Determination of 2, 4-epibrassinolide residues in Dendrobium officinale by solid phase extraction-high performance liquid chromatography-mass spectrometry

doi:10.3969/j.issn.1004-1524.20221050

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (8): 1896-1903.DOI: 10.3969/j.issn.1004-1524.20221050

• Quality and Safety of Agricultural Products • Previous Articles Next Articles

Determination of 2, 4-epibrassinolide residues in Dendrobium officinale by solid phase extraction-high performance liquid chromatography-mass spectrometry

LU Lanfei¹^,²(), ZHAO Xueping², MA Zheng¹, FANG Nan², LUO Yuqin², WANG Xiaomei¹^,², YE Hui², LEI Yuan², WANG Qiang¹^,²^,^*(), ZHANG Changpeng²^,^*()

1. College of Life Sciences, China Jiliang University, Hangzhou 310018, China
2. Institute of Agro-Products Safety and Nutrition, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2022-07-15 Online:2023-08-25 Published:2023-08-29

Abstract

Abstract:

In order to establish an efficient, sensitive and accurate residue analysis method for 2,4-epibrassinolide in Dendrobium officinale, the samples of Dendrobium officinale were extracted by ultrasonic and vortex coupling, and then cleaned by Cleanert Pesti Carb/NH₂and determined by HPLC-MS/MS. This method is clear and colorless after purification, matrix effect is reduced, easy to operate and fast. The limits of quantification and limits of detection of 2,4-epibrassinolide in Dendrobium officinale were 0.005 mg·kg^-1and 0.000 5 mg·L^-1, respectively. The average recoveries of 2,4-epibrassinolide in Dendrobium officinale were 94.70%-102.25% with the relative standard deviations (RSDs) of 1.3%-3.4% in the spiked range of 0.005-0.5 mg·kg^-1. It showed that the sensitivity, accuracy and precision of the method met the requirements for the analysis of pesticide residues of 2,4-epibrassinolide in Dendrobium officinale. The samples of Dendrobium officinale were obtained by indoor pot test, and the method was used to determine the 2, 4-epibrassinolide in the samples of Dendrobium officinale. The digestion dynamics of 2,4-epibrassinolide in Dendrobium officinale conformed to the first-order kinetics, and the half-lives of stem and leaf were 10.35-22.36 d and 8.89-11.00 d, respectively. It showed that this method can be applied to the detection of actual samples of Dendrobium officinale.

Key words: solid phase extraction (SPE), high performance liquid chromatograph-mass spectrometer (HPLC-MS/MS), plant growth regulators, Dendrobium officinale

CLC Number:

S567

LU Lanfei, ZHAO Xueping, MA Zheng, FANG Nan, LUO Yuqin, WANG Xiaomei, YE Hui, LEI Yuan, WANG Qiang, ZHANG Changpeng. Determination of 2, 4-epibrassinolide residues in Dendrobium officinale by solid phase extraction-high performance liquid chromatography-mass spectrometry[J]. Acta Agriculturae Zhejiangensis, 2023, 35(8): 1896-1903.

