全 文 ：　　　　 天然产物研究与开发　 　　　　　
　　　　　　　　　　 NATU RAL PRODUCT RESEARCH AND DEVELOPMENT　 2003　Vo1.15　No.6
　
　
　
　
　　收稿日期:2003-07-02　　　修回日期:2003-09-17
＊Zheng Min-yan , lecturer , master.Major study di rection:natu-
ral organic chemist ry.E-mail:zmy1011@sina.com
STUDIES ON THE CONTENTOF FLAVONOIDS
IN JASMINUM NUDIFLORUM LINDL.
ZHENG Min-yan ,WEI Yong-sheng
(Department of chemistry , X ianyang Normal College , Xianyang　712000 , China)
Abstract　The flav onoids in Jasminum nudi florum Lindl.extracted by Soxhle t ex tractor with 75% ethanol
w as studied by colo rimetry and HPLC.Determined by colorimetry w ith rutin as standard sample , the content
of total flavonoids in its flowers , leaves and stems were 13.7%, 16.2% and 6.77%, RSD were 2.63%,
2.57% and 2.96% respectively , w hile determined by HPLC with quercetin , kaempferol , isorhamnetin as
standard samples , the content of total flav onoids in its flow ers , leaves and stems counted by ag lycone were
4.205%, 3.371%, 1.703%, RSD were 1.76%, 1.58% and 1.98% respectiv ely.
Key words　Jasminum nudi florum Lindl.;flavonoids;HPLC;colo rimetry
Introduction
Jasminum nudi f lorum Lindl.(Chinese name Ying-
chun), a kind of defoliation shrub , belongs to family
Oleaceae and genus Jasminum.In China , it s original
planted areas were in middle and north provinces , and
i t is planted in every place at present.Compendium
of Materia Medica recorded it could be used as
medicine.It tasting bitter and innocuous is alw ays
used for dispelling heat , sw eating , diuresising , activ-
iting blood , detumescence , obtunding etc[ 1] .But
there are few reports about it s chemical constituents.
In this paper , abundant flavonoids were found in this
plant by qualitative analysis , colorimetry and HPLC.
Present research has proved that f lavonoids have the
eff icacies of low ering blood fat and sugar , resisting
arteriosclerosis , oxidation , tumour and inf lammation
etc[ 2] .S tudying the content of flavonoids in J .nud-
if lorum Lindl.can provide scientif ic attestation to
exploit this natural plant.
1　Materials , reagents and apparatus
1.1　Experimental materials
J .nudi f lorum Lindl.picked in campus of Qinghai
Normal University w as identified by Li Jian-min , a
botany associate professo r of Biology department of
this university .The flowers , leaves and stems w ere
picked up , dried naturally , crushed respectively.Af-
ter being passed by 20 screen mesh , these materials
were dried to constant weight in the vacuum condi-
tion at 50 ℃.
1.2　Principal reagents
Rutin , quercetin and kaempferol w ere produced by
Chinese Medicine Checking Institute;Isorhamnetin
w as produced by Edward Gurr L td.;Other reagents
(A.R.)
1.3　Principal apparatus
UV265UV-V IS photometer(Simadzu);LC-6A liquid
chromatog raph(SPD6AV UVdetector;C-R3A data
processing computer)(Simadzu).
2　Experimental
2.1　Extracting flavonoids
By studying the results of orthogonal experiment , the
optimal rate of ex tracting flavonoids in this plant was
w ith 75% ethanol as solvent.Accurately w eighting
494
DOI :10.16333/j.1001-6880.2003.06.004
specimen 1.500 g was ext racted by Soxhlet ext rac-
tor , ho t reflux w ith benzinum to remove grease and
w ax until there w as no any color in ci rculation ref lux
liquid.Get ting rid of benzinum , the residue w as
bathed by 75% ethanol , the w ashing solutions were
conflated and constant volume to 250 mL , then the
ex traction of J .nudif lorum Lindl.(test solution)
used for qualitative and quantitative analysis w as pre-
pared.
2.2　The qual itative determination of flavonoids
Taking some test solution reacted w ith HCl-Mg , 4%
NaOH and 1%FeCl3 respectively , observing the
change of colo r[ 3] .
2.3　Determining the content of total flavonoids by
colorimetry
2.3.1　Determining method
Adopt ing NaNO2-Al(NO3)3-NaOH coloration system
determined the content of total f lavonoids with rutin
as standard sample[ 4] .Test solution accurately quan-
tified to 25 mL measuring f lask was added 30%
ethanol to 12.5 ml , 5%NaNO2 0.7 mL , shook up ,
laid 5 min , then added 10%Al(NO3)3 0.7 mL ,
shook up , laid 6min , added 10%NaOH 5 mL , finally
made the solution constant volume w ith 30%
ethanol , laid 10 min , determined the absorbency at
510 nm w ith corresponding reagents as blank solu-
tion.
