二次纳流电喷雾电离耦合超高分辨质谱检测人体呼出气中相对高分子量化合物

doi:10.7503/cjcu20160821

摘要/Abstract

摘要：

二次电喷雾电离源耦合超高分辨质谱(SESI-UHRMS)有望检出人体呼出气中分子量大于300的相对高分子量化合物, 这些化合物的发现将有助于更准确地理解呼出气中挥发性有机化合物(VOCs)的来源、产生机制以及SESI源电离机理, 更好地实现SESI-UHRMS的转化应用. 本研究自组装nanoSESI源(尚无商业产品)耦合四极杆-静电场轨道阱质谱(最高质量分辨率1.2×10⁵), 考察了该装置对健康人体呼出气中分子量为300~500化合物的检出情况. 结果表明, 所搭建的nanoSESI-UHRMS装置检测人体呼出气的重现性好、灵敏度高, 可检出数十种分子量为300~500的化合物. 根据这些化合物在单次呼气过程中信号强度随时间变化的趋势, 推断其来源分别为内源性和外源性; 各化合物的元素组成主要包括C, H, N和O, 环-双键当量(RDB)的均值为(4.5±3.1), 表明检出的化合物可能为醛酮或不饱和脂肪酸, 容易在SESI源中被电离. 本研究初步验证了自组建nanoSESI-UHRMS检测人体呼出气中相对高分子量化合物的可行性, 为后续进一步开展方法应用奠定了基础.

关键词: 人体呼出气, 相对高分子量, 直接质谱分析, 二次电喷雾电离, 超高分辨率

Abstract:

Human breath contains thousands of volatile organic compounds(VOCs), which would be potentially helpful for studying disease diagnosis and environmental exposure. However, owing to limitations in current analytical methods, only the compounds with molecular weights <300 have been widely reported in exhaled human breath. In this study, a high performance homemade secondary nanoelectrospray ionization source coupled with ultrahigh resolution mass spectrometer(nanoSESI-UHRMS) was set up and applied to investigating the presence of compounds with relatively high molecular weight(RHMW)(300—500) in healthy human breath. The results indicated that 29 RHMW compounds have been detected in real time by nanoSESI-UHRMS. These organic compounds mostly consist of C, H, N and O atoms, and the average of ring-double bond equivalent(RDB) values is (4.5±3.1), implying they are most likely to be ketones, aldehydes and fatty acids. These unsaturated compounds are characterized with high ionization efficiency in SESI, and thus readily detected by nanoSESI-UHRMS.

Key words: Human breath, Relative high molecular weight compound, Direct mass spectrometric analysis, Secondary electrospray ionization, Ultrahigh resolution

中图分类号:

O657.63

TrendMD:

黄磊, 李雪, 徐萌, 黄正旭, 周振. 二次纳流电喷雾电离耦合超高分辨质谱检测人体呼出气中相对高分子量化合物. 高等学校化学学报, 2017, 38(5): 752.

HUANG Lei, LI Xue, XU Meng, HUANG Zhengxu, ZHOU Zhen. Identification of Relatively High Molecular Weight Compounds in Human Breath Using Secondary Nano Electrospray Ionization Ultrahigh Resolution Mass Spectrometry^†. Chem. J. Chinese Universities, 2017, 38(5): 752.

图/表 6

Fig.1 Extracted ion chromatograms of three unknown compounds shown peaks at m/z 391.2829(a), 380.3375(b) and 336.2032(c) for five replicated measurements

Fig.2 Background-subtracted mass spectra(m/z 300—500) of exhaled breath of a subject under positive(A) and negative(B) ion detection modes

Fig.3 Background-subtracted mass spectra of isobaric ions m/z 300.1806, 300.2170, 300.2535 and 300.2896 at R values of 1.2×105(solid line) and 1.5×104(short dash line)

Fig.4 Six replicate measurements of m/z 300.2170 at resolutions of 1.2×105(A) and 1.5×104(B)

