三叶青不同部位无机营养元素的ICP-MS/MS分析与评价

doi:10.3969/j.issn.1004-1524.20240174

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (9): 2122-2131.DOI: 10.3969/j.issn.1004-1524.20240174

三叶青不同部位无机营养元素的ICP-MS/MS分析与评价

朱思蓓¹^,²(), 聂晶²^,³, 张进军⁴, 李春霖²^,³, 张永志²^,³, 王平¹, 陶益¹^,^*(), 袁玉伟²^,³^,^*()

1.浙江工业大学药学院,浙江杭州 310014
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
3.农业农村部农产品信息溯源重点实验室,浙江杭州 310021
4.浙江武义汇美中药材有限公司,浙江武义 321200

收稿日期:2024-03-01 出版日期:2024-09-25 发布日期:2024-09-30
作者简介:*陶益,E-mail:taoyi1985812@126.com;袁玉伟,E-mail:ywytea@163.com
朱思蓓(1999—),女,浙江东阳人,硕士研究生,研究方向为中药资源与道地识别。E-mail:zsb19990228@163.com
通讯作者: *陶益,E-mail:taoyi1985812@126.com; 袁玉伟,E-mail:ywytea@163.com
基金资助:
浙江省基础公益研究计划(LGN22H280010);浙江省自然科学基金(LY21H280008);国家自然科学基金(82304654)

ICP-MS/MS analysis and evaluation of inorganic nutrient elements in different parts of Tetrastigma hemsleyanum Diels et Gilg

ZHU Sibei¹^,²(), NIE Jing²^,³, ZHANG Jinjun⁴, LI Chunlin²^,³, ZHANG Yongzhi²^,³, WANG Ping¹, TAO Yi¹^,^*(), YUAN Yuwei²^,³^,^*()

1. College of Pharmacy, Zhejiang University of Technology, Hangzhou 310014, China
2. Institute of Agro-Products Safety and Nutrition, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
3. Key Laboratory of Information Traceability for Agricultural Products, Ministry of Agriculture and Rural Affairs, Hangzhou 310021, China
4. Zhejiang Wuyi Huimei Traditional Chinese Medicine Co. Ltd., Wuyi 321200, Zhejiang, China

Received:2024-03-01 Online:2024-09-25 Published:2024-09-30

摘要/Abstract

摘要：

为利用电感耦合等离子体串联质谱法(ICP-MS/MS)建立同时测定三叶青中Na、Mg、Al、P、S、K、Ca、Fe、Ni、Cu、Zn、Se共12种无机营养元素含量的方法,并探究不同部位(茎叶、块根、须根)的12种元素含量分布,通过ICP-MS/MS中的单杆和串联四级杆模式下的多种反应气体(O₂、H₂、NH₃)形式建立同时分析三叶青中12种营养元素的方法,测定三叶青中不同部位的无机营养元素含量,运用单因素方差分析(ANOVA)比较药材不同部位无机营养元素含量的差异,采用偏最小二乘-判别分析(PLS-DA)构建不同部位的鉴别模型。结果表明,ICP-MS/MS方法的线性、检测限、精密度、重复性和稳定性均良好,实际药材的加样回收率为90.66%~108.50%;不同部位的三叶青无机营养元素含量差异较大,地下部的无机营养元素含量显著高于地上部,Ca、Al、K、Na这4种元素是三叶青3个部位的主要差异元素。本研究通过电感耦合等离子体串联质谱法建立了快速鉴别三叶青不同部位中无机营养元素含量的方法,有效鉴定了块根、须根、茎叶中的无机营养元素含量,三叶青地下部的无机营养元素含量显著高于地上部,研究结果可以为三叶青质量控制、原产地溯源和种质资源的评价提供技术依据。

关键词: 三叶青, 无机元素, 电感耦合等离子体串联质谱, 偏最小二乘-判别分析

Abstract:

