Name: Using Multiple Light Scattering to Examine the Stability of Phyllanthus emblica L. Extracts Obtained with Different Extraction Methods (Video) | JoVE Journal
Uploaded: 2023-04-14T12:30:00.000+00:00
Duration: 6 min 12 s
Description: Learn multiple light scattering to evaluate Phyllanthus emblica L. extract stability rapidly. This method reduces evaluation time 200-fold while providing detailed instability analysis for optimizing extraction processes.

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This study was supported by grants from the National Natural Science Foundation of China (81973493); National Interdisciplinary Innovation Team of Traditional Chinese Medicine (ZYYCXTD-D-202209); Sanajon Pharmaceutical Group Chengdu University of TCM production, study, and Research Joint Laboratory Project (2019-YF04-00086-JH); and Sichuan Province Science and Technology Plan Funded Project (2021YFN0100). The authors thank the Innovative Institute of Chinese Medicine and Pharmacy of Chengdu University of TCM for its technical support in the mass spectrometry work.

1. Extract preparation
<ol>
	<li>Accurately weigh an appropriate amount of PE, and add 10x (weight) of deionized water for reflux extraction.</li>
	<li>Set five samples for reflux extraction for 0 h (E1), 0.5 h (E2), 1 h (E3), 1.5 h (E4), and 2 h (E5) after weighing.</li>
	<li>After extraction, cool the samples to room temperature, and weigh to make up for the lost weight to ensure consistency with the pre-extraction weights.</li>
	<li>Centrifuge the samples at 8,581 &#215; g for 10 min to ensure the removal of insoluble material and herbal residues from the sample solution.</li>
	<li>Use a pipette to add 20 mL of sample solution into the sample bottle to ensure that the solution added each time is at the same height. 
	&#8203;NOTE: Avoid contamination, such as fingerprints, on the scanning part of the sample bottle, ensure that the sample bottle is clean, and check whether there are visible scratches on the bottle surface. When adding the sample solution, be careful not to spill or splash on the sample bottle, and ensure that the liquid level is at the same height in each bottle.</li>
</ol>
2. Instrument operation
<ol>
	<li>Turn on the MLS detection instrument, and warm it up for 30 min.</li>
	<li>Click on the Create file button in the top menu (or click on the File | New file function) to create a new test file.</li>
	<li>Click on the Show Turbiscan Lab Temperature button in the top menu to set the instrument target temperature to 25 &#176;C. 
	NOTE: The set temperature of the instrument must be higher than the room temperature; otherwise, the sample temperature will be affected by the room temperature.</li>
	<li>Click on Program Scan in the top menu to enter the setup analysis program. Add the program to the list, and in the taskbar, add 5 min as a cycle, scan for 48 h to the analysis sequence, and set the balance time to 20 min. Select this analysis program for all the subsequent measurements.</li>
	<li>Move the prepared sample bottle into the MLS detection system. After setting up the program, click on Start to start the measurement. 
	&#8203;NOTE: Be careful not to shake the glass bottle when loading the sample. The measurement can only be started after the sample temperature and setting temperature are balanced.</li>
</ol>
3. Multiple light scattering analysis program setting
<ol>
	<li>After the data collection, click on the calculation parameters list to set the optical parameters to calculate the stability index (SI), particle size, and particle migration speed.</li>
	<li>Set the optical parameters as follows: the continuous phase light transmission intensity (T0) as 99.99% (water), the dispersed phase refractive index (np) as 1.36, and the continuous phase refractive index (nf) as 1.33.</li>
</ol>

<ol><li>Huang, H. -Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Exploration+on+the+approaches+of+diverse+sedimentations+in+polyphenol+solutions%3A+An+integrated+chain+of+evidence+based+on+the+physical+phase%2C+chemical+profile%2C+and+sediment+elements%5BTitle%5D)+AND+%22Frontiers+in+Pharmacology%22%5BJournal%5D)">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/pubmed?term=((High+or+low+temperature+extraction%2C+which+is+more+conducive+to+Triphala+against+chronic+pharyngitis%5BTitle%5D)+AND+%22Biomedicine+and+Pharmacotherapy%22%5BJournal%5D)">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/pubmed?term=((Hepatoprotective+effects+of+different+extracts+from+Triphala+against+CCl%284%29-induced+acute+liver+injury+in+mice%5BTitle%5D)+AND+%22Frontiers+in+Pharmacology%22%5BJournal%5D)">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/pubmed?term=((Study+on+the+stability+control+strategy+of+Triphala+solution+based+on+the+balance+of+physical+stability+and+chemical+stabilities%5BTitle%5D)+AND+%22Journal+of+Pharmaceutical+and+Biomedical+Analysis%22%5BJournal%5D)">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/pubmed?term=((Antioxidant+activity+of+active+tannoid+principles+of+Emblica+officinalis+%28amla%29%5BTitle%5D)+AND+%22Indian+Journal+of+Experimental+Biology%22%5BJournal%5D)">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/pubmed?term=((Anti-inflammatory+and+anti-coagulatory+activities+of+caffeic+acid+and+ellagic+acid+in+cardiac+tissue+of+diabetic+mice%5BTitle%5D)+AND+%22Nutrition+and+Metabolism%22%5BJournal%5D)">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/pubmed?term=((Emblica+officinalis+corrects+functional%2C+biochemical+and+molecular+deficits+in+experimental+diabetic+neuropathy+by+targeting+the+oxido-nitrosative+stress+mediated+inflammatory+cascade%5BTitle%5D)+AND+%22Phytotherapy+Research%22%5BJournal%5D)">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/pubmed?term=((Amla+%28Emblica+officinalis+Gaertn%29%2C+a+wonder+berry+in+the+treatment+and+prevention+of+cancer%5BTitle%5D)+AND+%22European+Journal+of+Cancer+Prevention%22%5BJournal%5D)">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/pubmed?term=((Studies+on+the+chemical+constituents+of+Phyllanthus+emblica%5BTitle%5D)+AND+%22Natural+Product+Research%22%5BJournal%5D)">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/pubmed?term=((Characterisation+and+storage+stability+of+aronia+anthocyanins+encapsulated+with+combinations+of+maltodextrin+with+carboxymethyl+cellulose%2C+gum+Arabic%2C+and+xanthan+gum%5BTitle%5D)+AND+%22Food+Chemistry%22%5BJournal%5D)">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/pubmed?term=((Novel+phenylalanine-modified+magnetic+ferroferric+oxide+nanoparticles+for+ciprofloxacin+removal+from+aqueous+solution%29%5BTitle%5D)+AND+%22Journal+of+Colloid+and+Interface+Science%22%5BJournal%5D)">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>
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The authors have no conflicts of interest to disclose.

