. 2019 Mar 7;10:169. doi: 10.3389/fphar.2019.00169

Table 2.

Intraday accuracy (acc.) and precision (prec.) for the extraction of a subset of SPMs from human plasma (500 μL) and human serum (500 μL).

Analyte	Mass transition			0.1 nM in plasma^a				0.3 nM in plasma^a				1.0 nM in plasma		3.0 nM in plasma		3.0 nM in serum
				5 μL		10 μL		5 μL		10 μL		5 μL		5 μL		5 μL
		Q1	Q3	acc. [%]	prec. [%]	acc. [%]	prec. [%]	acc. [%]	prec. [%]	acc. [%]	prec. [%]	acc. [%]	prec. [%]	acc. [%]	prec. [%]	acc. [%]	prec. [%]
6(R)-LXA₄	(1)	351.2	115.2	94	14	93	5	100	10	92	4	98	2	99	4	97	3
	(2)	351.2	235.0	72	11	83	9	88	9	79	6	94	2	99	4	94	4
	(3)	351.2	217.1	<LOD		75	22	76	7	94	11	99	5	98	5	98	3
6(S)-LXA₄	(1)	351.2	115.1	82	17	87	4	93	4	86	5	85	0	88	5	88	1
	(2)	351.2	235.2	71	22	73	8	86	5	83	4	87	2	88	2	87	5
	(3)	351.2	217.1	<LOD		102	11	95	5	87	2	85	8	87	5	87	2
LXB₄	(1)	351.2	221.0	105	5	106	9	96	8	97	4	98	5	100	3	95	3
	(2)	351.2	233.1	109	14	119	10	103	8	97	7	96	3	100	4	92	2
	(3)	351.2	251.0	<LOD		<LOD		102	9	101	15	93	2	100	5	95	6
LXA₅	(1)	349.1	114.9	114	7	119	7	110	5	114	4	111	4	112	3	102	4
	(2)	349.1	215.0	97	9	105	5	106	3	112	1	105	5	111	4	102	4
	(3)	349.1	233.1	86	9	100	7	93	4	109	7	106	5	113	4	103	6
RvE1	(1)	349.3	195.0	68	8	78	4	73	6	81	6	69	8	71	7	108	2
	(2)	349.3	161.0	81	17	73	15	74	11	79	6	72	11	68	5	105	6
	(3)	349.3	205.0	<LOD		72	7	62	12	79	7	73	5	69	6	108	5
RvD1	(1)	375.3	141.0	94	9	96	4	98	3	91	3	96	7	100	1	97	1
	(2)	375.3	215.0	94	8	93	4	91	7	93	4	93	5	98	4	98	4
	(3)	375.3	233.3	80	5	82	14	89	9	91	8	95	6	101	1	98	1
RvD2	(1)	375.3	175.0	101	8	98	3	96	5	104	4	98	7	100	4	105	3
	(2)	375.3	141.0	99	10	98	7	91	6	94	5	94	9	97	3	108	2
	(3)	375.3	277.0	105	13	93	5	105	2	98	5	98	9	96	2	105	3
RvD3	(1)	375.3	147.0	105	8	103	6	109	5	103	4	106	7	103	3	113	2
	(2)	375.3	137.0	82	35	103	15	102	9	103	11	106	10	105	4	115	4
	(3)	375.3	181.0	<LOD		128	11	101	17	105	6	112	14	106	4	116	6
RvD5	(1)	359.1	199.1	109	3	122	11	111	4	121	4	109	10	107	4	106	3
	(2)	359.1	141.0	111	6	110	12	107	16	116	8	102	11	104	6	107	5
	(3)	359.1	261.0	<LOD		<LOD		115	7	98	8	108	13	108	3	100	5
MaR1	(1)	359.1	250.2	110	19	119	14	109	10	107	4	105	8	104	5	105	5
	(2)	359.1	177.0	88	24	122	12	99	7	107	6	93	11	100	6	101	3
	(3)	359.1	221.0	<LOD		<LOD		110	11	113	5	92	11	104	2	104	7
7(S)-MaR1	(1)	359.1	250.1	97	6	94	12	99	4	102	2	91	9	96	4	103	2
	(2)	359.1	177.0	105	9	90	18	99	2	103	7	99	5	97	6	103	3
	(3)	359.1	221.0	<LOD		<LOD		110	13	108	3	95	11	97	4	104	3
(N)PD1	(1)	359.0	153.0	87	17	114	9	91	11	95	3	98	6	92	4	98	3
	(2)	359.0	206.0	95	8	99	15	109	8	96	6	98	7	89	3	105	5
PDX	(1)	359.1	153.1	96	5	106	3	98	2	99	4	94	6	93	5	102	3
	(2)	359.1	206.1	103	6	96	6	95	6	95	5	89	8	90	4	101	2

SPMs were spiked into plasma at four concentration levels of 0.1, 0.3, 1.0, and 3.0 nM and additionally into serum at the highest concentration level (3.0 nM in serum). Accuracy was calculated based on the determined concentration in sample after sample preparation utilizing SPE extraction in comparison to the concentration in the spiking standard solution. Precision was expressed as relative standard deviation of the sample set (n = 4). For concentration levels 0.1 and 0.3 nM acc. and prec. for 5 and 10 μL injection volume are shown. Quantification was carried out for the quantifier (most sensitive transition) as well as for the alternative transitions if concentration was >LOD. No calculation of accuracy and precision if analyte was in ≥50% of the samples <LOD.

concentrations <LLOQ and >LOD were quantified to calculate accuracy and precision.