Fig.5 Six replicate measurements of m/z 300.2896 at resolutions of 1.2×105(A) and 1.5×104(B)

Table 1 Identification of 29 compounds in the range of m/z 300—500 from breath samples of 4 subjects

Compound	m/z	Detection mode	Composition	RDB^a	S1^b	S2^b	S3^b	S4^b
1	300.2896	Positive	C₁₈H₃₈NO₂	0.5	Det.^c	ND^d	Det.	Det.
2	301.2385	Negative	$C 1 7 H 3 3$ O₄	1.5	Det.	Det.	Det.	Det.
3	312.1806	Positive	C₁₆H₂₆NO₅	4.5	Det.	Det.	Det.	ND
4	322.1865	Positive	C₁₄H₂₈NO₇	1.5	ND	ND	ND	Det.
5	324.2170	Positive	C₁₈H₃₀NO₄	4.5	Det.	Det.	Det.	Det.
6	332.9061	Negative			ND	ND	Det.	ND
7	334.9033	Negative			ND	ND	Det.	ND
8	336.2032	Positive	C₁₅H₃₀NO₇	1.5	Det.	Det.	Det.	Det.
9	338.1675	Positive			Det.	ND	ND	Det.
10	338.2328	Positive	C₁₉H₃₂NO₄	4.5	Det.	Det.	Det.	Det.
11	339.5948	Negative			Det.	ND	Det.	ND
12	346.0331	Positive			ND	ND	Det.	ND
13	346.1862	Positive	C₁₆H₂₈NO₇	3.5	Det.	ND	ND	ND
14	348.2170	Positive	C₂₀H₃₀NO₄	6.5	Det.	Det.	Det.	Det.
15	352.2483	Positive	C₂₀H₃₄NO₄	4.5	Det.	ND	ND	Det.
16	356.2797	Positive	C₂₀H₃₈NO₄	2.5	Det.	Det.	Det.	Det.
17	358.2016	Positive	C₂₁H₂₈NO₄	8.5	ND	Det.	Det.	Det.
18	363.3107	Positive	C₂₀H₄₃O₅	-0.5	Det.	Det.	Det.	Det.
			C₁₉H₃₇N₇	5.0	Det.	Det.	Det.	Det.
19	373.8173	Negative			Det.	ND	Det.	Det.
20	378.9106	Negative			ND	Det.	Det.	ND
21	380.3375	Positive	C₁₉H₄₀N₈	4.0	Det.	Det.	Det.	Det.
22	380.9078	Negative			ND	Det.	Det.	ND
23	382.9049	Negative			ND	Det.	ND	ND
24	386.0830	Positive	C₁₂H₁₄N₆O₉	9.0	Det.	ND	ND	Det.
25	388.0789	Positive	C₁₉H₁₆O₉	12.0	Det.	ND	ND	Det.
26	391.2829	Positive	C₂₄H₃₉O₄	5.5	Det.	Det.	Det.	Det.
27	452.3952	Positive	C₂₃H₄₈N₈O	4.0	Det.	Det.	Det.	Det.
28	462.1468	Positive	C₁₂H₄₀NO₆Si₆	-0.5	ND	ND	Det.	ND
29	464.1280	Positive	C₁₅H₂₂N₅O₁₂	7.5	ND	ND	Det.	ND

Table 1 Identification of 29 compounds in the range of m/z 300—500 from breath samples of 4 subjects

