).</a> Journal of Colloid and Interface Science. 632, 345-356 (2023).</li><li>Jiang, T., Charcosset, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Encapsulation+of+curcumin+within+oil-in-water+emulsions+prepared+by+premix+membrane+emulsification:+Impact+of+droplet+size+and+carrier+oil+on+the+chemical+stability+of+curcumin.">Encapsulation of curcumin within oil-in-water emulsions prepared by premix membrane emulsification: Impact of droplet size and carrier oil on the chemical stability of curcumin.</a> Food Research International. 157, 111475 (2022).</li></ol>

快速、准确地评估中药稳定性一直是中药研究的重点。本研究采用近红外无损技术分析了样品的稳定性和不稳定性机理，为指导提取工艺的改进提供更多有用的信息。
在该协议中，重要的稳定性参数是基于准确的MLS扫描数据计算的。MLS扫描可以实时采集样品的透射率（T%）和反向散射（BS%），并计算稳定性指数（SI）、粒径、粒子迁移速度和其他重要的物?...

An erratum was issued for:&#160;<a href="https://www.jove.com/t/65130">Using Multiple Light Scattering to Examine the Stability of Phyllanthus emblica L. Extracts Obtained with Different Extraction Methods</a>. The Authors section was updated from:
Haozhou Huang1 
Mengqi Li2 
Chuanhong Luo3 
Sanhu Fan4 
Taigang Mo4 
Li Han3 
Dingkun Zhang3 
Junzhi Lin5 
1Innovative Institute of Chinese Medicine and Pharmacy/Academy for Interdiscipline, Chengdu University of Traditional Chinese Medicine 
2Sichuan Nursing Vocational College 
3School of Pharmacy/School of Modern Chinese Medicine Industry, State Key Laboratory of Characteristic Chinese Medicine Resources in Southwest China 
4Sanajon Pharmaceutical Group 
5TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province, Hospital of Chengdu University of Traditional Chinese Medicine
to
Haozhou Huang1,2 
Mengqi Li3 
Chuanhong Luo4 
Sanhu Fan5 
Taigang Mo5 
Li Han4 
Dingkun Zhang4 
Junzhi Lin6 
1State Key Laboratory of Southwestern Chinese Medicine Resources, Innovative Institute of Chinese Medicine and Pharmacy/Academy for Interdiscipline, Chengdu University of Traditional Chinese Medicine 
2Meishan Hospital of Chengdu University of Traditional Chinese Medicine 
3Sichuan Nursing Vocational College 
4State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine 
5Sanajon Pharmaceutical Group 
6TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province, Hospital of Chengdu University of Traditional Chinese Medicine

An erratum was issued for:&#160;<a href="https://www.jove.com/t/65130">Using Multiple Light Scattering to Examine the Stability of Phyllanthus emblica L. Extracts Obtained with Different Extraction Methods</a>. The Authors section was updated.

Erratum: Using Multiple Light Scattering to Examine the Stability of Phyllanthus emblica L. Extracts Obtained with Different Extraction Methods

在中药（TCM）的制造中，中药提取中间体和相关液体制剂的稳定性一直是检查的重点1.医药产品的临床疗效，特别是以多酚为主要活性成分的药物，由于显着的稳定性问题而受到影响2,3。Sanajon口服液和诺迪康口服液是本期典型案例4。因此，学习如何使用高效的工具快速准确地评估和优化中药生产过程中液体中间体的稳定性至关重要。余甘子（PE）是一种在东南亚广泛分布的药用植物，被认为具有良好的抗氧化特性5，以及抗炎6，抗菌7和抗肿瘤作用8。在热提取过程中，PE中的单宁剧烈变化9。在高温催化下，这些单宁迅速水解生成没食子酸和鞣花酸等分子，由于其溶解度差导致不稳定或沉淀1。目前用于评估TCM稳定性的方法，例如加速测试或离心，通常很麻烦4，这限制了相关制备过程的进一步发展。
基于多重光散射（MLS）原理，建立了PEF提取物的快速稳定性评价方法，并分析了其失稳机理。MLS是一种基于近红外光源扫描的测量方法。任何解决方案系统的变化都会导致光强度的变化。入射光在被样品颗粒吸收或穿透时被散射。系统记录通过样品时的透射光信号;如果样品的透光率差，系统会记录反向散射光信号。与目视观察相比，这样可以节省大量时间1 ，可以快速准确地详细分析失稳现象，从而为指导提取过程的优化提供更多有用的信息。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Adjustable electric heating jacket</td>
			<td>Beijing Kewei Yongxing Instrument Co., Ltd</td>
			<td>MH-1000&#160;</td>
			<td>www.keweiyq.com</td>
		</tr>
		<tr>
			<td>Analytical balance(1/10000)</td>
			<td>Sartorious, Germany</td>
			<td>BSA224S&#160;</td>
			<td>www.sartorius.com.cn</td>
		</tr>
		<tr>
			<td>CNC ultrasonic instrument</td>
			<td>Kunshan Ultrasonic Instrument Co., Ltd</td>
			<td>KQ-500DE</td>
			<td>www.ks-csyq.com</td>
		</tr>
		<tr>
			<td>GL-16 high-speed centrifuge</td>
			<td>&#160;Sichuan Shuke Instrument Co., Ltd</td>
			<td>18091403</td>
			<td>www.sklxj.com</td>
		</tr>
		<tr>
			<td>Phyllanthus emblica L.</td>
			<td>Hehuachi medicinal materials market</td>
			<td>&#160;YJL2004</td>
			<td>Produced in Yunnan</td>
		</tr>
		<tr>
			<td>Turbisoft Lab multiple light scattering instrument</td>
			<td>French Formulaction Company</td>
			<td>Turbisoft Lab 2.3.1.125 Fanalyser 1.3.5</td>
			<td>www.formulaction.com</td>
		</tr>
		<tr>
			<td>UPR-II-5T ultra-pure water device</td>
			<td>Sichuan ULUPURE&#160; Ultrapure Technology Co., Ltd</td>
			<td>Z16030559</td>
			<td>www.ccdup.com</td>
		</tr>
	</tbody>
</table>

using multiple light scattering to examine the stability of phyllanthus emblica l. extracts obtained with different extraction methods

