Objective: To establish a real-time direct analysis mass spectrometry (DART-MS) method for the rapid characterisation and determination of Strychnos alkaloids in processed Strychni Semen and its preparations. Methods: The DART-MS characteristic spectrum of processed Strychni Semen was collected in the positive ion mode by full scan, using direct analysis in real time ionization source combined with ion trap mass spectrometry. And comparing the exact mass with the reported chemical constituents of Strychni Semen to confirm the Strychnos alkaloids of processed Strychni Semen. Internal standard method and monocrotaline was used as the internal standard to determine the content of brucine and strychnine in traditional Chinese patent medicines containing Strychni Semen. The distance between the ceramic tube of DART ion source to the MS inlet was 7.5 mm, the sample was centered, the capillary temperature was set to 350 ℃, the gas heating temperature was 350 ℃, the capillary voltage was 10 V, and the tube lens voltage was 80 V. Results: Different alkaloids were identified by analyzing the DART-MS spectrum of processed Strychni Semen. Brucine showed good linear relationships within the ranges of 62.80-3 140.00 ng·mL-1, the r was 0.999 5, and the average recovery rate was 93.8%. The linear range of strychnine was 68.00-3 400.00 ng·mL-1, the r was 0.998 8, and the average recovery rate was 89.1%. The limit of detection (LOD) and limit of quantitation (LOQ) of brucine were 1.57 and 3.14 ng·mL-1, respectively. The LOD and LOQ of strychnine were 1.70 and 3.40 ng·mL-1, respectively. The contents of brucine and strychnine in processed Strychni Semen, Yaotongning capsules, Pingxiao capsules, Fengshi Maqian tablets, Shangke Jiegu tablets, Shufeng Dingtong pills, Shujin pills, Shenjindan capsules, Shenjin Huoluo pills, Shufeng Huoluo pills, Fenghan Shuangliguai capsules were 0.03-9.07 mg·g-1, 0.14-20.25 mg·g-1, respectively. Conclusion: The method is simple, rapid, high in sensitivity and low in reagent consumption, can be used for the rapid characterisation and determination of brucine and strychnine in various traditional Chinese patent medicines containing Strychni Semen.
[1] 孙子洲, 曹兰秀. 基于药理作用的马钱子七情配伍规律研究[J]. 中药药理与临床, 2020, 36(2):257
SUN ZZ, CAO XL. Study on the Qiqing compatibility of Strychni Semen based on pharmacological effect[J]. Pharmacol Clin Chin Mater Med, 2020, 36(2):257
[2] 吴小娟, 马凤森, 郑高利, 等. 马钱子吲哚类生物碱毒性研究进展[J]. 中药药理与临床, 2016, 32(6):231
WU XJ, MA FS, ZHENG GL, et al. Toxicity research progress on indole alkaloids of Strychni Semen[J]. Pharmacol Clin Chin Mater Med, 2016, 32(6):231
[3] 巫坚, 张磊, 冯小兵. 服用马钱子自杀中毒死亡法医学分析1例[J]. 广东公安科技, 2020, 28(4):74
WU J, ZHANG L, FENG XB. Forensic analysis of one case of suicide by Strychni Semen poisoning[J]. Guangdong Public Secur Technol, 2020, 28(4):74
[4] AHMAD S, SALEEM K, KAMAL A, et al. Simultaneous HPTLC determination of strychnine and brucine in strychnos nux-vomica seed[J]. J Pharm Bioallied Sci, 2012, 4(2):134
[5] ZHANG J, WANG S, CHEN X, et al. Capillary electrophoresis with field-enhanced stacking for rapid and sensitive determination of strychnine and brucine[J]. Anal Bioanal Chem, 2003, 376(2):210
[6] 张云静, 张自品, 张雪燕, 等. 制马钱子及其制剂中士的宁和马钱子碱的含量测定[J]. 广州化工, 2019, 47(14):121
ZHANG YJ, ZHANG ZP, ZHANG XY, et al. Determination of strychnine and brucine in processed Strychni Semen and its preparation[J]. Guangzhou Chem Ind, 2019, 47(14):121
[7] 段存贤, 郭承军. HPLC法测定神农丸中士的宁与马钱子碱的含量[J]. 药物分析杂志, 2018, 38(2):331
DUAN CX, GUO CJ. Determination of strychnine and brucine in Shennong pills by HPLC[J]. Chin J Pharm Anal, 2018, 38(2):331
[8] MADDISETTY PNP, DOSS VA, MOHANASUNDARAM S, et al. HPLC and GC-MS: Identification and quantification of strychnine and brucine in Strychnos nux-vomica leaves[J]. Pak J Pharm Sci, 2017, 30[6(Suppl)]: 2369
