全 文 ：第 51 卷 第 8 期
2 0 1 5 年 8 月
林 业 科 学
SCIENTIA SILVAE SINICAE
Vol. 51，No. 8
Aug．，2 0 1 5
doi:10．11707 / j．1001-7488．20150811
Ｒeceived date: 2014 － 06 － 30; Ｒevised date: 2014 － 12 － 15．
Foundation project: National Science and Technology Infrastructure Program (2012BAD23B03) ．
* Yue Yongde is corresponding author．
高效液相色谱法同时测定竹叶中 13种黄酮类化合物*
魏 琦1，2 王淑英1 汤 锋1 张华新2 喻 谨1 岳永德1
(1．国际竹藤中心竹藤科学与技术重点实验室 北京 100102; 2．国家林业局盐碱地研究中心 北京 100091)
摘 要: 【目的】建立一种同时测定竹叶中 13 种黄酮类化合物(异荭草苷、荭草苷、异牡荆苷、牡荆苷、芹菜素、
木犀草 素、苜 蓿 素、7 － 甲 氧 基 － 苜 蓿 素、苜 蓿 素 － 7 － O － 葡 萄 糖 苷、芹 菜 素 － 7 － O － 葡 萄 糖 苷、
Demethyltorosaflavone、芹菜素 － 7 － O －葡萄糖 － 6″ － O －鼠李糖苷、6 － 反式 － (2″ － O － α － 鼠李糖基)乙烯基 －
5，7，3，4 －四羟基黄酮)含量的高效液相色谱法，并用此方法对牡竹属 10 种竹种(麻竹、龙竹、牡竹、黄竹、梁山
慈竹、花吊丝竹、云南龙竹、福贡龙竹、勃氏甜龙竹、版纳甜龙竹)竹叶黄酮类化合物进行分析比较，以期为黄酮类
化合物的定性定量检测、牡竹属竹叶的开发利用及提取竹叶黄酮时的竹种选择提供参考。【方法】采用 HPLC
梯度洗脱分离黄酮类化合物，通过仪器精密度、LOD 值、LOQ 值、日内稳定性、日间稳定性及添加回收率对方法进
行验证。竹叶样品用 70%乙醇超声提取，经石油醚萃取后采用 HPLC 进行检测。【结果】各黄酮类化合物标准
品分离效果良好，保留时间在 13. 23 ～ 54. 67 min 之间。13 种黄酮类化合物标准品在 0. 01 ～ 500 mg·L － 1的线性
范围内呈良好的线性关系，相关系数 Ｒ2 在 0. 999 6 ～ 1. 000 0 之间。仪器精密度 ＲSD 在1. 06% ～ 2. 55% 之间，
LOD 值在 0. 01 ～ 0. 10 mg·kg － 1之间，LOQ 值在 0. 03 ～ 0. 34 mg·kg － 1之间。方法准确性、日间与日内精密度良好，
日内稳定性 ＲSD 在 0. 15% ～ 0. 67%之间，日间稳定性在 0. 44% ～ 5. 61%之间，13 种黄酮类化合较稳定。7 种黄
酮类化合物的添加回收率高于 70%，其余 6 种黄酮类化合物的添加回收率在 39. 76% ～ 68. 75% 之间。采用此方
法对牡竹属 10 种竹种竹叶黄酮类化合物进行分析，结果显示各竹种竹叶黄酮类化合物含量存在差异，除木犀草
素、6 －反式 － (2″ － O － α －鼠李糖基)乙烯基 － 5，7，3，4 －四羟基黄酮和 Demethyltorosaflavone 未在部分竹种中
检测到外，其余 10 种黄酮类化合物均在 10 种竹叶中检测到。13 种黄酮类化合物总含量在 419. 94 ～ 5 155. 84
mg·kg － 1之间，花吊丝竹叶中总黄酮含量最高，福贡龙竹含量最低。【结论】本文建立的同时测定竹叶中 13 种黄
酮类化合物含量的高效液相色谱法简便、快速、准确，牡竹属 10 种竹种竹叶中黄酮类化合物含量较丰富，有利于
开发利用。
关键词: HPLC; 黄酮类化合物; 竹叶; 牡竹属
中图分类号:S781. 41 文献标识码:A 文章编号:1001 － 7488(2015)08 － 0081 － 07
Simultaneous Determination of 13 Flavonoids in Bamboo Leaves by HPLC
Wei Qi1，2 Wang Shuying1 Tang Feng1 Zhang Huaxin2 Yu Jin1 Yue Yongde1
(1. Key Laboratory of Bamboo and Ｒattan，International Centre for Bamboo and Ｒattan Beijing 100102;
2. Ｒesearch Center of Saline and Alkali Land of State Forestry Administration Beijing 100091)
Abstract: 【Objective】Most flavonoids possess a variety of medical and biological activities such as anti-free radical，
antioxidant，antiaging，anti-inflammatory，etc． They are widely used in pharmaceutical，health products and cosmetic
fields． The objective of this paper was to establish a HPLC method for simultaneously qualitative-quantitative analysis of 13
flavonoids ( isoorientin， orientin， vitexin， isovitexin， apigenin， luteolin， tricin， 7-methoxy-tricin， tricin-7-O-
glucopyranoside， apigenin-7-O-glucopyranoside， demethyltorosaflavone， apigenin-7-O-glucopyranoside-6″-O-rhamnoside
and 6-trans-(2″-O-α-rhamnopyranosyl) ethenyl-5，7，3，4-tetrahydroxyflavone ) in bamboo leaves，and leaf flavonoids in
10 species of Dendrocalamus (D． latiflorus，D． giganteus，D． strictus，D． membranceus，D． farinosus，D． minor var．
amoenus，D． yunnanicus，D． fugongensis，D． brandisii and D． hamiltonii ) were comparatively analyzed by the
established method with the aims to provide a guidance for qualitative and quantitative analysis of flavonoids， the
