Simultaneous Determination of Pyrrolidine Alkaloids and Their Nitrogen Oxides in Farfarae Flos by UPLC-MS/MS

Yue QIAO; Li-hua CHEN; Hong-xiu HU; Ran-ran GAO; Zhao-hui GAO; Jian-yong SI; Yong-hong LIAO; Di-an SUN

doi:10.13422/j.cnki.syfjx.20192013

您当前的位置：

首页 >

文章列表页 >

Simultaneous Determination of Pyrrolidine Alkaloids and Their Nitrogen Oxides in Farfarae Flos by UPLC-MS/MS

Resource and Quality Evaluation | 更新时间：2021-02-09

- Simultaneous Determination of Pyrrolidine Alkaloids and Their Nitrogen Oxides in Farfarae Flos by UPLC-MS/MS
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 1, Pages: 155-162(2020)
- 作者机构：
  
  中国医学科学院&北京协和医学院药用植物研究所，天然药物化学研究中心，北京　 100193
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20192013
  CLC： R289; R284.1; R22; R2-031
- Received：28 April 2019，
  
  Published Online：10 July 2019，
  
  Published：05 January 2020
- 稿件说明：
移动端阅览
Yue QIAO, Li-hua CHEN, Hong-xiu HU, et al. Simultaneous Determination of Pyrrolidine Alkaloids and Their Nitrogen Oxides in Farfarae Flos by UPLC-MS/MS[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(1): 155-162.
DOI：

Yue QIAO, Li-hua CHEN, Hong-xiu HU, et al. Simultaneous Determination of Pyrrolidine Alkaloids and Their Nitrogen Oxides in Farfarae Flos by UPLC-MS/MS[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(1): 155-162. DOI： 10.13422/j.cnki.syfjx.20192013.

摘要

目的：

建立一种同时测定15个吡咯里西啶生物碱(pyrrolidine alkaloids，PAs)及其氮氧化物的超高效液相色谱-串联质谱法(UPLC/MS/MS)，完成15批不同来源的款冬花样品的含量测定。从而初步了解不同来源款冬花中PAs的分布现状，为药材的安全合理使用提供相关参考。

方法：

采用Agilent Eclipse Plus C

色谱柱(3.0 mm×150 mm，1.8 μm)，流动相为含0.05%甲酸和2.5 mmol·L

-1

甲酸铵的水溶液(A)以及含0.05%甲酸和2.5 mmol·L

-1

甲酸铵的甲醇(B)，梯度洗脱，流速0.4 mL·min

-1

，进样量2 μL，柱温40 ℃。采用Agilent 1290-6470 QQQ型超高效液相色谱-三重四级杆质谱联用仪，离子化模式为ESI

＋

，多反应监测模式(MRM)。利用外标法计算药材样品中所测成分的含量，通过计算不同成分的相对标准偏差(RSD)对样品间的差异进行分析。

结果：

建立的测定方法灵敏度高，分离度良好，方法学考察结果良好。样品测定结果表明，15批药材均含有PAs及其氮氧化物，且所含PAs种类基本相同。克氏千里光碱(senkirkine)是款冬花中最主要的PAs类成分。药材款冬花中不同PAs的含量与分布有较大差异。

结论：

款冬花中普遍含有吡咯里西啶生物碱及其氮氧化物，且以具有显著肝毒性的克氏千里光碱为其中最主要种类。针对PAs的含量测定为确保款冬药材的用药安全提供了有效的保证。

Abstract

Objective:

To establish an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the simultaneous determination of 15 pyrrolidine alkaloids (PAs) and their nitrogen oxides

and determine the content of the 15 PAs in the 15 batches of Farfarae Flos samples obtained from different sources

in order to understand the distribution status of these 15 PAs in Farfarae Flos from different sources

and provide relevant references for the safe and rational use of this medicinal materials.

