AJAC  Vol.13 No.9 , September 2022
LC-MS/MS Analysis of Lycorine and Galantamine in Human Serum Using Pentafluorophenyl Column
Abstract: Lycorine and galantamine are natural alkaloids found in Amaryllidaceae plants, such as narcissus. Narcissus leaves and roots are sometimes accidentally ingested because they resemble vegetables. Lycorine and galantamine are toxic and cause such effects as nausea, vomiting, and abdominal pain, when accidentally ingested. In a case of narcissus poisoning, the detection of lycorine and galantamine in biological samples is vital to determine whether they have been ingested. This study establishes a liquid chromatography-tandem mass spectrometry (LC/MS/MS) method to measure the lycorine and galantamine content of human serum, which can be used for mild to fatal poisoning cases. A serum pretreatment procedure was performed using acetonitrile and QuEChERS AOAC powder. The separation of the compounds was conducted using a pentafluorophenyl column, CAPCELL CORE PFP (2.1 mm I.D. × 100 mm, 2.7 μm). Lycorine, galantamine, and galantamine-d6 (internal standard) were identified by the transitions of m/z 288 → 147, m/z 288 → 213, and m/z 294 → 216, respectively. The calibration curves were linear in the ranges of 0.05 to 5 ng/mL and 5 to 100 ng/mL, with R2 > 0.999. The precision and accuracy were within the permissible range. The matrix effects of lycorine and galantamine were 94.3% - 98.4% and 87.8% - 91.1%, respectively. The extraction recovery rates of lycorine and galantamine were 101.9% - 112.7% and 95.6% - 107.1%, respectively. The present method detected lycorine and galantamine in the sera of three patients with mild poisoning that had accidentally ingested. This method is applicable in cases of lycorine and galantamine poisoning.
Cite this paper: Sasaki, C. , Shinozuka, T. , Yoshimura, K. , Maruhashi, T. , Asari, Y. and Satoh, F. (2022) LC-MS/MS Analysis of Lycorine and Galantamine in Human Serum Using Pentafluorophenyl Column. American Journal of Analytical Chemistry, 13, 300-313. doi: 10.4236/ajac.2022.139021.

[1]   Kretzing, S., Abraham, G., Seiwert, B., Ungemach, F.R., Krügel, U. and Regenthal, R. (2011) Dose-Dependent Emetic Effects of the Amaryllidaceous Alkaloid Lycorine in Beagle Dogs. Toxicon, 57, 117-124.

[2]   Kretzing, S., Abraham, G., Seiwert, B., Ungemach, F.R., Krügel, U., Teichert, J. and Regenthal, R. (2011) In Vivo Assessment of Antiemetic Drugs and Mechanism of Lycorine-Induced Nausea and Emesis. Archives of Toxicology, 85, 1565-1573.

[3]   Miyazaki, S. (2014) Narcissus. Japanese Journal of Clinical Toxicology, 27, 17-19.

[4]   Campbell, A. (1988) Poisoning in Small Animals from Commonly Ingested Plants. In Practice, 20, 587-591.

[5]   Saxon, B.S. (2004) Daffodil Toxicosis in an Adult Cat. Canadian Veterinary Journal, 45, 248-250.

[6]   Toda, M., Uneyama, C. and Kasuga, F. (2014) Trends of Plant Toxin Food Poisonings during the Past 50 Years in Japan. Food Hygiene and Safety Science, 55, 55-63.

[7]   Ageta, K., Yakushiji, H., Kosaki, Y., Obara, T., Nojima, T., Gochi, A., Naito, H. and Nakao, A. (2020) A Family Intoxicated by Daffodil Bulbs Mistaken for Onions. Acute Medicine & Surgery, 7, Article No. e595.

[8]   Matulkova, P., Gobin, M., Evans, M., Parkyn, P.C., Palmer, C. and Oliver, I. (2012) Gastro-Interstinal Poisoning Due to Consumption of Daffodils Mistaken for Vegetables at Commercial Markets, Bristol, United Kingdom. Clinical Toxicology, 50, 788-790.

[9]   Niitsu, H., Fujita, Y., Kumagai, R., Fujita, S., Nakamura, Y., Sujiura, T., Yoshida, T. and Dewa, K. (2017) An Autopsy Case of Death Following Accidental Ingestion of Narcissus. Research and Practice in Forensic Medicine, 60, 47-53.

[10]   Katoch, D. and Sharma, U. (2019) Simultaneous Quantification and Identification of Amaryllidaceae Alkaloids in Narcissus tazetta by Ultra Performance Liquid Chromatography-Diode Array Detector-Electrospray Ionization Tandem Mass Spectrometry. Journal of Pharmaceutical and Biomedical Analysis, 175, Article No. 112750.