development and utilization of Dendrocalamus bamboo leaves and the selection of bamboo species for flavonoids extraction．
林 业 科 学 51 卷
【Method】The flavonoids were separated by HPLC gradient elution method，and the validation was guaranteed by accuracy
of equipment，LOD，LOQ，intraday / interday precision and recovery rate． Bamboo leaves were extracted with 70% ( v / v)
ethanol-water by ultrasonic extraction，and then tested by HPLC after petroleum ether extraction． 【Ｒesult】A good
separation was found for each flavonoid，the retention times were ranged from 13． 23 to 54． 67 min． The standard curves
showed a good linearity in the corresponding ranges between 0． 01 and 500 mg·L － 1 ( Ｒ2≥0． 999 6) ． The equipment
accuracy ＲSD were 1． 06% － 2． 55%，LOD and LOQ ranged from 0． 01 to 0． 10 mg·kg － 1，and from 0． 03 to 0． 34 mg·
kg － 1，respectively． The ＲSD values of instrument accuracy， and intraday / interday precision indicated that it was
reasonable to analyze the samples within 5 days． The ＲSD of intraday and interday precisions were 0． 15% － 0． 67% and
0． 44% － 5． 61%，respectively． Ｒecovery rate for 7 kinds of flavonoids in spiked D． latiflorus were higher than 70%，and
that for the other 6 kinds of flavonoids were ranged from 39． 76% to 68． 75% ． The established HPLC method was valid in
application for 10 species of Dendrocalamus． The results exhibited differences in flavonoids content among the bamboo
leaves． As luteolin，6-trans-(2″-O-α-rhamnopyranosyl) ethenyl-5，7，3，4-tetrahydroxyflavone and demethyltorosaflavone
were observed in some bamboo species，the other 10 flavonoids were detected in all 10 kinds of bamboo leaves． The
concentrations of total 13 flavonoids were in the range of 419． 94 to 5 155． 84 mg·kg － 1 ． The maximum content of total
flavonoids was found in D． minor var． amoenus，and the minimum was in D． fugongensis． 【Conclusion】The established
HPLC method for simultaneously qualitative-quantitative analysis of 13 flavonoids in bamboo leaves was simple，sensitive
and accurate． The flavonoids content was rich in leaves of 10 bamboo species in Dendrocalamus genus which are potential
resources for exploitation and utilization．
Key words: HPLC; flavonoids; bamboo leaves; Dendrocalamus
Most flavonoids are found to have a variety of medical
and biological activities such as anti-free radical，
antioxidant， antitumor， antiviral， anti-inflammatory，
antibacterial，etc． activities and are widely used in food
and pharmaceutical fields ( Zhang，2008; Wang et al．，
2011) ． Bamboo is a renewable natural resource and a
vital component of forest resources ( Jiang， 2007 ) ．
Bamboo leaf flavonoids were found in many bamboo
species ( Guo，2007; Sun，2010; 2012 ) ． According to
reports，bamboo leaf flavonoids have an obvious fresh-
keeping effect on soybean milk (Wei et al．，2012) ． The
compound preservative of bamboo leaf flavonoids can
prolong the shelf life of chilled meat and improve
preservation effect (Shi et al．，2013) ．
Many methods such as spectrophotometry (SP)，high
