基于HPLC-UV-MS技术的甘草和马钱子大鼠口服入血成分分析

张敏, 邓阳, 文雅洁, 方平飞, 罗世林, 王超

中国药学杂志 ›› 2017, Vol. 52 ›› Issue (9) : 731-737.

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中国药学杂志 ›› 2017, Vol. 52 ›› Issue (9) : 731-737. DOI: 10.11669/cpj.2017.09.004
论著

基于HPLC-UV-MS技术的甘草和马钱子大鼠口服入血成分分析

  • 张敏1,2, 邓阳1,2, 文雅洁3, 方平飞1,2*, 罗世林1,2, 王超1,2
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Characteristic Ingredients Profiling of Glycyrrhiza Radix and Semen Strychni in Rat Plasma Using HPLC-UV-MS

  • ZHANG Min1,2, DENG Yang1,2, WEN Ya-jie3, FANG Ping-fei1,2*, LUO Shi-lin 1,2, WANG Chao1,2
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摘要

目的 建立解毒剂量甘草和中毒剂量马钱子药材提取液混合物的高效液相色谱-紫外-串联质谱(HPLC-UV-MS)分析方法,并运用于甘草和马钱子大鼠口服入血成分分析。方法 采用Sino Chrome ODS-BP (4.6 mm×250 mm, 5 μm)色谱柱,水(含0.1%甲酸)(A)-乙腈(B) (V/V)为流动相,梯度洗脱:0.01 min 8% B,2 min 8% B,30 min 25% B,32 min 30% B,50 min 45% B,53 min 85% B,60 min 85% B,61 min 8% B,77 min停止,流速0.3 mL·min-1。UV检测波长254 nm,柱温40 ℃。质谱扫描采用正负离子模式同时检测的全扫描模式,正负离子模式锥孔电压分别30、25 V,毛细管电压分别为2.5、3.5 V。待测血浆用乙腈沉淀蛋白处理后测定。结果 建立了同时测定甘草和马钱子药材提取物及入血成分的HPLC-UV-MS方法,比较了甘草和马钱子体内外物质变化。结论 采用简便,可重复的HPLC-UV-MS方法对甘草和马钱子提取物体内外成分进行确证和分析,为甘草对马钱子解毒机制的物质基础研究提供实验依据。

Abstract

OBJECTIVE To establish a high performance liquid chromatography coupled with tandem mass spectrometry and ultraviolet detector (HPLC-UV-MS) method for determination of the mixture of Glycyrrhiza Radix extract at detoxification dosage and Semen Strychni extract at poisonous dosage, which will be applied to the characteristic ingredients profiling in rat plasma. METHODS Sino Chrome ODS-BP column (4.6 mm×250 mm, 5 μm) was employed, which was maintained at 40 ℃, and the mobile phase was composed of water (0.1% formic acid) (A) and acetonitrile (B). The gradient elution program was as follows: 0.01 min 8% B, 2 min 8% B, 30 min 25% B, 32 min 30% B, 50 min 45% B, 53 min 85% B, 60 min 85% B, 61 min 8% B, and terminated at 77 min, eluted at a flow rate of 0.3 mL·min-1. A UV detector was equipped and the detection wavelength was 254 nm. ESI+ mode was selected for strychnos alkaloids with cone voltage of 30 V and capillary voltage of 2.5 V, while ESI- mode for glycyrrhiza with cone voltage of 25 V and capillary voltage of 3.5 V. Acetonitrile was used as protein precipitation agent to determine the characteristic ingredients in rat plasma. RESULTS An HPLC-UV-MS method for simultaneous determination of liquorice and Semen Strychni extract was established and applied to measure the components absorbed into blood. The material changes of liquorice and Semen Strychni in vivo and vitro were also compared. CONCLUSION A simple, repeatable HPLC-UV-MS method for analysis of Glycyrrhiza Radix and Semen Strychni in vivo and vitro was established. The related components are speculates, which provided the experimental basis to research the detoxification mechanism of Glycyrrhiza Radix against strychninization.

关键词

甘草 / 马钱子 / 解毒 / 高效液相色谱-紫外-串联质谱 / 入血成分

Key words

Glycyrrhiza Radix / Strychni Semen / detoxification / HPLC-UV-MS / characteristic ingredients profiling

引用本文

导出引用
张敏, 邓阳, 文雅洁, 方平飞, 罗世林, 王超. 基于HPLC-UV-MS技术的甘草和马钱子大鼠口服入血成分分析[J]. 中国药学杂志, 2017, 52(9): 731-737 https://doi.org/10.11669/cpj.2017.09.004
ZHANG Min, DENG Yang, WEN Ya-jie, FANG Ping-fei, LUO Shi-lin, WANG Chao. Characteristic Ingredients Profiling of Glycyrrhiza Radix and Semen Strychni in Rat Plasma Using HPLC-UV-MS[J]. Chinese Pharmaceutical Journal, 2017, 52(9): 731-737 https://doi.org/10.11669/cpj.2017.09.004
中图分类号: R917   

