全 文 ：Effect of Different Size of Ground Cyanotis arach-
noidea Particles on the Release of β-ecdysone
ZHU Guo-dong，ZHANG Yi-xiang * ，YE Jin-yun，WU Cheng-long，YU Lan，YANG Xia
School of Life Sciences，Huzhou Teachers College，Huzhou 313000
Abstract ［Objective］The aim was to investigate the influence of different size of ground Cyanotis arachnoidea C． B． Clarke particles on the re-
lease of β-ecdysone． ［Method］The content of β-ecdysone extracted from Cyanotis arachnoidea powders of different sizes was detected through
HPLC，under the chromatographic conditions:GRACE ODS C18 column (250 mm × 4． 6 mm，5 μm) ，mobile phase methanol∶water = 40∶60
(V/V) ，column temperature 26 ℃，flow rate 1． 0 ml /min and detection wavelength 248 nm． ［Result］The extracted β-ecdysone from ultra-fine
Cyanotis arachnoidea powder (particle size 10，30 and 50 μm)was as more than two folds as common powder(particle size 180 μm)．［Conclu-
sion］The β-ecdysone in ultra-fine powder of Cyanotis arachnoidea was more easily soluble in methanol． The extraction rate of β-ecdysone was
significantly higher than that of common powder． The result showed that the technology of ultra-fine pulverization markedly improved the dissolu-
tion of the β-ecdysone for increasing the cell breakage rate，and then the active ingredients could be extracted more easily．
Key words HPLC;Cyanotis arachnoidea C． B． Clarke;β-ecdysone
Received:June 24，2011 Accepted:August 20，2011
Supported by Major Project for International Cooperation in Zhejiang
(2009C14019); 948 Project of Ministry of Agriculture (2010-Z40);
Major Project from Department of Education of Zhejiang (ZD2009009);
Huzhou Major Science and Technology Project (2010ZD1002);the Na-
tional Natural Science Foundation of China (20977027)，Science and
Technology Innovation Program for Undergraduate Students in Zhejiang
Province (2011R425009)．
* Corresponding author． E-mail:yxzhang@hutc． zj． cn
20-hydroxy ecdysterone，also known as β-ecdysone，is a
kind of insect metamorphosis hormones，can be used to con-
trol pests and increase crab and shrimp production［1］． Cyano-
tis arachnoidea C． B． Clarke，an herb of Commelinaceae，is
one of the fine plants for extracting insect molting hormones．
In the dried Cyanotis arachnoidea，the insect molting hormone
content reaches as high as 1． 2% in whole plant，and 2．9% in
the underground part［2 －4］．
Ultrafine grinding technology can process herb into ultra-
fine particles smaller than 10 μm． As the ground herb particles
is ultra-fine，the active ingredients in them can be released
very easily; in addition， the small powder is more easily
absorbed in the small intestine wall，which can accelerate the
absorption of the active ingredients in the small intestine［5］．
In this study，ultrafine grinding technology was used to
process the Cyanotis arachnoidea into powers of different sizes，
in which the β-ecdysone contents were detected with high-per-
formance liquid chromatography，and thus to investigate the re-
lease of β-ecdysone in the Cyanotis arachnoidea powders pro-
