权帆(1997年生),女,硕士研究生,研究方向为植物资源开发及利用研究。E-mail:
为了解药用植物大叶藤黄(
To study the chemical constituents of
大叶藤黄(
现代植物化学研究表明,大叶藤黄的化学成分为苯甲酮、黄酮、双黄酮、三萜和多异戊烯基呫吨酮等化合物[
AVANCE-600型核磁共振波谱仪(德国Bruker公司);紫外光谱仪Evolution 300紫外-可见光谱仪(Thermo Fisher Scientific);TLC (薄层层析)硅胶板和柱色谱硅胶(青岛海洋化工有限公司);MCI GEL (日本三菱公司);Sephadex LH-20葡聚糖凝胶(美国安玛西公司);超高液相-四极杆飞行时间质谱联用仪(UPLC-QTOF-MS, Xevo G2, 美国沃特世科技有限公司);Synergy MX酶标仪(BIOTEK公司);色谱纯甲酸和乙腈(美国Thermo Fisher公司);其余试剂均为分析纯分析级试剂(广州化学试剂厂)。
本研究所用大叶藤黄采自云南西双版纳,经华南农业大学羊海军博士鉴定为藤黄科藤黄属植物大叶藤黄(
将叶片用80%体积分数甲醇提取得到的浸膏, 均匀分散在适量纯水中,依次用等体积石油醚、氯仿和乙酸乙酯萃取后,减压浓缩获得各个萃取层的提取物。其中,乙酸乙酯层浸膏(450 g)进行硅胶柱色谱分离,以不同比例的二氯甲烷-甲醇(100:0→ 0:100)进行梯度洗脱,得到11个组分Fr.1~Fr.11。取组分Fr.4 (16 g)过MCI柱,得到组分Fr.4-1,Fr.4-1经硅胶柱层析,二氯甲烷-甲醇体系(40:1→8:1)梯度洗脱,得到29个子馏分(Fr.4-1-1~Fr.4-1-29)。馏分Fr.4-1-12经Sephadex LH-20纯化得化合物
将分离得到的化合物配制成质量浓度为5、10、20、40、60、80、100
利用UPLC-QTOF-MS对大叶藤黄叶片进行化学成分分析,得到叶片提取物中主要成分的保留时间和准分子离子峰,此外,还从MSE数据分析模式中获得所有化合物的碎片离子或二级质谱。通过准确分子量,MSE二级质谱与公共质谱数据库(METLIN,MassBank,ReSpect等)及藤黄属植物化合物的文献比较,鉴定化合物的可能结构或类型。初步从大叶藤黄叶片中鉴定了19个化合物,包括双黄酮类、黄酮类和间苯三酚类化合物(
化合物
Structures of compounds
大叶藤黄叶片化合物UPLC-QTOF-MS检测结果
Chemical constituent information of
峰号 |
保留时间 |
[M + H]+/[M-H]– | 离子碎片MSE fragment ions | 化合物 |
分类 |
|||||
M.F. | ppm | M.F. | ppm | |||||||
1 | 0.490 | 191.056 4 | C7H11O6 | 4.2 | - | - | - | 3-Furanacetic acid, tetrahydro- |
- | |
- | - | - | - | - | - | |||||
2 | 0.656 | - | - | - | - | - | - | Eucryphin[ |
黄酮 | |
339.072 6 | C15H15O9 | 2.9 | - | - | - | Flavonoid | ||||
3 | 2.200 | - | - | - | - | - | - | Xanthochymusside[ |
双黄酮 | |
719.160 9 | C36H31O16 | –0.4 | 593.129 3 | C30H25O13 | –0.3 | Biflavonoid | ||||
431.075 7 | C24H15O8 | –2.3 | ||||||||
295.024 6 | C16H7O6 | 1.0 | ||||||||
4 | 2.269 | 719.157 0 | C36H31O16 | –5.8 | 557.108 0 | C30H21O11 | –0.7 | 褔木苷[ |
双黄酮 | |
431.071 0 | C24H15O8 | 0.9 | Biflavonoid | |||||||
403.082 6 | C23H15O7 | 2.0 | ||||||||
717.145 9 | C36H29O16 | 0.4 | 565.133 2 | C29H25O12 | –2.5 | |||||
429.060 0 | C24H13O8 | –2.3 | ||||||||
5 | 2.435 | 721.172 6 | C36H33O16 | –6.0 | - | - | - | Xanthochymusside[ |