2.3.2　Standard curve
Accurately weighting rutin that had been constant
w eight in the vacuum condition at 105 ℃27.90 mg
w as constant volume to 100 mL by ethanol to prepare
standard solution , accurately measuring different
quantity of standard solution detected absorbency ac-
co rding to the method in 2.3.1 , the regression equa-
tion of concentration C(mg/mL)against absorbency A
w as C =0.07853A +0.0002804 , R=0.9997 , the
linear range w as 0.01 ～ 0.06 mg/mL.
2.3.3　Experiment of standard addition recovery
The results of adding rutin to test solution of known
concentration w ere listed in table 1 , the average re-
covery w as103.3%, RSD = 3.70%.Through ana-
lyzing the results , we could see that the recovery w as
elevated as the accreting concentration of solution ,
these also show ed , in higher concentration , the both-
ers w ere bad , the results on the high side and the er-
rors bigger.
Table 1 　The recovery of standard addition obtained by col-
orimetry
S ample
quant ity(mL)
Rutin
Added(mL) Absorbency Recovery(%)
0.2 1.0 0.224 101.0
0.4 1.0 0.307 100.2
0.6 1.0 0.394 102.2
0.8 1.0 0.480 103.5
1.0 1.0 0.573 109.8
2.4 　Determining the content of flavonoids by
HPLC
2.4.1　Chromatog raphic conditions
Column , YWG C18(10 μm , 4.6 mm ×250 mm);
Flowing phase , V(methanol):V(0.4% o rthophos-
pho ric acid)=60∶40;Detecting w avelength 360 nm ;
Flow rate , 1 mL/min;Column temperature , room
temperature;The quantity of injecting sample ,10 μL.
Fig.1 　HPLC chromatog ram of standard samples(A)and
the extractive from Jasminum nudi florum Lindl.
(B)
2.4.2　Standard curve
Accurately w eighting Quercetin , Kaempferol ,
Isorhamnetin ,which had been constant w eight in the
vacuum condition at 105 ℃, 13.07 mg ,12.91 mg and
8.63 mg , were constant volume to 50 mL w ith
methanol to prepare their standard solutions respec-
tively.Accurately measuring each standard solut ion
0.2 ,0.5 ,1.0 ,2.0 and 4.0mL to five 10 mL measur-
4952003　Vo1.15　No.6 郑敏燕等:迎春花中黄酮类化合物含量的研究 　　　　　　　　　
ing flasks w as constant volume by methanol , each
w as sucked off 10 μL to carry out HPLC analysis in
terms of the condition in 2.4.1 , the figure of stan-
dard sample were show ed in fig.1(A), the peak a , b
and c belonged to Quercetin , Kaempferol and
Iso rhamnet in respectively , by reg ression analysis of
each peak area(A) against concentration(C , mg/
mL), the regression equation obtained w as listed in
table 2.
2.4.3　Preparing samples fo r HPLC analy sis
Through analy zing of T LC and HPLC , the ex traction
of J .nudi f lorum Lindl.had many flavonoids of dif-
ferent structures.These flavonoids had bigger
molecule and more complex structures , so i t w as dif fi-
cult to find standard samples to analyze one by one.
Present report showed[ 5～ 6] that the flavonoids in
plants w hich hydrolyzed by acid can produce a few
kinds of aglycones , we could analy ze these ag lycones
by HPLC to gain the content of flavonoids.From the
figure w e could see that the ag lycones obtained by hy-
drolyzing the ext ract ion of J .nudi f lorum Lindl.
mainly were quercetin , kaempferol , isorhamnetin.
By referring to concerned literature
[ 5～ 6]
and acco rd-
ing to the experimental fact , we selected the hy-
droly tic condiction below to prepare the HPLC ana-
ly zing samples of J .nudif lorum Lindl.:Taking
25mL test solution dried solvent before added 25%
HCl 5mL and methanol 30mL in it , then ref lux in
w ater bathe to remain acid hydrolysis one hour , af ter
being cold , the solution w as constant to 50 mL by
methanol , and then the analy zing sample was ob-
tained;f ilt rat ing the solution w ith 0.45μm microfil-
t ration membrane w as used to do HPLC , the figure of
samples w as fig.1(B)
2.4.4　Recovery experiment
Accurately sucking of f 10 mL test solution of known
content w as added 2.0 mL stardard samples of
quercet in , kaempferol and isorhamnetin respectively ,
then acco rding to 2.4.3 , we prepared them to be the
analyzing sample for HPLC , finally w e detected the
recovery .The results were showed in table 2.
