改进的固相萃取-高效液相色谱法测定土壤中15种醛酮类化合物

doi:10.3724/SP.J.1123.2022.05021

摘要/Abstract

摘要：

建立了改进的固相萃取-高效液相色谱法测定土壤中甲醛、乙醛、丙烯醛、丙酮、丙醛、丁烯醛、丁醛、苯甲醛、异戊醛、正戊醛、邻-甲基苯甲醛、间-甲基苯甲醛、对-甲基苯甲醛、正己醛和2,5-二甲基苯甲醛等15种醛酮类化合物的分析方法。利用乙腈对土壤进行超声提取,样品提取液与2,4-二硝基苯肼(2,4-DNPH)进行衍生化反应,生成稳定的腙类化合物;随后利用装有亲水亲脂平衡的N-乙烯基吡咯烷酮/二乙烯基苯共聚物填料的固相萃取小柱(Welchrom^® BRP)对衍生后的溶液进行净化;采用Ultimate^® XB-C18色谱柱(250 mm×4.6 mm, 5 μm)进行分离,以乙腈-水(65∶35, v/v)为流动相进行等度洗脱,于波长360 nm处进行检测,利用外标法对土壤中15种醛酮类化合物进行定量。该方法改进了环境标准HJ 997-2018《土壤和沉积物醛、酮类化合物的测定高效液相色谱法》中试样的处理方法,优化后得到土壤的最佳提取条件,即:提取溶剂为乙腈,提取温度为30 ℃,提取时间为10 min。结果表明:采用BRP小柱的净化效果明显优于普通硅胶基质C18小柱,15种醛酮类化合物在各自的范围内线性关系良好,线性相关系数均在0.996以上;平均加标回收率为84.6%~115.9%,相对标准偏差(RSD)为0.2%~5.1%;检出限为0.02~0.06 mg/L。该方法简便,灵敏度高,准确性好,适用于HJ 997-2018中规定的土壤和沉积物中15种醛酮类化合物的准确定量分析,为研究土壤中醛酮类化合物的残留状况和环境行为提供了可靠的技术支持。

关键词: 固相萃取, 高效液相色谱, 醛酮类化合物, 土壤

Abstract:

An improved solid phase extraction (SPE)-high performance liquid chromatography method was established to determine 15 carbonyl compounds, namely, formaldehyde (FOR), acetaldehyde (ACETA), acrolein (ACR), acetone (ACETO), propionaldehyde (PRO), crotonaldehyde (CRO), butyraldehyde (BUT), benzaldehyde (BEN), isovaleraldehyde (ISO), n-valeraldehyde (VAL), o-methylbenzaldehyde (o-TOL), m-methylbenzaldehyde (m-TOL), p-methylbenzaldehyde (p-TOL), n-hexanal (HEX), and 2,5-dimethylbenzaldehyde (DIM), in soil. The soil was ultrasonically extracted with acetonitrile, and the extracted samples were derivatized with 2,4-dinitrophenylhydrazine (2,4-DNPH) to generate stable hydrazone compounds. The derivatized solutions were cleaned using an SPE cartridge (Welchrom^® BRP) packed with N-vinylpyrrolidone/divinylbenzene copolymer. Separation was performed on an Ultimate^® XB-C18 column (250 mm×4.6 mm, 5 μm), isocratic elution was performed with acetonitrile-water (65∶35, v/v) as the mobile phase, and detection was performed at a wavelength of 360 nm. The 15 carbonyl compounds in the soil were then quantified using an external standard method. The proposed method improves the sample processing method described in the environmental standard HJ 997-2018: Soil and sediment-Determination of carbonyl compounds-High performance liquid chromatography. A series of experiments revealed the following optimal conditions for soil extraction: acetonitrile as the extraction solvent, extraction temperature of 30 ℃, and extraction time of 10 min. The results showed that the purification effect of the BRP cartridge was significantly better than that of the conventional silica-based C18 cartridge. The 15 carbonyl compounds showed good linearities, and all correlation coefficients were above 0.996. The recoveries ranged from 84.6% to 115.9%, the relative standard deviations (RSDs) ranged from 0.2% to 5.1%, and the detection limits were 0.02-0.06 mg/L. The method is simple, sensitive, and suitable for the accurate quantitative analysis of the 15 carbonyl compounds in soil specified in HJ 997-2018. Thus, the improved method provides reliable technical support for studying the residual status and environmental behavior of carbonyl compounds in soil.