双黄酮 |
|
719.160 5 | C36H31O16 | –1.0 | 593.127 7 | C30H25O13 | –3.0 | |||||
431.075 3 | C24H15O8 | –3.2 | ||||||||
295.024 3 | C16H7O6 | 0.0 | ||||||||
6 | 2.469 | 575.117 7 | C30H23O12 | –2.3 | 431.076 2 | C24H15O8 | –1.2 | Buchananiflavanone[ |
双黄酮 | |
403.081 8 | C23H15O7 | 0.0 | Biflavonoid | |||||||
297.041 2 | C16H9O6 | 4.4 | ||||||||
573.103 7 | C30H21O12 | 0.7 | 447.070 6 | C24H15O9 | 0.2 | |||||
429.060 3 | C24H13O8 | –2.3 | ||||||||
295.024 0 | C16H7O6 | –1.0 | ||||||||
7 | 2.653 | 559.123 0 | C30H23O11 | –1.8 | - | - | - | GB-2a[ |
双黄酮 | |
557.108 5 | C30H21O11 | 0.2 | 447.072 8 | C24H15O9 | 2.7 | Biflavonoid | ||||
429.060 6 | C24H13O8 | –0.9 | ||||||||
8 | 2.733 | 557.108 9 | C30H21O11 | 0.9 | 431.077 6 | C24H15O8 | 2.1 | 藤黄双黄酮[ |
双黄酮 |
|
403.082 9 | C23H15O7 | 2.7 | ||||||||
555.092 4 | C30H19O11 | –0.5 | - | - | - | |||||
9 | 2.785 | 559.122 6 | C30H23O11 | –2.5 | - | - | - | GB-2a[ |
双黄酮 | |
557.107 7 | C30H21O11 | –1.3 | 429.059 7 | C24H13O8 | –2.1 | Biflavonoid | ||||
401.065 2 | C23H13O7 | –2.2 | ||||||||
295.024 0 | C16H7O6 | –0.3 | ||||||||
10 | 2.917 | 543.127 0 | C30H23O10 | –3.9 | - | - | - | GB 1a[ |
双黄酮 | |
541.112 9 | C30H21O10 | –1.1 | 413.064 3 | C24H13O7 | –4.4 | Biflavonoid | ||||
385.069 8 | C23H13O6 | –3.6 | ||||||||
267.028 8 | C15H7O5 | –1.9 | ||||||||
11 | 2.969 | 539.097 0 | C30H19O10 | –1.5 | 415.082 1 | C24H15O7 | 0.7 | 穗花杉双黄酮[ |
双黄酮 | |
387.087 9 | C23H15O6 | 2.6 | Biflavonoid | |||||||
537.081 4 | C30H17O10 | –1.5 | 443.039 2 | C24H11O9 | –2.5 | |||||
12 | 3.382 | 553.112 3 | C31H21O10 | –2.2 | 431.075 2 | C24H15O8 | –3.5 | 苏铁双黄酮[ |
双黄酮 | |
403.044 5 | C22H11O8 | –2.2 | Biflavonoid | |||||||
551.097 7 | C31H19O10 | –0.2 | 429.060 2 | C24H13O8 | –1.9 | |||||
331.062 1 | C20H11O5 | 4.5 | ||||||||
13 | 3.48 | 595.142 8 | C30H27O13 | –4.0 | - | - | - | 银椴甙[ |
黄酮 | |
593.127 4 | C30H25O13 | –3.5 | - | - | - | Flavonoid | ||||
14 | 3.864 | 295.192 6 | C17H27O4 | 5.8 | - | - | - | Tanacetol A[ |
苯甲酮 | |
