基于液质联用的莱茵衣藻极性甘油酯组定性定量分析

doi:10.11983/CBB17179

植物学报 ›› 2018, Vol. 53 ›› Issue (6): 812-828.DOI: 10.11983/CBB17179

基于液质联用的莱茵衣藻极性甘油酯组定性定量分析

杨淼^1,^2,³, 孟迎迎^1,³, 褚亚东¹, 薛松^1,^*()

¹中国科学院大连化学物理研究所, 大连 116023
²中国科学院大学, 北京 100049
³大连理工大学生命科学与技术学院, 大连 116024

收稿日期:2017-09-24 出版日期:2018-11-01 发布日期:2018-12-05
通讯作者: 薛松
作者简介:
作者简介:白克智, 1959年开始在中国科学院植物研究所工作, 先后任助理研究员、研究员, 长期从事植物生长发育及其调控的研究。1986年,其主持的“满江红生物学特性研究”荣获中国科学院科技进步二等奖。曾任《植物生理学报》编委、《植物学报》常务编委、中国植物生长调节剂协会主任等职。
基金资助:
国家自然科学基金(No.21576253)

Qualitative and Quantitative Analyses of Polar Glycerolipids in Chlamydomonas reinhardtii based on LC-MS Techniques

Yang Miao^1,^2,³, Meng Yingying^1,³, Chu Yadong¹, Xue Song^1,^*()

¹Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
²University of Chinese Academy of Sciences, Beijing 100049, China
³School of Life Sciences and Biotechnology, Dalian University of Technology, Dalian 116024, China

Received:2017-09-24 Online:2018-11-01 Published:2018-12-05
Contact: Xue Song

摘要/Abstract

摘要： 以模式藻株莱茵衣藻(Chlamydomonas reinhardtii)为材料, 基于液质联用技术对其极性甘油酯组进行定性定量分析。通过综合利用UPLC-ESI-Q-Trap/MS的一级质谱扫描(中性丢失或母离子扫描)及UPLC-ESI-Orbitrap/MS²的二级碎片信息扫描, 共鉴定出109种极性甘油酯分子; 再通过外标法利用UPLC-ESI-Q-Trap/MS在多级反应监测模式下对各分子进行靶向定量分析。结果表明, 莱茵衣藻的极性脂以糖脂MGDG、DGDG及甜菜碱脂DGTS为主, 所有极性脂的分子组成表明, DGDG、SQDG、DGTS及PI是C18脂肪酸的去饱和载体。该研究利用液质联用技术建立了莱茵衣藻极性甘油酯组的结构图谱及定量分析技术平台, 为微藻极性脂生物学功能及脂质代谢研究奠定了基础。

关键词: 莱茵衣藻, 液质联用技术, 极性甘油酯组, 定性定量分析

Abstract: The polar glycerolipidome of the model algal strain Chlamydomonas reinhardtii was analyzed qualitatively and quantitatively by using liquid chromatography coupled with mass spectrometry (LC-MS). Based on UPLC-ESI-Q-Trap/MS and UPLC-ESI-Orbitrap/MS², MS scanning, including neutral loss or precursor ion scanning, and MS/MS scanning were used to identify 109 molecular species possessing distinct mass-to-charge ratios and acyl chains. Targeted quantitative analyses based on external standards with UPLC-ESI-Q-Trap/MS in multiple reactions monitoring (MRM) mode was used to analyze individual molecular species. The results indicated that the glycerolipids of C. reinhardtii mainly included glycolipids (i.e., MGDG and DGDG) and betaine lipid, DGTS. In addition, analysis of fatty acyl constituents of all polar membrane lipids revealed that DGDG, SQDG, DGTS and PI acted as carriers for desaturation of C18 fatty acids. This study established a platform for characterizing the structure spectra and quantitative analysis with LC-MS techniques for C. reinhardtii, which provides a useful tool for studies of the biological function of polar membrane lipids and lipid metabolism in microalgae.

Key words: Chlamydomonas reinhardtii, LC-MS technique, polar glycerolipids, qualitative and quantitative analyses

杨淼, 孟迎迎, 褚亚东, 薛松. 基于液质联用的莱茵衣藻极性甘油酯组定性定量分析. 植物学报, 2018, 53(6): 812-828.

