Serum Untargeted UHPLC-HRMS-Based Lipidomics to Discover the Potential Biomarker of Colorectal Advanced Adenoma

Figures & data

Table 1 Information of Clinical Characteristics for Study Subjects

Group	Gender (Female/Male)	Age (Year)	Position	Vienna Classification
NR	21/29	53 ± 8	–	–
			–	–
CAA	18/28	57 ± 11	Rectum (19)	High (24)
			Colon (27)	Low (22)

Table 2 53 Differential Lipids Between NR and CAA Groups in Both ESI Modes

ESI Mode	Name	Formula	RT [min]	P value	VIP	Fold Change	Detective m/z	AUC	Adduct	Class
ESI+	2-Arachidonoyl glycerol	C23 H38 O4	5.26	0.000	1.04	↓	379.28220		H	FAs
	Arachidonic acid ethyl ester	C22 H36 O2	10.03	0.003	1.01	↓	339.29010	0.866	H	FAs
	Cer-NS d40:1	C40 H79 N O3	10.56	0.000	1.08	↓	622.62395	0.882	H	SLs
	Cer-NS d41:1	C41 H81 N O3	10.74	0.000	1.01	↓	636.63171	0.859	H	SLs
	DAG 32:1	C35 H66 O5	9.59	0.000	1.10	↓	584.52533	0.856	NH4	GLs
	DAG 34:1	C37 H70 O5	10.02	0.003	1.01	↓	612.55823	0.830	NH4	GLs
	DAG 34:3	C37 H66 O5	9.34	0.002	1.00	↓	608.52734	0.830	NH4	GLs
	DAG 34:5	C37 H62 O5	8.56	0.024	1.01	↓	604.49463		NH4	GLs
	DAG 36:5	C39 H66 O5	9.00	0.001	1.09	↓	632.52502	0.825	NH4	GLs
	DAG 38:6	C41 H68 O5	9.23	0.017	1.01	↓	658.54144	0.800	NH4	GLs
	DAG 40:8	C43 H68 O5	9.05	0.004	1.00	↓	682.54205	0.762	NH4	GLs
	Docosanamide	C22 H45 N O	6.91	0.000	1.43	↑	340.35806	0.917	H	FAs
	LPC 17:0	C25 H52 N O7 P	4.23	0.000	1.12	↓	510.35626	0.920	H	GPs
	LPC 18:0	C26 H54 N O7 P	4.89	0.000	1.04	↓	524.37170	0.928	H	GPs
	LPC 19:0	C27 H56 N O7 P	5.26	0.000	1.09	↓	538.38757	0.895	H	GPs
	Methyl palmitate	C17 H34 O2	1.36	0.000	1.37	↑	288.29022	0.930	NH4	FAs
	Palmitic Acid	C16 H32 O2	1.19	0.000	1.14	↑	274.27435	0.989	NH4	FAs
	PC 18:0e	C26 H54 N O7 P	4.61	0.000	1.12	↓	524.37177	0.941	H	GLs
	PC 31:2	C39 H74 N O8 P	8.60	0.000	1.35	↑	716.56000	0.975	H	GPs
	PC 35:6e	C43 H76 N O7 P	8.44	0.000	1.68	↑	750.55353	1.000	H	GPs
	PC 37:7	C45 H76 N O8 P	7.81	0.000	1.15	↑	790.53979	0.974	H	GPs
	PC 38:7	C46 H78 N O8 P	8.40	0.000	1.09	↓	804.55536	0.896	H	GPs
	PC 42:9	C50 H82 N O8 P	8.23	0.000	1.30	↓	856.58630	0.964	H	GPs
	PC 44:5	C52 H94 N O8 P	10.44	0.000	1.61	↑	892.67596	1.000	H	GPs
	TAG 44:1	C47 H88 O6	11.32	0.000	1.06	↓	766.68427	0.868	NH4	GLs
	TAG 46:0	C49 H94 O6	12.42	0.000	1.01	↓	796.74023	0.803	NH4	GLs
	TAG 49:0	C52 H100 O6	13.55	0.003	1.02	↓	838.78638	0.810	NH4	GLs
	TAG 49:1	C52 H98 O6	12.90	0.003	1.06	↓	835.77142	0.759	NH4	GLs
	TAG 51:1	C54 H102 O6	13.45	0.002	1.12	↓	864.80219	0.743	NH4	GLs
	TAG 53:3	C56 H102 O6	12.97	0.001	1.04	↓	888.80316	0.802	NH4	GLs
	TAG 53:5	C56 H98 O6	11.80	0.002	1.01	↓	884.76721	0.766	NH4	GLs
	TAG 54:1	C57 H108 O6	14.32	0.001	1.10	↓	906.84503	0.836	NH4	GLs
	TAG 54:2	C57 H106 O6	13.86	0.006	1.03	↓	904.83417	0.763	NH4	GLs
	TAG 54:3	C57 H104 O6	13.76	0.005	1.01	↓	902.81677	0.791	NH4	GLs
	TAG 54:7	C57 H96 O6	11.38	0.004	1.01	↓	899.70984	0.730	Na	GLs
	TAG 55:2	C58 H108 O6	14.23	0.011	1.02	↓	918.84888	0.831	NH4	GLs
	TAG 55:4	C58 H104 O6	12.69	0.000	1.14	↑	914.81598	0.883	NH4	GLs
	TAG 56:7	C59 H100 O6	12.12	0.003	1.03	↓	922.78705	0.776	NH4	GLs
	TAG 57:1	C60 H114 O6	16.07	0.000	1.12	↓	948.89667	0.940	NH4	GLs
	TAG 58:1	C61 H116 O6	15.55	0.013	1.00	↓	962.90228	0.784	NH4	GLs
	TAG 58:8	C61 H102 O6	12.38	0.007	1.00	↓	948.80267	0.722	NH4	GLs
	TAG 60:5	C63 H112 O6	14.54	0.015	1.02	↓	982.88098	0.770	NH4	GLs
	TAG 48:1	C51 H96 O6	12.60	0.034	1.01	↓	822.75555	0.636	NH4	GLs
	TAG 56:2	C59 H110 O6	14.58	0.021	1.01	↓	932.86536	0.762	NH4	GLs
ESI-	Cer-AP t42:1	C42 H83 N O5	10.78	0.042	1.09	↓	680.62024	0.653	H	SLs
	Cer-AS d42:1	C42 H83 N O4	10.79	0.020	1.06	↓	710.63586	0.688	FA-H	SLs
	LPC 17:1	C25 H50 N O7 P	3.10	0.038	1.02	↓	552.33105	0.669	FA-H	GPs
	PE 36:3	C41 H76 N O8 P	9.00	0.013	1.16	↓	740.52527	0.692	H	GPs
	PE 36:3e	C41 H78 N O7 P	9.72	0.007	1.14	↓	726.54504	0.723	H	GPs
	PE 36:5e	C41 H74 N O7 P	9.15	0.006	1.24	↓	722.51422	0.704	H	GPs
	PE 38:5e	C43 H78 N O7 P	9.63	0.004	1.22	↓	750.54559	0.705	H	GPs
	PG 34:1	C40 H77 O10 P	9.00	0.010	1.18	↓	747.51691	0.694	H	GPs
	PI 38:4	C47 H83 O13 P	8.69	0.038	1.00	↑	885.54956		H	GPs

