Development of an Analytical Method for Kasugamycin Residue in Herbal Medicine, Achyranthes japonica Nakai

Jeong Yoon Choi(1), Hun Ju Ham(2), Min-woo Kim(3), Abd Elaziz Sulieman Ahmed Ishag(4), Jang-Hyun Hur(5*)

(1) Department of Biological Environment, Kangwon National University, Chuncheon 24341, Republic of Korea
(2) Environment Friendly Agricultural Products Safety Center, Chuncheon 24341, Republic of Korea
(3) Department of Biological Environment, Kangwon National University, Chuncheon 24341, Republic of Korea
(4) Department of Biological Environment, Kangwon National University, Chuncheon 24341, Republic of Korea Department of Crop Protection, University of Khartoum, Khartoum North, Shambat 13314, Sudan
(5) Department of Biological Environment, Kangwon National University, Chuncheon 24341, Republic of Korea
(*) Corresponding Author


This study developed a suitable analytical method for kasugamycin residues in Achyranthes japonica using LC/MS/MS equipped with an amide column for polar substances. Extraction and cleanup processes were done at pH 4.5–5. Purification efficiency was assessed and confirmed step by step by selecting silica, hydrophilic-lipophilic balance (HLB), strong cation exchange (SCX), and double (HLB and SCX) cleanup SPE cartridges. The results indicated that silica SPE cartridge exhibited overloading tendency, while HLB SPE cartridge had low cleaning efficiency. Among SPE cartridges used, double cleanup and SCX were found sufficient with respective matrix effects of –15% and +14%, respectively. The LOD and LOQ were 0.008 ng and 0.04 mg/kg, respectively. The correlation coefficient (R2) was higher than 0.99, recovery rate ranges were 86.3–97.2%, and the RSD was below 8.8%. All methods are consistent with the Codex guidelines criteria. This study developed an appropriate LC/MS/MS analytical method for kasugamycin residue analysis in A. japonica with optimized, efficient extraction and purification conditions using a single SCX SPE cartridge, which is simple and time-efficient. In addition, the HLB and SCX SPE cartridges of the double cleanup methods were identified as primary methods that can be applied for the cleanup of other medicinal herbs.


LC-MS/MS analysis; Kasugamycin residues; Achyranthes japonica; SPE-cartridges


[1] Na, E.S., Kim, S.S., Hong, S.S., Kim, K.J., Lee, Y.J., Lee, B.C., and Lee, K.S., 2020, Development of Multi-residue analytical method for 261 pesticides in herbal medicines using GC-MS/MS and LC-MS/MS, Korean J. Environ. Agric., 39 (2), 142–169.

[2] Kang, M.Y., Lee, S.H., Lee, S.W., Cha, S.W., Song, J.L., and Lee, S.C., 2015, Effect of Achyranthis radix and Drynariae rhizoma extracts on antioxidant activity and antioxidant enzymes, Korean J. Plant Resour., 28 (5), 600–607.

[3] Rural Development Administration, 2021, Pesticide Safety Information System (PSIS), Rural Development Administration, Jeonju, Korea,

[4] Kim, S.H., and Park, Y.H., 2008, Antimicrobial resistance and food safety, Safe Food, 3 (1), 30–36.

[5] Park, J.S., Do, J.A., Lee, H. S., Park, S.M., Cho, S.M., Shin, H.S., Jang, D.E., Cho, M.S., Jung, Y.H., and Lee, K., 2019, Development of analytical method for detection of fungicide validamycin A residues in agricultural products using LC-MS/MS, J. Food Hyg. Saf., 34 (1), 22–29.

[6] Kim, T.J., Kim, K.S., Yoon, C.H., Joo, J.B., and Kim, C.H., 1997, Analysis of kasugamycin in pesticides formulations by reversed-phase ion pair liquid chromatography, Anal. Sci. Technol., 10 (5), 343–349.

[7] Li, W., Dai, X., Pu, E., Bian, H., Chen, Z., Zhang, X., Guo, Z., Li, P., Li, H., Yong, Y., Wang, C., Zhang, Y., and Han, L., 2020, HLB-MCX-based solid-phase extraction combined with liquid chromatography-tandem mass spectrometry for the simultaneous determination of four agricultural antibiotics (kasugamycin, validamycin A, ningnanmycin, and polyoxin b) residues in plant-origin foods, J. Agric. Food Chem., 68 (47), 14025–14037.

[8] Zhang, H., Wang, C., Li, H., Nie, Y., Fang, L., and Chen, Z., 2017, Simultaneous determination of kasugamycin and validamycin-A residues in cereals by consecutive solid-phase extraction combined with liquid chromatography-tandem mass spectrometry, Food Addit. Contam., Part A, 35 (3), 487–497.

[9] Isoherranen, N., and Soback, S., 1999, Chromatographic methods for analysis of aminoglycoside antibiotics, J. AOAC Int., 82 (5), 1017–1045.