Method:

The method was achieved by Agilent Eclipse Plus C

column (3.0 mm×150 mm

1.8 μm) using a mobile phase made up of 0.05%formic acid and 2.5 mmol·L

-1

ammonium formate in water (A)-0.05%formic acid and 2.5 mmol·L

-1

ammonium formate in methanol(B). The flow rate and the injection volume were 0.4 mL·min

-1

and 2 μL

respectively. The column temperature was 40 ℃. The instrument was Agilent 1290-6470 QQQ ultra high performance liquid chromatography-triple quaternary bar mass spectrometer. The components were detected in multiple reaction monitoring mode by mass spectrometry with ionizationmode of ESI

. The content of the components measured in the samples was calculated by using the external standard method

and the difference between samples was analyzed based on RSD of different components.

Result:

The established method had a high sensitivity and good separation degree. The results of methodological investigation met the requirements. The results showed that all of the 15 batches of Farfarae Flos contained PAs and their nitrogen oxides. These PAs had almost the same types of structure. There were significant differences in the content and distribution of PAs in Farfarae Flos obtained from different sources.

Conclusion:

In general

Farfarae Flos contains pyrrolidine alkaloids and their nitrogen oxides. Senkirkine with a significant hepatotoxicity is the main compound. The content determination of PAs will provide scientific fundaments for the safe and effective use of Farfarae Flos.

关键词

Keywords

references

国家药典委员会．中华人民共和国药典：一部 [M]．北京：中国医药科技出版社， 2015 ： 332 .

H GAO ， Y N HUANG ， B GAO ， et al . α -Glucosidase inhibitory effect by the flower buds of Tussilago farfara L． [J]． Food Chem ， 2008 ， 106 ( 3 )： 1195 - 1201 .

H R Park ， M Y Yoo ， J H Seo ， et al . Sesquiterpenoids isolated from the flower buds of Tussilago farfara L. inhibit diacylglycerol acyltransferase [J]． J Agric Food Chem ， 2008 ， 56 ( 22 )： 10493 - 10497 .

L Joohee ， S Kwangho ， H Eugene ， et al . Neuroprotection against 6-OHDA toxicity in PC12 cells and mice through the Nrf2 pathway by a sesquiterpenoid from， Tussilago farfara [J]． Redox Biol ， 2018 ， 18 ： 6 - 15 .

P A G M D Smet ， K Keller , R Hänsel ， et al . Adverse Effects of Herbal Drugs [M]. Berlin ： Springer Berlin Heidelberg ， 1997 .

S Y XUE ， Z Y LI ， H J ZHI ， et al . Metabolic fingerprinting investigation of Tussilago farfara L. by GC-MS and multivariate data analysis [J]. Biochem Syst Ecol ， 2012 ， 41 ： 6 - 12 .

A Nedelcheva ， N Kostova ， A Sidjimov ． Pyrrolizidine alkaloids in Tussilago farfara from Bulgaria [J]． Biotechnol Biotechnol Equip ， 2015 ， 29 ( sup1 )： 1 - 7 .

C M Pabreiter . Co-occurrence of 2-pyrrolidineacetic acid with the pyrrolizidines tussilaginic acid and isotussilaginic acid and their 1-epimers in Arnica species and Tussilago farfara [J]． Phytochemistry ， 1992 ， 31 ( 12 )： 4135 - 4137 .

吴笛，张朝凤，张勉，等．中药款冬花的化学成分研究 [J]．中国药学杂志， 2008 ， 43 ( 4 )： 260 - 263 .

赵显国，王峥涛，张勉，等．肝毒吡咯里西啶生物碱与中草药 [J]．中草药， 1998 ， 29 ( 5 )： 341 - 346 .

G LIN ， J Y WANG ， N LI ， et al . Hepatic sinusoidal obstruction syndrome associated with consumption of Gynura segetum [J]． J Hepatol ， 2011 ， 54 ( 4 )： 666 - 673 .

