QuEChERS-液相色谱-串联质谱法同时测定果蔬中15种植物生长调节剂

doi:10.16178/j.issn.0528-9017.20240212

摘要/Abstract

摘要：

采用液相色谱-串联质谱(LC-MS/MS)仪器,建立了同时测定果蔬中15种植物生长调节剂的残留量方法。以葡萄为代表基质,样品使用QuEChERS法进行预处理,选用1%乙酸乙腈溶液为提取液,无水硫酸镁和氯化钠为盐析试剂。对吸附剂种类和用量进行系统优化,选取50 mg C18和150 mg MgSO₄对提取液进行净化。最终样品以Inertsil ODS-3 2 μm(75 mm×2.1 mm)柱为色谱分析柱,0.5 mmol·L^-1甲酸铵水溶液和甲醇作为流动相,进行梯度洗脱,采用液相色谱-串联质谱法进行分析;系统的方法验证结果表明,在6种果蔬中,除氯苯胺灵和2, 3, 5-三碘苯甲酸外,其余13种植物生长调节剂在0.5~200.0 μg·L^-1的浓度范围内具有良好的线性关系(R²>0.98);当添加浓度为10、50、200 μg·L^-1时,各分析物在空白果蔬样品中的添加回收率为72.3%~105.0%,相对标准偏差(RSD)小于9.3%,方法定量限为10 μg·kg^-1。该方法简便、快速、灵敏、准确,可同时满足多种水果和蔬菜中15种生长调节剂的残留量检测。

关键词: QuEChERS, 液相色谱-串联质谱法, 植物生长调节剂, 果蔬

Abstract:

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of 15 plant growth regulators residues in fruits and vegetables. With grapes as the representative substrate, the samples were pretreated by QuEChERS method. 1% formic acid acetonitrile solution was used as extraction solution, and anhydrous magnesium sulfate and sodium chloride were used as salt-out reagents. The type and dosage of adsorbent were optimized, and 50 mg C18 and 150 mg MgSO₄ were selected to purify the extraction solution. The final samples were analyzed on Inertsil ODS-3 2 μm (75 mm×2.1 mm) column. Mobile phase was 0.5 mmol·L^-1 ammonium formate aqueous solution and methanol by gradient elution, and then samples analyzed by LC-MS/MS. The results of systematic method verification showed that except for chloroaniline and 2,3,5-triodobenzoic acid, the other 13 plant growth regulators showed good linear relationships (R²>0.98) within the concentration range of 0.5-200.0 μg·L^-1. When the concentration was 10, 50, 200 μg·L^-1, the recoveries rate of each analyte added to blank fruit and vegetable samples were 72.3%-105.0%, with the RSD less than 9.3%. The limit of quantitation was 10 μg·kg^-1. The method is simple, rapid, sensitive and accurate, and can simultaneously satisfy the determination of 15 kinds of growth regulator residues in various fruits and vegetables.

Key words: QuEChERS, liquid chromatography-tandem mass spectrometry (LC-MS/MS), plant growth regulator, fruit and vegetable

中图分类号:

S482.8

范晓民, 王娇, 刘之炜, 刘真真, 王新全, 狄珊珊, 赵慧宇, 汪志威, 顾成波, 齐沛沛. QuEChERS-液相色谱-串联质谱法同时测定果蔬中15种植物生长调节剂[J]. 浙江农业科学, 2025, 66(5): 1237-1243.

FAN Xiaomin, WANG Jiao, LIU Zhiwei, LIU Zhenzhen, WANG Xinquan, DI Shanshan, ZHAO Huiyu, WANG Zhiwei, GU Chengbo, QI Peipei. Simultaneous determination of 15 plant growth regulators in fruits and vegetables by QuEChERS-liquid chromatography-tandem mass spectrometry[J]. Journal of Zhejiang Agricultural Sciences, 2025, 66(5): 1237-1243.