Yang Miao, Meng Yingying, Chu Yadong, Xue Song. Qualitative and Quantitative Analyses of Polar Glycerolipids in Chlamydomonas reinhardtii based on LC-MS Techniques. Chinese Bulletin of Botany, 2018, 53(6): 812-828.

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Lipids	Classes	Adduct ion	Scanning mode	Mass of characteristic fragment (m/z)	Chemical formula of characteristic fragment	Collision energy (eV)
Glycolipids	MGDG	[M+NH₄]⁺	NL	179	[C₆H₁₃NO₅]	20
	DGDG	[M+NH₄]⁺	NL	341	[C₁₂H₂₃NO₁₀]	20
	SQDG	[M+NH₄]⁺	NL	261	[C₆H₁₅NO₈S]	32
Phospholipids	PE	[M+H]⁺	NL	141	[C₂H₈NO₄P]	28
	PG	[M+NH₄]⁺	NL	189	[C₃H₁₂NO₆P]	15
	PI	[M+NH₄]⁺	NL	277	[C₆H₁₆NO₉P]	27
Betaine lipids	DGTS	[M+H]⁺	PIS	236	[C₁₀H₂₂NO₅]⁺	43

Lipids	Classes	Adduct ion	Scanning mode	Mass of characteristic fragment (m/z)	Chemical formula of characteristic fragment	Collision energy (eV)
Glycolipids	MGDG	[M+NH₄]⁺	NL	179	[C₆H₁₃NO₅]	20
	DGDG	[M+NH₄]⁺	NL	341	[C₁₂H₂₃NO₁₀]	20
	SQDG	[M+NH₄]⁺	NL	261	[C₆H₁₅NO₈S]	32
Phospholipids	PE	[M+H]⁺	NL	141	[C₂H₈NO₄P]	28
	PG	[M+NH₄]⁺	NL	189	[C₃H₁₂NO₆P]	15
	PI	[M+NH₄]⁺	NL	277	[C₆H₁₆NO₉P]	27
Betaine lipids	DGTS	[M+H]⁺	PIS	236	[C₁₀H₂₂NO₅]⁺	43