cessed through ultrafine grinding technology，so as to provide
reference for the application of Cyanotis arachnoidea．
Materials and Methods
The equipments，reagents and samples
The main equipments included electronic balance，Soxhlet
extractor，rotary evaporator，high-performance liquid chroma-
tography system (Waters Alliance 2695)with a diode array de-
tector (Waters 2996)． The β-ecdysone standard preparation
was purchased from Sigm-Aldrich Corporation;methanol rea-
gent was HPLC grade． Cyanotis arachnoidea was purchased
from Yunnan，identified as an herb of Commelinaceae by Pro-
fessor GAI Guo-sheng． The root was dried，and then sent to
Qinghua Powder and New Materials Processing Center in
Zhejiang Changxing to be processed in to ultra-fine powder
and common powder．
Methods［1］
Extraction of β-ecdysone 1 g of Cyanotis arachnoidea
samples with particle size of 10，30 and 50 μm was respec-
tively weighed，added into 100 ml of methanol，extracted by
using the Soxhlet extractor for 4 h． After suck filtration，the
extract obtained was concentrated and dried in a rotary evapo-
rator． The residue was dissolved in methanol，and then the
solution volume was set to 10 ml，filtered through micropore
film of 0． 45 μm and then tested through HPLC system． The
control sample was common powder of 180 μm．
Preparation of standard solution 0． 020 0 g of β-ecdysone
standard preparation was accurately weighed and added into
methanol to prepare standard stock solution of 0． 80 mg/ml，
which could be diluted by adding methanol to different concen-
trations in later use．
Drawing standard curve 0． 1，0． 2，0． 3，0． 4，0． 5，0． 6，
0．7 and 0． 8 ml of β-ecdysone standard stock solutions were
precisely measured and transferred into volumetric flasks，di-
luted with methanol to 1 ml，to obtain a series of standard
solutions of 0． 10，0． 20，0． 30，0． 40，0． 50，0． 60，0． 70 and
0．80 mg/ml． Finally，10 μl of each solution was injected into
HPLC system as sample．
Chromatographic conditions GRACE ODS C18 column
(250 mm × 4．6 mm，5 μm)was used in this study;the column
temperature was 26 ℃;the mobile phase was methanol∶water =
40∶60 (V/V);the flow rate was 1． 0 ml /min;and the detection
wavelength was 248 nm．
Results
Linear relationship
With peak area of β-ecdysone as the Y-axis，concentra-
tion as the X-axis，the linear equation Y =20 555X + 279． 55
(R2 = 0． 999 8)was obtained． The results showed that the
β-ecdysone concentration in the range of 0． 1 － 0． 8 mg/ml
shared a good linear relationship with the peak area (Fig． 1)．
Agronomy and Forestry
Agricultural Science ＆ Technology，2011，12(9):1318 －1319，1326
Copyright 2011，Information Institute of HAAS． All rights reserved．
DOI:10.16175/j.cnki.1009-4229.2011.09.028
Fig． 1 The linear relationship between β-ecdysone content and
its peak area integral value
The β-ecdysone content
As shown in Fig． 2，under the chromatographic conditions
mentioned above， the β-ecdysone content extracted from
Cyanotis arachnoidea powders of different sizes (10，30 and