performance liquid chromatography ( HPLC )， high
performance thin layer chromatography ( HPTLC ) have
been reported for the study of bamboo leaf flavonoids (Guo
et al．，2007; Wang et al．，2010; Sun et al．，2010a;
2010b) ． Most of the researches were focused on a few
flavonoids． Bamboo leaves contain a variety of flavonoids
(Guo，2007; Sun，2010; 2012; Wang et al．，2013 )，
and those methods of detecting a few flavonoids have
proved to be unsuitable for determination of other
flavonoids founded simultaneously． It was necessary for a
sensitive analytical technique for determining more
flavonoids content levels simultaneously． In this paper，
thirteen flavonoids ( isoorientin， orientin， vitexin，
isovitexin， luteolin， apigenin-7-O-glucoside-6″-O-
rhamnose， apigenin-7-O- glucopyranoside， tricin-7-O-
glucoside，6-trans-( 2″-O-α-rhamnopyranosyl ) ethenyl-5，
7， 3， 4-tetrahydroxyflavone， apigenin， tricin，
demethyltorosaflavone， 7-methoxytricin ) were studied．
According to reports， these flavonoids were founded in
bamboo leaves，and some of them are common ingredients
in bamboo leaves． We presented a simple， rapid and
sensitive HPLC method with UV detection for qualitative
and quantitative analysis of thirteen flavonoids in bamboo
leaves．
1 Materials and methods
1. 1 Plant materials
Bamboo leaves from ten species of Dendrocalamus，
Dendrocalamus latiflorus，D． giganteus，D． strictus，D．
membranaceus，D． farinosus，D． minor var． amoenus，D．
yunnanicus， D． fugongensis， D． brandisii and D．
hamiltonii，were collected in Changning Century Bamboo
Garden and Xishuangbanna Tropical Botanical Garden in
China． They were dried in shade and smashed with a
grinder． The comminution granularity was 60．
1. 2 Instrumental equipment
KQ-250B-type ultrasonic cleaner ( Kunshan
28
第 8 期 魏 琦等: 高效液相色谱法同时测定竹叶中 13 种黄酮类化合物
Ultrasonic Instrument Co．，Ltd) were used． HPLC was
performed with a Waters Corp． ( Milford，MA ) 2695
separations module and 2487 dual wavelength absorbance
detector．
1. 3 Flavonoids standards
Soorientin，orientin，vitexin，isovitexin and apigenin
( purity ＞ 98% ) were obtained from Shanghai Winherb
Medical Technology Co．， Ltd． ( Shanghai， China ) ．
Luteolin， apigenin-7-O-glucopyranoside-6″-O-rhamnoside，
apigenin-7-O-glucopyranoside，tricin-7-O-glucopyranoside，
6-trans-( 2″-O-α-rhamnopyranosyl ) ethenyl-5，7，3，4-
tetrahydroxyflavone， tricin，demethyltorosaflavone and 7-
methoxy-tricin were separated and prepared from bamboo
leaves in the State Forestry Administration Key Open
Laboratory，International Centre for Bamboo and Ｒattan
(Beijing，China) ．
1. 4 Ｒeagents
Analysis grade solvents were obtained from the Beijing
Chemical Works (Beijing，China) ． HPLC grade solvents
were obtained from Fisher Scientific (Pittsburgh，PA) ．
1. 5 Standard solutions preparation
The stock solution of flavonoids were prepared by
putting 3. 5 mg isoorientin， 3 mg 6-trans-( 2″-O-α-
rhamnopyranosyl ) ethenyl-5，7，3，4-tetrahydroxyflavone
and 5 mg 11 other flavonoids in 10 mL volumetric flasks
respectively，dissolving with different ratios of methanol，