Lipid species (C:N)	Theoretical mass (m/z)	Chemical formula	No.	Molecular species (sn-1/sn-2)	Precursor ion (m/z)	Product ion (m/z)
MGDG 34:7	762.5156	C₄₃H₇₂NO₁₀	1	MGDG 18:3/16:4	762.5	335.2
MGDG 34:6	764.5312	C₄₃H₇₄NO₁₀	2	MGDG 18:3/16:3	764.5	335.2
			3	MGDG 18:2/16:4	764.5	337.2
			4	MGDG 18:4/16:2	764.5	309.2
MGDG 34:5	766.5469	C₄₃H₇₆NO₁₀	5	MGDG 18:3/16:2	766.5	335.2
			6	MGDG 18:2/16:3	766.5	337.2
			7	MGDG 18:1/16:4	766.5	339.2
			8	MGDG 18:4/16:1	766.5	333.2
MGDG 34:4	768.5625	C₄₃H₇₈NO₁₀	9	MGDG 18:1/16:3	768.5	339.2
			10	MGDG 18:2/16:2	768.5	337.2
			11	MGDG 18:3/16:1	768.5	311.2
			12	MGDG 18:4/16:0	768.5	313.2
			13	MGDG 18:0/16:4	768.5	341.2
MGDG 34:3	770.5781	C₄₃H₈₀NO₁₀	14	MGDG 18:1/16:2	770.5	339.2
			15	MGDG 18:2/16:1	770.5	337.2
			16	MGDG 18:3/16:0	770.5	335.2
MGDG 34:2	772.5938	C₄₃H₈₂NO₁₀	17	MGDG 18:1/16:1	772.5	339.2
			18	MGDG 18:0/16:2	772.5	341.2
			19	MGDG 18:2/16:0	772.5	313.2
MGDG 34:1	774.6094	C₄₃H₈₄NO₁₀	20	MGDG 18:1/16:0	774.6	339.2
MGDG 34:1	774.6094	C₄₃H₈₄NO₁₀	21	MGDG 18:0/16:1	774.6	341.2
DGDG 32:3	904.5997	C₄₇H₈₆NO₁₅	22	DGDG 16:0/16:3	904.5	313.2
DGDG 32:3	904.5997	C₄₇H₈₆NO₁₅	23	DGDG 16:1/16:2	904.5	311.2
DGDG 34:7	924.5684	C₄₉H₈₂NO₁₅	24	DGDG 18:3/16:4	924.5	335.2
DGDG 34:6	926.5840	C₄₉H₈₄NO₁₅	25	DGDG 18:3/16:3	926.5	335.2
			26	DGDG 18:2/16:4	926.5	337.2
			27	DGDG 18:4/16:2	926.5	333.2
DGDG 34:5	928.5997	C₄₉H₈₆NO₁₅	28	DGDG 18:2/16:3	928.5	337.2
DGDG 34:5	928.5997	C₄₉H₈₆NO₁₅	29	DGDG 18:3/16:2	928.5	335.2
DGDG 34:4	930.6153	C₄₉H₈₈NO₁₅	30	DGDG 18:3/16:1	930.6	335.2
			31	DGDG 18:2/16:2	930.6	337.2
			32	DGDG 18:1/16:3	930.6	339.2
DGDG 34:3	932.6309	C₄₉H₉₀NO₁₅	33	DGDG 18:1/16:2	932.6	339.2
			34	DGDG 18:3/16:0	932.6	313.2
			35	DGDG 18:2/16:1	932.6	337.2
DGDG 34:2	934.6466	C₄₉H₉₂NO₁₅	36	DGDG 18:1/16:1	934.6	339.2
DGDG 34:2	934.6466	C₄₉H₉₂NO₁₅	37	DGDG 18:2/16:0	934.6	337.2
DGDG 34:1	936.6622	C₄₉H₉₄NO₁₅	38	DGDG 18:1/16:0	936.6	339.2
DGDG 34:0	938.6779	C₄₉H₉₆NO₁₅	39	DGDG 18:0/16:0	938.6	341.2
DGDG 36:4	958.6466	C₅₁H₉₂NO₁₅	40	DGDG 18:1/18:3	958.6	339.2
DGDG 36:4	958.6466	C₅₁H₉₂NO₁₅	41	DGDG 18:2/18:2	958.6	337.2
DGDG 36:3	960.6622	C₅₁H₉₄NO₁₅	42	DGDG 18:0/18:3	960.6	341.2
SQDG 32:1	810.5400	C₄₁H₈₀NO₁₂S	43	SQDG 16:0/16:1	810.5	313.2
SQDG 32:0	812.5557	C₄₁H₈₂NO₁₂S	44	SQDG 16:0/16:0	812.5	313.2
SQDG 34:3	834.5400	C₄₃H₈₀NO₁₂S	45	SQDG 18:3/16:0	834.5	313.2
表2 (续) Table 2 (continued)
Lipid species (C:N)	Theoretical mass (m/z)	Chemical formula	No.	Molecular species (sn-1/sn-2)	Precursor ion (m/z)	Product ion (m/z)
SQDG 34:2	836.5557	C₄₃H₈₂NO₁₂S	46	SQDG 18:2/16:0	836.5	313.2
SQDG 34:1	838.5713	C₄₃H₈₄NO₁₂S	47	SQDG 18:1/16:0	838.5	313.2
SQDG 34:0	840.587	C₄₃H₈₆NO₁₂S	48	SQDG 18:0/16:0	840.5	341.2
PE 32:2	688.4916	C₃₇H₇₁NO₈P	49	PE 16:1/16:1	688.4	547.4
PE 32:1	690.5073	C₃₇H₇₃NO₈P	50	PE 16:0/16:1	690.5	549.5
PE 32:0	692.5229	C₃₇H₇₅NO₈P	51	PE 16:0/16:0	692.5	551.5
PE 34:4	712.4916	C₃₉H₇₁NO₈P	52	PE 16:0/18:3	714.5	573.5
PE 34:2	716.5229	C₃₉H₇₅NO₈P	53	PE 34:2	716.5	575.5
PE 34:1	718.5386	C₃₉H₇₇NO₈P	54	PE 16:0/18:1	718.5	577.5