50 μm)were 0． 739 4%，0．658 3% and 0．739 4% respective-
ly，while that from control powder (180 μm)was 0． 326 7%．
Such result indicated that extraction of β-ecdysone from ultra-
fine powder was easier than from common Cyanotis arachnoidea
powder;the β-ecdysone extracted from ultra-fine powder was as
two folds as from common powder on average．
Conclusions and Discussions
Ultra-fine grinding technology is rapidly developed in
recent years，which can process raw materials into powders
of micron or even nanometer． It is an important part in powder
technology，includes ultra-fine grinding，high-precision grad-
ing，changing surface activity，and so on． Using traditional
methods，it is difficult to crush solid material to such a fine de-
gree． After the solid material is processed into powder of mi-
cron or even nanometer by ultra-fine grinding，its physical and
chemical properties are changed dramatically［5］． The dissolu-
tion and extraction ratio of active ingredients in Rhubarb［6］，
licorice［7］，Liuwei Dihuang［8］，dandelion［9］ and Panax［10］ are
significantly improved after the plants are treated with ultra-
fine grinding; in addition， the particle size has significant
effect on the dissolution of glycyrrhizin，and the ultra-fine pow-
der is more easily absorbed by animals than common powder．
In this study，the content of the β-ecdysone extracted from
Cyanotis arachnoidea powders of different sizes (10，30 and
50 μm)and common powder (180 μm)was measured，and
the results showed that the β-ecdysone in ultra-fine powder
was more easily dissolved in methanol，in which，the extrac-
tion rate was significantly higher than in common powder，indi-
cating that the technology of ultra-fine pulverization markedly
A:Standard preparation;B:Sample (10 μm)
Fig． 2 HPLC map of β-ecdysone standard and the sample
improved the dissolution of the β-ecdysone for increasing the
cell breakage rate，and then the active ingredients could be
extracted more easily，which creates better conditions for utili-
zation of organism．
The Cyanotis arachnoidea used in this experiment was col-
lected from Yunnan，and its root was processed into powder，
but the β-ecdysone content was significantly lower than the
2．9% reported before［4］，which may be related to the samples
origin，harvesting，transport and storage and other aspects．
References
［1］XU H(徐慧)． Determination of β －Ecdysone in Cyanotis vaga(Lour．)
Roem．et Schult by HPLC(高效液相色谱法测定蓝耳草中 β －脱皮激素
含量)［J］． Journal of Guangxi Academy of Sciences(广西科学院学
报)，2010，6(3):239 － 241．
［2］NIE RL(聂瑞麟)，XU XY(许祥誉)，HE M(何敏)，et al． Isolation and
identification of Ecdysone from Cyanotis arachnoidea plants(露水草植
物中蜕皮激素的分离与鉴定)［J］． Acta Chimica Sinica(化学学报)，
1978，36(2):137 －140．
［3］CHEN ZL(陈宗莲)． A preliminary study on the cultivation of Cyanotis
erechnoidea(露水草家化栽培研究简报)［J］． Acta Botanica Yunnani-
ca(云南植物研究)，1983，5(3):319 －322．
［4］NIE RL(聂瑞麟)，YUE YZ(岳远征)． Determination to the cultivation
of β －Ecdysone from Cyanotis arachnoidea(栽培露水草 β －蜕皮激素
含量的测定)［J］． 云南植物研究，1983，5(3):317 －318．
