全 文 ：Determination of Total Flavones Content in Cerato-
carpus arenarius L． by Spectrophotometry
LIU Shan-shan，MA Xian-mei，YANG Hong-bing *
Xinjiang Bingtuan Key Laboratory for Green Processing of Chemical Engineering /School of Chemistry and Chemical Engineering，Shihezi Uni-
versity，Shihezi 832003
Abstract ［Objective］The aim was to establish the method for determining total flavoens content in Ceratocarpus arenarius L．［Method］Fla-
vonoids were extracted from C． arenarius by heating refluxing，and determined with rutin as the standard．［Result］In the range of 0． 019 －0． 102
mg/ml，linear relationship was good ( r = 0． 999 1) ． This method had higher precision and accuracy with RSD of 0． 226% and recoveries of
99． 36%． The total flavones content from C． arenarius was 10． 12 mg/g． ［Conclusion］The method is simple and reliable，which could be used
to control the quality of C． arenariu．
Key words Ceratocarpus arenarius L． ; Flavonoids; Spectrophotometry
Received: July 15，2011 Accepted: September 13，2011
Supported by International Science and Technology Cooperation
Programme of Xinjiang Crops ( 2011BC003) ．
* Corresponding author． E-mail: yhb-tea@shzu． edu． cn
Xinjiang is the largest distribution area of Chenopodiaceae
plants in China［1］ as its complex geographical environment
and unique climatic conditions． Chenopodiaceae plants are
closely related to human life． For example，Beta vulgaris is
the raw material for sugar manufacture; Chenopodium ambro-
sioides and Salsola collina are pharmaceutical raw materials;
Anabasis aphylla is no raw materials for agricultural insecticide
manufacture． Ceratocarpus arenarius L． is an annual herb be-
longing to Genus Ceratocarpus of Chenopodiaceae plants and
it is only distributed in Xinjiang in China． C． arenarius which is
not favored by animals grown in desert and wasteland rich in
resources． Currently，the researches on C． arenarius mainly
focused on ecological studies［2 －4］． However，the researches
on its chemical composition have not been reported． In the
previous study of YANG Hong-bing et al．［5］，it was found that
C． arenarius had higher biological activity． In this study，fla-
vonoids were extracted from C． arenarius by heating reflu-
xing，and total flavoens content in C． arenarius was detected
with NaNO2 － Al ( NO3 ) 3 － NaOH coloration method so as to
provide theoretical basis for the rational development and utili-
zation of plant resources in Xinjiang．
Materials and Methods
Materials
Experimental materials C． arenarius was collected in Man-
asi of Xinjiang in September 2010 and identified by Professor
YANG Ping of Shihezi University． The C． arenarius was dried
and crushed．
Reagents Rutin and biochemical reagents were purchased
from National Institute for the Control of Pharmaceutical and
Biological Products; ethanol， sodium nitrite， aluminum
nitrate，sodium hydroxide were of analytical grade． Distilled
water was used for the experiment．
Main instruments UV-Vis spectrophotometer ( Beijing
LabTech Instrument Co．，Ltd． ) ; 722S-visible spectrophotom-
eter ( Shanghai Jinghua Instrument Co．，Ltd． ) ; R-200 /205
rotary evaporator ( Buchi Company，Switzerland ) ; SHZ-C-
Multi-purpose circulating water pump ( Gongyi Yuhua Instru-
ment Factory) ．
Methods
Preparation of standard solution of the reference sub-
stance 20． 2 mg rutin dried at 120 ℃ to constant weight was
weighed and placed into a 100 ml flask，and then methanol
was added to dissolve the rutin by ultrasonic treatment． After
cooled，methanol was added till the mark，subsequently sha-
king，and then the reference substance solution ( with the
concentration of 0． 202 mg/ml) was obtained．
Preparation of sample solution 5． 0 g powder of C． arena-
rius was accurately weighed and placed in a 250 ml flask with
100 ml 95% ethanol solution，after 2 h of reflux extraction at
80 ℃ for three times，the filtrates were combined and concen-
trated． Then，they were metered volume to 100 ml，and 25 ml
of which was drawn to a 50 ml flask and metered volume to 50
ml as the sample solution．
Qualitative identification of samples ① Fluorescence
test: a small amount of sample solution was dropped on the
filter paper，after dried，the fluorescence was observed under