PE 36:6	736.4916	C₄₁H₇₁NO₈P	55	PE 18:3/18:3	736.4	595.4
PE 36:5	738.5073	C₄₁H₇₃NO₈P	56	PE 18:2/18:3	738.5	597.5
PE 36:4	740.5229	C₄₁H₇₅NO₈P	57	PE 36:4	740.5	599.5
PE 36:3	742.5386	C₄₁H₇₇NO₈P	58	PE 18:0/18:3	742.5	601.5
PE 36:2	744.5543	C₄₁H₇₉NO₈P	59	PE 18:1/18:1	744.5	603.5
PG 32:2	736.5128	C₃₈H₇₅NO₁₀P	60	PG 16:1/16:1	736.5	547.5
PG 32:1	738.5284	C₃₈H₇₇NO₁₀P	61	PG 16:0/16:1	738.5	549.5
PG 32:0	740.5441	C₃₈H₇₉NO₁₀P	62	PG 16:0/16:0	740.5	551.5
PG 34:4	760.5128	C₄₀H₇₅NO₁₀P	63	PG 18:3/16:1	760.5	571.5
PG 34:3	762.5284	C₄₀H₇₇NO₁₀P	64	PG 34:3	762.5	573.5
PG 34:2	764.5441	C₄₀H₇₉NO₁₀P	65	PG 34:2	764.5	575.5
PG 34:1	766.5598	C₄₀H₈₁NO₁₀P	66	PG 18:1/16:0	766.5	577.5
PG 34:0	768.5754	C₄₀H₈₃NO₁₀P	67	PG 18:0/16:0	768.5	579.5
PI 34:3	850.5445	C₄₃H₈₁NO₁₃P	68	PI 18:3/16:0	850.5	573.5
PI 34:2	852.5601	C₄₃H₈₃NO₁₃P	69	PI 34:2	852.5	575.5
PI 34:1	854.5757	C₄₃H₈₅NO₁₃P	70	PI 18:1/16:0	854.5	577.5
PI 34:0	856.5914	C₄₃H₈₇NO₁₃P	71	PI 18:0/16:0	856.5	579.5
DGTS 32:4	704.5464	C₄₂H₇₄NO₇	72	DGTS 16:0/16:4	704.5	474.4
			73	DGTS 16:3/16:1	704.5	472.4
			74	DGTS 16:2/16:2	704.5	470.4
DGTS 32:3	706.5621	C₄₂H₇₆NO₇	75	DGTS 16:0/16:3	706.5	474.4
DGTS 32:3	706.5621	C₄₂H₇₆NO₇	76	DGTS 16:2/16:1	706.5	470.4
DGTS 32:2	708.5778	C₄₂H₇₈NO₇	77	DGTS 16:0/16:2	708.5	474.4
DGTS 32:1	710.5934	C₄₂H₈₀NO₇	78	DGTS 16:0/16:1	710.5	474.4
DGTS 32:0	712.6091	C₄₂H₈₂NO₇	79	DGTS 16:0/16:0	712.6	474.4
DGTS 34:7	726.5308	C₄₄H₇₂NO₇	80	DGTS 16:4/18:3	726.5	466.4
DGTS 34:7	726.5308	C₄₄H₇₂NO₇	81	DGTS 16:3/18:4	726.5	468.4
DGTS 34:6	728.5464	C₄₄H₇₄NO₇	82	DGTS 16:3/18:3	728.5	468.4
			83	DGTS 16:4/18:2	728.5	466.4
			84	DGTS 16:2/18:4	728.5	494.4
DGTS 34:5	730.5621	C₄₄H₇₆NO₇	85	DGTS 16:4/18:1	730.5	500.4
			86	DGTS 16:1/18:4	730.5	472.4
			87	DGTS 16:3/18:2	730.5	498.4
			88	DGTS 16:2/18:3	730.5	470.4
DGTS 34:4	732.5778	C₄₄H₇₈NO₇	89	DGTS 16:1/18:3	732.5	496.4
			90	DGTS 16:2/18:2	732.5	498.4
			91	DGTS 16:3/18:1	732.5	500.4
			92	DGTS 16:0/18:4	732.5	474.4
表2 (续) Table 2 (continued)
Lipid species (C:N)	Theoretical mass (m/z)	Chemical formula	No.	Molecular species (sn-1/sn-2)	Precursor ion (m/z)	Product ion (m/z)
			93	DGTS 16:4/18:0	732.5	502.4
DGTS 34:3	734.5934	C₄₄H₈₀NO₇	94	DGTS 16:0/18:3	734.5	474.4
DGTS 34:2	736.6091	C₄₄H₈₂NO₇	95	DGTS 16:0/18:2	736.6	474.4
DGTS 34:1	738.6247	C₄₄H₈₄NO₇	96	DGTS 16:0/18:1	738.6	474.4
DGTS 34:1	738.6247	C₄₄H₈₄NO₇	97	DGTS 16:1/18:0	738.6	502.4
DGTS 36:7	754.5621	C₄₆H₇₆NO₇	98	DGTS 18:3/18:4	754.5	496.4
DGTS 36:6	756.5778	C₄₆H₇₈NO₇	99	DGTS 18:3/18:3	756.5	496.4
DGTS 36:6	756.5778	C₄₆H₇₈NO₇	100	DGTS 18:4/18:2	756.5	498.4
DGTS 36:5	758.5934	C₄₆H₈₀NO₇	101	DGTS 18:2/18:3	758.5	498.4
DGTS 36:5	758.5934	C₄₆H₈₀NO₇	102	DGTS 18:1/18:4	758.5	500.4
DGTS 36:4	760.6091	C₄₆H₈₂NO₇	103	DGTS 18:2/18:2	760.6	498.4
			104	DGTS 18:1/18:3	760.6	500.4
			105	DGTS 18:0/18:4	760.6	502.4
DGTS 36:3	762.6247	C₄₆H₈₄NO₇	106	DGTS 18:0/18:3	762.6	502.4
DGTS 36:3	762.6247	C₄₆H₈₄NO₇	107	DGTS 18:1/18:2	762.6	500.4
DGTS 36:2	764.6404	C₄₆H₈₆NO₇	108	DGTS 18:1/18:1	764.6	500.4
DGTS 36:2	764.6404	C₄₆H₈₆NO₇	109	DGTS 18:0/18:2	764.6	502.4