［5］YANG LW(杨连威)，ZHAO XY(赵晓燕)，LI T(李婷)，et al． Ultrafine
grinding and its effects on traditional Chinese Medicine(中药超微粉碎
后对其性能的影响研究)［J］． World Science and Technology －Mod-
ernization of Traditional Chinese Medicine and Materia Medica(世界科
学技术———中医药现代化)，2008，10(6):77 －81．
［6］LI XM(李晓明)，WANG YS(王跃生)． Influence of Semen cassia by
Supermicro －pulverization on its dissolved matter of Chrysophanol(超
微粉碎决明子对其大黄酚溶出量的影响)［J］． Chinese Journal of Ex-
perimental Traditional Medical Formulae(中国实验方剂学杂志)，2001，
7(6):6 －8．
［7］LI ZM(李志猛)，WANG YS(王跃生)，LI XM(李晓明)，et al． Com-
parative Study of dissolved matter of Glycyrrhizic Acid before and after
Supermicro －pulverization of Radix Glycyrrhizae(甘草饮片超微粉碎前
后甘草酸溶出行为的比较研究)［J］． China Journal of Chinese Materia
Medica(中国中药杂志)，2003，28(11):1030 －1033．
［8］ZHANG QM(张启明)，HE Y(何颖)，SONG LL(宋丽丽)． Influence of
ultramicro －porphyrizing on dissolving － out characteristics of Ursolic
acid in Liuwei Dihuang powder(超微粉碎对六味地黄粉中熊果酸溶出
特性的影响)［J］． Journal of Henan University:Medical Science(河南
大学学报:医学科学版)，2002，21(3):26 －28．
［9］SONG LL(宋丽丽)，FAN BY(范丙义)，JIANG SZ(蒋士忠)，et al．
Probe into characteristics of Taraxacummongolicum Ultramicro －pow-
er (蒲公英超微细粉体特性探讨)［J］． China Journal of Chinese Mate-
ria Medica(中国中药杂志)，2002，27(1):12 －15．
［10］CHEN X(陈旭)． Preliminary studies on the effect of ultramicro －pul-
verization on absorption of Sanqi in vivo(三七超微粉碎对生物体内吸
收影响的初步试验)［J］． Northwest Pharmaceutical Journal(西北药
学杂志)，2002，17(5):203．
Responsible editor:YIN Qing-qing Responsible proofreader:WU Xiao-yan
(Turn to page 1326)
9131ZHU Guo-dong et al． Effect of Different Size of Ground Cyanotis arachnoidea Particles on the Release of β-ecdysone
［5］CAI WG(蔡文贵)，LIN Q(林钦)，JIA XP(贾晓平)，et al． Integrat-
ed assessment on the level of the food organisms in kaozhou bay
(考洲洋饵料生物水平综合评价) ［J］． Acta Hydrobiologica Sinica
(水生生物学报)，2006(30):269 －275．
［6］DU N(杜宁)，YANG NS (杨宁生)，SUN YZ (孙英泽)GIS －
based evaluation of aquaculture suitability in Henan Province(基于
GIS的池塘养殖适宜性评价———以河南省为例) ［J］． Journal of
Fishery Sciences of China(中国水产科学)，2008(15):476 －482．
［7］DUAN JR(段金荣)，ZHANG HY(张红燕)，LIU K(刘凯)，et al．
Evaluation of suitability of aquatic animals habitat in Lihu(蠡湖水生
动物栖息地适宜性评估)［J］． Journal of Shanghai Ocean University
(上海海洋大学学报)，2010(19):116 －119．
［8］DUAN JR(段金荣)，ZHANG HY(张红燕)，LIU K(刘凯)，et al．
Biodiversity study of phytoplankton community from dianchi lake
based on GIS(基于地理信息系统的滇池浮游植物生物多样性的研
究)［J］． Journal of Zhejiang Ocean University(浙江海洋学院学
报)，2006(25):322 －326．
［9］LI DH，YANG S，FANG T，et al． Recovery of aquatic macrophytes
by use of water level regulation method in eutrophicated lakes － a
case study of uuli lake［J］． Environmental Science ＆ Technology，
2008(31):59 －62．
［10］ PENG HJ． Evaluation system of scientific and technological
achievements［D］． Hangzhou:University of Zhenjiang，2005．
［11］CHEN ZZ，LIN ZJ，QIU YS． Evaluation of sustainability of fisher-
ies resources for south China sea based on the AHP(基于 AHP
的南海海域渔业资源可持续利用评价)［J］． Journal of Natural Re-
