固相萃取-高效液相色谱-质谱联用法测定铁皮石斛中2,4-表芸苔素内酯残留

doi:10.3969/j.issn.1004-1524.20221050

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (8): 1896-1903.DOI: 10.3969/j.issn.1004-1524.20221050

固相萃取-高效液相色谱-质谱联用法测定铁皮石斛中2,4-表芸苔素内酯残留

陆兰菲¹^,²(), 赵学平², 马正¹, 方楠², 骆玉琴², 王晓梅¹^,², 叶会², 雷圆², 王强¹^,²^,^*(), 张昌朋²^,^*()

1.中国计量大学生命科学学院,浙江杭州 310018
2.浙江省农业科学院农产品质量安全与营养研究所,浙江杭州 310021

收稿日期:2022-07-15 出版日期:2023-08-25 发布日期:2023-08-29
作者简介:陆兰菲(1988—),女,陕西西安人,硕士研究生,主要研究方向为药品、食品质量与安全。E-mail:lfyx216@163.com
通讯作者: *王强,E-mail: wq13575733860@126.com;张昌朋,E-mail: cpzhang1215@zaas.ac.cn
基金资助:
浙江省基础公益研究计划(LGN21C140006);“一县一品一策”标准化生产创建(ZJNY2022001-014)

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

摘要/Abstract

摘要：

为建立铁皮石斛中2, 4-表芸苔素内酯的高效、灵敏和准确的残留分析方法,利用超声与涡旋联用方法对铁皮石斛样品进行了提取,然后采用固相萃取小柱(Cleanert Pesti Carb/NH₂)方法净化且经HPLC-MS/MS测定。经过方法学验证,2, 4-表芸苔素内酯在铁皮石斛中的定量限和检出限分别为0.005 mg·kg^-1和0.000 5 mg·L^-1。在0.005~0.5 mg·kg^-1的添加范围下,2, 4-表芸苔素内酯在铁皮石斛中的平均回收率为94.70%~102.25%,相对标准偏差为1.3%~3.4%,表明该方法的灵敏度、准确度和精密度均满足铁皮石斛中2, 4-表芸苔素内酯的农药残留分析要求。通过室内盆栽试验,获得了铁皮石斛样品,并利用该方法测定了铁皮石斛样品中2, 4-表芸苔素内酯残留。2, 4-表芸苔素内酯在铁皮石斛中的消解动态符合一级动力学,茎、叶半衰期分别为10.35~22.36 d和8.89~11.00 d,表明该方法能够应用于铁皮石斛实际样品的检测。

关键词: 固相萃取, 高效液相色谱-质谱联用仪, 植物生长调节剂, 铁皮石斛

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

中图分类号:

S567

陆兰菲, 赵学平, 马正, 方楠, 骆玉琴, 王晓梅, 叶会, 雷圆, 王强, 张昌朋. 固相萃取-高效液相色谱-质谱联用法测定铁皮石斛中2,4-表芸苔素内酯残留[J]. 浙江农业学报, 2023, 35(8): 1896-1903.

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.

图/表 7

参考文献 22

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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

固相萃取-高效液相色谱-质谱联用法测定铁皮石斛中2,4-表芸苔素内酯残留

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

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