-
饮用水消毒技术的广泛应用,使得霍乱和伤寒等介水传染病的发生率大大降低。但是,消毒剂与水中天然有机物和溴/碘离子等反应,生成有毒的消毒副产物 (DBPs),如三卤甲烷、卤乙酸和溴酸盐等[1-5]。然而,水中仍有大量DBPs因为检测技术的限制而无法被鉴定识别[2, 6-11]。
cis-2-丁烯-1,4-二醛 (BDA)是近年报道的一种新型DBPs[12-15]。BDA是一种不饱和的脂肪族醛类化合物,具有强亲核性,易与氨基酸、多肽、蛋白质和DNA等生物分子快速反应[16-19]。BDA会导致氨基酸之间的交联,造成生物体中毒[18-20]。同时,BDA是DNA直接活性诱变剂,可通过与DNA反应形成诱变加合物,例如:BDA可以与肝脏核苷酸和肾脏核苷酸形成加合物,产生量为 (33 ± 21) 个加合物/108个肝脏核苷酸和 (13 ± 5) 加合物/108个肾脏核苷酸,导致DNA单链断裂和DNA交联,增加致癌风险。另有报道,BDA会刺激肝细胞增殖,导致肿瘤的形成,例如,BDA与肝脏蛋白质共价结合,增加小鼠的肿瘤发病率[18, 20-21]。
在水体消毒过程中,酚类和呋喃类化合物有较大的BDA生成潜能,例如,实际水体中酚类物质的最高浓度可达到10 μg·L−1,经过氯消毒后BDA的产率高达20%[12, 22- 23]。同时,大气中芳香类化合物的氧化过程[24, 25]和食品中的呋喃在生物体的代谢过程也可以产生BDA [20]。鉴于BDA的生物毒性强,筛查鉴定消毒后生成的BDA非常重要。
目前有关水体中BDA的分析检测还未能引起人们广泛的关注。Churchwell等[20]利用生物标记物2-脱氧胞苷,采用四极杆飞行时间质谱 (Quadrupole-Time-of-Flight mass spectrometer) 技术分析了肝细胞中产生的BDA,衍生生物样品中BDA用到缓冲盐、水解酶和核酸外切酶等多种试剂,操作繁琐且价格昂贵,不适用于水体中BDA的检测。Prasse等[15]采用高效液相色谱-串联质谱 (HPLC-MS/MS) 技术,用标准加入法定量了紫外/过氧化氢 (UV/H2O2) 及氯消毒过程中产生的BDA,但是该方法操作繁琐,不适用于大批量样品的测试,并且其检测范围在1—10 μmol·L−1之间,定量限较高。本文拟建立适用于大批量样品测试的外标定量分析方法。在众多分析方法中,HPLC-MS/MS由于其灵敏度高和使用方便等优点,广泛应用于未知有机化合物的筛查和已知有机化合物的定量分析等[26-33]。
鉴于BDA是反应性亲电试剂,与蛋白质中的亲核部分,包括半胱氨酸、赖氨酸和组氨酸以及DNA中的伯胺或硫醇基团,发生特异性反应[12-15]。例如,1当量的BDA与1当量的氨基或硫醇反应 (当有两种氨基酸为衍生剂时,1分子BDA中的两个羰基均会参与亲核反应,形成2分子氨基酸衍生物),产生吡咯环结构加合物[15]。BDA与N-乙酰基赖氨酸 (NAL) 形成的吡咯结构的衍生物较稳定,且在HPLC-MS/MS具有较高的响应[12, 15]。
针对近年引起关注的饮用水消毒副产物BDA分析方法不完善的问题,本文选择NAL为目标生物分子,对BDA进行衍生化,再进一步选用HPLC-MS/MS在正离子条件下,应用多反应监测 (MRM) 模式进行定量分析,建立了BDA的HPLC-MS/MS分析方法,并进行了方法学验证。
新型消毒副产物cis-2-丁烯-1,4-二醛的检测方法建立与评估
Establishment and evaluation of the determination method of an emerging disinfection by-product of cis-2-butene-1,4-diformaldehyde
-
摘要: 饮用水消毒对病原微生物的预防起到至关重要的作用,但是有毒有害消毒副产物的产生,对人类健康造成潜在威胁。cis-2-丁烯-1,4-二醛 (BDA) 是近年检出的新型消毒副产物,生物毒性强,具有严重的潜在危害。本文建立了BDA的高效液相色谱-串联质谱的分析方法,旨在定量筛查出水体中的BDA。结果表明:BDA的纯溶剂标准曲线和基质匹配标准曲线的线性较好,相关系数 (R2) 均大于0.99。方法检出限为0.035 μmol·L−1,定量限值为0.105 μmol·L−1。回收率在83.6% ‒ 112%之间,相对标准偏差小于10% (n = 7)。该方法能够快速高效地完成水体中BDA的分析。
-
关键词:
- 消毒 /
- 消毒副产物 /
- cis-2-丁烯-1,4-二醛 /
- 高效液相色谱-串联质谱 (HPLC-MS/MS) /
- N-乙酰基赖氨酸衍生
Abstract: Disinfection plays a crucial role in prevention of pathogenic microorganisms in drinking water. However, the formation of toxic disinfection by-products posed a potential threat to human health. An emerging disinfection by-product, cis-2-butene-1,4-dialdehyde (BDA), possesses strong biological toxicity and serious potential hazards. This study established an analytical method for BDA by using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), which aimed to screen and quantify BDA in water. The results showed that the linearity of standard curves in both pure water and matrix-match solution were good, with the correlation coefficients (R2) higher than 0.99. The detection limit of the method was 0.035 μmol·L−1, and the quantification limit was 0.105 μmol·L−1. The recovery rate ranged from 83.6% to 112%, and the relative standard deviation was less than 10% (n = 7). This method is able to accomplish quick and efficient analysis of BDA in water. -
表 1 MRM模式下BDA的质谱参数
Table 1. Mass spectrometry parameters of BDA in MRM mode
母离子/Da
Q1 Mass/Da子离子/Da
Q3 Mass/Da停留时间/ms
Dwell time/ms去簇电压/V