图/表 7

参考文献 16

[1]	ZHANG Z X, SHAO S C, PAN D D, et al. Role, residues and microbial degradation of plant growth regulators (PGRs): a scoping review[J]. Advanced Agrochem, 2024, 3(1): 43-46.
[2]	WANG M W, YANG C L, CAO J K, et al. Dual-template hydrophilic imprinted resin as an adsorbent for highly selective simultaneous extraction and determination of multiple trace plant growth regulators in red wine samples[J]. Food Chemistry, 2023, 411: 135471.
[3]	LI N, WU D, LI X T, et al. Effective enrichment and detection of plant growth regulators in fruits and vegetables using a novel magnetic covalent organic framework material as the adsorbents[J]. Food Chemistry, 2020, 306: 125455.
[4]	LIU L J, XIA L, WU C X, et al. Zirconium (Ⅳ)-based metal organic framework (UIO-67) as efficient sorbent in dispersive solid phase extraction of plant growth regulator from fruits coupled with HPLC fluorescence detection[J]. Talanta, 2016, 154: 23-30.
[5]	CAO J P, SHI L, WANG Y F, et al. Novel ferrofluid based on water-based deep eutectic solvents: application in dispersive liquid-liquid microextraction of naphthalene-derived plant growth regulators in edible oil[J]. Journal of Hazardous Materials, 2024, 465: 133234.
[6]	陈卫军, 张耀海, 李云成, 等. 果蔬中常用植物生长调节剂分析方法研究进展[J]. 食品科学, 2012, 33(11): 283-289.
[7]	高红顺, 谷博颖. 植物生长调节剂在作物中的应用现状及发展前景[J]. 科技创新与应用, 2024, 14(5): 176-180.
[8]	王欢欢, 王久照, 姜继元, 等. 植物生长调节剂在苹果疏花疏果上的应用研究进展[J]. 现代农业科技, 2023(23): 77-82.
[9]	王莹, 刘芫汐, 郑尊涛, 等. 《GB 2763食品安全国家标准食品中农药最大残留限量》中药品种限量标准转化原则初探[J]. 中国药学杂志, 2023, 58(15): 1416-1421.
[10]	何成军, 钟恋, 耿昭, 等. 超高效液相色谱-串联质谱法快速测定麦冬种植土壤中25种植物生长调节剂[J]. 分析科学学报, 2023, 39(6): 665-671.
[11]	林俊燕, 马恩耀, 周劲松, 等. 高效液相色谱-串联质谱法测定金银花中植物生长调节剂的残留量[J]. 中南农业科技, 2023, 44 (11): 57-60.
[12]	邓秀清, 郭燕, 伍淑明, 等. HPLC-MS/MS同时测定豆芽中30种植物生长调节剂和抗生素类药物[J]. 中国测试, 2023, 49 (S1): 97-105.
[13]	褚悟男, 李翔, 韩玲玲, 等. UHPLC-MS/MS同时测定白芷中34种植物生长调节剂残留量[J]. 中国现代应用药学, 2023, 40(6):783-791.
[14]	QI P P, WANG J, LIU Z Z, et al. Fabrication of magnetic magnesium oxide cleanup adsorbent for high-throughput pesticides residue analysis coupled with supercritical fluid chromatography-tandem mass spectrometry[J]. Analytica Chimica Acta, 2023, 1265: 341266.
[15]	QI P P, WANG J, LIU Z Z, et al. Fabrication of poly-dopamine-modified magnetic nanomaterial and development of integrated QuEChERS method for 122 pesticides residue analysis in fruits[J]. Journal of Chromatography A, 2023, 1708: 464336.
[16]	QI P P, WANG J, LIU Z Z, et al. Integrated QuEChERS strategy for high-throughput multi-pesticide residues analysis of vegetables[J]. Journal of Chromatography A, 2021, 1659: 462589.

时间/min	甲醇/水(V/V)比例
1.0	20∶1
7.0	90∶1
10.0	90∶1
10.5	20∶1
13.0	20∶1

时间/min	甲醇/水(V/V)比例
1.0	20∶1
7.0	90∶1
10.0	90∶1
10.5	20∶1
13.0	20∶1

分析物	ESI模式	母离子(m/z)	子离子(m/z)	碰撞能量/V
抗倒酯	ESI+	253.10	69.10^*; 207.10	-23;-12
噻苯隆	ESI+	221.20	102.00^*; 128.00	-16;-17
烯效唑	ESI+	292.10	70.10^*; 125.00	-24;-28
甲基嘧啶磷	ESI+	306.05	108.00^*; 95.00	-31;-29
多效唑	ESI+	294.10	70.05^*;125.05	-21;-40
氯苯胺灵	ESI+	214.15	172.00^*;154.00	-9;-17
氯吡脲	ESI+	248.10	129.10^*;93.10	-17;-34
吲哚乙酸	ESI+	176.00	130.15^*;77.10	-16;-43
6-苄基腺嘌呤	ESI+	226.00	91.10^*;65.10	-25;-54
脱落酸	ESI-	263.30	153.35^*;219.40	11;14
赤霉酸	ESI-	345.20	239.40^*;143.35	15;31
2,4-滴丁酯	ESI-	218.95	161.00^*;125.00	13;26
2,3,5-三碘苯甲酸	ESI-	498.75	454.75^*;127.00	12;24
对氯苯氧乙酸	ESI-	185.00	127.10^*	15
调果酸	ESI-	199.00	127.10^*	14