ethanol and water，and diluting to volume with methanol．
Heated the flasks and dissolved with ultrasound in order to
promote dissolution． The mixed standard solution of the 13
flavonoids were prepared with 13 flavonoide stock solutions．
The concentrations of 13 flavonoids in the mixed standard
solution were isoorientin 26. 25 mg·L － 1，6-trans-(2″-O-α-
rhamnopyranosyl ) ethenyl-5，7，3，4-tetrahydroxyflavone
30 mg·L － 1 and 11 other flavonoids 37. 5 mg·L － 1 ． The
mixed standard solution was diluted to 0. 01 － 37. 5 mg·L － 1
with methanol． Solutions were stored at 4 ℃ ．
1. 6 HPLC analysis
HPLC was performed with a Waters Corp．
(Milford，MA ) 2695 separations module and 2487
dual wavelength absorbance detector． Compounds were
separated on a YMC-Pack Ｒ＆D ODS-A ＲP C18
column (4. 6 mm × 250 mm，5 μm particle size) ． The
mobile phases gradient consisted of acetonitrile and
0. 5% phosphoric acid， see Tab． 1． The injection
volume was 10 μL，the flow rate 1. 0 mL·min － 1，UV
detection wavelength at 340 nm．
Tab． 1 Method of gradient elution
Time /min Acetonitrile (% )
0. 5% phosphoric
acid (% )
Curve
0 15 85
15 20 80 11
20 20 80 6
50 45 55 6
51 80 20 11
58 15 85 11
65 15 85 11
1. 7 Validation of the method
A series concentration of the mixed standard solutions
were prepared to assess the linearity． Each of the different
concentrations of the solutions was injected using the
chromatographic conditions to generate corresponding
regression equations． The LOD and LOQ for the flavonoids
were measured by duplicate injections of the standard
solutions based on S /Ns of 3 and 10，respectively． The
precision of the method was checked by 5 replicate
injections of the mixed standard solutions and was
expressed as ＲSD． The intraday precision was studied by
injecting the mixed standard solutions five times on the
same day ( 0，2，4，8 and 12 h )，and the interday
precision was studied by injecting the mixed standard
solutions five times on 5 consecutive days． The results
were expressed as ＲSD． The accuracy of the method was
tested by obtaining recovery rates． A standard addition
recovery test of the method was carried out with a 2 g
sample of D． latiflorus， to which was added three
concentrations of mixed standard solutions． Three replicate
spiked samples were extracted，purified and analyzed by
HPLC． Average recovery rates were calculated． A blank
control was also analyzed．
1. 8 Sample preparation
Samples of 2. 0 g dry bamboo leaves were extracted
with 70% ( v / v) ethanol-water (3 × 30 mL; each time for
30 min) by ultrasonic extraction in 50 mL centrifuge tubes
at 60 ℃ ． The power of the ultrasonic instrument is
250 W． The extracts were filtered after extracting each
time． The filtrates were combined and evaporated to
dryness by vacuum distillation． The residues were
suspended with 50 mL water in separation funnels． After
being extracted with petroleum ether (3 × 50 mL)，the
aqueous solutions were evaporated to dryness with rotary
evaporator． The residues were dissolved with 70% ( v / v)
ethanol-water in 10 mL volumetric flasks and adjusted to
