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壮瑶药材马蹄蕨多糖的提取工艺及其3种单糖的含量测定方法研究
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| 篇名： | 壮瑶药材马蹄蕨多糖的提取工艺及其3种单糖的含量测定方法研究 |
| TITLE： | |
| 摘要： | 目的：对壮瑶药材马蹄蕨的多糖成分进行提取分离纯化，并建立同时测定其中鼠李糖、果糖和D-无水葡萄糖3种单糖含量的方法。方法：优化水提醇沉法提取马蹄蕨多糖，比较不同型号大孔树脂除蛋白效果，并考察DEAE-52纤维素柱纯化所用不同洗脱液。采用高效液相色谱-蒸发光散射法进行含量测定。色谱柱为ZORBAX NH2，流动相为乙腈-水（72 ∶ 28，V/V），流速为0.8 mL/min，柱温为40 ℃，进样量为10 μL。漂移管温度为50 ℃，载气流速为2.0 L/min，增益为1。结果：采用水提75％乙醇沉淀提取马蹄蕨粗多糖，D101大孔树脂除蛋白质，DEAE-52纤维素柱纯化，并依次用蒸馏水、0.2 mol/L NaCl、0.2 mol/L NaOH溶液进行洗脱，冷冻干燥，即得马蹄蕨多糖。鼠李糖、果糖和D-无水葡萄糖检测质量浓度线性范围分别为49.10～982.02、54.47～1 089.42、62.15～1 242.96 μg/mL（r≥0.999 6）；检测限分别为7.4、3.2、4.4 μg/mL，定量限分别为24.4、10.6、14.5 μg/mL；精密度、稳定性（15 h）试验的RSD均＜2％（n＝6）；重复性试验的RSD＜3％（n＝6）；平均加样回收率分别为96.91％、97.43％、99.46％，RSD分别为1.3％、1.7％、2.2％（n＝6）。结论：建立的多糖提取分离工艺简单，易操作；高效液相色谱-蒸发光散射法操作简单，结果准确度高，可用于马蹄蕨多糖的提取纯化及其中3种单糖的定量分析。 |
| ABSTRACT： | OBJECTIVE： To extract， separate and purify the polysaccharides from Zhuang and Yao medicine Angiopteris fokiensis， and to establish a method for simultaneous determination of isodulcite， fructose and D-anhydrous glucose in 3 kinds of monosaccharide. METHODS： Optimization of water extraction and alcohol precipitation for extracting A. fokiensis polysaccharide，compare the effect of different types of macroporous resin on protein removal， and investigate the different eluents used in the purification of DEAE-52 cellulose column. HPLC-ELSD method was used. The determination was performed on ZORBAX NH2 column with mobile phase consisted of acetonitrile-water （72 ∶ 28， V/V） at the flow rate of 0.8 mL/min. The column temperature was 40 ℃ and sample size was 10 μL. The drift tube temperature was 50 ℃， the carrier gas velocity was 2.0 L/min， and the gain was 1. RESULTS： Extraction of crude polysaccharides from the horseshoe fern by water extraction with 75％ ethanol， D101 macroporous resin in addition to protein，DEAE-52 cellulose column for purification， and eluted sequentially with distilled water， 0.2 mol/L NaCl， and 0.2 mol/L NaOH solution， freeze-dried to obtain the A. fokiensis polysaccharide. The linear range of isodulcite， fructose and D-anhydrous glucose were 49.10-982.02 μg/mL， 54.47-1 089.42 μg/mL， 62.15-1 242.96 μg/mL （r≥0.999 6）. The detection limits were 7.4， 3.2 and 4.4 μg/mL， and the quantitation limits were 24.4， 10.6， 14.5 μg/mL. RSDs of precision and stability （15 h） test results were all lower than 2％ （n＝6）. RSDs of reproducibility tests were all lower than 3％ （n＝6）. The average recoveries were 96.91％， 97.43％ and 99.46％； RSD were 1.3％， 1.7％， 2.2％（n＝6）， respectively. CONCLUSIONS： The extraction and isolation technology of polysaccharides are simple and simple to operate. The HPLC-ELSD method is simple， accurate and suitable for the separation and purification of polysaccharides from A. fokiensis and quantitative analysis of 3 kinds of monosaccharides. |
| 期刊： | 2018年第29卷第19期 |
| 作者： | 张赟赟，杨海船，周萍，何春欢，严赟 |
| AUTHORS： | ZHANG Yunyun，YANG Haichuan，ZHOU Ping，HE Chunhuan，YAN Yun |
| 关键字： | 马蹄蕨；壮瑶药材；多糖；提取分离；含量测定；高效液相色谱-蒸发光散射法 |
| KEYWORDS： | Angiopteris fokiensis； Zhuang and Yao medicine； Polysaccharide； Extraction and separation； Content determination； HPLC-ELSD |
| 阅读数： | 294 次 |
| 本月下载数： | 4 次 |
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