Table 2　The average recovery of standard addition obtained by HPLC
S tandard
Sample
Regression equat ion R
Linear range
(μg/mL)
Average recovery
(%)(n=5) RSD(%)
Quercet in A=2.9014×106-1194 0.9998 5.228～ 104.6 98.1 1.45
Kaempferol A=2.7208×106-2969 0.9996 5.164～ 103.3 98.5 1.21
Isorhamnetin A=3.9514×106-5723 0.9997 3.452～ 69.04 101.2 1.69
3　Results
3.1　The results of qualitative test of flavonoids
The results show ed that the test solution of f low er
and stem of J .nudi f lorum Lindl.produced mauve
foam and the solution turned to mauve as reacted with
M g-HCl , the leaf produced o range foam and solution
as reacted w ith Mg-HCl , the three reacted with
NaOH turned the solution to yellow and w ith FeCl3
turned to dark green.These results all proved there
w ere flavones , flavonols and dihydrogen flavones etc
in J .nudi f lorum Lindl.[ 3].
3.2　Results of spectrophotometry
Taking the f low ers , leaves and stems determined the
content of f lavonoids according to the method in 2.3 ,
each sample w as determined three times.The results
were showed in table 3.
Table 3 　The content of to tal flavonoids in J.nudi florum
Lindl.determined by colorimetry
S ample Determining(%) Average(%) RSD(%)
Flow ers 13.4 , 13.6 , 14.1 13.7 2.63
Leaves 15.7 , 16.3 , 16.5 16.2 2.57
S tems 6.56 , 6.79 , 6.96 6.77 2.96
3.3　Results of HPLC analysis
Taking the flowers , leaves and stems determined the
content of f lavonoids according to the method in 2.4 ,
each sample w as determined three times.The results
were showed in table 4.
496　　　　　　　　　 天然产物研究与开发 2003　Vo1.15　No.6
Table 4　The average content o f flavonoid ag lycones in J.nudiflorum Lindl.determined by HPLC(n=3)
Sample Quercetin(%) Kaempferol(%) Isorhamnet in(%) Total aglycones(%) RSD(%)
Flow ers 2.573 0.2521 1.380 4.205 1.76
Leaves 0.2667 2.937 0.1676 3.371 1.58
Stems 0.6632 0.7663 0.2732 1.703 1.98
4　Discussion
4.1　The content w as higher by colorimetry than by
HPLC , the content of flavones detected by HPLC
was only 20%～ 30% of that detected by colorime-
try.The possible reasons were as follow s:① There
w ere many disturbs in colorimetry determination , in
alkalescence medium , many poly atomic phenols also
reacted w ith Al
3+
to change colors of solution so that
affected the accuracy of result , so the content deter-
mined by this method w as alw ays on the high side.
②For this plant maybe had other flavonoid ag lycones
besides quercetin , kaempferol , isorhamnetin , these
aglycones w ere difficult to be analyzed by HPLC fo r
the lack of standard samples.③What HPLC detected
w as the content of aglycones that w ere produced by
hydrolyzing flavonoids.If it was w nverted to the con-
tent of f lavonoids , the latter value w as almost tw ice
higher than the former[ 6] .
4.2 　 Results of HPLC showed quercetin and
isorhamnetin were higher in flowers , while kaempfer-
ol w as higher in leaves.
4.3 　The f low ers and leaves of J .nudi f lorum
Lindl.had abundance of flavonoids , whose content
w ere far higher than that of Ginkgo bi loba L ., Hip-
pophae rhamnoides , Glycine max(L .)Merr., their
content of flavonoids had repo rted in some literature.
For the many health pro tection and medical treatment
ef ficacies of flavonoids , it is need to do mo re research
to this natural source , J .nudif lorum Lindl.
References
1　 Ning Ke-fu.Eating Yingchunhua can cure some diseases.
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2　 Wang Qiu-an.The function and application of flavonoids.
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～ 33
3　 Phytochemistry Room of Shanghai Institute of Materia
Medica , CAS.Identification Handbook of Flavonoids.Bei-
jing:Science Press , 1981.389～ 292
4　 Xiang Zhao-bao , Ren Shao-guang , Shi Yi-song , et al.Ab-
sorptiophotometric determination of to tal flavones in stems
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605:41 ～ 48
迎春花中黄酮类化合物含量的研究
郑敏燕　魏永生
(咸阳师范学院化学系　咸阳　712000)
摘　要　以 75%乙醇为溶剂 ,索氏提取器提取 , 采用比色法和 HPLC 法研究了迎春花中黄酮类化合物
的含量。以芦丁为对照物 ,比色法测定出其花 、叶 、茎中总黄酮的含量分别为 13.7%、16.2%、6.77%,
RSD分别为 2.63%、2.57%、2.96%;以槲皮素 、山奈素 、异鼠李素为对照物 ,HPLC 法测定出其花 、叶 、茎
中的黄酮总量 ,以甙元计分别为 4.205%、3.371%、1.703%, RSD分别为 1.76%、1.58%、1.98%。
关键词　迎春花;黄酮;HPLC;比色法
4972003　Vo1.15　No.6 郑敏燕等:迎春花中黄酮类化合物含量的研究