UV light． ② Hydrochloric acid-Mg reaction: appropriate a-
mount of sample solution was placed in a test tube，followed
by small amount of Mg powder and 1 －2 drops of concentrat-
ed hydrochloric acid to observe the reaction phenomenon． ③
Aluminum salt color reaction: a small amount of sample solu-
tion was dropped on the filter paper，after dried and sprayed
with 1% AlCl3 ethanol solution，the reaction phenomenon was
observed and the fluorescence was observed under ultraviolet
light． ④ Alkaline reagent color reaction: a small amount of
sample solution was dropped on the filter paper，after dried
and sprayed with 1% NaOH solution，the reaction phenome-
non was observed and the fluorescence was observed under
ultraviolet light． ⑤ Ultraviolet absorption spectrum: after the
sample solution was diluted with ethanol，the absorption peak
was measured at a wavelength of 200 －450 nm．
Selection of determination wavelength 0． 1 ml of Rutin
standard solution and 2． 0 ml C． arenarius sample solution
were drawn and placed in 10 ml of volumetric flask，with 3． 0
ml of 60% ethanol，followed by the addition of 0． 3 ml of 5%
NaNO2 solution ( after shaken，placed for 6 min) ，0． 3 ml of
Storage and Processing
Agricultural Science ＆ Technology，2011，12(11) : 1612 －1614
Copyright2011，Information Institute of HAAS． All rights reserved．
DOI:10.16175/j.cnki.1009-4229.2011.11.028
10% Al ( NO3 ) 3 solution ( after shaken，placed for 6 min) and
4．0 ml of 4% NaOH solution ( after shaken，placed for 6 min) ．
Then，60% ethanol was added to the mark，after shaken，
placed for 15 min． The UV-Vis spectrophotometer at 200 －600
nm wavelengths was used to determine the best determination
wavelength．
Linear relationship The calibration curve was made accord-
ing to the Chinese Pharmacopoeia［6］． Rutin standard solution
( 0，1．0，2．0，3．0，4．0 and 5．0 ml) was added into the 10． 0
ml flask，respectively． And then the coloration was carried out
according to method of Selection of determination wave-
length ． The absorbance was measured at the wavelength of
406 nm with blank as the control． The calibration curve was
made with the absorbance as vertical axis and the concentra-
tion as abscissa．
Sample assay 2． 0 ml of sample solution was drawn in 10． 0
ml of flask，and then the coloration was carried out according to
Selection of determination wavelength ． The absorbance was
measured at 406 nm，after substituted into the regression equa-
tion，the total flavonoid content in the sample was calculated．
Precision test Five copies of C． arenarius samples were
accurately weighed，with five times of determination for each
sample． Then，the mean value was taken．
Recovery experiment 0． 5，1．0 and 1． 5 ml of sample solu-
tion were placed into 10． 0 ml of flask，with 1． 0 ml of rutin
standard solution in each flask． The coloration was carried out
according to Selection of determination wavelength，and
then the recovery rate was calculated．
Results and Analysis
Qualitative identification of flavonoids
Phenolic hydroxyl group and γ-pyrone ring contained in fla-
vonoids appear colorable reaction with many reagents． The fluo-
rescent test，hydrochloric acid-Mg reaction，aluminum salt color
reaction and alkaline reagent color reaction carried out on the
sample solution showed the yellow-green fluorescence，orange
solution，yellow spots with the bright yellow fluorescence，deep
yellow spots with yellow fluorescence，respectively．
The results of color reaction suggested that there were
flavonoids in C． arenarius samples． Fig． 1 showed that the
sample solution had the characteristic absorption peaks of fla-
vonoids at 230，268 and 367 nm，which further confirming that
there were flavonoids in C． arenarius samples．
Selection of the optimal determination wavelength
Fig．2 showed that the sample solution had the maximum
absorption at 406 nm，while the rutin standard also had the
absorption here，with no absorption for the blank． Therefore，
406 nm was selected as the determination wavelength．
Linear relationship results
The standard curve made according to Linear relation-
ship was shown in Fig． 3． Linear regression equation was: y
=11．867x +0． 019 ( r =0． 999 1) ，with the linear concentra-
tion range of 0． 019 －0．102 mg/ml．