To establish a method for the simultaneous determination of 12 inorganic nutrient elements (Na, Mg, Al, P, S, K, Ca, Fe, Ni, Cu, Zn, Se) in Tetrastigma hemsleyanum Diels et Gilg (T. hemsleyanum) by inductively coupled plasma tandem mass spectrometry (ICP-MS/MS), and to explore the content distribution of these 12 inorganic nutrient elements in different parts (stem and leaf, tuberous root, fibrous root). The method for simultaneous analysis of 12 nutrient elements in T. hemsleyanum was established by ICP-MS/MS in the form of multiple reaction gases (O₂, H₂, NH₃) under the mode of single rod and series four rod, and the contents of inorganic nutrient elements in different parts of T. hemsleyanum were determined, and the differences of inorganic nutrient elements in different parts of medicinal materials were compared by one-way analysis of variance (ANOVA). Partial least squares-discriminant analysis (PLS-DA) was used to construct identification models for different parts. The results showed that the linearity, detection limit, precision, repeatability and stability test results of ICP-MS/MS method were good, and the recoveries ranged from 90.66% to 108.50%. There were significant differences in contents of inorganic nutrient elements in different parts of T. hemsleyanum, and the contents of inorganic nutrients in root were significantly higher than those in aboveground. Ca, Al, K and Na were the main difference elements among the three parts of T. hemsleyanum. In this study, the rapid analysis and determination of T. hemsleyanum inorganic elements was established, and the contents of inorganic elements in tuberous root, fibrous root, stem and leaf were effectively identified, which provided technical basis for quality control, origin tracing and germplasm resource evaluation of T. hemsleyanum.

Key words: Tetrastigma hemsleyanum Diels et Gilg, inorganic element, inductively coupled plasma tandem mass spectrometry, partial least squares discriminant analysis

中图分类号:

S567.239

朱思蓓, 聂晶, 张进军, 李春霖, 张永志, 王平, 陶益, 袁玉伟. 三叶青不同部位无机营养元素的ICP-MS/MS分析与评价[J]. 浙江农业学报, 2024, 36(9): 2122-2131.

ZHU Sibei, NIE Jing, ZHANG Jinjun, LI Chunlin, ZHANG Yongzhi, WANG Ping, TAO Yi, YUAN Yuwei. ICP-MS/MS analysis and evaluation of inorganic nutrient elements in different parts of Tetrastigma hemsleyanum Diels et Gilg[J]. Acta Agriculturae Zhejiangensis, 2024, 36(9): 2122-2131.

图/表 8

表1 不同分析模式下各元素的背景等效浓度

Table 1 The background equivalent concentration of each element under different analysis models

元素 Elements	潜在干扰离子 Potentially interfering ions	背景等效浓度 Background equivalent concentration/(μg·L^-1)
		SQ模式SQ model			MS/MS模式MS/MS model
		No gas	He	HEHe	H₂	O₂	NH₃
²³Na		16.255	15.850	14.985	18.220	17.335	18.280
²⁴Mg	¹² $C_{2}^{+}$	1.454	1.461	1.385	1.833	1.746	1.725
²⁷Al		1.984	1.173	1.842	3.579	4.179	3.909
³¹P	³⁰Si¹H⁺、¹⁴N¹H¹⁶O⁺、¹⁴N¹⁷O⁺、¹⁵H¹⁶O⁺、¹²C¹⁹F⁺	24.230	6.929	2.437	67.860	0.955	3.817
³²S	¹⁶O¹⁶O⁺、¹⁶O¹H¹⁵N⁺、¹⁸O¹⁴N⁺、¹H³¹P⁺、¹²C³⁰Ne⁺、¹³C¹⁹F⁺	4 046	2 109	849.8	2 004	4.985	4 104
³⁹K	³⁸Ar¹H⁺	51.010	16.800	49.760	19.410	25.260	24.580
⁴⁴Ca	¹²C¹⁶ $O_{2}^{+}$ 、¹⁴ $N_{2}^{16}$ O、²⁸Si¹⁶O	21.210	18.240	19.990	19.860	19.720	26.190
⁵⁶Fe	⁴⁰Ar¹⁶O⁺、⁴⁰Ca¹⁶O⁺、³⁷Cl¹⁷O⁺、¹¹²Sn²⁺、⁴⁰Ar¹⁵N¹H⁺、	86.090	1.341	1.357	1.223	1.593	4.499
	³⁸Ar¹⁸O⁺、³⁸Ar¹⁷O¹H⁺、³⁷Cl¹⁸O¹H⁺
⁵⁸Ni	⁵⁸Fe、⁴⁰Ar¹⁸O⁺、⁴⁰Ca¹⁸O⁺、⁴²Ca¹⁶O⁺、¹¹⁶Sn²⁺、	0.091	0.203	0.176	0.094	0.136
	⁴⁰Ar¹⁷O¹H⁺、⁴⁰Ca¹⁷O¹H⁺	0.471
⁶³Cu	⁴⁷Ti¹⁶O⁺、⁴⁶Ca¹⁶O¹H⁺、³⁶Ar¹²C¹⁴N¹H⁺、¹⁴N¹²C³⁷Cl⁺、	0.113	0.087	0.166	0.504	0.140	0.223
	¹⁶O¹²C³⁵Cl⁺
⁶⁶Zn	⁵⁰Ti¹⁶O⁺	1.045	0.893	1.200	0.930	1.239	1.643
⁷⁸Se	⁷⁸Kr、⁴⁰Ar³⁸Ar⁺、³⁸Ar⁴⁰Ca²⁺	5.520	0.130	0.015	0.012	0.016	0.026