Declustering potential/V碰撞能/V
Collision energy/V255.1 167.1 10 100 24 255.1 209.1 10 147 18 表 2 实际消毒水样结果验证总结
Table 2. Summary of result verificationof real disinfection water samples
加标梯度
Spiking
level峰面积
Area加标浓度/
(μmol·L−1)
Spiking
concentrition实测浓度/
(μmol·L−1)
Experimental
concentration标准偏差
Standard
deviation平均值
Mean valueRSD/% 回收率/%
Recovery空白
Blank1.09×106 0 0.15 0 0.15 3.05 — 1.09×106 0 0.16 — 1.02×106 0 0.14 — 低浓度加标
Low level spiking1.27×106 0.2 0.18 0.01 0.18 5.56 83.6 低浓度加标
Low level spiking1.24×106 0.2 0.18 0.01 0.18 5.56 97.1 1.26×106 0.2 0.18 83.6 中浓度加标
Medium level spiking7.62×106 1 1.09 0.01 1.11 1.16 93.6 7.78×106 1 1.11 96 7.85×106 1 1.12 97 高浓度加标
High level spiking3.81×107 5 5.45 0.13 5.64 2.33 106 4.03×107 5 5.76 112 4.01×107 5 5.73 112 表 3 SPE对低浓度BDA测定准确度的影响
Table 3. The influence of SPE on the accuracy of low-concentration BDA analysis
SPE前加标浓度/ (μmol·L−1)
Spiking concentration before SPESPE后理论浓度/ (μmol·L−1)
Theory concentration after SPE实测值/ (μmol·L−1)
Experimental concentration加标回收率/ %
Spiking recovery0.1 10.0 9.25 92.5 0.01 1.0 1.08 108 0.001 0.1 0.08 80 -
[1] GHANBARI F, MORADI M. Application of peroxymonosulfate and its activation methods for degradation of environmental organic pollutants: Review [J]. Chemical Engineering Journal, 2017, 310: 41-62. doi: 10.1016/j.cej.2016.10.064 [2] BOND T, HUANG J, TEMPLETON M R, et al. Occurrence and control of nitrogenous disinfection by-products in drinking water - A review [J]. Water Research, 2011, 45(15): 4341-4354. doi: 10.1016/j.watres.2011.05.034 [3] RICHARDSON S D, PLEWA M J, WAGNER E D, et al. Occurrence, genotoxicity, and carcinogenicity of regulated and emerging disinfection by-products in drinking water: A review and roadmap for research [J]. Mutation Research/Reviews in Mutation Research, 2007, 636(1/2/3): 178-242. [4] BOND T, TEMPLETON M R, GRAHAM N. Precursors of nitrogenous disinfection by-products in drinking water: A critical review and analysis [J]. Journal of Hazardous Materials, 2012, 235/236: 1-16. doi: 10.1016/j.jhazmat.2012.07.017 [5] DEBORDE M, von GUNTEN U. Reactions of chlorine with inorganic and organic compounds during water treatment—Kinetics and mechanisms: A critical review [J]. Water Research, 2008, 42(1/2): 13-51. [6] ESCHER B I, ALLINSON M, ALTENBURGER R, et al. Benchmarking organic micropollutants in wastewater, recycled water and drinking water with in vitro bioassays [J]. Environmental Science & Technology, 2014, 48(3): 1940-1956. [7] LIU X K, LIU R R, ZHU B, et al. Characterization of carbonyl disinfection by-products during ozonation, chlorination, and chloramination of dissolved organic matters [J]. Environmental Science & Technology, 2020, 54(4): 2218-2227. [8] LIU X K, LIN Y F, RUAN T, et al. Identification of N-nitrosamines and nitrogenous heterocyclic byproducts during chloramination of aromatic secondary amine precursors [J]. Environmental Science & Technology, 2020, 54(20): 12949-12958. [9] XIANG Y Y, GONSIOR M, SCHMITT-KOPPLIN P, et al. Influence of the UV/H2O2 advanced oxidation process on dissolved organic