[10] Lu, L., Zhao, S., Deng, L., Chen, Y., Liu, X., and Li., D., 2012, Residues and dynamics of kasugamycin in chilli and soil, Bull. Environ. Contam. Toxicol., 89 (3), 649–653.

[11] Alechaga, É., Moyano, E., and Galceran, M.T., 2015, Simultaneous analysis of kasugamycin and streptomycin in vegetables by liquid chromatography-tandem mass spectrometry, Anal. Methods, 7 (8), 3600–3607.

[12] Lee, H.S., Do, J.A., Park, J.S., Cho, S.M., Shin, H.S., Jang, D.E., Jung, Y.H., and Lee, K., 2019, Development of analytical method for kasugamycin in agricultural products using LC-MS/MS, J. Food Hyg. Saf., 34 (3), 235–241.

[13] Ministry of Food and Drug Safety Guidelines (MFDSG), 2016, Standard Procedure for Preparing Testing Methods for Food, etc. Test Guidelines, South Korea’s Ministry of Food and Drug Safety.

[14] Turner, J.A., 2015, The Pesticide Manual, 17th Ed., BPC Publications, Alton, Hampshire, UK.

[15] Zhou, W., Yang, S., and Wang, P.G., 2017, Matrix effects and application of matrix effect factor, Bioanalysis, 9 (23), 1839–1844.

[16] Ministry of Food and Drug Safety (MFDS), 2013, Pesticide Analytical Residues Manual in Food Code, 13–14.

[17] Chemicalize, 2021, Kasugamycin,, accessed on 23 August 2021.

[18] Arsand, J.B., Jank, L., Martins, M.T., Hoff, R.B., Barreto, F., Pizzolato, T.M., and Sirtori., C., 2016, Determination of aminoglycoside residues in milk and muscle based on a simple and fast extraction procedure followed by liquid chromatography coupled to tandem mass spectrometry and time of flight mass spectrometry, Talanta, 154, 38–45.

[19] Wang, C., Li, H., Wang, N., Li, H., Fang, L., Dong, Z., Du, H., Guan, H., Zhu, Q., Chen, Z., and Yang, G., 2017, Simultaneous analysis of kasugamycin and validamycin-1 A in fruits and vegetables using liquid chromatography-tandem mass spectrometry and consecutive solid-phase extraction, Anal. Methods, 9 (4), 634–642.

[20] New, L.S. and Chan, E.C.Y., 2008, Evaluation of BEH C18, BEH HILIC, and HSS T3 (C18) column chemistries for the UPLC–MS–MS analysis of glutathione, glutathione disulfide, and ophthalmic acid in mouse liver and human plasma, J. Chromatogr. Sci., 46 (3), 209–214.

[21] Asakawa, D., Uemura, M., Sakiyama, T., and Yamano, T., 2017, Sensitivity enhancement of aminoglycosides in hydrophilic interaction liquid chromatography with tandem mass spectrometry by post-column addition of trace sodium acetate in methanol, Food Addit. Contam., Part A, 35 (6), 1116–1126.

[22] Ministry of Food and Drug Safety (MFDS), 2017, Guideline for Pesticide Residue Analysis Practice in the Korean Food Public Code, 4th Ed, Osong, Republic of Korea.

[23] Kim, J.H., Moon, S.E., Kim, K.Y., Jung, Y.J., Lee, C.H., Ku, E.J., Yoon, M.H., and Lee, J.B., 2016, Simultaneous analysis for veterinary drug residues in honey by HPLC/MS/MS, J. Food Hyg. Saf., 31 (2), 94–98.

[24] Fontanals, N., Zohar, J., Borrull, F., Ronka, S., and Marcé, R.M., 2021, Development of a maleic acid-based material to selectively solid-phase extract basic compounds from environmental samples, J. Chromatogr. A, 1647, 462165.

[25] Fontanals, N., Marcé, R.M., and Borrull, F., 2019, Materials for solid-phase extraction of organic compounds, Separations, 6 (4), 56.

[26] Choi, J.Y., Lee, Y.J., Ham, H.J., Ishag, A.E.S.A., and Hur, J.H., 2021, A study on improvement of the analytical method of chlorantraniliprole residue in herbal medicine (Rehmannia glutinosa Libosch) using HPLC-UVD, Korean J. Pestic. Sci., 25 (3), 196–211.

[27] Bierman, S., and Campognone, M.V., 2009, Simplifying solid-phase extraction method development: exploring the use of software, LCGC, 27 (8), 24–26.

[28] Joint FAO/WHO Codex Alimentarius Commission, 2003, Codex Alimentarius: Guidelines on Good Laboratory Practice in Pesticide Residue Analysis, World Health Organization: Food and Agriculture Organization of the United Nations, Rome, CAC/GL 40-1993, 1–36.


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