分析物	ESI模式	母离子(m/z)	子离子(m/z)	碰撞能量/V
抗倒酯	ESI+	253.10	69.10^*; 207.10	-23;-12
噻苯隆	ESI+	221.20	102.00^*; 128.00	-16;-17
烯效唑	ESI+	292.10	70.10^*; 125.00	-24;-28
甲基嘧啶磷	ESI+	306.05	108.00^*; 95.00	-31;-29
多效唑	ESI+	294.10	70.05^*;125.05	-21;-40
氯苯胺灵	ESI+	214.15	172.00^*;154.00	-9;-17
氯吡脲	ESI+	248.10	129.10^*;93.10	-17;-34
吲哚乙酸	ESI+	176.00	130.15^*;77.10	-16;-43
6-苄基腺嘌呤	ESI+	226.00	91.10^*;65.10	-25;-54
脱落酸	ESI-	263.30	153.35^*;219.40	11;14
赤霉酸	ESI-	345.20	239.40^*;143.35	15;31
2,4-滴丁酯	ESI-	218.95	161.00^*;125.00	13;26
2,3,5-三碘苯甲酸	ESI-	498.75	454.75^*;127.00	12;24
对氯苯氧乙酸	ESI-	185.00	127.10^*	15
调果酸	ESI-	199.00	127.10^*	14

分析物	线性范围/ (μg·L^-1)	回归方程	R²	检出限LOD/ (μg·kg^-1)	回收率(RSD)/%
分析物	线性范围/ (μg·L^-1)	回归方程	R²	检出限LOD/ (μg·kg^-1)	10 μg·kg^-1	50 μg·kg^-1	200 μg·kg^-1
抗倒酯	0.5~200.0	y=18 862x+127 219	0.999 4	0.36	85.6(7.1)	95.1(1.6)	80.1(0.4)
	0.5~200.0	y = 24 609x-18 228	0.999 7
噻苯隆	0.5~200.0	y = 6 377.4x + 5 192	0.999 7	0.26	72.3(1.4)	96.0(1.1)	84.2(0.6)
	0.5~200.0	y=6 479x-3 466	0.999 6
烯效唑	0.5~200.0	y = 8 519x + 7 724.6	0.999 4	0.26	83.8(1.5)	95.2(0.3)	79.4(0.6)
	0.5~200.0	y = 9 592.9x + 1 897.1	0.999 9
甲基嘧啶磷	0.5~200.0	y = 72 407x + 52 709	0.999 5	0.49	76.7(0.5)	90.3(0.6)	75.3(1.4)
	0.5~200.0	y = 80 937x + 42 872	0.999 8
多效唑	0.5~200.0	y = 24 017x + 23 212	0.998 9	0.13	84.1(1.1)	97.2(1.3)	87.8(1.0)
	0.5~200.0	y = 23 390x + 26 756	0.999 3
氯苯胺灵	5.0~200.0	y = 72.378x - 382.43	0.995 5	1.59	94.6(5.1)	85.6(6.7)	83.5(2.5)
	5.0~200.0	y = 82.991x + 44.995	0.998 7
氯吡脲	0.5~200.0	y = 28 662x + 39 256	0.998 8	0.25	82.7(0.5)	99.7(2.2)	92.2(0.8)
	0.5~200.0	y = 29 567x + 19 624	0.999 7
吲哚乙酸	0.5~200.0	y = 24 479x + 11 875	0.999 9	0.46	78.7(1.0)	92.4(1.1)	78.2(0.4)
	0.5~200.0	y = 19 686x + 9 354.6	0.999 9
6-苄基腺嘌呤	0.5~200.0	y = 127 118x + 149 139	0.998 8	0.32	74.8(1.2)	85.4(0.5)	72.8(0.9)
	0.5~200.0	y = 124 736x + 145 616	0.999 3
脱落酸	0.5~200.0	y = 1 120.8x + 2 401.9	0.999 4	0.13	93.3(2.8)	103.0(3.3)	83.4 (4.5)
	0.5~200.0	y = 1 150.4x + 43 377	0.995 1
赤霉酸	0.5~200.0	y = 431.28x - 11.239	0.999 3	0.13	89.4 (9.2)	100.0(5.6)	96.2 (2.9)
	0.5~200.0	y = 436.93x + 897.89	0.997 8
2,4-滴丁酯	0.5~200.0	y = 818.98x + 618.39	0.999 1	0.14	77.3 (1.3)	100.0(2.9)	79.0(0.5)
	0.5~200.0	y = 700.91x + 2 131.1	0.991 7
2,3,5-三碘苯甲酸	10.0~200.0	y = 16.447x - 43.618	0.991 7	4.60	86.3(9.3)	93.3(2.9)	88.0(4.6)
	10.0~200.0	y = 15.869x - 41.213	0.982 2
对氯苯氧乙酸	0.5~200.0	y = 771.63x + 608.1	0.998 1	1.46	85.9(8.5)	97.7(5.8)	87.4(0.9)
	0.5~200.0	y = 695.09x - 24.301	0.998 1
调果酸	0.5~200.0	y = 1 299.2x + 361.36	0.999 9	0.46	78.0(2.3)	105.0(2.3)	91.2(1.6)
	0.5~200.0	y = 1 148.2x + 497.47	0.999 7