matter and the connection between elemental composition and disinfection byproduct formation [J]. Environmental Science & Technology, 2020, 54(23): 14964-14973. [10] KRASNER S W, WEINBERG H S, RICHARDSON S D, et al. Occurrence of a new generation of disinfection byproducts [J]. Environmental Science & Technology, 2006, 40(23): 7175-7185. [11] RICHARDSON S D, TERNES T A. Water analysis: Emerging contaminants and current issues [J]. Analytical Chemistry, 2018, 90(1): 398-428. doi: 10.1021/acs.analchem.7b04577 [12] PRASSE C, von GUNTEN U, SEDLAK D L. Chlorination of phenols revisited: Unexpected formation of α, β-unsaturated C4-dicarbonyl ring cleavage products [J]. Environmental Science & Technology, 2020, 54(2): 826-834. [13] MARRON E L, van BUREN J, CUTHBERTSON A A, et al. Reactions of α, β-unsaturated carbonyls with free chlorine, free bromine, and combined chlorine [J]. Environmental Science & Technology, 2021, 55(5): 3305-3312. [14] van BUREN J, PRASSE C, MARRON E L, et al. Ring-cleavage products produced during the initial phase of oxidative treatment of alkyl-substituted aromatic compounds [J]. Environmental Science & Technology, 2020, 54(13): 8352-8361. [15] PRASSE C, FORD B, NOMURA D K, et al. Unexpected transformation of dissolved phenols to toxic dicarbonyls by hydroxyl radicals and UV light [J]. Proceedings of the National Academy of Sciences of the United States of America, 2018, 115(10): 2311-2316. doi: 10.1073/pnas.1715821115 [16] KELLERT M, WAGNER S, LUTZ U, et al. Biomarkers of furan exposure by metabolic profiling of rat urine with liquid chromatography-tandem mass spectrometry and principal component analysis [J]. Chemical Research in Toxicology, 2008, 21(3): 761-768. doi: 10.1021/tx7004212 [17] CHEN L J, HECHT S S, PETERSON L A. Characterization of amino acid and glutathione adducts of cis-2-butene-1,4-dial, a reactive metabolite of furan [J]. Chemical Research in Toxicology, 1997, 10(8): 866-874. [18] PETERSON L A, CUMMINGS M E, CHAN J Y, et al. Identification of a cis-2-butene-1, 4-dial-derived glutathione conjugate in the urine of furan-treated rats [J]. Chemical Research in Toxicology, 2006, 19(9): 1138-1141. doi: 10.1021/tx060111x [19] GATES L A, LU D, PETERSON L A. Trapping of cis-2-butene-1, 4-dial to measure furan metabolism in human liver microsomes by cytochrome P450 enzymes [J]. Drug Metabolism and Disposition:the Biological Fate of Chemicals, 2012, 40(3): 596-601. doi: 10.1124/dmd.111.043679 [20] CHURCHWELL M I, SCHERI R C, von TUNGELN L S, et al. Evaluation of serum and liver toxicokinetics for furan and liver DNA adduct formation in male Fischer 344 rats [J]. Food and Chemical Toxicology, 2015, 86: 1-8. doi: 10.1016/j.fct.2015.08.029 [21] CHEN L J, HECHT S S, PETERSON L A. Identification of cis-2-butene-1, 4-dial as a microsomal metabolite of furan [J]. Chemical Research in Toxicology, 1995, 8(7): 903-906. doi: 10.1021/tx00049a001 [22] ACERO J L, PIRIOU P, von GUNTEN U. Kinetics and mechanisms of formation of bromophenols during drinking water chlorination: Assessment of taste and odor development [J]. Water Research, 2005, 39(13): 2979-2993. doi: 10.1016/j.watres.2005.04.055 [23] PIRIOU P, SOULET C, ACERO J L, et al. Understanding medicinal taste and odour formation in drinking waters [J]. Water Science and Technology, 2007, 55(5): 85-94. doi: 10.2166/wst.2007.166 [24] WANG L M, WU R R, XU C. Atmospheric oxidation mechanism of benzene. Fates of alkoxy radical intermediates and revised mechanism [J]. The Journal of Physical Chemistry. A, 2013, 117(51): 14163-14168. doi: 10.1021/jp4101762 [25] WU R R, PAN S S, LI Y, et al. Atmospheric oxidation mechanism of toluene [J]. The Journal of Physical Chemistry. A, 2014, 118(25): 4533-4547. doi: 10.1021/jp500077f [26] 黄晓梅, 吴杨, 崔君涛, 等. 高分辨质谱在氯化石蜡分析方法中的应用 [J]. 分析化学, 2019, 47(3): 323-334. doi: 10.1016/S1872-2040(19)61144-8 HUANG X M, WU Y, CUI J T, et al. Applications of high-resolution mass spectrometry in determination of chlorinated paraffins [J]. Chinese Journal of Analytical Chemistry, 2019, 47(3): 323-334(in Chinese). doi: 10.1016/S1872-2040(19)61144-8
[27] 黄林艳, 鲁炳闻, 赵彦辉, 等. 高效液相色谱分析四溴双酚A标准样品方法优化及应用 [J]. 环境化学, 2020, 39(12): 3524-3530. HUANG L Y, LU B W, ZHAO Y H, et al. Optimization and application of HPLC method for tetrabromobisphenol A reference material analysis [J]. Environmental Chemistry, 2020, 39(12): 3524-3530(in Chinese).
[28] 孙腾飞, 向垒, 陈雷, 等. 环境水样及固相样品中全氟化合物分析方法研究进展 [J]. 分析化学, 2017, 45(4): 601-610. doi: 10.11895/j.issn.0253-3820.160817 SUN T F, XIANG L, CHEN L, et al. Research progresses of determination of perfluorinated compounds in environmental water and solid samples [J]. Chinese Journal of Analytical Chemistry, 2017, 45(4): 601-610(in Chinese). doi: 10.11895/j.issn.0253-3820.160817
[29] 康春莉, 王英, 杜尧国, 等. 水体中挥发酚测定方法的改进 [J]. 分析化学, 2000, 28(7): 872-875. doi: 10.3321/j.issn:0253-3820.2000.07.019 KANG C L, WANG Y, DU Y G, et al. Improvement on the determination of volatile phenols in water [J]. Chinese Journal of Analytieal Chemistry, 2000, 28(7): 872-875(in Chinese). doi: 10.3321/j.issn:0253-3820.2000.07.019
[30] 高婷婷, 杜鹏, 徐泽琼, 等. 污水中常见违禁药物分析方法优化及验证 [J]. 环境科学, 2017, 38(1): 201-211. GAO T T, DU P, XU Z Q, et al. Optimization and validation of the analytical method to detect common illicit drugs in wastewater [J]. Environmental Science, 2017, 38(1): 201-211(in Chinese).
[31] 寇弘儒, 刘士峰, 孙艳超, 等. 基于两种前处理对苹果叶片中虫酰肼残留的液相分析方法 [J]. 环境化学, 2020, 39(1): 179-187. doi: 10.7524/j.issn.0254-6108.2019061201 KOU H R, LIU S F, SUN Y C, et al. Liquid phase analysis of tebufenozide residues in apple leaves based on two pretreatments [J]. Environmental Chemistry, 2020, 39(1): 179-187(in Chinese). doi: 10.7524/j.issn.0254-6108.2019061201
[32] ZHANG Z P, ZHANG R J, XIAO H, et al. Development of a standardized food model for studying the impact of food matrix effects on the gastrointestinal fate and toxicity of ingested nanomaterials [J]. NanoImpact, 2019, 13: 13-25. doi: 10.1016/j.impact.2018.11.002 [33] CAPPIELLO A, FAMIGLINI G, PALMA P, et al. Overcoming matrix effects in liquid chromatography-mass spectrometry [J]. Analytical Chemistry, 2008, 80(23): 9343-9348. doi: 10.1021/ac8018312