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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2016, Vol. 42 Issue (1): 23-29    DOI: 10.3785/j.issn.1008-9209.2015.03.211
生物科学与技术     
栗蓬抗菌成分提取工艺的优化与联合测定
刘淑萍, 王蕾
华北理工大学化学工程学院,河北 唐山063009
Optimization of the extraction process and combined determination
of  antibacterial ingredients of Li Peng
LIU Shuping, WANG Lei
(College of Chemical Engineering, North China University of Science and Technology, Tangshan 063009, Hebei, China)
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摘要： 正交试验优化栗蓬抗菌成分的提取工艺,进一步对提取液进行分级醇沉及去除蛋白操作后,采用反相高效液相色谱(reversed-phase high-performance liquid chromatography,RP-HPLC)法对所提抗菌成分进行定性及联合定量测定。结果表明,在最优条件下,一次的提取率可达88.2%。采用Lichrospher C18色谱柱(150 mm×4.6 mm,5 μm),以甲醇0.1%的磷酸水溶液为流动相,梯度洗脱程序,检测波长为280 nm,流速1 mL/min,柱温30 ℃可较好地测定栗蓬中没食子酸、原儿茶酸、槲皮素的含量。各成分质量浓度与峰面积在测定范围内均呈良好的线性关系(r＞0.999),平均加样回收率(n=9)分别为98.9%、96.0%和97.1%,相对标准偏差(relative standard deviation,RSD)分别为2.36%、0.77%和4.73%。
Abstract: Li Peng is the Chaetomium of chestnut (involucre of plant chestnut), also called the involucre of chestnut, thorn shell of chestnut, or Sarg. of chestnut etc. In the areas rich in Chinese chestnut,  large amounts of waste Li Peng was produced, besides a small amount of which can be used for firewood, the rest will be discarded to rot or burned; however, it is not only a waste of resources, but also cause serious environmental pollution. Reports showed that  Li Peng could be used as a raw material to cultivate chestnut mushrooms, but it was only an accessory ingredient, and the main ingredient was cotton seed shell of chestnut tree, so Li Peng has yet to be fully used. In some areas, the phenomenon of hyphae overgrowth occurred in chestnut mushrooms cultivated by Li Peng, the reason was due to its antibacterial effect. The purpose of this paper was to establish a simple and low-cost technique for extraction of antibacterial components from Li Peng. At the same time, a high sensitive, good repeatable and stable, simple and rapid method was used to determine the antibacterial ingredients of Li Peng. This paper used the single factor experiment and the orthogonal experiment optimization for the extraction process of antibacterial components in Li Peng. Furthermore, polysaccharide in the samples was precipitated by ethanol and the qualitative and quantitative combined determination of antibacterial components were carried out using reversed-phase high-performance liquid chromatography (RP-HPLC) method after protein removal. The results showed that the best way to extract the antibacterial composition of the 60-mesh fine slag in Li Peng was under 1∶40 material liquid ratio, 70 W ultrasonic reflux 20 min. Using Lichrospher C18 column (150 mm × 4.6 mm, 5 μm), methanol-0.1% phosphoric acid solution as the mobile phase, the flow rate of 1 mL/min, the detection wavelength at 280 nm, column temperature of 30 ℃ with the gradient elution program, the gallic acid, protocatechuic acid, and quercetin content in Li Peng could be better determined. The quality concentrations of each component with peak area showed a good linear relationship (r>0.999) in the measurement range. The average sample recovery (n=9) rate were 98.9%, 96.0%, 97.1%. The relative standard deviation (RSD) were 2.36%, 0.77%, 4.73%. We optimized the extraction  of antibacterial ingredients in Li Peng. By simple operation, low energy consumption and high extract efficiency method increased the feasibility of this technology, and provides theoretical basis of mass industrial production for the extraction of antibacterial ingredients in Li Peng. Under the optimum conditions, we can extract 88.2% antibacterial components from chestnut at one time. The antibacterial component of gallic acid, protocatechuic acid, quercetin content in Li Peng was 0.176 0 mg/g (RSD=0.23%), 0.053 7 mg/g (RSD=0.16%), 0.019 6 mg/g (RSD=0.21%), respectively. The RP-HPLC  is a high sensitive, good repeatable and stable method, which is also simple, convenient and accurate. This paper enriched the reference basis for the combined determination of the antibacterial components of  Li Peng.
出版日期: 2016-01-20
CLC:  Q 5-3  
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王蕾
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引用本文:

刘淑萍, 王蕾. 栗蓬抗菌成分提取工艺的优化与联合测定[J]. 浙江大学学报(农业与生命科学版), 2016, 42(1): 23-29.

LIU Shuping, WANG Lei . Optimization of the extraction process and combined determination
of  antibacterial ingredients of Li Peng. Journal of Zhejiang University (Agriculture and Life Sciences), 2016, 42(1): 23-29.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2015.03.211    或    http://www.zjujournals.com/agr/CN/Y2016/V42/I1/23

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