Compound	m/z	Detection mode	Composition	RDB^a	S1^b	S2^b	S3^b	S4^b
1	300.2896	Positive	C₁₈H₃₈NO₂	0.5	Det.^c	ND^d	Det.	Det.
2	301.2385	Negative	$C 1 7 H 3 3$ O₄	1.5	Det.	Det.	Det.	Det.
3	312.1806	Positive	C₁₆H₂₆NO₅	4.5	Det.	Det.	Det.	ND
4	322.1865	Positive	C₁₄H₂₈NO₇	1.5	ND	ND	ND	Det.
5	324.2170	Positive	C₁₈H₃₀NO₄	4.5	Det.	Det.	Det.	Det.
6	332.9061	Negative			ND	ND	Det.	ND
7	334.9033	Negative			ND	ND	Det.	ND
8	336.2032	Positive	C₁₅H₃₀NO₇	1.5	Det.	Det.	Det.	Det.
9	338.1675	Positive			Det.	ND	ND	Det.
10	338.2328	Positive	C₁₉H₃₂NO₄	4.5	Det.	Det.	Det.	Det.
11	339.5948	Negative			Det.	ND	Det.	ND
12	346.0331	Positive			ND	ND	Det.	ND
13	346.1862	Positive	C₁₆H₂₈NO₇	3.5	Det.	ND	ND	ND
14	348.2170	Positive	C₂₀H₃₀NO₄	6.5	Det.	Det.	Det.	Det.
15	352.2483	Positive	C₂₀H₃₄NO₄	4.5	Det.	ND	ND	Det.
16	356.2797	Positive	C₂₀H₃₈NO₄	2.5	Det.	Det.	Det.	Det.
17	358.2016	Positive	C₂₁H₂₈NO₄	8.5	ND	Det.	Det.	Det.
18	363.3107	Positive	C₂₀H₄₃O₅	-0.5	Det.	Det.	Det.	Det.
			C₁₉H₃₇N₇	5.0	Det.	Det.	Det.	Det.
19	373.8173	Negative			Det.	ND	Det.	Det.
20	378.9106	Negative			ND	Det.	Det.	ND
21	380.3375	Positive	C₁₉H₄₀N₈	4.0	Det.	Det.	Det.	Det.
22	380.9078	Negative			ND	Det.	Det.	ND
23	382.9049	Negative			ND	Det.	ND	ND
24	386.0830	Positive	C₁₂H₁₄N₆O₉	9.0	Det.	ND	ND	Det.
25	388.0789	Positive	C₁₉H₁₆O₉	12.0	Det.	ND	ND	Det.
26	391.2829	Positive	C₂₄H₃₉O₄	5.5	Det.	Det.	Det.	Det.
27	452.3952	Positive	C₂₃H₄₈N₈O	4.0	Det.	Det.	Det.	Det.
28	462.1468	Positive	C₁₂H₄₀NO₆Si₆	-0.5	ND	ND	Det.	ND
29	464.1280	Positive	C₁₅H₂₂N₅O₁₂	7.5	ND	ND	Det.	ND