volume with 70% ( v / v) ethanol-water． All the samples
were filtrated by 0. 45 μm microporous membrane (Bonna-
38
林 业 科 学 51 卷
Agela Technologies ) before injection． Each bamboo
species were set three repetitions．
2 Ｒesults and discussion
2. 1 Optimization of the HPLC
The structures and polarities of some flavonoids are
similar，so it is difficult to separate those flavonoids． Of
the various mobile phases tried，acetonitrile and 0. 5%
phosphoric acid with gradient elution ( Tab． 1 ) gave the
best resolution of the 13 flavonoids． These flavonoids were
also separated from the other components in the sample
extracts． A typical chromatogram obtained from the
flavonoids standards is shown in Fig． 1．
Fig． 1 Chromatogram of the mixed standards
2. 2 Validation
The HPLC method was validated for linearity，
accuracy，LOD，LOQ，intraday and interday precision and
recovery． The linear relationships are given in Tab． 2． The
standard curves in the corresponding ranges had good
linearity． Accuracy，LOD，LOQ and intraday and interday
precision are presented in Tab． 3． LOD was in the range of
0. 01 to 0. 10 mg·kg － 1，and LOQ 0. 03 to 0. 34 mg·kg － 1 ．
The values of ＲSD for the accuracy and intraday and
interday precision indicated it was safe to analyze the
samples within 5 days． Ｒecoveries of flavonoids from spiked
D． latiflorus are shown in Tab． 4． According to the results，
the average recoveries of orientin， 6-trans-( 2″-O-α-
rhamnopyranosyl ) ethenyl-5，7，3，4-tetrahydroxyflavone，
apigenin， tricin， demethyltorosaflavone and 7-methoxy-
tricin were 39. 76% － 68. 75% ． The average recoveries of
the other 7 flavonoids were higher than 70% ． The method
was found to be feasible for qualitative and quantitative
determination of flavonoids in bamboo leaves．
Tab． 2 Linear relationships between peak area and concentration①
Names of standards
Ｒetention
time /min
Ｒegression
equation Ｒ
2 Linear range /
(mg·L － 1 )
Isoorientin 13. 23 Y = 30 059X － 567. 86 1. 000 0 0. 01 － 350. 00
Orientin 15. 43 Y = 25 947X － 720. 17 0. 999 9 0. 02 － 125. 00
Vitexin 20. 38 Y = 28 472X + 834. 72 0. 999 9 0. 02 － 500. 00
Isovitexin 20. 68 Y = 29 984X + 1275. 2 0. 999 9 0. 02 － 500. 00
Luteolin 22. 19 Y = 28 333X + 326. 6 0. 999 9 0. 02 － 37. 50
Apigenin-7-O-glucopyranoside-6″-O-rhamnoside 25. 60 Y = 20 559X + 382. 8 0. 999 9 0. 02 － 125. 00
Apigenin-7-O-glucopyranoside 27. 67 Y = 19 966X + 387. 47 0. 999 9 0. 02 － 37. 50
Tricin-7-O-glucopyranoside 28. 65 Y = 22 939X + 782. 65 0. 999 9 0. 02 － 37. 50
6-trans-(2″-O-α-rhamnopyranosyl)
ethenyl-5，7，3，4-tetrahydroxyflavone
32. 20 Y = 10 168X － 1 293. 7 0. 999 6 0. 02 － 30. 00
Apigenin 42. 45 Y = 45 372X － 1 039 1. 000 0 0. 02 － 37. 50
Tricin 43. 07 Y = 30 388X － 574. 26 1. 000 0 0. 02 － 37. 50
Demethyltorosaflavone 44. 83 Y = 28 411X － 1 396. 9 1. 000 0 0. 02 － 37. 50
7-methoxy-tricin 54. 67 Y = 36 008X － 1 071. 5 0. 999 9 0. 02 － 18. 75
①Y = Peak area and X = compound concentration．
48
第 8 期 魏 琦等: 高效液相色谱法同时测定竹叶中 13 种黄酮类化合物
Tab． 3 LOD，LOQ and precision for the 13 flavonoids
Names of standards