Abbreviations: FAs, fatty acids; SLs, sphingolipids; GPs, glycerophospholipids; GLs, glycerides.

Figure 1 The TIC chromatography of normal (NR) and colorectal advanced adenoma (CAA) groups in both ESI modes. (A and B) TIC chromatography of NR in ESI+ and ESI- modes, respectively; (C and D) TIC chromatography of CAA in ESI+ and ESI- modes, respectively.

Figure 2 Multivariate statistical analysis for lipid profile between the NR and CAA groups. (A and B) 3D PCA analysis of the two groups in ESI+ and ESI- modes, respectively; (C and D) 3D OPLS-DA analysis of the two groups in ESI+ and ESI- modes, respectively. R2X[cum] = 0.379 and 0.522, R2Y[cum] = 0.981 and 0.951, Q2[cum] = 0.976 and 0.870; (E and F) Permutation test (200 permutations) corresponding to OPLS-DA model in ESI+ and ESI- modes, respectively.

Figure 3 The proportion of differential lipids between two groups.

Abbreviations: FA, fatty acid; SL, sphingolipid; DAG, diacylglycerol; LPC, lysophosphatidylcholine, PC, phosphatidylcholine; TAG, triacylglycerol; PE, phosphatidylethanolamine; PG, phosphatidylglycerol; PI, phosphatidylinositol.

Figure 4 Heat map analysis of 53 differential lipids between NR and CAA groups. The color bars represent the log10 value of the ratio for each lipid species and only statistically significant changes are shown (VIP > 1, P < 0.05).

Figure 5 Normalization of the differential lipids between the NR and CAA groups. (A and B) Sample normalization of two groups in ESI+ and ESI- modes, respectively; (C and D) Lipids normalization of two groups in ESI+ and ESI- modes, respectively.

Figure 6 Performance evaluation of differential lipids between NR and CAA groups. (A) PC 35:6e; (B) PC 44:5; (C) Palmitic acid; (D) PC 31:2; (E) PC 37:7; (F) PC 42:9; (G) PC 18:0e; (H) TAG 57:1; (I) Methyl palmitate; (J) LPC 18:0; (K) LPC 17:0; (L) Docosanamide.

Figure 7 The identification of 12 differential lipids with good diagnosis between NR and CAA groups. (A) PC 35:6e; (B) PC 44:5; (C) Palmitic acid; (D) PC 31:2; (E) PC 37:7; (F) PC 42:9; (G) PC 18:0e; (H) TAG 57:1; (I) TAG 57:1; (J) Methyl palmitate; (K) LPC 18:0; (L) LPC 17:0; (M) Docosanamide.

Figure 8 The trend of differential lipids in NR and CAA groups. (A) Docosanamide; (B) LPC 17:0; (C) LPC 18:0; (D) Methyl palmitate; (E) Palmitic Acid; (F) PC 18:0e; (G) PC 31:2; (H) PC 35:6e; (I) PC 37:7; (J) PC 42:9; (K) PC 44:5; (L) TAG 57:1. *Means the significant difference of levels of differential lipid between NR and CAA. The levels of differential lipids were displayed with mean ± SEM.