s 15 contact s W Si 3 N μm N / m 4 AFM 95% AFM WPOJ UPOJ WPOJ UPOJ 260 ~ 280 nm WPOJ-DS UPOJ-DS 1 cm 1 cm DEAE-ce

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1 Journal of Chinese Electron Microscopy Society Vol. 32 No * AFM SEM WPOJ-DS UPOJ-DS WPOJ-DS β- UPOJ-DS AFM WPOJ-DS UPOJ-DS SEM UPOJ-DS WPOJ-DS S TG TG A doi / j ~ DEAE-cellulose52 Whatman Sephacryl S-300 Pharmacia 95% 5 ~ 8 SPM-9500J3 TU Avatar360E. S. P. FTIR RE-52AA FD-1A 9 10 Quanta 200 FEI WPOJ-DS UPOJ-DS % g WPOJ-DS UPOJ-DS h 2 80 g No No. SJ08A wxm @ 163. com *. sunrunguang@ snnu. edu. cn

2 s 15 contact s W Si 3 N μm N / m 4 AFM 95% AFM WPOJ UPOJ WPOJ UPOJ 260 ~ 280 nm WPOJ-DS UPOJ-DS 1 cm 1 cm DEAE-cellulose52 20 kv SEM 6 s / 15 min / 2 WPOJ-DS UPOJ-DS nm SEM DEAE-cellulose52 2 WPOJ-D UPOJ-D WPOJ-D UPOJ-D 2. 1 Sephacryl S WPOJ-DS UPOJ-DS 5 s / 15 min - WPOJ- DS UPOJ-DS cm cm - 1 O H cm cm mg WPOJ-DS UPOJ- DS 100 mg KBr cm cm - 1 N H ~ 400 cm ~ cm - 1 C H ~ cm mg WPOJ-DS UPOJ-DS 50 μg / ml 820 cm h cm - 1 α- 400 ~ 600 cm μg / ml 10 μg / ml 5 μg / ml 1 10 μg / ml CH 2 C H C O 934 cm cm μl WPOJ-DS 872 cm - 1 UPOJ-DS SPM WPOJ-DS β J3 1 Fig. 1 a WPOJ-DS b UPOJ-DS IR spectrum of WPOJ-DS a and UPOJ-DS b.

56 J. Chin. Electr. Microsc. Soc. 32 2. 2 90 nm WPOJ-DS UPOJ-DS 1 nm 1 AFM UPOJ-DS 2 25μg / ml AFM WPOJ-DS 30 ~ 70 nm 8 ~ 20 nm UPOJ-DS 3 10 μg / ml WPOJ- Fig.

3 56 J. Chin. Electr. Microsc. Soc nm WPOJ-DS UPOJ-DS 1 nm 1 AFM UPOJ-DS 2 25μg / ml AFM WPOJ-DS 30 ~ 70 nm 8 ~ 20 nm UPOJ-DS 3 10 μg / ml WPOJ- Fig μg / ml WPOJ-DS a ~ c UPOJ-DS a' ~ c' AFM a ~ c Bar = 5 μm 1 μm 200 nm a' ~ c' Bar = 5 μm 5 μm 200 nm AFM images of WPOJ-DS a - c and UPOJ-DS a' - c' on the concentration of 25 μg / ml. DS 1. 3 nm ~ nm UPOJ-DS 80 nm 10 nm β- 4 5 μg / m WPOJ-DS UPOJ-DS 2 ~ 4 AFM AFM WPOJ-DS 50 nm 1 mm a 5b WPOJ-DS UPOJ- DS 50 nm WPOJ-DS nm 150 5a' 5b' UPOJ-DS

μm 200 nm AFM images of WPOJ-DS a - c and UPOJ-DS a' - c' on the concentration of 10 μg / ml. Fig.

4 1 57 Fig μg / ml WPOJ-DS a ~ c UPOJ-DS a' ~ c' AFM a ~ c Bar = 5 μm 1 μm 500 nm a' ~ c' Bar = 5 μm 1 μm 200 nm AFM images of WPOJ-DS a - c and UPOJ-DS a' - c' on the concentration of 10 μg / ml. Fig μg / ml WPOJ-DS a ~ c UPOJ-DS a' ~ c' AFM a ~ c Bar = 5 μm 2 μm 500 nm a' ~ c' Bar = 5 μm 1 μm 200 nm AFM images of WPOJ-DS a - c and UPOJ-DS a' - c' on the concentration of 5 μg / ml.