Fig． 1 Absorption spectrum of the samples
Fig． 2 Absorption spectrum of rutin control substance and
sample
Fig． 3 The standard curve of rutin
Results of sample assay
Sample absorbance was measured at 406 nm and then
substituted into the regression equation． The obtained fla-
vonoid content was 10．12 mg/g．
Results of precision test
The relative standard deviation of the experimental meth-
od of Precision test was 0． 226%，indicating a good preci-
sion of this method．
Results of recovery test
Table 1 showed the results of recovery test． The average
recovery rate was 99． 36% with rutin as reference to measure
the total flavonoid content，while the relative standard deviation
was 0．65%，indicating that the method is stable and reliable．
Table 1 The average recovery
Amount of sample
solution
Content in
sample∥mg
Added
amount∥mg Absorbance
Measured
quantity∥mg
Recovery
rate∥%
Average recovery
rate∥% RSD∥%
0．5 0． 126 5 0． 202 0． 406 0． 327 0 99． 26
1． 0 0． 253 0 0． 202 0． 557 0． 454 3 99． 65 99． 36 0． 65
1． 5 0． 379 5 0． 202 0． 706 0． 579 8 99． 16
3161LIU Shan-shan et al． Determination of Total Flavones Content in Ceratocarpus arenarius L． by Spectrophotometry
Conclusion
In this study，flavonoids were extracted from C． arenari-
us by heating refluxing，and total flavoens content in C． are-
narius was detected with NaNO2 － Al ( NO3 ) 3 － NaOH colora-
tion method． The total flavones content from C． arenarius was
10．12 mg/g． This method had higher precision and accuracy
with RSD of 0． 226% and recoveries of 99． 36%．
References
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Analysis on the niche of the main plant species of oasis － deseart
ecotone in Fukang of Xinjiang( 新疆阜康绿洲荒漠过渡带主要植物种
的生态位分析) ［J］． Acta Ecologica Sinica( 生态学报) ，2002，22
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Responsible editor: LI Ting-ting 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
分光光度法测定角果藜中总黄酮含量( 摘要)
刘珊珊，马先梅，杨红兵* ( 新疆兵团化工绿色过程重点实验室，石河子大学化学化工学院，新疆石河子 832003)
［目的］建立角果藜中总黄酮含量的测定方法。
［方法］采用加热回流法提取角果藜中的黄酮类化合物，以芦丁为标准品测定总黄酮含量。
［结果］采用紫外分光光度法测定总黄酮含量时，在 0． 019 ～0． 102 mg /ml浓度范围内线性关系良好( r = 0． 999 1) 。方法的精密度和准确度均
较高，相对标准偏差为 0． 226%，平均回收率为 99． 36%。测得角果藜中总黄酮含量为 10． 12 mg /g。
［结论］该方法操作简单，科学准确，适合用于角果藜中总黄酮含量的测定。
关键词 角果藜;黄酮类化合物;分光光度法
基金项目 新疆兵团国际科技合作计划( 2011BC003) 。
作者简介 刘珊珊( 1987 － ) ，女，山东省泰安人，硕士研究生，研究方向:为天然产物活性成分及功能研究，E-mail: lshan87@ 163． com。* 通讯作者，教
授，硕士，硕导，天然产物分析，E-mail: yhb-tea@ shzu． edu． cn。
收稿日期 2011-07-15 修回日期
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2011-09-13
( From page 1611)
［15］DONG YB(董轶博) ，KONG H(孔华) ，PENG YF(彭于发) ，et al．
Flowering and reproduction habits of Oryza rufipogon in Wanning
City of Hainan Province( 海南万宁普通野生稻居群开花习性和生殖
特性研究) ［J］． Journal of Plant Genetic Resources( 植物遗传资源
学报) ，2008，9( 2) : 218 －222．
［16］ LANGEVIN SA． The incidence and effects of hybridization be-
tween cultivated rice and its related weed red rice ( Oryza sativa
L．) ［J］． Evolution，1990，44: 1000 －1008．
［17］LI MT(栗茂腾) ，CAI DT(蔡得田) ，HUANG LM(黄利民) ，et al．
Comparison of anther structer between cleistogamous rice and an-
thesis rice(开颖授粉和闭颖授粉水稻的耐湿性和花药结构比较)
［J］． Journal of Huazhong Agricultural University(华中农业大学学
报) ，2001，20( 5) : 407 －412．
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
高州普通野生稻在海南陵水的开花习性和育性研究( 摘要)
余昌花，袁潜华* ，徐立新，何美丹，李厚奇，曹树威 ( 海南大学农学院，海南海口 570228)
［目的］探讨高州普通野生稻( Oryza rufipogon Griff． ) 在海南陵水的开花习性和育性。
［方法］对广东高州普通野生稻在海南陵水的开花习性、花粉育性和结实率进行了研究，并与海南普通野生稻开花特性进行了比较。
［结果］广东高州普通野生稻在陵水 1年开花 2次，第一开花期在 4月中旬至 6月中旬，历时约 60 d，开花高峰期为 5 月 13 ～ 22日，日开花历
时主要集中在 11: 20 ～13: 00，日开花高峰在 11: 30左右;第二开花期在 8月下旬至 10月下旬，历时 45 d，开花高峰期在 9月 8 ～ 18日，日开花
历时在 11: 15 ～12: 50，日开花高峰在11: 30左右。2次开花期间的颖花开放特性基本相同，柱头外露表现为单露率 46． 23%，双露率 49． 14%，
柱头总外露率达 95． 37%。单个颖花从开颖到闭颖一般需 1． 5 ～ 2． 5 h，开颖角度为 29． 9° ～ 32． 4°。上、下半年 2 个季次的花粉育性分别为
89． 67%和 85． 62%，平均结实率分别为 58． 81%和 55． 79%。海南普通野生稻一般 1年仅开花 1次，开花期在 10 月中旬至 12 月中旬，这与高
州野生稻在陵水 1年开花 2次的表现有明显差异。
［结论］为研究转基因水稻向普通野生稻基因漂流频率及与普通野生稻的开花习性相互关系奠定了基础。
关键词 高州普通野生稻;开花习性;花粉育性;结实率;陵水
基金项目 转基因专项( 2009ZX08011-010B，2008ZX08011-001) 。
作者简介 余昌花( 1984 － ) ，女，江西宜春人，硕士研究生，研究方向:作物遗传育种，E-mail: fengxi2008@ yeah． net。* 通讯作者。
鸣 谢 试验期间陈向东、郑金冲，2009级研究生尹昭坤、张蕾，农学系 2006级本科生钟坤、范官俊等参加部分工作，谨此致谢。
收稿日期 2011-07-10 修回日期 2011-09-26
4161 Agricultural Science ＆ Technology Vol． 12，No．11，2011