LOD /
(mg·kg － 1 )
LOQ /
(mg·kg － 1 )
Accuracy
［ＲSD (% )，
n = 5］
Intraday
precision
［ＲSD (% )，
n = 5］
Interday
precision
［ＲSD (% )，n = 5］
Isoorientin 0. 03 0. 09 1. 90 0. 15 0. 45
Orientin 0. 02 0. 06 2. 23 0. 17 0. 44
Vitexin 0. 01 0. 04 1. 93 0. 29 3. 74
Isovitexin 0. 01 0. 04 1. 53 0. 25 5. 61
Luteolin 0. 01 0. 03 1. 91 0. 20 0. 59
Apigenin-7-O-glucopyranoside-6″-O-rhamnoside 0. 02 0. 07 1. 34 0. 19 1. 99
Apigenin-7-O-glucopyranoside 0. 01 0. 04 1. 06 0. 18 1. 86
Tricin-7-O-glucopyranoside 0. 01 0. 03 1. 79 0. 16 0. 98
6-trans-(2″-O-α-rhamnopyranosyl) ethenyl-
5，7，3，4-tetrahydroxyflavone
0. 10 0. 34 2. 16 0. 24 3. 15
Apigenin 0. 01 0. 04 1. 82 0. 37 0. 88
Tricin 0. 01 0. 04 1. 49 0. 67 1. 72
Demethyltorosaflavone 0. 01 0. 04 1. 08 0. 24 1. 50
7-methoxy-tricin 0. 01 0. 03 2. 55 0. 49 3. 02
Tab． 4 Ｒecovery of 13 flavonoids from spiked D． latiflorus (n = 3)
Names of standards
Original
quantity /
(mg·kg － 1 )
Addition
quantity /
(mg·kg － 1 )
Amount
found /
(mg·kg － 1 )
Ｒecovery
(% )
ＲSD
(% )
Isoorientin 8. 06 26. 25 31. 72 82. 75 7. 21
8. 06 19. 69 28. 13 88. 53 10. 20
8. 06 13. 13 22. 49 89. 21 11. 56
Orientin 99. 17 37. 50 115. 59 62. 06 22. 14
99. 17 28. 13 112. 79 63. 76 25. 83
99. 17 18. 75 115. 16 68. 75 3. 95
Vitexin 45. 45 37. 50 80. 32 76. 00 4. 28
45. 45 28. 13 69. 25 71. 91 1. 68
45. 45 18. 75 61. 41 70. 65 3. 23
Isovitexin 136. 55 37. 50 211. 20 87. 15 3. 06
136. 55 28. 13 205. 76 88. 33 5. 16
136. 55 18. 75 208. 32 93. 17 3. 84
Luteolin 6. 38 37. 50 48. 28 102. 57 4. 16
6. 38 28. 13 25. 29 67. 09 1. 10
6. 38 18. 75 20. 13 71. 06 7. 83
Apigenin-7-O-glucopyranoside-6″-O-rhamnoside 244. 69 37. 50 308. 25 76. 20 2. 22
244. 69 28. 13 291. 63 73. 80 3. 38
244. 69 18. 75 281. 68 73. 01 2. 63
Apigenin-7-O-glucopyranoside 67. 22 37. 50 107. 50 77. 72 2. 80
67. 22 28. 13 95. 64 74. 17 6. 21
67. 22 18. 75 89. 08 74. 50 9. 20
Tricin-7-O-glucopyranoside 37. 23 37. 50 75. 39 80. 76 2. 35
37. 23 28. 13 65. 09 77. 52 10. 82
37. 23 18. 75 54. 63 73. 23 19. 43
6-trans-(2″-O-α-rhamnopyranosyl) ethenyl- 12. 19 30. 00 27. 03 55. 97 2. 62
5，7，3，4-tetrahydroxyflavone 12. 19 22. 50 21. 87 53. 63 37. 52
12. 19 15. 00 29. 56 88. 80 24. 13
Apigenin 0. 91 37. 50 18. 42 47. 40 1. 37
0. 91 28. 13 14. 24 48. 28 3. 67
0. 91 18. 75 9. 80 48. 73 8. 19
Tricin 5. 38 37. 50 23. 06 50. 59 11. 49
5. 38 28. 13 17. 40 48. 06 7. 02
5. 38 18. 75 18. 35 68. 40 13. 08
Demethyltorosaflavone 0. 00 37. 50 19. 89 53. 05 2. 53
0. 00 28. 13 15. 70 55. 84 11. 54
0. 00 18. 75 8. 33 44. 44 5. 47
7-methoxy-tricin 5. 97 37. 50 23. 61 50. 82 7. 97
5. 97 28. 13 14. 74 39. 76 12. 92
5. 97 18. 75 17. 54 63. 30 35. 04
58
林 业 科 学 51 卷
2. 3 Determination of flavonoids in bamboo leaves
The 13 flavonoids in 10 bamboo leaf species of
Dendrocalamus were compared by use of the proposed method．
The results are shown in Tab． 5． The chromatogram of an extract
of D． latiflorus is shown in Fig． 2． The results obtained reveal
differences in the flavonoid composition of the different bamboo
species． Luteolin，6-trans- (2″-O-α-rhamnopyranosyl)ethenyl-