Figures/Tables 7

References 22

[1]	国家药典委员会. 中华人民共和国药典:2020年版第一部[M]. 北京: 中国医药科技出版社, 2020.
[2]	XU X F, DAI D C, YAN H, et al. Chemical constituents from the Dendrobium officinale and their chemotaxonomic significance[J]. Biochemical Systematics and Ecology, 2022, 102: 104420.
[3]	LIANG J, LI H L, CHEN J Q, et al. Dendrobium officinale polysaccharides alleviate colon tumorigenesis via restoring intestinal barrier function and enhancing anti-tumor immune response[J]. Pharmacological Research, 2019, 148: 104417.
[4]	YANG J R, CHEN H H, NIE Q X, et al. Dendrobium officinale polysaccharide ameliorates the liver metabolism disorders of type Ⅱ diabetic rats[J]. International Journal of Biological Macromolecules, 2020, 164: 1939-1948.
[5]	XIE H L, FANG J Y, FARAG M A, et al. Dendrobium officinale leaf polysaccharides regulation of immune response and gut microbiota composition in cyclophosphamide-treated mice[J]. Food Chemistry: X, 2022, 13: 100235.
[6]	TANG H X, ZHAO T W, SHENG Y J, et al. Dendrobium officinale Kimura et Migo: a review on its ethnopharmacology, phytochemistry, pharmacology, and industrialization[J]. Evidence-Based Complementary and Alternative Medicine, 2017, 2017: 7436259.
[7]	陈志, 汪一婷, 吕永平, 等. 铁皮石斛无菌播种成苗体系优化[J]. 浙江农业学报, 2018, 30(11): 1870-1878.
	CHEN Z, WANG Y T, LYU Y P, et al. Optimization of aseptic seeding system of Dendrobium officinale Kimura et Migo[J]. Acta Agriculturae Zhejiangensis, 2018, 30(11): 1870-1878. (in Chinese with English abstract)
[8]	ZHANG Z H, SONG J J, ZHANG H P, et al. Analysis method development and health risk assessment of pesticide and heavy metal residues in Dendrobium candidum[J]. RSC Advances, 2022, 12(11): 6869-6875.
[9]	VARDHINI B V, ANJUM N A. Brassinosteroids make plant life easier under abiotic stresses mainly by modulating major components of antioxidant defense system[J]. Frontiers in Environmental Science, 2015, 2: 67.
[10]	陆剑飞, 黄晓华, 殷琛. 0.0016%丙酰芸苔素内酯AS对茶叶生产的影响[J]. 浙江农业学报, 2011, 23(4): 763-765.
	LU J F, HUANG X H, YIN C. Effects of 0.0016% propionyl brassins AS on tea production[J]. Acta Agriculturae Zhejiangensis, 2011, 23(4): 763-765. (in Chinese with English abstract)
[11]	ZHU T, TAN W R, DENG X G, et al. Effects of brassinosteroids on quality attributes and ethylene synthesis in postharvest tomato fruit[J]. Postharvest Biology and Technology, 2015, 100: 196-204.
[12]	YASSER-LORENTE G, RODRÍGUEZ-HERNÁNDEZ D, CAMACHO-RAJO L, et al. Biobras-16 application effect on the growth and quality of pineapple fruits ‘MD-2’[J]. Cultivos Tropicales, 2021, 2:42-49.
[13]	曹立冬, 杨晶, 郑丽, 等. 高效液相色谱-蒸发光散射检测器检测24-表芸苔素内酯原药含量[J]. 农药学学报, 2014, 16(1): 72-77.
	CAO L D, YANG J, ZHENG L, et al. Quantitative determination of 24-epibrassinolide by high-performance liquid chromatography with evaporative light scattering detector[J]. Chinese Journal of Pesticide Science, 2014, 16(1): 72-77. (in Chinese with English abstract)
[14]	潘波, 方佳, 林勇, 等. 丁硫克百威原药和制剂对蚯蚓的毒性及其降解产物克百威的动态变化[J]. 热带作物学报, 2013, 34(11): 2272-2277.
	PAN B, FANG J, LIN Y, et al. A major degradation product’s dynamic changes and toxicity to earthworms of carbosulfan[J]. Chinese Journal of Tropical Crops, 2013, 34(11): 2272-2277. (in Chinese with English abstract)
[15]	徐玫, 黄艳刚, 彭超美, 等. 95%菊乙胺酯(WD-5)原药调节小麦生长田间药效试验[J]. 农药, 2003, 42(9): 39.
	XU M, HUANG Y G, PENG C M, et al. Field efficacy test of 95% pyrethroid (WD-5) in regulating wheat growth[J]. Pesticides, 2003, 42(9): 39. (in Chinese)
[16]	褚倩倩, 刘海龙, 李静朴, 等. 24-表芸苔素内酯可溶液剂组合物: CN114532341A[P]. 2022-05-27.
	CHU Q Q, LIU H L, LI J P, et al. 24-epibrassinolide soluble concentrate composition: CN114532341A[P]. 2022-05-27. (in Chinese)
[17]	FU Y W, ZHANG J, QIN J A, et al. Representative matrices for use in matrix-matched calibration in gas chromatography-mass spectrometry for the analysis of pesticide residues in different types of food-medicine plants[J]. Journal of Food Composition and Analysis, 2022, 111: 104617.
[18]	ZHOU J L, KANG Y H. Matrix effect in high-performance liquid chromatography-tandem mass spectrometry analysis of antibiotics in environmental water samples[J]. Journal of Separation Science, 2013, 36(3): 564-571.
[19]	FARLIN J, GALLÉ T, BAYERLE M, et al. Using the long-term memory effect of pesticide and metabolite soil residues to estimate field degradation half-life and test leaching predictions[J]. Geoderma, 2013, 207/208: 15-24.
[20]	CUI Z L, ZHAO Z R, YAO L, et al. Determination and analysis of solubility of brassinolide in different solvent systems at different temperatures (T=278.15-323.15 K)[J]. Journal of Molecular Liquids, 2021, 340: 117316.
[21]	诸力, 陈红平, 柴云峰, 等. 分散固相萃取-超高效液相色谱-串联质谱法测定不同茶类中2, 4-表芸苔素内酯残留[J]. 茶叶科学, 2018, 38(6): 589-594.
	ZHU L, CHEN H P, CHAI Y F, et al. Determination of 2, 4-epibrassinolide in different teas by ultra performance liquid chromatography coupled to tandem mass spectrometry with dispersive solid phase extraction[J]. Journal of Tea Science, 2018, 38(6): 589-594. (in Chinese with English abstract)
[22]	许振岚, 陈丽萍, 徐明飞, 等. 氯虫苯甲酰胺和吡唑醚菌酯在铁皮石斛中的残留及消解动态[J]. 农药学学报, 2018, 20(2): 223-231.
	XU Z L, CHEN L P, XU M F, et al. Residues and dissipation dynamics of chlorantraniliprole and pyraclostrobin in Dendrobium officinale[J]. Chinese Journal of Pesticide Science, 2018, 20(2): 223-231. (in Chinese with English abstract)

