Sugar cane is the most important sugar crop in China. It is not only an important raw material for the sugar industry, but also a raw material for energy, chemicals, brewing and feed. The sugarcane internode contains a large amount of sugar, mainly three kinds of carbohydrates such as glucose, fructose and sucrose. Whether it is sensory or research, there is a big difference in sugar content between sugarcane in different growth stages and different internodes. Therefore, the sugar content of sugarcane in different periods and different internodes was analyzed and analyzed, and the growth and accumulation of various sugars were analyzed. The relationship between sucrose accumulation and enzyme activity related to glucose metabolism was analyzed by statistical analysis. Study the growth process of sugarcane and the rule of sugar accumulation, and provide theoretical basis for sugarcane high-yield and high-sugar cultivation and genetic improvement.
The sugar content is determined by chemical colorimetry, optical rotation, gas chromatography, and liquid chromatography. The general chemical method can only measure the total amount of various sugars and cannot separately measure the content of various sugars. It has been shown that the measured values â€‹â€‹of glucose and fructose are slightly higher than the actual values â€‹â€‹and there is a deviation, which is highly likely to affect the quality of sugar cane. judgment. Although gas chromatography can measure the content of different sugars separately, the sensitivity is high. However, since the boiling point of carbohydrate compounds is high and it is difficult to vaporize, it is necessary to carry out derivatization reaction before detection. The sample processing, analysis and detection steps are cumbersome, and the interference error is considered to be large. . Shandong Luchuang Analytical Instrument Co., Ltd. chromatographic laboratory, using the LC-3000 high performance liquid chromatograph produced by our company to analyze the carbohydrate content of different sugars in honey, pineapple, potato, etc., has achieved good analytical results in practical applications. , get the customer's affirmation. At present, the determination of sugar carbohydrate content in sugarcane internodes is currently reported to be based on general chemical methods. Luchuang Analytical Instruments Co., Ltd. fully absorbs relevant research programs at home and abroad, further optimizes the analysis conditions, and uses the special column for carbohydrate analysis to separate and analyze the sugar content of sugarcane internodes, and establishes a simple, fast, accurate and reliable method for simultaneous determination of fructose. , HPLC, sucrose content by high performance liquid chromatography separation method.
1. Instruments and reagents
High Performance Liquid Chromatograph LC-3000 (Lu Chuang Instrument), Refractive Index Detector, Pinhole Sampler. Glucose, fructose and sucrose standards are domestically pure chromatographically pure (purity not less than 99.5%), acetonitrile is chromatographically pure, and experimental water is ultrapure water.
2. Chromatographic conditions
Column: Agilent Zorbax cellulose column (4.6 mm Ã— 250 mm, 5.0 Î¼m); mobile phase was acetonitrile-water (75:25, V/V); column temperature 30 Â° C; detection cell temperature 35 Â° C; flow rate 1.0 mL / min, The injection volume was 20 Î¼L.
3. Preparation of sugar standard solution
Accurately weigh 2.0g of fructose, glucose and sucrose (accurate to 0.0001g), transfer to a 100mL volumetric flask, and dilute to the mark with high-purity water to obtain a mixed solution with a mass concentration of 20mg/mL. Dilute to 10, 5, 2.5, 1.25, 0.625 mg/mL step by step. It was filtered through a 0.45 Î¼m microporous membrane before the injection was analyzed. Each sample was repeated 3 times.
4. Sample processing
The frozen inter-segment samples were peeled, chopped, and ground into powder in liquid nitrogen. The sample powder was accurately weighed to 1.00 g (accurate to 0.0001 g), 80% ethanol was added to 5 mL, and the temperature was heated at 80 Â° C for 30 min, 12000 r. Centrifuge at /min for 20 min, take the supernatant, add the residue to 5 mL of 80% ethanol solution and repeat the extraction once. Combine the supernatant, and evaporate the ethanol in a 90 Â°C water bath. Add high-purity water to a volume of 5 mL. Mix well and use once. The sample was taken out from the syringe and filtered through a 0.45 Î¼m microporous membrane filter for analysis by injection. Each sample was repeated 3 times.
Lu Chuang analysis showed that the high-performance liquid chromatograph was used to analyze fructose, glucose and sucrose in sugarcane internodes, and the retention time was good. The linear relationship between the content of the three and the peak area ratio was good, and the relative standard deviation was better. The accuracy of the spiked recovery rate is high, which is a simple, rapid and accurate method for detecting and analyzing the sugar composition and content of sugarcane internodes. If you have any relevant analysis requirements, please contact Shandong Luchuang Analytical Instrument Co., Ltd. 0632-5080618, or visit the company's website http:// for an instant online consultation.
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