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

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 ens...

<ol>
	<li>Huang, H. -. 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S., Dsouza, J. 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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 border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
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		<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>
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		<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>
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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&#160;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 transm...

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

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

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 ...

Stability evaluation methods are often time-consuming and labor-intensive. This technique can save approximately 200X time and collect many stability parameters for Phyllanthus emblica L.extracts. This technique can quickly and accurately analyze the instability phenomenon in detail, thus providing more useful information for guiding the optimism of the extraction process.To begin, accurately weigh an appropriate amount of Phyllanthus emblica L.and add deionized water 10 times the weight of the plant for reflux extraction. After weighing, set five samples for reflux extraction, E1 at 0 hours, E2 at 0.5 hours, E3 at 1 hour, E4 at 1.5 hours, and E5 at 2 hours. Use a pipette to add 20 milliliters of sample solution to the sample bottle to ensure that the solution added each time is at the same height.Turn on the multiple light scattering, or MLS, detection instrument, and warm it up for 30 minutes. Click on the Create File button in the top menu to create a new test file, and then click on the Show Turbiscan lab temperature button to set the instrument target temperature to 25 degrees Celsius. Click on Program Scan to enter the setup analysis program, and add the program to the list.Set the balance time to 20 minutes. In the task bar, add Scan for 48 hours to the analysis sequence, and 5 minutes as a cycle. Select this analysis program for all the subsequent measurements.Place the prepared sample bottle into the MLS detection system, and click on Start to start the measurement. After data collection, click on the calculation parameters list. Set the dispersed phase refractive index to 1.36 and the continuous phase refractive index to 1.33 to calculate the stability index, particle size, and particle migration speed, and set the volume fraction to 1, and the continuous phase light transmission intensity to 99.99%MLS spectra of E1 to E5 samples are shown in this figure.The spectral data suggests that the E2 sample fluctuated less, indicating greater sample stability, while E1 may have had turbidity due to the overall decline in transmission light. E3 to E5 samples were quite unstable, and the spectral data of the samples at different heights were different, indicating that the stratification occurred in the later period. The T value increases with time, making the sample more unstable.For E3 and E4, the delta-T level returned to that of the earlier stage in the end, indicating that aggregation and precipitation occurred in these extracts. The delta-T of E5 remained low after turbidity, indicating that E5 may have had a large amount of sedimentation. The trend in the photon free path can reflect the changes in the transmitted light of the sample.The stability of various extracts fluctuated continuously over time. The dynamic changes in particle size revealed that the particle sizes of all the samples changed considerably within 8 to 20 hours, with the particle size of E3 and E5 even exceeding the measurement range. The instability of Phyllanthus emblica L.extracts obtained by different extraction methods is shown here.The chromaticity band at the top of each result represents the light intensity values corresponding to different colors, where the blue part represents transmission, and the brown part represents backscattering intensity. Selecting a proper analysis program for the measurements, and choosing the parameters list for calculating the stability index, particle size, and the particle migration speed, are crucial for this particle, as these two steps will directly affect the currency of the results. Some significant information, such as layer thickness, can also be obtained, and the formation rules of emulsification or precipitation can be analyzed.This technique will facilitate the development of a stability prediction model based on extensive medication instability data. Additionally, this technique could be combined and enhanced with other detection methods, further expanding the research possibilities.

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Cutaneous pharmacokinetics (cPK) after topical formulation application has been a research area of particular interest for regulatory and drug development ...

Muscle Function Obtained with Motion Mode Ultrasound and Surface Electromyography during Core Endurance Exercise

