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Determination of Crude Polysaccharides by Ultraviolet Spectrophotometry
Key words: UV spectrophotometry; Lycium barbarum polysaccharide; Ganoderma lucidum polysaccharide; Aesthetic analysis instrument
The content of crude polysaccharides in food was determined using the sulfuric acid-phenol colorimetric method, with glucose as the standard and dextran as a reference. The results showed that the former method yielded slightly higher values, approximately 4.8% higher. This method is simple, rapid, accurate, and highly reproducible, making it suitable for the determination of various types of crude polysaccharides.
Polysaccharides are carbohydrates composed of ten or more monosaccharides linked by glycosidic bonds. They are typically natural polymers and can be categorized into active polysaccharides and dietary fiber. Active polysaccharides, such as those found in fungi, plants, and chitosan, exhibit complex physiological functions like immune regulation, anti-cancer effects, anti-aging properties, and blood fat reduction. These compounds have gained increasing attention due to their wide-ranging health benefits.
Currently, there are no nationally standardized methods for determining polysaccharides. Most literature reports use either the phenol-sulfuric acid or anthrone-sulfuric acid method. However, most standard curves are prepared using glucose as the standard rather than dextran, which is difficult to obtain and costly in China. Despite this, research has shown that water-soluble polysaccharides from mushrooms, Flammulina, Yunzhi, Dongcao, Xia Cao, Ganoderma lucidum, Poria, and Polyporus possess significant biological activities such as immune enhancement, tumor suppression, sedation, and anti-inflammatory effects.
In this study, both Lycium barbarum polysaccharide and Ganoderma lucidum polysaccharide were used to compare the differences between using glucose and dextran as standards. The experimental procedure involved sample preparation, standard curve development, and sample analysis.
**Materials and Methods**
- **Sample Materials**: Lycium barbarum polysaccharide (40–60% concentration), Ganoderma lucidum polysaccharide (40–60% concentration), and dextran (MW 500,000).
- **Reagents**: Copper stock solution, copper reagent solution, detergent, ethanol solution (80%), sodium hydroxide solution (10%), sulfuric acid solution, phenol solution (5%), and glucose/dextran standard solutions.
- **Instruments**: UV spectrophotometer UV-1100, UV-1200 centrifuge rotary mixer.
**Experimental Procedure**
- **Standard Curve Preparation**: Glucose and dextran standard solutions were prepared, and absorbance values were measured at specific wavelengths (490 nm for glucose, 485 nm for dextran).
- **Sample Extraction and Precipitation**: Crude polysaccharides were extracted, precipitated using ethanol, and dissolved for further analysis.
- **Colorimetric Analysis**: The samples were analyzed using the phenol-sulfuric acid method, with absorbance measured at 490 nm.
- **Stability and Recovery Tests**: The stability of the color reaction and the recovery rate of the method were evaluated.
The results demonstrated that the method is reliable and efficient, with good linearity and accuracy. The recovery rate ranged from 93.15% to 99.7%, confirming its effectiveness. Additionally, the method does not require extensive purification or decolorization of the sample, making it ideal for routine analysis of crude polysaccharides in food products.
In conclusion, the ultraviolet spectrophotometric method provides a practical and accurate approach for determining crude polysaccharide content. Although dextran is a more appropriate standard, glucose can be used as an alternative due to its availability and cost-effectiveness. This technique offers a valuable tool for researchers and quality control professionals in the food and pharmaceutical industries.
Key words: ultraviolet spectrophotometry; alfalfa polysaccharide; ganoderma lucidum polysaccharide; aesthetic analyzer; UV-1100; UV-1200
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