Lipid species (C:N)	Theoretical mass (m/z)	Chemical formula	No.	Molecular species (sn-1/sn-2)	Precursor ion (m/z)	Product ion (m/z)
MGDG 34:7	762.5156	C₄₃H₇₂NO₁₀	1	MGDG 18:3/16:4	762.5	335.2
MGDG 34:6	764.5312	C₄₃H₇₄NO₁₀	2	MGDG 18:3/16:3	764.5	335.2
			3	MGDG 18:2/16:4	764.5	337.2
			4	MGDG 18:4/16:2	764.5	309.2
MGDG 34:5	766.5469	C₄₃H₇₆NO₁₀	5	MGDG 18:3/16:2	766.5	335.2
			6	MGDG 18:2/16:3	766.5	337.2
			7	MGDG 18:1/16:4	766.5	339.2
			8	MGDG 18:4/16:1	766.5	333.2
MGDG 34:4	768.5625	C₄₃H₇₈NO₁₀	9	MGDG 18:1/16:3	768.5	339.2
			10	MGDG 18:2/16:2	768.5	337.2
			11	MGDG 18:3/16:1	768.5	311.2
			12	MGDG 18:4/16:0	768.5	313.2
			13	MGDG 18:0/16:4	768.5	341.2
MGDG 34:3	770.5781	C₄₃H₈₀NO₁₀	14	MGDG 18:1/16:2	770.5	339.2
			15	MGDG 18:2/16:1	770.5	337.2
			16	MGDG 18:3/16:0	770.5	335.2
MGDG 34:2	772.5938	C₄₃H₈₂NO₁₀	17	MGDG 18:1/16:1	772.5	339.2
			18	MGDG 18:0/16:2	772.5	341.2
			19	MGDG 18:2/16:0	772.5	313.2
MGDG 34:1	774.6094	C₄₃H₈₄NO₁₀	20	MGDG 18:1/16:0	774.6	339.2
MGDG 34:1	774.6094	C₄₃H₈₄NO₁₀	21	MGDG 18:0/16:1	774.6	341.2
DGDG 32:3	904.5997	C₄₇H₈₆NO₁₅	22	DGDG 16:0/16:3	904.5	313.2
DGDG 32:3	904.5997	C₄₇H₈₆NO₁₅	23	DGDG 16:1/16:2	904.5	311.2
DGDG 34:7	924.5684	C₄₉H₈₂NO₁₅	24	DGDG 18:3/16:4	924.5	335.2
DGDG 34:6	926.5840	C₄₉H₈₄NO₁₅	25	DGDG 18:3/16:3	926.5	335.2
			26	DGDG 18:2/16:4	926.5	337.2
			27	DGDG 18:4/16:2	926.5	333.2
DGDG 34:5	928.5997	C₄₉H₈₆NO₁₅	28	DGDG 18:2/16:3	928.5	337.2
DGDG 34:5	928.5997	C₄₉H₈₆NO₁₅	29	DGDG 18:3/16:2	928.5	335.2
DGDG 34:4	930.6153	C₄₉H₈₈NO₁₅	30	DGDG 18:3/16:1	930.6	335.2
			31	DGDG 18:2/16:2	930.6	337.2
			32	DGDG 18:1/16:3	930.6	339.2
DGDG 34:3	932.6309	C₄₉H₉₀NO₁₅	33	DGDG 18:1/16:2	932.6	339.2
			34	DGDG 18:3/16:0	932.6	313.2
			35	DGDG 18:2/16:1	932.6	337.2
DGDG 34:2	934.6466	C₄₉H₉₂NO₁₅	36	DGDG 18:1/16:1	934.6	339.2
DGDG 34:2	934.6466	C₄₉H₉₂NO₁₅	37	DGDG 18:2/16:0	934.6	337.2
DGDG 34:1	936.6622	C₄₉H₉₄NO₁₅	38	DGDG 18:1/16:0	936.6	339.2
DGDG 34:0	938.6779	C₄₉H₉₆NO₁₅	39	DGDG 18:0/16:0	938.6	341.2
DGDG 36:4	958.6466	C₅₁H₉₂NO₁₅	40	DGDG 18:1/18:3	958.6	339.2