Key words: solid phase extraction (SPE), high performance liquid chromatography (HPLC), carbonyl compounds, soil

中图分类号:

O658

薛昆鹏, 俞灵钰, 任兴发, 屠炳芳, 陈超, 徐婷, 何欢, 胡帅. 改进的固相萃取-高效液相色谱法测定土壤中15种醛酮类化合物[J]. 色谱, 2023, 41(3): 265-273.

XUE Kunpeng, YU Lingyu, REN Xingfa, TU Bingfang, CHEN Chao, XU Ting, HE Huan, HU Shuai. Determination of 15 carbonyl compounds in soil using improved solid phase extraction-high performance liquid chromatography[J]. Chinese Journal of Chromatography, 2023, 41(3): 265-273.

图/表 11

图1 HJ 997-2018标准中15种醛酮-DNPH衍生物的色谱图[23]

Fig. 1 Chromatograms of the 15 carbonyl compound- dinitrophenylhydrazine (DNPH) derivatives in the national standard HJ 997-2018[23] Peak identifications: 1. formaldehyde (FOR); 2. acetaldehyde (ACETA); 3. acrolein (ACR); 4. acetone (ACETO); 5. propionaldehyde (PRO); 6. crotonaldehyde (CRO); 7. butanal (BUT); 8. benzaldehyde (BEN); 9. isovaleraldehyde (ISO); 10. valeraldehyde (VAL); 11. o-tolualdehyde (o-TOL); 12. m-tolualdehyde (m-TOL); 13. p-tolualdehyde (p-TOL); 14. hexanal (HEX); 15. 2,5-dimethylbenzaldehyde (DIM).

图2 15种醛酮-DNPH衍生物标准溶液的色谱图

Fig. 2 Chromatograms of 15 carbonyl compound-DNPH derivatives in the standard solution Experimental conditions: mixed standard solution of the 15 carbonyl compounds at 0.1 mg/L; separation column, Ultimate? XB-C18 (250 mm×4.6 mm, 5 μm); mobile phase: acetonitrile-water; flow rate: 1.0 mL/min; column temperature: 30 ℃; injection volume: 20 μL; detection wavelength: 360 nm.

图3 不同溶剂体系对15种醛酮类化合物的衍生效果

Fig. 3 Derivative effects of different solvents on the 15 carbonyl compounds The sample solution was prepared from a 10.0 g sample spiked with a mixed standard solution of the 15 carbonyl compounds at 0.1 mg/L.Conditions: a. 100 mL of sample solution, extracted with 0.065 mol/L acetic acid sodium-acetate buffer solution, added with 4 mL pH 3.0 citric acid-sodium citrate buffer solution and 6 mL 2,4-DNPH acetonitrile solution, derivatized at 40 ℃ for 1 h; b. 100 mL of sample solution, extracted with 0.065 mol/L acetic acid-sodium acetate buffer, added with 4 mL acetonitrile and 6 mL 2,4-DNPH acetonitrile solution, derivatized at 40 ℃ for 1 h; c. 100 mL of sample solution, extracted with acetonitrile, added with 6 mL 2,4-DNPH acetonitrile solution and 1.0 mL 20% phosphoric acid solution, derivatized at 4 ℃ for 6 h.

图4 提取溶剂对15种醛酮类化合物提取效率的影响

Fig. 4 Effect of extraction solvent on the recoveries of the 15 carbonyl compounds

图5 提取温度对15种醛酮类化合物提取效率的影响

Fig. 5 Effect of extraction temperature on the recoveries of the 15 carbonyl compounds

图6 提取时间对15种醛酮类化合物提取效率的影响

Fig. 6 Effect of extraction time on the recoveries of the 15 carbonyl compounds The extracted solution was prepared from a 10.0 g sample spiked with a mixed standard solution of the 15 carbonyl compounds at 0.1 mg/L.

表1 3种反相固相萃取小柱的填料理化参数

Table 1 Physicochemical properties of the packing materials of the three reversed-phase SPE cartridges

SPE column type	Stationary phase group	Morphology	Specific surface area/(m²/g)	Particle size/μm	Pore size/nm
C18	octadecyl	amorphous	500	40-63	4
PS-DVB	polystyrene-divinyl polymer	spherical	700	50-70	20
BRP	N-vinylpyrrolidone-divinyl polymer	spherical	700	50-70	20

图7 3种固相萃取小柱填料的SEM形貌图

Fig. 7 SEM images of three SPE cartridge packing materials a. C18; b. PS-DVB; c. BRP.