5，7，3，4-tetrahydroxyflavone and demethyltorosaflavone were
not found in some bamboo species，and the other 10 flavonoids
were found in all of 10 bamboo species． 12 flavonoids were
found in D． latiflorus，D． farinosus，D． yunnanicus and D．
hamiltonii; 10 flavonoids were found in D． brandisii; 11
flavonoids were found in the other 5 bamboo species． The
concentrations of isoorientin，orientin，vitexin，isovitexin and
apigenin-7-O-glucoside-6″-O-rhamnose were more than 100 mg·
kg －1，which were higher than the contents of other flavonoids
in 10 bamboo leaves． The contents of apigenin-7-O-
glucopyranoside and tricin-7-O-glucoside in most bamboo
species were 20 －100 mg·kg －1 ． The concentrations of luteolin，
6-trans-( 2″-O-α-rhamnopyranosyl ) ethenyl-5， 7， 3， 4-
tetrahydroxyflavone， apigenin， tricin， demethyltorosaflavone
and 7-methoxytricin were less than 20 mg·kg －1 ．
The content of apigenin-7-O-glucoside-6″-O-rhamnose
was higher than other flavonoids in the leaves of D．
latiflorus，at a level of 263. 67 mg·kg － 1 ． In the leaves of
D． giganteus， D． strictus， D． membranceus， D．
fugongensis， D． brandisii and D． hamiltonii， the
concentrations of vitexin were maximum， they were
893. 34，702. 81，1 862. 60，98. 63，357. 69 and 330. 12
mg·kg － 1，respectively． In the leaves of D． farinosus，D．
minor var． amoenus and D． yunnanicus，the concents of
isovitexin were maximum，they were 352. 37，1 996. 40，
233. 87 mg·kg － 1， respectively． In the 10 species of
bamboo leaves，the concentrations of total flavonoids were
in the range of 419. 94 to 5 155. 84 mg·kg － 1 ; the
maximum content was found in D． minor var． amoenus．
Tab． 5 The contents of flavonoids in the dry leaves of Dendrocalamus①
Compounds
Flavonoids contents in the leaves of Dendrocalamus /(mg·kg － 1 )
D．
latiflorus
D．
giganteus
D．
strictus
D．
membranceus
D．
farinosus
D． minor var．
amoenus
D．
yunnanicus
D．
fugongensis
D．
brandisii
D．
hamiltonii
Isoorientin
9. 23 ±
0. 85a
148. 60 ±
11. 79b
9. 79 ±
0. 08a
55. 55 ±
1. 22c
58. 39 ±
0. 20c
1 582. 60 ±
2. 73d
35. 04 ±
0. 11e
55. 45 ±
0. 30c
28. 23 ±
1. 13e
217. 15 ±
1. 47f
Orientin
102. 47 ±
6. 73a
406. 19 ±
5. 64b
20. 81 ±
0. 58c
162. 85 ±
3. 97d
31. 84 ±
0. 64e
389. 98 ±
0. 81f
81. 55 ±
0. 60g
31. 72 ±
0. 57e
28. 67 ±
0. 33ce
97. 10 ±
2. 68a
Vitexin
42. 00 ±
2. 00a
893. 34 ±
2. 79b
702. 81 ±
0. 14c
1 862. 60 ±
1. 06d
44. 75 ±
0. 08a
286. 22 ±
2. 62e
62. 09 ±
0. 31f
98. 63 ±
0. 56g
357. 69 ±
0. 29h
330. 12 ±
1. 23i
Isovitexin
141. 85 ±
8. 71a
143. 23 ±
2. 12a
122. 82 ±
1. 14b
239. 72 ±
2. 28c
352. 37 ±
0. 98d
1 996. 40 ±
2. 27e
233. 87 ±
0. 64c
81. 12 ±
0. 40f
144. 63 ±
0. 39a
187. 07 ±
0. 28g
Luteolin
6. 38 ±
0. 54a
33. 62 ±
1. 68b
9. 93 ±
0. 17aef
58. 76 ±
0. 89c
8. 98 ±
0. 29af
98. 93 ±
2. 45d
13. 49 ±
0. 18e
/ /
11. 37 ±
0. 63ef
Apigenin-7-O-
glucopyranoside-6″-
O-rhamnoside
263. 67 ±
15. 44a
264. 19 ±
4. 98a
54. 82 ±
0. 74b
37. 53 ±
3. 26c
201. 18 ±
1. 46d
598. 12 ±
6. 30e
206. 51 ±
0. 33d
75. 34 ±
1. 74f
15. 38 ±
0. 15g
24. 02 ±
1. 07cg
Apigenin-7-O-
glucopyranoside
67. 22 ±
2. 18a
46. 20 ±
