TABLE 1.

Method characteristics for the quantification of eicosanoid metabolites in urine by LC-MS/MS

Method	Compound	RT (min)	Transition	LOQ (pg on-column)	r	Recovery (%)			Intra-Day Precision (%)			Inter-Day Precision (%)			Matrix Effect (%)	Stabilitya
						Low (n = 6)	Medium (n = 6)	High (n = 6)	Low (n = 6)	Medium (n = 6)	High (n = 6)	Low (n = 6)	Medium (n = 6)	High (n = 6)		t = 0 Months (n = 6)	t = 15 Months (n = 6)
I	Tetranor PGEM	3.43	327 > 143	13	0.996	106	99	103	10	7	5	5	5	5	101	31.1 ± 5.3	40.8 ± 1.4
	Tetranor PGDM	3.91	327 > 143	1	0.999	96	95	100	9	11	13	12	11	16	109	8.1 ± 0.3	7.9 ± 0.3
	2,3-Dinor-8-iso PGF2α	6.81	325 > 237	1	0.998	105	99	104	4	6	3	7	5	6	84	4 ± 0.5	3.8 ± 0.2
	2,3-Dinor-11β-PGF2α	7.15	325 > 145	1	0.996	100	97	102	10	2	1	16	18	12	82	4.7 ± 0.5	3.3 ± 0.2
	6-Keto-PGF1α	7.24	369 > 163	1	0.998	104	98	98	0	3	4	9	4	4	119	2.3 ± 0.1	2.8 ± 0.2
	13,14-Dihydro-15- keto-tetranor-PGE2	7.51	297 > 109	5	0.996	92	107	103	11	7	9	ND	18	12	92	5.5 ± 0.2	4 ± 0.2
	13,14-Dihydro-15- keto-tetranor-PGD2	8.12	297 > 109	3	0.999	102	98	103	8	5	4	6	2	7	92	8.1 ± 0.4	8.8 ± 0.3
	8-Iso-PGF2α	9.08	353 > 193	2	0.999	100	97	95	11	4	5	14	6	7	96	2.4 ± 0.3	1.8 ± 0.1
	11β-PGF2α	9.49	353 > 291	2	0.999	96	91	94	8	3	4	1	4	1	117	11.8 ± 0.1	15.3 ± 0.3
	5-iPF2α-VI	9.85	353 > 115	4	0.999	94	93	99	9	3	3	7	2	2	109	8.9 ± 0.3	8.5 ± 0.8
	PGF2α	10.91	353 > 291	2	0.999	103	86	88	5	5	2	5	2	2	123	17.5 ± 1.2	15.8 ± 0.8
	11-Dehydro-TXB2	11.33	367 > 161	3	0.999	97	92	95	8	2	4	6	3	1	121	7.7 ± 0.3	8.7 ± 0.7
	PGE2	11.40	351 > 271	1	0.998	100	91	94	13	6	4	3	2	1	114	3.5 ± 0.2	3.6 ± 0.3
	PGD2	12.19	351 > 271	2	0.996	ND	79	98	ND	11	10	ND	13	5	122	2.1 ± 0.2	5.3 ± 0.2
	PGE1	12.25	353 > 273	10	0.999	ND	85	91	ND	5	4	ND	6	2	113	12.2 ± 0.8	12.4 ± 0.3
	13,14-Dihydro-15-keto-PGE2	13.98	351 > 175	2	0.999	90	90	95	8	4	2	10	2	1	113	9.1 ± 0.3	10 ± 0.5
	8,12-Iso-iPF2α-VI	14.12	353 > 115	1	0.999	100	91	91	6	4	4	2	1	1	126	11.6 ± 0.4	11.3 ± 0.7
	13,14-Dihydro-15-keto-PGF2α	14.19	353 > 291	3	0.999	101	90	94	8	4	5	2	2	2	125	20.2 ± 0.5	21.2 ± 0.7
	13,14-Dihydro-15-keto-PGE1	14.86	353 > 209	2	0.997	93	92	88	7	5	4	27	10	4	124	10.1 ± 0.2	10.5 ± 0.6
	13,14-Dihydro-15-keto-PGD2	15.04	351 > 175	3	0.998	97	90	90	11	6	1	6	5	3	102	12.1 ± 0.5	16.4 ± 0.9
II	2,3-Dinor-6-keto-PGF1α	3.12	370 > 232	10	0.999	106	104	104	6	4	6	12	14	11	117	82.1 ± 5.8	68.2 ± 7.8
	2,3-Dinor-TXB2	3.45	370 > 155	3	0.999	108	97	100	3	3	3	5	2	2	115	21.7 ± 2.3	21.3 ± 0.9
	TXB2	3.81	398 > 169	1	0.999	97	92	95	7	3	3	3	3	3	118	3.5 ± 0.3	3.4 ± 0.1
III	20-Carboxy-LTB4	0.85	365 > 195	3	0.996	ND	89	86	ND	6	5	ND	25	27	5	0.2 ± 0.1	4.7 ± 0.4b
	20-Hydroxy-LTB4	0.90	351 > 195	3	0.997	85	82	82	0	6	6	9	18	18	5	0.2 ± 0.1	0.1 ± 0.1
	EXC4	1.55	624 > 306	4	0.999	90	69	80	6	15	13	19	12	4	76	2.2 ± 0.3	0.9 ± 0.1
	LTD4	1.68	495 > 187	3	0.997	74	65	68	8	9	8	16	8	9	136	1.3 ± 0.1c	3 ± 0.2
	EXE4	1.87	438 > 333	3	0.999	74	75	76	13	6	6	2	5	3	173	2.7 ± 0.2	3 ± 0.2
	LTC4	2.29	624 > 306	4	0.999	93	75	83	10	8	14	9	7	7	80	1.6 ± 0.2	0.8 ± 0.1
	LTE4	2.35	438 > 333	3	0.999	82	81	83	2	3	4	3	3	1	131	5.1 ± 0.3	4.6 ± 0.4
	6-trans-LTB4	3.23	335 > 195	2	0.997	84	82	83	2	1	0	4	5	5	140	2.7 ± 0.1	2.8 ± 0.2
	LTB4	3.46	335 > 195	2	0.999	77	76	77	2	1	1	5	5	6	129	4.3 ± 0.2	3.4 ± 0.1
RT, retention time; r, linearity; Low, low concentration; Medium, medium concentration; High, high concentration; ND, not detected.
a Data are shown as the concentration (nanograns per milligram) of the indicated compound in the fortified laboratory reference material at the time of preparation (t = 0 months) or after 15 months storage at −80°C (t = 15 months).
b The observed reduced stability of this compound is most likely a combination of large matrix effects, low precision, and low recovery for quantification.
c The initial timepoint is t = 4 months for this compound.