参考文献 27

[1]	Pleil J. D., Stiegel M. A., Anal. Chem., 2013, 85, 9984—9990
[2]	de Lacy C. B., Amann A., Al-Kateb H., Flynn C., Filipiak W., Khalid T., Osborne D., Ratcliffe N. M., J. Breath Res., 2014, 8, 911—922
[3]	Phillips M., Gleeson K., Hughes J.M. B., Greenberg J., Cataneo R. N., Baker L., McVay W. P., Lancet, 1999, 353, 1930—1933
[4]	Peng G., Tisch U., Adams O., Hakim M., Shehada N., Broza Y. Y., Billan S., Abdah-Bortnyak R., Kuten A., Haick H., Nature Nanotech., 2009, 4, 669—673
[5]	Pleil J. D., Fisher J. W., Lindstrom A. B., Environ. Health Perspect., 1998, 106, 573—580
[6]	Pleil J. D., Stiegel M. A., Anal. Chem., 2013, 85, 9984—9990
[7]	Vereb H., Dietrich A. M., Alfeeli B., Agah M., Environ. Sci. Technol., 2011, 45, 8167—8175
[8]	Bajtarevic A., Ager C., Pienz M., Klieber M., Schwarz K., Ligor M., Ligor T., Filipiak W., Denz H., Fiegl M., Hilbe W., Weiss W., Lukas P., Jamnig H., Hackl M., Haidenberger A., Buszewski B., Miekisch W., Schubert J., Amann A., BMC Cancer, 2009, 9, 1—16
[9]	Shen C. Y., Li J. Q., Wang H. Z., Zhi Z. H., Wang H. M., Huang C. Q., Liu S., Jiang H. H., Chu Y. N., Chinese J. Anal. Chem., 2012, 40(5), 773—777
	(沈成银, 李建权, 王宏志, 志中华, 王鸿梅, 黄超群, 刘升, 江海河, 储焰南.分析化学, 2012,40(5), 773—777)
[10]	Shen C. Y., Wang H. M., Huang C. Q., Lu Y., Xia L., Chen X. J., Wang H. Z., Chu Y. N., Chem. J. Chinese Universities, 2015, 36(2), 236—240
	(沈成银, 王鸿梅, 黄超群, 陆燕, 夏磊, 陈小景, 王宏志, 储焰南.高等学校化学学报, 2015,36(2), 236—240)
[11]	Endre Z. H., Pickering J. W., Storer M. K., Hu W. P., Moorhead K. T., Allardyce R., McGregor D. O., Scotter J. M., Physiol. Meas., 2011, 32, 115—130
[12]	Smith D., Wang T. S., Pysanenko A., Španěl P., Rapid Commun. Mass Spectrom., 2008, 22, 783—789
[13]	Turner C., Španěl P., Smith D., Physiol. Meas., 2006, 27, 321—337
[14]	Zeng Q., Li P. H., Cai Y. F., Zhou W., Wang H. D., Luo J., Ding J. H., Chen H. W., J. Breath Res., 2016, 10, 016008
[15]	Chen H. W., Wortmann A., Zhang W. H., Zenobi R., Angew. Chem. Int. Ed., 2007, 46, 580—583
[16]	Li X., Sinues P. M. L., Dallmann R., Bregy L., Hollmén M., Proulx S., Brown S. A., Detma M., Kohler M., Zenobi R., Angew. Chem. Int. Ed., 2015, 54, 7815—7818
[17]	Martinez-Lozano Sinues P., Kohler M., Zenobi R., Anal. Chem., 2013, 85, 369—373
[18]	Benoit F. M., Davidson W. R., Lovett A. M., Nacson S., Ngo A., Int. Arch. Occup. Environ. Health, 1985, 55, 113—120
[19]	Benoit F. M., Davidson W. R., Lovett A. M., Ngo A., Anal. Chem., 1983, 55, 805—807
[20]	Bregy L., Sinues P. M. L., Nudnova M. M., Zenobi R., J. Breath Res., 2014, 8, 027102
[21]	Li X., Huang L., Zhu H., Zhou Z., Rapid Commun. Mass Spectrom., 2016, 31, 301—308
[22]	Martinez-Lozano Sinues P., Malcolm K., Brown S. A., Zenobi R., Dallmann R., Chem. Commun., 2017, 53, 2264—2267
[23]	García-Gómez D., Gaisl T., Bregy L., Cremonesi A., Martinez-Lozano Sinues P., Kohler M., Zenobi R., Clin. Chem., 2016, 62, 1230—1237
[24]	García-Gómez D., Gaisl T., Bregy L., Martinez-Lozano Sinues P., Kohler M., Zenobi R., Chem. Commun., 2016, 52, 8526—8528
[25]	Gaugg M. T., García-Gómez D., Collado C. B., de Miguel G. V., Kohler M., Zenobi R., J. Breath Res., 2016, 10, 016010
[26]	Phillips M., Herrera J., Krishnan S., Zain M., Greenberg J., Cataneo R. N., J. Chromatogr. B, 1999, 729, 75—88
[27]	Poli D., Carbognani P., Corradi M., Goldoni M., Acampa O., Balbi B., Bianchi L., Rusca M., Mutti A., Respir. Res., 2005, 6, 1—10