0. 91b
48. 40 ±
0. 79b
20. 73 ±
1. 34c
85. 51 ±
0. 52d
96. 39 ±
3. 29e
58. 49 ±
0. 21f
30. 36 ±
0. 51g
19. 33 ±
0. 16c
18. 36 ±
0. 97c
Tricin-7-
O-glucopyranoside
37. 23 ±
1. 52a
35. 43 ±
1. 55a
21. 17 ±
1. 28bf
117. 51 ±
2. 46c
48. 08 ±
0. 36d
89. 23 ±
4. 72e
43. 05 ±
0. 21d
18. 95 ±
0. 86b
16. 64 ±
0. 05b
25. 55 ±
1. 51f
6-trans-(2″-O-α-
rhamnopyranosyl)
ethenyl-5，7，3，4-
tetrahydroxyflavone
12. 19 ±
0. 41a
/ / /
7. 06 ±
0. 31b
/
23. 51 ±
0. 86c
13. 78 ±
0. 22a
/ /
Apigenin
0. 91 ±
0. 11a
4. 96 ±
0. 14b
3. 53 ±
0. 01c
5. 25 ±
0. 15d
1. 65 ±
0. 04e
3. 73 ±
0. 05c
0. 83 ±
0. 02a
1. 05 ±
0. 04a
2. 37 ±
0. 03f
0. 88 ±
0. 10a
Tricin
5. 38 ±
0. 62a
7. 84 ±
0. 29b
7. 57 ±
0. 28b
18. 58 ±
0. 52c
3. 88 ±
0. 08d
7. 86 ±
0. 23b
4. 19 ±
0. 09d
5. 71 ±
0. 16a
11. 95 ±
0. 77e
8. 06 ±
0. 13b
Demethyltorosaflavone / / / / / / / / /
4. 00 ±
0. 04
7-methoxy-tricin
5. 97 ±
0. 50af
11. 89 ±
0. 39b
8. 23 ±
0. 19c
17. 19 ±
0. 29d
3. 76 ±
0. 12e
6. 38 ±
0. 13f
7. 44 ±
0. 02g
7. 83 ±
0. 07cg
11. 75 ±
0. 09b
5. 32 ±
0. 01a
Total flavonoids
①Values are mean ± S． E． ( n = 3) ． Values in a low followed by same letters are not significantly different (P = 0. 05) according to Duncan’s
multiple comparison (DMＲT) ． “ /”: Not detected．
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第 8 期 魏 琦等: 高效液相色谱法同时测定竹叶中 13 种黄酮类化合物
Fig． 2 Chromatogram of an extract of D． latiflorus
3 Conclusions
An HPLC method for simultaneous qualitative and
quantitative analysis of 13 flavonoids was established． The
method was simple， sensitive and accurate． The results
obtained showed that the standard curves in the corresponding
ranges had good linear relationships． LOD was in the range of
0. 01 to 0. 10 mg·kg － 1，and LOQ 0. 03 to 0. 34 mg·kg － 1 ． The
values of ＲSD for the accuracy and intraday and interday
precision indicated it was safe to analyze the samples within 5
days． The average recoveries of orientin，6-trans-(2″-O-α-
rhamnopyranosyl ) ethenyl-5，7，3，4-tetrahydroxyflavone，
apigenin，tricin，demethyltorosaflavone and 7-methoxy-tricin
from D． latiflorus were 39. 76% － 68. 75% ; the average
recoveries of the other 7 flavonoids were higher than 70% ．
Bamboo leaves of ten species of Dendrocalamus were
extracted and analyzed． The results obtained reveal
differences in the flavonoids of the different bamboo species．
In 10 dry bamboo leaves，the concentrations of isoorientin，
orientin，vitexin，isovitexin and apigenin-7-O-glucoside-6″-
O-rhamnose were generally higher; followed with apigenin-7-
O- glucopyranoside and tricin-7-O-glucoside; luteolin，6-
trans-( 2″-O-α-rhamnopyranosyl ) ethenyl-5， 7， 3， 4-
tetrahydroxyflavone，apigenin，tricin，demethyltorosaflavone
and 7-methoxy-tricin had lower contents． The maximum
concentration of total flavonoids was found in D． minor var．
amoenus，the minimum in D． fugongensis． The proposed
method would be useful for the research on flavonoids and
chemical utilization of bamboo leave resources．
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(责任编辑 石红青)
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