表2 3种SPE填料对15种醛酮类化合物基质效应的影响(n=6)

Table 2 Matrix effects of the SPE packing materials on the 15 carbonyl compounds (n=6)

Compound	C18		PS-DVB		BRP
Compound	ME/%	RSD/%	ME/%	RSD/%	ME/%	RSD/%
FOR	110.4	1.8	108.7	1.2	90.5	1.3
ACETA	125.6	3.4	129.0	3.6	115.9	3.1
ACR	113.9	2.8	110.2	2.1	102.7	4.6
ACETO	129.0	2.7	118.0	3.5	98.4	1.3
PRO	112.8	2.4	116.1	2.4	105.2	2.3
CRO	127.2	4.6	125.4	3.6	107.8	1.0
BUT	145.4	5.6	132.1	5.3	109.7	2.2
BEN	128.0	3.7	127.1	2.0	104.4	1.3
ISO	120.9	4.1	123.7	4.7	108.3.	2.8
VAL	130.5	4.3	124.3	4.4	109.1	3.0
o-TOL	115.7	3.2	118.6	3.5	92.0	1.7
m-TOL	113.3	4.0	113.7	3.0	101.7	2.5
p-TOL	117.3	4.2	118.5	3.7	103.6	1.3
HEX	111.2	1.6	116.4	1.9	103.8	1.3
DIM	115.6	1.8	118.9	2.3	107.6	1.6

表3 15种醛酮类化合物的线性关系、检出限和定量限

Table 3 Linear relationships, limits of detection (LODs) and limits of quantification (LOQs) of the 15 carbonyl compounds

Compound	Linear range/ (μg/mL)	Linear equation	r²	LODs/(mg/L)		LOQs/(mg/L)
Compound	Linear range/ (μg/mL)	Linear equation	r²	This method	HJ 997-2018	This method	HJ 997-2018
FOR	0.1-20.0	y=13.613x+0.052	0.997	0.02	0.02	0.05	0.08
ACETA	0.2-20.0	y=10.242x+0.045	0.996	0.03	0.04	0.10	0.16
ACR	0.2-20.0	y=9.355x+0.038	0.996	0.03	0.04	0.10	0.16
ACETO	0.2-20.0	y=7.977x+0.040	0.997	0.03	0.04	0.10	0.16
PRO	0.2-20.0	y=7.419x+0.017	0.996	0.03	0.04	0.10	0.16
CRO	0.2-20.0	y=6.648x+0.017	0.997	0.04	0.04	0.14	0.16
BUT	0.5-20.0	y=4.789x+0.005	0.998	0.04	0.04	0.13	0.16
BEN	0.2-20.0	y=4.633x+0.024	0.996	0.04	0.06	0.13	0.24
ISO	0.5-20.0	y=4.037x+0.015	0.996	0.05	0.06	0.18	0.24
VAL	0.5-20.0	y=3.558x+0.067	0.997	0.05	0.06	0.18	0.24
o-TOL	0.5-20.0	y=3.865x+0.012	0.996	0.05	0.05	0.10	0.20
m-TOL	0.5-20.0	y=4.113x+0.014	0.999	0.05	0.06	0.18	0.24
p-TOL	0.5-20.0	y=3.588x+0.025	0.999	0.05	0.06	0.18	0.24
HEX	0.5-20.0	y=3.931x-0.047	0.998	0.06	0.06	0.20	0.24
DIM	0.5-20.0	y=3.294x-0.003	0.999	0.06	0.06	0.20	0.24

表4 15种醛酮类化合物的加标回收率及RSD (n=6)

Table 4 Spiked recoveries and RSDs of the 15 carbonyl compounds (n=6)

Compound	0.2 mg/kg		0.5 mg/kg		2.0 mg/kg
Compound	Recovery/ %	RSD/ %	Recovery/ %	RSD/ %	Recovery/ %	RSD/ %
FOR	84.6	1.7	88.3	2.6	90.5	1.3
ACETA	112.7	4.1	110.2	4.1	115.9	4.1
ACR	99.3	5.0	106.9	2.8	102.7	4.6
ACETO	104.4	1.2	101.1	1.7	98.4	1.3
PRO	105.1	3.5	106.1	2.7	105.2	2.3
CRO	107.2	1.6	108.4	2.5	107.8	1.0
BUT	100.2	3.5	99.8	3.6	109.7	0.2
BEN	106.3	3.9	105.4	2.7	104.4	1.3
ISO	107.0	4.3	110.6	3.0	108.3	2.8
VAL	105.2	1.9	98.6	1.6	109.1	3.0
o-TOL	87.5	0.6	89.7	3.6	92.0	1.7
m-TOL	85.4	2.8	90.2	2.7	101.7	2.5
p-TOL	91.2	5.1	96.6	2.9	103.6	1.3
HEX	104.0	2.9	106.5	2.5	103.8	0.3
DIM	97.6	2.0	96.1	1.8	107.6	0.6

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