5 58 J. Chin. Electr. Microsc. Soc. 32 UPOJ-DS WPOJ-DS WPOJ-DS UPOJ-DS 6 a ~ d WPOJ-DS 5 WPOJ-DS a b UPOJ-DS a' b' SEM a b a' b' Bar = 20μm Fig. 5 SEM images of mica samples of WPOJ-DS a b and UPOJ-DS a' b'. UPOJ-DS WPOJ-DS WPOJ-DS 6 a' ~ d' UPOJ-DS β- UPOJ-DS WPOJ-DS UPOJ-DS β-

6 1 59 Fig. 6 6 WPOJ-DS a ~ d UPOJ-DS a' ~ d' SEM a ~ d Bar = μm a' ~ d' Bar = μm SEM images of powder samples of WPOJ-DS a - d and UPOJ-DS a' - d'.

7 60 J. Chin. Electr. Microsc. Soc. 32 japonicus J. Carbohydrate Polymers Ying Zhi Han Xiaoxiang Li Jianrong. Ultrasoundassisted extraction of polysaccharides from mulberry β- leaves J. Food Chemistry J J Camesano T A Wilkinson K J. Single molecule study of xanthan. conformation using atomic force microscopy 1 Li Y N Zhu D N Qi J et al. Characterization of homoisoflavonoids in different cultivation regions of Ophiopogon japonicus and related antioxidant activity J. Journal of Pharmaceutical and Biomedical Analysis Xiong Shuangli Li Anlin Huang Ni et al. Antioxidant and immunoregulatory a ctivity of different polysaccharide fractions from tuber of ophiopogon japonicus J. Carbohydrate Polymers Chen X M Jin J Tang J et al. Extraction purification characterization and hypoglycemic activity of a polysaccharide isolated from the root of ophiopogon J. Biomacromolecules Ding Xiang Feng Su Cao Mei et al. Structure characterization of polysaccharide isolated from the fruiting bodies of Tricholoma matsutake J. Carbohydrate Polymers J Bel7402 J Ding Xiang Tang Jie Cao Mei et al. Structure elucidation and antioxidant activity of a novel polysaccharide isolated from Tricholoma matsutake J. International Journal of Biological Macromolecules

8 1 61 Comparative study on the structure and aggregation behavior of ophiopogon japonicus polysaccharides extracted by two methods WANG Xiao-mei 1 SUN Run-guang 1* ZHANG Jing 2 HAO Chang-chun 1 ZHANG Li-ni 1 1. College of Physics and Information Technology 2. College of Food Engineering and Nutritional Science Shaanxi Normal University Xi'an Shaanxi China Abstract In this paper different extraction methods including hot water and ultrasound were used to extract polysaccharides in ophiopogon japonicus. The functional group distribution and surface morphology of polysaccharides extracted by the two methods were studied by FT-IR spectroscopy atomic force microscopy AFM and scanning electron microscopy SEM. The results showed that the difference of functional group between ophiopogon japonicus polysaccharides extracted by hot water WPOJ-DS and ultrasound UPOJ-DS was obvious. β-glucopyranoside was detected in WPOJ-DS using infrared spectroscopy but not in UPOJ-DS. The results of AFM showed that the dispersion of UPOJ-DS was significantly higher than that of WPOJ-DS. Single molecule chain and helical structure were observed in UPOJ-DS by AFM indicating that intermolecular and intramolecular hydrogen bonds of polysaccharides were affected by ultrasound. The results of SEM showed that less of bigger schistose structures were observed in UPOJ-DS compared to WPOJ-DS and a lot of rod-shaped and small globular aggregates were observed in UPOJ-DS indicating that supersonic treatment created degradation on ophiopogon japonicus polysaccharides. Therefore ultrasonic extraction has an effect on molecular structure and aggregation behavior of ophiopogon japonicus polysaccharides. Keywords ophiopogon japonicus polysaccharides ultrasound atomic force microscope AFM scanning electron microscope SEM * Corresponding author

H 2 SO ml ml 1. 0 ml C 4. 0 ml - 30 min 490 nm 0 ~ 100 μg /ml Zhao = VρN 100% 1 m V ml ρ g

H 2 SO ml ml 1. 0 ml C 4. 0 ml - 30 min 490 nm 0 ~ 100 μg /ml Zhao = VρN 100% 1 m V ml ρ g 16 6 2018 11 Chinese Journal of Bioprocess Engineering Vol. 16 No. 6 Nov. 2018 doi 10. 3969 /j. issn. 1672-3678. 2018. 06. 004 1 2 1 1 1. 330200 2. 330006 4 Box-Behnken 1 25 g /ml 121 92 min 2 6. 284%