sources(自然资源学报)，2010(25):249 －257．
［12］HUANG QH，YANG YG，CHEN YH． Hydrological process factors
analysis of Heihe River mountain basin based on GIS［J］． Agri-
cultural Science ＆ Technology，2010，11(3):147 －150．
［13］CHEN J(陈静)． Research on primary productivity and fishery po-
tential of Guocun Reservoir(郭村水库初级生产力及渔产潜力研
究)［J］． Journal of Anhui Agricultural Sciences(安徽农业科学)，
2011，39(7):4032 －4033．
［14］CHEN Q，WUWZ，ZHOU Q，et al． Synthesis analysis of soil ero-
sion for Three-River headwater region based on GIS［J］． Agri-
cultural Science ＆ Technology，2010，11(5):155 －158．
［15］ LIU ZQ(刘智琦)． Research on distributed logistics distribution
system of agricultural products(分布式农产品物流配送系统研究)
［J］． Journal of Anhui Agricultural Sciences(安徽农业科学)，
2011，39(21):13096 －13097．
［16］FEI L，TIAN QY，LI QJ． The anylasis of fitness on maize cultiva-
tion based on RS and GIS［J］． Agricultural Science ＆ Technolo-
gy，2010，11(6):75 －78．
［17］ZHANG DL(张德利)，FENG XM(冯雪梅)． Analysis on the varia-
tion of Jiangsu coastal tidal flat based on GIS(基于GIS的江苏沿
海滩涂变化分析)［J］． Journal of Anhui Agricultural Sciences(安
徽农业科学)，2010，38(33):19220 －19222．
［18］ XIAO Y． Research of comprehensive evaluation of agricultural
land suitability in typical area of Huanghuaihai Plain based on
GIS［J］． Agricultural Science ＆ Technology，2010，11(6):125 －
129，178．
Responsible editor:YIN Jian-li Responsible proofreader:
檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪檪
WU Xiao-yan
基于 GIS湖泊渔产潜力评价 －以蠡湖为例(摘要)
段金荣，张红燕，刘凯，徐东坡，张敏莹，施炜纲* (农业部长江下游渔业资源环境重点野外科学观测试验站，中国水产科学研
究院内陆渔业生态环境和资源重点开放实验室，中国水产科学研究院淡水渔业研究中心，江苏无锡 214081)
［目的］利用 GIS技术对蠡湖的渔产潜力进行科学的评价。
［方法］根据湖泊渔产特点和相关资源，采用层次分析法和 GIS技术建立综合评价蠡湖渔产潜力的数量化方法，该评价体系包括水质条件、生
物多样性和水文条件等 3个层次的 10个指标。通过咨询专家并构建判断矩阵，确定选择因子对于渔产潜力评价的贡献率，利用 ARC/INFO
软件内嵌的栅格数据分析模块生成单因子评价图层，结合栅格计算、空间叠加和分级模块，得出蠡湖渔产潜力等级的分布图。
［结果］渔产潜力评价为较高等级的区域面积为 2． 643 3 km2、渔产潜力评价为高等级的区域为 3． 844 8 km2、渔产潜力评价为一般等级的区域
面积为 1． 121 4 km2 和渔产潜力评价为低等级的区域面积为 0． 400 5 km2。
［结论］该研究为蠡湖的生态重建提供了理论依据。
关键词 渔产潜力;评价;GIS;蠡湖
基金项目 中央级公益性科研院所基本科研业务费专项资金(2011JBFC04)、公益性行业(农业)科研专项(200903048-03)和农业部财政专项
(6125005)。
作者简介 段金荣(1978 － )，男，江苏海门人，助理研究员，硕士，从事渔业 GIS研究。* 通讯作者，研究员，本科，从事渔业 GIS研究。
收稿日期 2011-06-28 修回日期
欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁欁
2011-08-29
(From page 1319)
露水草加工粒径和其 β-蜕皮激素释放的关系(摘要)
朱国栋，张易祥* ，叶金云，吴成龙，于岚，杨霞 (湖州师范学院生命科学学院，浙江湖州 313000)
［目的］比较不同粒径露水草(Cyanotis arachnoidea C． B． Clarke)对所含 β-蜕皮激素释放的影响。
［方法］采用 GRACE ODS C18色谱柱(250 mm ×4． 6 mm，5 μm) ，流动相甲醇∶水 =40∶ 60(V/V，%)洗脱。柱温 26 ℃，流速 1． 0 ml /min，检测波
长 248 nm。
［结果］相同来源的露水草粉体(粒径 10、30、50 μm)β-蜕皮激素含量检测值和普通细粉(粒径 180 μm)相比，粉体中蜕皮激素激素的检测值
平均高 1倍多。
［结论］露水草粉体中的蜕皮激素更容易溶于甲醇，其提取率明显高于普通细粉。表明露水草经过超微粉碎后，细胞破壁率到得明显提高，更
有利于有效成分的溶出。
关键词 高效液相色谱法;露水草(Cyanotis arachnoidea C． B． Clarke) ;β-蜕皮激素
基金项目 浙江省重大国际合作项目(2009C14019);农业部“948”项目(2010-Z40);浙江省教育厅重大项目(ZD2009009);湖州市重大科技专项
(2010ZD1002);国家自然科学基金(20977027);浙江省大学生科技创新计划(新苗人才计划)(2011R425009)。
作者简介 朱国栋(1990 －)，男，浙江上虞人，本科，专业:生物工程。* 通讯作者，硕士生导师，副教授，E-mail:yxzhang@ hutc． zj． cn。
收稿日期 2011-06-24 修回日期 2011-08-20
6231 Agricultural Science ＆ Technology Vol． 12，No． 9，2011