DGDG 36:4	958.6466	C₅₁H₉₂NO₁₅	41	DGDG 18:2/18:2	958.6	337.2
DGDG 36:3	960.6622	C₅₁H₉₄NO₁₅	42	DGDG 18:0/18:3	960.6	341.2
SQDG 32:1	810.5400	C₄₁H₈₀NO₁₂S	43	SQDG 16:0/16:1	810.5	313.2
SQDG 32:0	812.5557	C₄₁H₈₂NO₁₂S	44	SQDG 16:0/16:0	812.5	313.2
SQDG 34:3	834.5400	C₄₃H₈₀NO₁₂S	45	SQDG 18:3/16:0	834.5	313.2
表2 (续) Table 2 (continued)
Lipid species (C:N)	Theoretical mass (m/z)	Chemical formula	No.	Molecular species (sn-1/sn-2)	Precursor ion (m/z)	Product ion (m/z)
SQDG 34:2	836.5557	C₄₃H₈₂NO₁₂S	46	SQDG 18:2/16:0	836.5	313.2
SQDG 34:1	838.5713	C₄₃H₈₄NO₁₂S	47	SQDG 18:1/16:0	838.5	313.2
SQDG 34:0	840.587	C₄₃H₈₆NO₁₂S	48	SQDG 18:0/16:0	840.5	341.2
PE 32:2	688.4916	C₃₇H₇₁NO₈P	49	PE 16:1/16:1	688.4	547.4
PE 32:1	690.5073	C₃₇H₇₃NO₈P	50	PE 16:0/16:1	690.5	549.5
PE 32:0	692.5229	C₃₇H₇₅NO₈P	51	PE 16:0/16:0	692.5	551.5
PE 34:4	712.4916	C₃₉H₇₁NO₈P	52	PE 16:0/18:3	714.5	573.5
PE 34:2	716.5229	C₃₉H₇₅NO₈P	53	PE 34:2	716.5	575.5
PE 34:1	718.5386	C₃₉H₇₇NO₈P	54	PE 16:0/18:1	718.5	577.5
PE 36:6	736.4916	C₄₁H₇₁NO₈P	55	PE 18:3/18:3	736.4	595.4
PE 36:5	738.5073	C₄₁H₇₃NO₈P	56	PE 18:2/18:3	738.5	597.5
PE 36:4	740.5229	C₄₁H₇₅NO₈P	57	PE 36:4	740.5	599.5
PE 36:3	742.5386	C₄₁H₇₇NO₈P	58	PE 18:0/18:3	742.5	601.5
PE 36:2	744.5543	C₄₁H₇₉NO₈P	59	PE 18:1/18:1	744.5	603.5
PG 32:2	736.5128	C₃₈H₇₅NO₁₀P	60	PG 16:1/16:1	736.5	547.5
PG 32:1	738.5284	C₃₈H₇₇NO₁₀P	61	PG 16:0/16:1	738.5	549.5
PG 32:0	740.5441	C₃₈H₇₉NO₁₀P	62	PG 16:0/16:0	740.5	551.5
PG 34:4	760.5128	C₄₀H₇₅NO₁₀P	63	PG 18:3/16:1	760.5	571.5
PG 34:3	762.5284	C₄₀H₇₇NO₁₀P	64	PG 34:3	762.5	573.5
PG 34:2	764.5441	C₄₀H₇₉NO₁₀P	65	PG 34:2	764.5	575.5
PG 34:1	766.5598	C₄₀H₈₁NO₁₀P	66	PG 18:1/16:0	766.5	577.5
PG 34:0	768.5754	C₄₀H₈₃NO₁₀P	67	PG 18:0/16:0	768.5	579.5
PI 34:3	850.5445	C₄₃H₈₁NO₁₃P	68	PI 18:3/16:0	850.5	573.5
PI 34:2	852.5601	C₄₃H₈₃NO₁₃P	69	PI 34:2	852.5	575.5
PI 34:1	854.5757	C₄₃H₈₅NO₁₃P	70	PI 18:1/16:0	854.5	577.5
PI 34:0	856.5914	C₄₃H₈₇NO₁₃P	71	PI 18:0/16:0	856.5	579.5
DGTS 32:4	704.5464	C₄₂H₇₄NO₇	72	DGTS 16:0/16:4	704.5	474.4