J TANG , T Akao , N Nakamura , et al . In vitro metabolism of isoline，a pyrrolizidine alkaloid from Ligularia duciformis ，by rodent liver microsomal esterase and enhanced hepatotoxicity by esterase inhibitors [J]. Drug Metab Dispos , 2007 , 35 ( 10 )： 1832 .

Q XIA ， M W CHOU ， F F Kadlubar ， et al . Human liver microsomal metabolism and DNA adduct formation of the tumorigenic pyrrolizidine alkaloid，riddelliine [J]. Chem Res Toxicol ， 2003 ， 16 ( 1 )： 66 - 73 .

张芳，王长虹，王峥涛．植物中吡咯里西啶生物碱的检测与分析 [J]．天然产物研究与开发， 2006 ， 18 ( 6 )： 1057 - 1063 .

E Roeder . Medicinal plants in China containing pyrrolizidine alkaloids [J]． Pharmazie ， 2000 ， 55 ( 10 )： 711 - 726 .

濮社班，徐德然，张勉，等 . 中药款冬中肝毒吡咯里西啶生物碱的LC/MS n 检测 [J]．中国天然药物， 2004 ， 2 ( 5 )： 293 - 297 .

王明芳，李坤，孟祥龙，等 . 款冬花炮制前后总生物碱含量比较 [J]．中国药事， 2015 ， 29 ( 2 )： 178 - 182 .

K CAO ， Y XU ， X MU ， et al . Sensitive determination of pyrrolizidine alkaloids in Tussilago farfara L. by field-amplified，sample-stacking，sweeping micellar electrokinetic chromatography [J]． J Sep Sci ， 2016 ， 39 ( 21 )： 4243 - 4250 .

BfR . Determination of pyrrolizidine alkaloids (PA) in plant material by SPE-LC-MS/MS [EB/OL]． https://www.bfr.bund.de/en/pyrrolizidine_alkaloids_pa_-192924.html ， 2014-10-24/2019-04-26 .

J Luethy ， U Zweifel ， C Schlatter ， et al . Pyrrolizidine alkaloids in colts ( Tussilago farfara L．) of various sources [J]． Mitt Geb Lebensmittelunters Hyg Bern ， 1980 ， 71 ( 1 )： 73 .

Z JIANG ， F LIU ， J J L Goh ， et al . Determination of senkirkine and senecionine in Tussilago farfara using microwave-assisted extraction and pressurized hot water extraction with liquid chromatography tandem mass spectrometry [J]． Talanta ， 2009 ， 79 ( 2 )： 539 - 546 .

回连强，高双荣，刘婷，等 . 款冬花及其总生物碱的肝脏毒性 [J]．中国实验方剂学杂志， 2012 ， 18 ( 4 )： 238 - 241 .

钟云青，许光兰，王秀峰，等 . 款冬花散治疗支气管扩张症急性加重期痰热郁肺证的临床观察 [J]．中国实验方剂学杂志， 2018 ， 24 ( 20 )： 169 - 174 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Herbal Textual Research on Farfarae Flos in Famous Classical Formulas

Relativity Analysis of Rutin Content and Traditional Specification Drade of Farfarae Flos

Optimization of Toxicity Attenuation Processing Technology for Farfarae Flos Being Processed with Glycyrrhizae Radix et Rhizoma by Orthogonal Test

Hepatotoxicity on Water Extracts and the Total Alkaloid of Farfarae Flos

Related Author

ZHANG Dan

ZHAN Zhilai

ZHENG Yuguang

ZHANG Huikang

XUE Zijing

AN Qi

LI Hengyang

DING Xiaoying

Related Institution

School of Pharmacy，Traditional Chinese Medicine Processing Technology Innovation Center of Hebei Province，Hebei University of Chinese Medicine

Hebei Chemical and Pharmaceutical College

National Resource Center for Chinese Materia Medica，China Academy of Chinese Medical Sciences

Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences

AI问答

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