母离子 Product ion(m/z)	子离子 Product ion(m/z)	碰撞能量 Collision energy/eV	碎裂电压 Fragmentor/V	保留时间 Retention time/min
481.1	315.2 *	17	152	3.48
481.1	445.2	9	152	3.48

母离子 Product ion(m/z)	子离子 Product ion(m/z)	碰撞能量 Collision energy/eV	碎裂电压 Fragmentor/V	保留时间 Retention time/min
481.1	315.2 *	17	152	3.48
481.1	445.2	9	152	3.48

组织 Tissue	添加水平 Spiked level/ (mg·kg^-1)	平均回收率 Average recovery/%	相对标准偏差 RSDs/%
茎Stem	0.005	97.75	3.1
	0.05	101.18	2.9
	0.5	102.25	2.1
叶Leaf	0.005	101.99	3.4
	0.05	101.64	3.3
	0.5	94.70	1.3

组织 Tissue	添加水平 Spiked level/ (mg·kg^-1)	平均回收率 Average recovery/%	相对标准偏差 RSDs/%
茎Stem	0.005	97.75	3.1
	0.05	101.18	2.9
	0.5	102.25	2.1
叶Leaf	0.005	101.99	3.4
	0.05	101.64	3.3
	0.5	94.70	1.3

组织 Tissue	t/d	残留 Residues/ (mg·kg^-1)		降解率 Degradation rate/%		回归方程 Regression equation		决定系数 Coefficient of determination (R²)		半衰期 Half-life/d
组织 Tissue	t/d	T₁	T₂	T₁	T₂	T₁	T₂	T₁	T₂	T₁	T₂
茎Stem	0	0.117±1.7	0.193±2.2	—	—	Ct=0.081 6e^-0.067^t	Ct =0.160 4e^-0.031^t	0.846 4	0.846 4	10.35	22.36
	1	0.083±1.6	0.174±0.5	28.56	9.75
	7	0.032±6.4	0.104±0.5	72.98	46.23
	14	0.029±0.3	0.081±4.4	75.06	58.26
	28	0.012±3.8	0.078±1.1	89.40	59.45
	35	0.009±0.6	0.055±1.8	92.32	71.34
叶Leaf	0	0.061±0.6	0.105±1.4	—	—	Ct=0.040 3e^-0.078^t	Ct=0.093 4e^-0.063^t	0.945 6	0.945 6	8.89	11.00
	1	0.058±4.9	0.096±1.4	5.04	8.45
	7	0.010±9.3	0.074±5.7	83.01	29.29
	14	0.009±2.4	0.021±3.1	85.59	80.02
	28	0.007±5.2	0.015±4.5	88.66	85.59
	35	—	0.013±1.9	—	87.33

Determination of 2, 4-epibrassinolide residues in Dendrobium officinale by solid phase extraction-high performance liquid chromatography-mass spectrometry

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