			73	DGTS 16:3/16:1	704.5	472.4
			74	DGTS 16:2/16:2	704.5	470.4
DGTS 32:3	706.5621	C₄₂H₇₆NO₇	75	DGTS 16:0/16:3	706.5	474.4
DGTS 32:3	706.5621	C₄₂H₇₆NO₇	76	DGTS 16:2/16:1	706.5	470.4
DGTS 32:2	708.5778	C₄₂H₇₈NO₇	77	DGTS 16:0/16:2	708.5	474.4
DGTS 32:1	710.5934	C₄₂H₈₀NO₇	78	DGTS 16:0/16:1	710.5	474.4
DGTS 32:0	712.6091	C₄₂H₈₂NO₇	79	DGTS 16:0/16:0	712.6	474.4
DGTS 34:7	726.5308	C₄₄H₇₂NO₇	80	DGTS 16:4/18:3	726.5	466.4
DGTS 34:7	726.5308	C₄₄H₇₂NO₇	81	DGTS 16:3/18:4	726.5	468.4
DGTS 34:6	728.5464	C₄₄H₇₄NO₇	82	DGTS 16:3/18:3	728.5	468.4
			83	DGTS 16:4/18:2	728.5	466.4
			84	DGTS 16:2/18:4	728.5	494.4
DGTS 34:5	730.5621	C₄₄H₇₆NO₇	85	DGTS 16:4/18:1	730.5	500.4
			86	DGTS 16:1/18:4	730.5	472.4
			87	DGTS 16:3/18:2	730.5	498.4
			88	DGTS 16:2/18:3	730.5	470.4
DGTS 34:4	732.5778	C₄₄H₇₈NO₇	89	DGTS 16:1/18:3	732.5	496.4
			90	DGTS 16:2/18:2	732.5	498.4
			91	DGTS 16:3/18:1	732.5	500.4
			92	DGTS 16:0/18:4	732.5	474.4
表2 (续) Table 2 (continued)
Lipid species (C:N)	Theoretical mass (m/z)	Chemical formula	No.	Molecular species (sn-1/sn-2)	Precursor ion (m/z)	Product ion (m/z)
			93	DGTS 16:4/18:0	732.5	502.4
DGTS 34:3	734.5934	C₄₄H₈₀NO₇	94	DGTS 16:0/18:3	734.5	474.4
DGTS 34:2	736.6091	C₄₄H₈₂NO₇	95	DGTS 16:0/18:2	736.6	474.4
DGTS 34:1	738.6247	C₄₄H₈₄NO₇	96	DGTS 16:0/18:1	738.6	474.4
DGTS 34:1	738.6247	C₄₄H₈₄NO₇	97	DGTS 16:1/18:0	738.6	502.4
DGTS 36:7	754.5621	C₄₆H₇₆NO₇	98	DGTS 18:3/18:4	754.5	496.4
DGTS 36:6	756.5778	C₄₆H₇₈NO₇	99	DGTS 18:3/18:3	756.5	496.4
DGTS 36:6	756.5778	C₄₆H₇₈NO₇	100	DGTS 18:4/18:2	756.5	498.4
DGTS 36:5	758.5934	C₄₆H₈₀NO₇	101	DGTS 18:2/18:3	758.5	498.4
DGTS 36:5	758.5934	C₄₆H₈₀NO₇	102	DGTS 18:1/18:4	758.5	500.4
DGTS 36:4	760.6091	C₄₆H₈₂NO₇	103	DGTS 18:2/18:2	760.6	498.4
			104	DGTS 18:1/18:3	760.6	500.4
			105	DGTS 18:0/18:4	760.6	502.4
DGTS 36:3	762.6247	C₄₆H₈₄NO₇	106	DGTS 18:0/18:3	762.6	502.4
DGTS 36:3	762.6247	C₄₆H₈₄NO₇	107	DGTS 18:1/18:2	762.6	500.4
DGTS 36:2	764.6404	C₄₆H₈₆NO₇	108	DGTS 18:1/18:1	764.6	500.4
DGTS 36:2	764.6404	C₄₆H₈₆NO₇	109	DGTS 18:0/18:2	764.6	502.4