293.176 5 | C17H25O4 | 4.1 | - | - | - | Benzophenone | ||||
15 | 5.080 | 301.143 4 | C18H21O4 | –2.0 | - | - | - | Bractebiphenyl B[ |
联苯 | |
- | - | - | - | - | - | Doitungbiphenyl A[ |
Biphenyl | |||
16 | 6.628 | 503.337 3 | C30H47O6 | 0.0 | - | - | - | Garcinielliptone E[ |
三萜 | |
501.322 0 | C30H45O6 | 0.8 | 389.195 5 | C22H29O6 | –2.3 | Triterpenoid | ||||
275.128 4 | C16H19O4 | 0.4 | ||||||||
233.081 0 | C13H13O4 | –1.7 | ||||||||
17 | 7.059 | 485.322 0 | C30H45O5 | –9.7 | - | - | - | Garcinielliptone F[ |
间苯三酚 | |
483.311 4 | C30H43O5 | 0.8 | 329.174 7 | C20H25O4 | –1.8 | Phloroglucinol | ||||
287.128 4 | C17H19O4 | 0.3 | ||||||||
18 | 7.294 | 517.353 0 | C31H49O6 | 0.2 | 539.333 6 | C31H48O6Na | –2.4 | Garcihombronane L[ |
弗里多烷 | |
375.217 2 | C22H31O5 | 0.3 | Friedolanostane | |||||||
291.161 1 | C17H23O4 | 5.2 | ||||||||
515.337 6 | C31H47O6 | 0.6 | 403.211 0 | C23H31O6 | –2.7 | |||||
289.144 0 | C17H21O4 | 0.0 | ||||||||
247.096 9 | C14H15O4 | –0.4 | ||||||||
19 | 7.690 | 499.341 8 | C31H47O5 | –1.0 | - | - | - | Garcinielliptone T[ |
间苯三酚 | |
497.326 6 | C31H45O5 | –0.2 | 343.190 6 | C21H27O4 | –0.9 | Phloroglucinol | ||||
301.143 9 | C18H21O4 | –0.3 |
大叶藤黄叶片提取物的负态总离子图。1~19见
TIC chromatography (negative mode) of leaf extracts of
通过与文献对比以及分析离子断裂途径,共鉴定出2个黄酮和10个双黄酮类化合物。峰2和13由准分子离子峰
褔木苷(峰4)的推断过程。A: 褔木苷质谱裂解途径; B: MSE质谱图谱(正态)。
Inference process of fukugiside (peak 4). A: Proposed fragmentation pathways for fukugiside, B: MSE spectra (positive mode).
在初步分离鉴定叶片中化合物的过程中,通过数据库比较及推断离子片段裂解途径,共鉴定出2个间苯三酚类化合物。峰17由准分子离子峰
garcinielliptone F (峰17)的推断过程。A: garcinielliptone F质谱裂解途径; B: MSE质谱图谱(负态)。
Inference process of garcinielliptone F (peak 17). A: Proposed fragmentation pathways for garcinielliptone F; B: MSE spectra (negative mode).
从大叶藤黄中分离得到的化合物
化合物1和5对DPPH自由基的清除作用
The scavenging effect of compounds 1 and 5 on DPPH
本研究采用UPLC-QTOF-MS对大叶藤黄叶片的主要化学成分进行分析,尝试性地鉴定出19个化合物,主要为双黄酮类、黄酮类和间苯三酚类化合物,其中,峰1、2、5、13~19为首次从该植物中报道。现代药理研究证明,黄酮类化合物具有抗肿瘤、抗自由基、抗氧化等作用[
据报道,化合物
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