Motion mode (M-mode) ultrasound allows researchers and clinicians to measure the change of muscle thickness across time. Muscle thickness can be measured between fascial borders at a given time point during an exercise. This selected time point produces a one-dimensional image resulting in real-time, live observation of anatomy. Ultrasound used during functional movement can be referred to as dynamic ultrasound; this is feasible and reliable with the use of a linear transducer, elastic belt, and foam block to secure consistent transducer placement. The lateral abdominal wall is commonly investigated using ultrasound due to the overlapping nature of the muscles. Surface electromyography (sEMG) can complement M-mode ultrasound imaging because it measures the electrical representation of muscle activation. There is minimal evidence using M-mode ultrasound and sEMG simultaneously during core exercise. Exercises that challenge the core musculature involve both isometric holds (e.g., side plank), as well as oscillatory extremity movements (e.g., dead bug). In this study, both instruments will be used simultaneously to measure core muscle function during exercise. Ultrasound measurements will be obtained using a linear transducer and ultrasound unit, and sEMG measurements will be acquired from a wireless sEMG system. To make comparisons between participants and exercises, normalization methods using static, exercise starting positions for both instruments will be used. An activation ratio will be used for ultrasound and calculated by dividing the contracted thickness (thickness during a time point of exercise) by the rested (starting position) thickness. Muscle thickness will be measured in centimeters from the superior inferior fascial border to inferior superior fascial border. These methods aim to offer an innovative and practical measurement of muscle function with M-mode ultrasound and sEMG during core endurance exercises.

This protocol uses motion mode ultrasound and surface electromyography simultaneously to measure muscle function of the core. Muscle thickness and activation of the local stabilizers (e.g., transverse abdominis, internal oblique) and global movers (e.g., external oblique) is achievable during specific time points of the side plank and dead bug exercises.

Motion mode (M-mode) ultrasound allows researchers and clinicians to measure the change of muscle thickness across time. Muscle thickness can be measured ...

Dynamic Light Scattering Analysis for the Determination of the Particle Size of Iron-Carbohydrate Complexes

Intravenously administered iron-carbohydrate nanoparticle complexes are widely used to treat iron deficiency. This class includes several structurally heterogeneous nanoparticle complexes, which exhibit varying sensitivity to the conditions required for the methodologies available to physicochemically characterize these agents. Currently, the critical quality attributes of iron-carbohydrate complexes have not been fully established. Dynamic light scattering (DLS) has emerged as a fundamental method to determine intact particle size and distribution. However, challenges still remain regarding the standardization of methodologies across laboratories, specific modifications required for individual iron-carbohydrate products, and how the size distribution can be best described. Importantly, the diluent and serial dilutions used must be standardized. The wide variance in approaches for sample preparation and data reporting limit the use of DLS for the comparison of iron-carbohydrate agents. Herein, we detail a robust and easily reproducible protocol to measure the size and size distribution of the iron-carbohydrate complex, iron sucrose, using the Z-average and polydispersity index.

Dynamic light scattering (DLS) has emerged as a suitable assay for evaluating the particle size and distribution of intravenously administered iron-carbohydrate complexes. However, the protocols lack standardization and need to be modified for each iron-carbohydrate complex analyzed. The present protocol describes the application and special considerations for the analysis of iron sucrose.

Intravenously administered iron-carbohydrate nanoparticle complexes are widely used to treat iron deficiency. This class includes several structurally ...

Standardized Tuina Protocol for Rat DRG Compression Model

Analgesic Effect of Tuina on Rat Models with Compression of the Dorsal Root Ganglion Pain

Neuropathic pain is a prevalent condition that affects 6.9%-10% of the population and results from nerve damage due to various etiologies, such as lumbar disc herniation, spinal canal stenosis, and intervertebral foramen stenosis. Although Tuina, a traditional Chinese manual therapy, has shown analgesic effects in clinical practice for the treatment of neuropathic pain, its underlying neurobiological mechanisms remain unclear. Animal models are essential for elucidating the basic principles of Tuina. In this study, we propose a standardized Tuina protocol for rats with compression of the dorsal root ganglion (DRG), which involves inducing DRG compression by inserting a stainless steel rod into the intervertebral foramen, performing Tuina manipulation with specific parameters of location, intensity, and frequency in a controlled environment, and assessing the behavioral and histopathological outcomes of Tuina treatment. This article also discusses the potential clinical implications and limitations of the study and suggests directions for future research on Tuina.

Author Spotlight: Analgesic Effect of Tuina on Rat Models with Compression of the Dorsal Root Ganglion Pain  

This article presents a manipulation for treating chronic compression of the dorsal root ganglion in rats using Tuina therapy, along with a method for evaluating its effectiveness based on pain behavior and histopathological results.

Neuropathic pain is a prevalent condition that affects 6.9%-10% of the population and results from nerve damage due to various etiologies, such as lumbar ...