[9] 陈玉华, 卢松. UPLC-QQQ-MS/MS法同时测定风湿安泰片中12种成分[J]. 中成药, 2021, 43(3):596
CHEN YH, LU S. Simultaneous determination of twelve constituents in Fengshi Antai tablets by UPLC-QQQ-MS/MS[J]. Chin Tradit Pat Med, 2021, 43(3):596
[10] 蓝晓庆, 韦瑀龙, 陆兰娟. LC-MS/MS法测定跌打酒中9种成分及主成分分析[J]. 实用药物与临床, 2021, 24(1):67
LAN XQ, WEI YL, LU LJ. Determination of 9 constituents in Die Da Jiu by LC-MS/MS and analysis of its principal component[J]. Pract Pharm Clin Remed, 2021, 24(1):67
[11] 韦立志, 李发娟, 巫丽丽. UPLC-MS/MS快速测定4种抗风湿类中成药中马钱子碱与士的宁的含量[J]. 中国医院药学杂志, 2020, 40(2):178
WEI LZ, LI FJ, WU LL. Rapid determination of strychnine and strychnine in 4 kinds of anti-rheumatic Chinese patent medicines by UPLC-MS/MS[J]. Chin J Hosp Pharm, 2020, 40(2):178
[12] 郭玉岩, 马文保, 肖洪彬, 等. 基于UPLC-MS技术分析马钱子-甘草药对配伍汤液不同相态中毒效物质的变化规律[J]. 中草药, 2017, 48(23):4880
GUO YY, MA WB, XIAO HB, et al. Changes of toxic substance in different phase of compatibility decoction Strychni Semen and Glycyrrhizae Radix et Rhizoma based on UPLC-MS technique[J]. Chin Tradit Herb Drugs, 2017, 48(23):4880
[13] GUO TY, YONG W, JIN Y, et al. Applications of DART-MS for food quality and safety assurance in food supply chain[J]. Mass Spectrom Rev, 2015, 36(2):161
[14] ABE H, TAKEI C, SAKAKURA M, et al. Comprehensive drug screening by thermal desorption and pyrolysis combined with direct analysis in real time-mass spectrometry(TDP/DART-MS)[J]. Methods Mol Biol, 2018, 1810: 115
[15] KARIN KN, POKLIS JL, PEACE MR. Evaluation of extraction methods for pharmacologically active compounds from anticonvulsant traditional Chinese medicines: Gou Teng, Tian Ma, Jiang Can using DART-TOF-MS[J]. Anal Methods, 2021, 13(7):884
[16] LEI YT, LU Y, ZHANG TC, et al. Rapid screening of testosterone in the aquatic environment using direct analysis in real-time (DART) mass spectrometry[J]. Environ Earth Sci, 2016, 75(12):1
[17] PAVLOVICH MJ, MUSSELMAN B, HALL AB. Direct analysis in real time-mass spectrometry (DART-MS) in forensic and security applications[J]. Mass Spectrom Rev, 2018, 37(2):171
[18] FORBES TP, VERKOUTEREN JR. Forensic analysis and differentiation of black powder and black powder substitute chemical signatures by infrared thermal desorption-DART-MS[J]. Anal Chem, 2019, 91(1):1089
[19] 李先强, 闫志, 张玲,等. 原位电离实时直接分析串联质谱法鉴别中药泽泻化学成分[J]. 化学分析计量, 2021, 30(3):14
LI XQ, YAN Z, ZHANG L, et al. Identification of chemical constituents of Alisma Orientailsin in real time by tandem mass spectroscopy[J]. Chem Anal Meterage, 2021, 30(3):14
[20] CHEN Z, YANG Y, TAO H, et al. Direct analysis in real-time mass spectrometry for rapid identification of traditional Chinese medicines with coumarins as primary characteristics[J]. Phytochem Anal, 2017, 28(3):137
[21] WANG Y, LI C, HUANG L, et al. Rapid identification of traditional Chinese herbal medicine by direct analysis in real time (DART) mass spectrometry[J]. Anal Chim Acta, 2014, 845: 70
[22] CHEN YL, LI LN, XIONG F, et al. Rapid identification and determination of pyrrolizidine alkaloids in herbal and food samples via direct analysis in real-time mass spectrometry[J]. Food Chem, 2021, 334: 127472
[23] GUO R, WANG T, ZHOU G, et al. Botany, phytochemistry, pharmacology and toxicity of Strychnos nux-vomica L.: a review[J]. Am J Chin Med, 2018, 46(1):1
[24] 张加余, 张倩, 张凡,等. HPLC-ESI-MS-MS鉴定马钱子中4类生物碱成分[J]. 中国实验方剂学杂志, 2013, 19(9):147
ZHANG JY, 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, 2013, 19(9):147
[25] TIAN JX, WANG M, XU L, et al. Metabolism of brucine: the important metabolic pathways of dihydroindole-type alkaloid for excretion in rats[J]. Bioanalysis, 2014, 6(2):137
[26] TIAN JX, 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