表1 不同分析模式下各元素的背景等效浓度

Table 1 The background equivalent concentration of each element under different analysis models

元素 Elements	潜在干扰离子 Potentially interfering ions	背景等效浓度 Background equivalent concentration/(μg·L^-1)
		SQ模式SQ model			MS/MS模式MS/MS model
		No gas	He	HEHe	H₂	O₂	NH₃
²³Na		16.255	15.850	14.985	18.220	17.335	18.280
²⁴Mg	¹² $C_{2}^{+}$	1.454	1.461	1.385	1.833	1.746	1.725
²⁷Al		1.984	1.173	1.842	3.579	4.179	3.909
³¹P	³⁰Si¹H⁺、¹⁴N¹H¹⁶O⁺、¹⁴N¹⁷O⁺、¹⁵H¹⁶O⁺、¹²C¹⁹F⁺	24.230	6.929	2.437	67.860	0.955	3.817
³²S	¹⁶O¹⁶O⁺、¹⁶O¹H¹⁵N⁺、¹⁸O¹⁴N⁺、¹H³¹P⁺、¹²C³⁰Ne⁺、¹³C¹⁹F⁺	4 046	2 109	849.8	2 004	4.985	4 104
³⁹K	³⁸Ar¹H⁺	51.010	16.800	49.760	19.410	25.260	24.580
⁴⁴Ca	¹²C¹⁶ $O_{2}^{+}$ 、¹⁴ $N_{2}^{16}$ O、²⁸Si¹⁶O	21.210	18.240	19.990	19.860	19.720	26.190
⁵⁶Fe	⁴⁰Ar¹⁶O⁺、⁴⁰Ca¹⁶O⁺、³⁷Cl¹⁷O⁺、¹¹²Sn²⁺、⁴⁰Ar¹⁵N¹H⁺、	86.090	1.341	1.357	1.223	1.593	4.499
	³⁸Ar¹⁸O⁺、³⁸Ar¹⁷O¹H⁺、³⁷Cl¹⁸O¹H⁺
⁵⁸Ni	⁵⁸Fe、⁴⁰Ar¹⁸O⁺、⁴⁰Ca¹⁸O⁺、⁴²Ca¹⁶O⁺、¹¹⁶Sn²⁺、	0.091	0.203	0.176	0.094	0.136
	⁴⁰Ar¹⁷O¹H⁺、⁴⁰Ca¹⁷O¹H⁺	0.471
⁶³Cu	⁴⁷Ti¹⁶O⁺、⁴⁶Ca¹⁶O¹H⁺、³⁶Ar¹²C¹⁴N¹H⁺、¹⁴N¹²C³⁷Cl⁺、	0.113	0.087	0.166	0.504	0.140	0.223
	¹⁶O¹²C³⁵Cl⁺
⁶⁶Zn	⁵⁰Ti¹⁶O⁺	1.045	0.893	1.200	0.930	1.239	1.643
⁷⁸Se	⁷⁸Kr、⁴⁰Ar³⁸Ar⁺、³⁸Ar⁴⁰Ca²⁺	5.520	0.130	0.015	0.012	0.016	0.026

图1 ICP-MS/MS分析无机营养元素消除干扰的原理图

Fig.1 Schematic diagram of eliminating interference by ICP-MS/MS analysis of inorganic nutrient elements

表2 三叶青中12种无机营养元素加标回收率

Table 2 Spike recovery rate of 12 inorganic nutrient elements in Tetrastigma hemsleyanum Diels et Gilg