The rapid and accurate assessment of TCM stability has always been a focus of TCM research. To provide more useful information for directing the improvement of the extraction process, this study analyzed the stability and instability mechanisms of a sample using a near-infrared non-destructive technology.
In this protocol, the important stability parameters are calculated based on accurate MLS scan data. MLS scans can collect the transmission (T%) and backscattering (BS%) of ...

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

In the manufacturing of traditional Chinese medicine (TCM), the stability of the TCM extraction intermediates and related liquid preparations has always been the focus of inspection1. The clinical efficacy of medicinal products, especially with polyphenols as the primary active ingredient, suffers due to significant stability issues2,3. Sanajon oral liquid and Nuodikang oral liquid are examples of typical cases of this issue4. Therefore, it is crucial to learn how to use efficient tools to rapidly and accurately evaluate and optimize the stability of liquid intermediates in the TCM production process. Phyllanthus emblica L. (PE), a widespread medicinal plant in Southeast Asia, is thought to have good antioxidant properties5, as well as anti-inflammatory6, antibacterial7, and antitumor actions8. During the thermal extraction procedure, the tannins in PE transform violently9. Under catalysis with high temperatures, these tannins hydrolyze quickly to produce molecules such as gallic acid and ellagic acid, which lead to instability or precipitation due to their poor solubility1. Current methods for evaluating TCM stability, such as accelerated testing or centrifugation, are usually cumbersome4, which limits the further development of pertinent preparation processes.
Based on the principle of multiple light scattering (MLS), we established a fast stability evaluation method for PEF extracts and analyzed the instability mechanism. MLS is a measurement method based on the scanning of near-infrared light sources. Any solution system change results in a change in the light intensity. The incident light is scattered when it is absorbed or penetrated by the particles of the sample. The system records the transmission light signal when it passes through the sample; if the light transmittance of the sample is poor, the system records the backscattering light signal. Compared with visual observation, this can save a lot of time1 and can quickly and accurately analyze the instability phenomenon in detail, thus providing more useful information for guiding the optimization of the extraction process.

<table><tbody><tr><td>Adjustable electric heating jacket</td><td>Beijing Kewei Yongxing Instrument Co., Ltd</td><td>MH-1000&amp;nbsp;</td><td>www.keweiyq.com</td></tr><tr><td>Analytical balance(1/10000)</td><td>Sartorious, Germany</td><td>BSA224S&amp;nbsp;</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>&amp;nbsp;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>&amp;nbsp;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&amp;nbsp; 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

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

The extraction intermediate of traditional Chinese medicine is the key intermediate in the preparation process, and its stability has an important impact on the effectiveness and quality of the final product. However, existing stability evaluation methods are often time-consuming and labor-intensive, requiring long-term observation and the operation of complex equipment (such as high-performance liquid chromatography), and it is difficult to obtain more physical information about the instability of the system. Therefore, there is an urgent need to establish a fast and accurate stability analysis technology for traditional Chinese medicine. Multiple light scattering is a cutting-edge analytical method that can accurately and rapidly evaluate the stability of traditional Chinese medicines in an environment-friendly manner without changing the nature or state of the sample or using organic reagents.
In this work, using the precise scanning data of multiple light scattering, the present protocol rapidly acquired the variation curves for layer thickness, particle migration speed, and average particle size over time. This enabled the precise identification of the mechanism and crucial characteristics causing the system&#39;s instability in its early stages. Of note, the research period for the extraction process can be considerably shortened by the detailed quantification of the system stability, which also allows for a quick, accurate, and in-depth analysis of the effects of various extraction processes on the stability of Phyllanthus emblica L.

Figure 1 shows the principle of multiple light measurement and the meaning of the collected results. In the MLS spectra results (Figure 2), the abscissa was the height of the sample cell, and the ordinate was the transmission (T%) and backscattering (BS%) intensity. By calculating the MLS spectra results, the system can obtain the changes in the key physical parameters of the sample during the measurement period, including the delta transmissio...

Here, we introduce a stability evaluation method based on multiple light scattering technology to evaluate the stability of traditional Chinese medicine extracts.

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

using-multiple-light-scattering-to-examine-stability-phyllanthus

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Learn multiple light scattering to evaluate Phyllanthus emblica L. extract stability rapidly. This method reduces evaluation time 200-fold while providing detailed instability analysis for optimizing extraction processes.

Using Multiple Light Scattering to Examine the Stability of Phyllanthus emblica L. Extracts Obtained with Different Extraction Methods (Video) | JoVE Journal

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

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