[11]   Goto, T., Ozeki, F., Aoyama, F., Ito, Y., Ueno, E. and Ikai, Y. (2015) The Simultaneous Analysis of Vegetable Natural Poison (Lycorine and Convallatoxin) That Causes Food Poisoning. In: Report of Aichi Prefectural Institute of Public Health, Aichi Prefectural Institute of Public Health, Aichi, No. 65, 31-38.

[12]   Oishi, A., Nagatomi, Y. and Suzuki, K. (2019) Simultameous LC-MS/MS Determination of 18 Plant Toxins in Beverages. Food Hygiene and Safety Science, 60, 108-112.

[13]   Liu, X., Hong, Y., He, Q. and Huang, K. (2015) Rapid and Sensitive HPLC-MS/MS Method for Quantitative Determination of Lycorine from the Plasma of Rats. Journal of Chromatography B, 974, 96-100.

[14]   Gray, N., Heaton, J., Musenga, A., Cowan, D.A., Plumb, R.S. and Smith, N.W. (2013) Comparison of Reversed-Phase and Hydrophilic Interaction Liquid Chromatography for the Quantification of Ephedrines Using Medium-Resolution Accurate Mass Spectrometry. Journal of Chromatography A, 1289, 37-46.

[15]   Kudo, K., Usumoto, Y., Usui, K., Hayashida, M., Kurisaki, E., Saka, K., Tsuji, A. and Ikeda, N. (2014) Rapid and Simultaneous Extraction of Acidic and Basic Drugs from Human Whole Blood for Reliable Semi-Quantitative NAGINATA Drug Screening by GC-MS. Forensic Toxicology, 32, 97-104.

[16]   (2014) Scientific Working Group for Forensic Toxicology (SWGTOX) Standard Practices for Method Validation in Forensic Toxicology. Journal of Analytical Toxicology, 37, 452-474.

[17]   Yıldırım, S., Karakoç, H.N., Yaşar, A. and Köksal, I. (2020) Determination of Levofloxacin, Ciprofloxacin, Moxifloxacin and Gemifloxacin in Urine and Plasma by HPLC-FLD-DAD Using Pentafluorophenyl Core-Shell Column: Application to Drug Monitoring. Biomedical Chromatography, 34, Article No. E4925.

[18]   Hussain, A., AlAjmi, M.F. and Ali, I. (2018) Supramolecular Separation Mechanism of Pentafluorophenyl Column Using Ibuprofen and Omeprazole as Markers: LC-MS and Simulation Study. Biomedical Chromatography, 32, Article No. E4206.

[19]   González-Rubio, S., Ballesteros-Gómez, A., Carreras, D., Munoz, G. and Rubio, S. (2021) A Comprehensive Study on the Performance of Different Retention Mechanisms in Sport Drug Testing by Liquid Chromatography Tandem Mass Spectrometry. Journal of Chromatography B, 1178, Article ID: 122821.

[20]   Ploumen, C., Marginean, I. and Lurie, I.S. (2020) The Utility of Silica Hydride-Based Stationary Phases for Dual-Mode Ultra High Performance Liquid Chromatography Separation of Synthetic Cathinone Positional Isomers. Journal of Separation Science, 43, 3449-3457.

[21]   Chidella, K.S., Dasari, V.B. and Anireddy, J. (2021) Simultaneous and Trace Level Quantification of Five Potential Genotoxic Impurities in Ranolazine Active Pharmaceutical Ingredient Using LC-MS/MS. American Journal of Analytical Chemistry, 12, 1-14.

[22]   Anastassiades, M. and Lehotay, S.J. (2003) Fast and Easy Multiresidue Method Employing Acetonitrile Extraction/Partitioning and “Dispersive Solid-Phase Extraction” for the Determination of Pesticide Residues in Produce. Journal of AOAC International, 86, 412-431.

[23]   Lehotay, S.J. (2007) Determination of Pesticide Residues in Foods by Acetonitrile Extraction and Partitioning With Magnesium Sulfate: Collaborative Study. Journal of AOAC International, 90, 485-520.

[24]   Usui, K., Hayashizaki, Y., Hashiyada, M. and Funayama, M. (2012) Rapid Drug Extraction from Human Whole Blood Using a Modified QuEChERS Extraction Method. Legal Medicine, 14, 286-296.

[25]   Usui, K., Hashiyada, M., Hayashizaki, Y., Igari, Y., Hosoya, T., Sakai, J. and Funayama, M. (2014) Application of Modified QuEChERS Method to Liver Samples for Forensic Toxicological Analysis. Forensic Toxicology, 32, 139-147.

[26]   Hasegawa, K., Wurita, A., Minakata, K., Gonmori, K., Yamagishi, I., Nozawa, H., Watanabe, K. and Suzuki, O. (2015) Identification and Quantitation of 5-Fluoro-ADB, One of the Most Dangerous Synthetic Cannabinoids, in the Stomach Contents and Solid Tissues of a Human Cadaver and in Some Herbal Products. Forensic Toxicology, 33, 112-121.