Lipid class	Adduct ion	Ion mode	CE (eV)	CID fragments
MGDG	[M+NH₄]⁺	+	50	[M+NH₄-17]⁺, [M+NH₄-35]⁺, [M+NH₄-179]⁺, [M+NH₄-197]⁺, [M+NH₄-179-R_xCOOH]⁺
DGDG	[M+NH₄]⁺	+	70	[M+NH₄-17]⁺, [M+NH₄-35]⁺, [M+NH₄-180]⁺, [M+NH₄-341]⁺, [M+NH₄- 359]⁺, [M+NH₄-341-R_xCOOH]⁺
SQDG	[M+NH₄]⁺	+	50	[M+NH₄-17]⁺, [M+NH₄-35]⁺, [M+NH₄-261]⁺, [M+NH₄-261-R_xCO+H]⁺
PE	[M-H]^-	-	35	[R_xCOO]^-, [M-H-R_xCOOH]^-, [M-H-R_xCO+H]^-
PG	[M-H]^-	-	35	[R_xCOO]^-, [M-H-R_xCOOH]^-, [M-H-R_xCO+H]^-, [PA-H-R_xCOOH]^-
PI	[M-H]^-	-	35	[R_xCOO]^-, [M-H-R_xCOOH]^-, [M-H-RC_xO+H]^-, [M-H-R₁COOH-R₂- COOH]^-, [PA-H-R_xCOOH]^-, 241
DGTS	[M+H]⁺	+	35	[M+H-R_xCOOH]⁺, [M+H-R_xCO+H]⁺, 236

基于液质联用的莱茵衣藻极性甘油酯组定性定量分析

Qualitative and Quantitative Analyses of Polar Glycerolipids in Chlamydomonas reinhardtii based on LC-MS Techniques

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Lipid standard	Linear equation	Correlation coefficient (R²)	Linear range (nmol·mL^-1)
MGDG 18:0/18:0	y=6.00E-05x+2.50E-01	0.9981	0.37-39.88
DGDG 18:0/18:0	y=6.45E-04x+7.98E-01	0.9989	1.26-41.38
SQDG 18:3/16:0	y=2.18E-04x-3.30E-01	0.9919	0.02-35.95
PE 17:0/17:0	y=2.39E-04x+4.26E-01	0.9994	1.02-27.76
PG 17:0/17:0	y=9.25E-05x+6.39E-01	0.9971	0.68-26.02
PI 16:0/16:0	y=1.49E-04x+1.65E-01	0.9993	0.23-22.61
DGTS 16:0/16:0	y=1.37E-05x+8.63E-02	0.9998	1.00-28.11