中药提取中间体是制备过程中的关键中间体，其稳定性对最终产品的有效性和质量有重要影响。然而，现有的稳定性评价方法往往费时费力，需要长期观察和复杂设备（如高效液相色谱）的操作，并且难以获得更多关于系统不稳定性的物理信息。因此，迫切需要为中药建立快速准确的稳定性分析技术。多重光散射是一种前沿的分析方法，可以在不改变样品性质或状态或使用有机试剂的情况下，以环保的方式准确快速地评估中药的稳定性。
在这项工作中，使用多次光散射的精确扫描数据，本协议快速获得了层厚度、颗粒迁移速度和平均粒径随时间的变化曲线。这使得能够精确识别导致系统早期不稳定的机制和关键特征。值得注意的是，通过系统稳定性的详细定量，可以大大缩短提取过程的研究周期，这也允许快速、准确和深入地分析各种提取过程对 余甘子 L稳定性的影响。

图1显示了多光测量的原理和收集结果的含义。在MLS光谱结果（图2）中，横坐标是样品池的高度，纵坐标是透射（T%）和反向散射（BS%）强度。通过计算MLS光谱结果，系统可以获得测量期间样品关键物理参数的变化，包括δ透射平均值（ΔT）（图3A）、光子自由程（图3B）、SI（图3C）和粒径（<strong class="xf...

Watch this Scientific Journal Video about Using Multiple Light Scattering to Examine the Stability of Phyllanthus emblica L. Extracts Obtained with Different Extraction Methods at JoVE.com

Using Multiple Light Scattering to Examine the Stability of Phyllanthus emblica L. Extracts Obtained with Different Extraction Methods

本文介绍了一种基于多重光散射技术的稳定性评价方法，用于评价中药提取物的稳定性。

使用多重光散射检查不同提取方法获得的余 甘子 L.提取物的稳定性

The extraction intermediate of traditional Chinese medicine is the key intermediate in the preparation process, and its stability has an important impact ...

稳定性评估方法通常既费时又费力。该技术可以节省约200倍的时间，并收集余甘子提取物的许多稳定性参数。该技术能够快速准确地详细分析失稳现象，从而为指导提取过程的乐观性提供更多有用的信息。首先，准确称取适量的余甘子L.，并加入植物重量10倍的去离子水进行回流提取。称量后，设置五个样品进行回流提取，E1在0小时，E2在0.5小时，E3在1小时，E4在1.5小时，E5在2小时。使用移液管向样品瓶中加入20毫升样品溶液，以确保每次添加的溶液处于相同的高度。打开多重光散射或MLS检测仪器，并将其预热30分钟。单击顶部菜单中的“创建文件”按钮以创建新的测试文件，然后单击“显示Turbiscan实验室温度”按钮将仪器目标温度设置为25摄氏度。单击程序扫描以进入设置分析程序，并将程序添加到列表中。将平衡时间设置为 20 分钟。在任务栏中，将扫描 48 小时添加到分析序列，将 5 分钟作为周期。为所有后续测量选择此分析程序。将准备好的样品瓶放入MLS检测系统，然后单击“开始”开始测量。数据收集后，单击计算参数列表。将分散相折射率设置为1.36，连续相折射率设置为1.33，以计算稳定性指数、粒径和粒子迁移速度，并将体积分数设置为1，E1至E5样品的连续相光透射强度为99.99%MLS光谱如图所示。光谱数据表明，E2样品波动较小，表明样品稳定性较高，而E1可能由于透射光的整体下降而出现浑浊。E3至E5样品相当不稳定，不同高度样品的光谱数据不同，表明分层发生在后期。T 值随时间增加，使样品更加不稳定。对于E3和E4，ΔT水平最终恢复到早期水平，表明这些提取物中发生了聚集和沉淀。E5的δT在浊度后保持较低水平，表明E5可能存在大量的沉降。光子自由程的趋势可以反映样品透射光的变化。各种提取物的稳定性随时间不断波动。粒径的动态变化表明，所有样品的粒径在8至20小时内发生了相当大的变化，E3和E5的粒径甚至超过了测量范围。这里显示了通过不同提取方法获得的余甘子L.提取物的不稳定性。每个结果顶部的色度带表示对应于不同颜色的光强度值，其中蓝色部分表示透射率，棕色部分表示反向散射强度。为测量选择合适的分析程序，并选择参数列表来计算稳定性指数、粒径和颗粒迁移速度，对于该颗粒至关重要，因为这两个步骤将直接影响结果的时效性。还可以获得一些重要信息，例如层厚，并且可以分析乳化或沉淀的形成规律。该技术将有助于开发基于大量药物不稳定数据的稳定性预测模型。此外，该技术可以与其他检测方法相结合和增强，进一步扩展研究的可能性。

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Using Multiple Light Scattering to Examine the Stability of Phyllanthus emblica L. Extracts Obtained with Different Extraction Methods

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使用多重光散射检查不同提取方法获得的余 甘子 L.提取物的稳定性

使用多重光散射检查不同提取方法获得的余甘子 L.提取物的稳定性