参考文献

[1] LI Z H, YAN M, ZHANG B K, et al. Advances in study on compatibility of licorice and its mechanism of detoxification based on pharmacokinetics [J]. Chin Tradit Herb Drugs(中草药), 2015,46(23):3611-3616.
[2] GU L, WANG X, LIU Z, et al. A study of Semen Strychni-induced renal injury and herb-herb interaction of Radix Glycyrrhizae extract and/or Rhizoma Ligustici extract on the comparative toxicokinetics of strychnine and brucine in rats [J]. Food Chem Toxicol, 2014,68(6):226-233.
[3] REN J J,ZHANG X S,CHU Z J. Research of decreasing toxicity and increasing efficacy on Semen Strychni [J]. J Liaoning Univ Tradit Chin Med(辽宁中医药大学学报), 2016,18(1):221-224.
[4] YANG Y, YIN X J, GUO H M, et al. Identification and comparative analysis of the major chemical constituents in the extracts of single Fuzi herb and Fuzi-gancao herb-pair by UFLC-IT-TOF/MS [J]. Chin J Nat Med(中国天然药物), 2014,12(7):542-553.
[5] ZHOU B, ZHANG J, WU S, et al. The influence of compatibility of processed Radix Aconiti Kusnezoffii on the pharmacokinetic of four components in Glycyrrhiza uralensis Fisch [J]. J Ethnopharmacol, 2015,169(7):1-7.
[6] SHEN L, FENG Y, XUD S,et al. Studies on changes of in vitro and in vivo material base of Shaoyao Gancao Decoction based on HPLC-DAD-ESI-MS [J]. China J Chin Mater Med (中国中药杂志), 2010,35(15):1947-1952.
[7] LIN M X. study on dose-effect relationship and toxicity of Nux Vomica [D]. Harbin:Heilongjiang University of Chinese Medicine, 2002.
[8] SUN J F. Methodology of Animal Experimental(动物实验方法学)[M]. Beijing:People's Medical Publishing House,2001:125.
[9] JIA X X, LI W, LI J S, et al. Progressive studies on toxity of Strychons nux vomica[J]. China J Chin Mater Med(中国中药杂志), 2009,34(18):2396-2399.
[10] TIAN J, TIAN Y, XU L, et al. Characterisation and identification of dihydroindole-type alkaloids from processed Semen Strychni by high-performance liquid chromatography coupled with electrospray ionisation ion trap time-of-flight mass spectrometry [J]. Phytochem Anal, 2014,25(1):36-44.
[11] ZHANG J Y, ZHANG Q, ZHANG F, et al. Characterization four kinds of alkaloids in Strychnos nux-vomica by HPLC-ESI-MS-MS [J]. Chin J Exp Tradit Med Form(中国实验方剂学), 2013,19(9):147-151.
[12] LIU X Y. Study about metabolites of stiychnos alkaloids in vivo and their analysis method [D]. Shanghai:Fudan University, 2014.
[13] MONTERO L, IBEZ E, RUSSO M, et al. Metabolite profiling of licorice (Glycyrrhiza glabra) from different locations using comprehensive two-dimensional liquid chromatography coupled to diode array and tandem mass spectrometry detection [J]. Anal Chim Acta, 2016,913(12):145-159.
[14] LENG J, ZHU Y X, CHEN L L,et al. Two new triterpenoid saponins from roots and rhizomes of Glycyrrhiza uralensis [J]. Chin Tradit Herb Drugs(中草药), 2015,46(11): 1576-1582.
[15] CHEN P D, ZHOU X, DING A W. Constituents in aqueous extract of honey fried licorice by HPLC-MS [J]. Chin Tradit Pat Med(中成药), 2014,39(11):2115-2120.
[16] LIU Y C, CHEN Y G, WANG D, et al. Studies on chemical constituents on roots of Glycyrrhiza uralensis [J]. Chin J Pharm Anal(药物分析杂志), 2011,31(7):1251-1255.
[17] ZHANG W, SAIF M W, DUTSCHMAN G E, et al. Identification of chemicals and their metabolites from PHY906, a Chinese medicine formulation, in the plasma of a patient treated with irinotecan and PHY906 using liquid chromatography/tandem mass spectrometry (LC-MS/MS) [J]. J Chromatogr A, 2010,1217(37):5785-5793.
[18] CHEN S D, LU C J, ZHAO R Z. Identification and quantitative characterization of PSORI-CM01, a Chinese medicine formula for psoriasis therapy, by liquid chromatography coupled with an LTQ orbitrap mass spectrometer [J]. Molecules, 2015,20(1):1594-1609.
[19] WANG Q, MIAO W J, XIANG C, et al. Chemical constituents from Glycyrrhiza uralensis [J]. Chin Tradit Herb Drugs(中草药), 2012,43(10):1886-1890.
[20] LIU Y. Study on antibacterial activity of extractive and active constituent of glycyrrhizae [D]. Jilin:Yanbian University, 2013.
[21] GUO J F, CHEN X Y, ZHONG D F. Identification of strychnine, brucine and their metabolites in body fluids by liquid chromatography-electrospray ion trap mass spectrometry [J]. Chin J Pharm Anal(药物分析杂志), 2001,21(3):167-170.
[22] WANG Y H. Studies on HPLC Fingerprints of Semen Strychni and Radix Glycyrrhizae [D]. Tianjin:Tianjin University, 2006.
[23] DUAN T X. Studies on HPLC Fingerprints of Licorice and its application [D]. Beijing:Beijing University of Chinese Medicine, 2006.
[24] SUN L, JIN Y, LIU X Q, et al. HPLC Fingerprint of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle[J]. China J Chin Mater Med (中国中药杂志), 2014,39(11):2056-2059.

基金

国家自然科学基金资助项目(81473411,81573686)
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