元素 Element	本底浓度 Background concentration/(mg·L^-1)	加标浓度 Standard concentration/ (mg·L^-1)	测定浓度 Determination of concentration/(mg·L^-1)	回收率 Recovery rate/%
²³Na	801.53	320.00	1 069.72	95.38
²⁴Mg	7 992.18	3 200.00	11 797.68	105.41
²⁷Al	343.40	120.00	445.70	96.18
³¹P	6 716.63	2 000.00	8 577.16	98.40
³²S	3 269.77	1 200.00	4 767.90	106.67
³⁹K	19 586.21	8 000.00	26 979.31	97.80
⁴⁴Ca	15 464.21	6 000.00	19 782.68	108.50
⁵⁶Fe	542.66	200.00	731.52	98.50
⁵⁸Ni	3.74	1.00	4.30	90.66
⁶³Cu	9.27	4.00	13.69	103.20
⁶⁶Zn	41.07	16.00	61.64	100.80
⁷⁸Se	0.05	0.02	0.07	101.25

表3 三叶青不同部位的无机营养元素含量(n=10)

Table 3 Contents of inorganic nutrient elements in different parts of Tetrastigma hemsleyanum Diels et Gilg (n=10)

元素 Elements	茎叶 Stem and leaf	块根 Tuberous root	须根 Fibrous root
²³Na	2 766.2±1 638.3 a	259.7±91.2 b	406.1±33.1 b
²⁴Mg	4 277.9±1 886.7 a	2 035.9±527.2 b	4 313.9±1 500.1 a
²⁷Al	1 776.8±827.5 b	1 153.9±640.5 b	6 738.2±3 219.9 a
³¹P	3 587.2±1 069.3 b	2 269.5±809.1 c	5 098.7±2 177.2 a
³²S	2 614.9±970.7 b	1 522.6±304.6 c	3 971.8±974.3 a
³⁹K	20 994.4±8 032.2 a	9 098.9±2 338.7 b	17 426.9±4 551.4 a
⁴⁴Ca	17 087.1±4 953.9 a	5 635.6±1 866.7 b	13 256.5±2 629.5 a
⁵⁶Fe	430.5±204.7 b	280.3±143.5 c	2 331.0±1 613.3 a
⁵⁸Ni	1.31±0.4 b	0.7±0.3 b	10.5±5.5 a
⁶³Cu	11.7±3.0 b	5.8±1.7 b	52.4±21.5 a
⁶⁶Zn	114.4±43.9 b	110.5±36.7 b	406.5±115.9 a
⁷⁸Se^*	133.5±102.3 b	55.0±32.4 b	300.0±143.3 a

表4 三叶青茎叶、块根、须根PLS-DA模型判别分析结果

Table 4 PLS-DA discriminant analysis results of stem and leaf, tuberou root and fibrous root of Tetrastigma hemsleyanum Diels et Gilg

数据集 Data set	部位 Part	数量 Number	测试结果Test result			判别准确率 Discriminant accuracy/%	总体准确率 Overall accuracy/%
数据集 Data set	部位 Part	数量 Number	茎叶 Stem and leaf	块根 Tuberous root	须根 Fibrous root	判别准确率 Discriminant accuracy/%	总体准确率 Overall accuracy/%
训练集	茎叶Stem and leaf	8	7	1	0	87.5	91.67
Training set	块根Tuberous root	8	0	8	0	100
	须根Fibrous root	8	1	0	7	87.5
验证集	茎叶Stem and leaf	2	2	0	0	100	100
Validation set	块根Tuberous root	2	0	2	0	100
	须根Fibrous root	2	0	0	2	100

图2 三叶青茎叶、块根、须根三部位PLS-DA模型得分图

Fig.2 PLS-DA model score diagram of stem and leaf, tuberous root and fibrous root of Tetrastigma hemsleyanum Diels et Gilg

图3 三叶青茎叶、块根、须根的PLS-DA模型置换检验图(n=200) R2为模型解释率,Q2为模型预测能力。

Fig.3 Permutation test plot of PLS-DA model for stem and leaf, tuberous root and fibrous root of Tetrastigma hemsleyanum Diels et Gilg (n=200) R2 was the model interpretation rate, Q2 was the model prediction ability.

图4 三叶青茎叶、块根、须根中无机营养元素的VIP

Fig.4 Variable importance in the projection (VIP) of inorganic nutrient elements in stem and leaf, tuberous root and fibrous root of Tetrastigma hemsleyanum Diels et Gilg

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三叶青不同部位无机营养元素的ICP-MS/MS分析与评价

ICP-MS/MS analysis and evaluation of inorganic nutrient elements in different parts of Tetrastigma hemsleyanum Diels et Gilg

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