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. Author manuscript; available in PMC: 2015 Mar 3.

Published in final edited form as: Biomed Chromatogr. 2013 Mar 12;27(8):994–1002. doi: 10.1002/bmc.2893

Table 2.

Method validation for 5-FU and MTX in mouse plasma, brain and urine

Matrix	Interday				Intraday
Matrix	Theoretical (ng/mL)	Observed (ng/mL)	CV (%)^a	RE (%)^b	Observed (ng/mL)	CV (%)^a	RE (%)^b
5-Fluorouracil
Plasma	15.6	17.2 ± 3.3	19.1	9.3	14.8 ± 1.7	11.5	−5.5
	31.3	33.9 ± 4.5	13.3	8.3	31.8 ± 1.9	5.9	1.8
	125	141.9 ± 21	14.8	13.5	139.7 ± 13.8	9.9	11.8
	500	501.2 ± 74.1	14.8	0.2	493.3 ± 31.6	6.4	−1.3
Brain	15.6	15.4 ± 2.4	15.6	−1.2	14.5 ± 1.5	10.1	−6.8
	31.3	29.5 ± 2.5	8.5	−6.0	29.9 ± 2.3	7.7	−4.2
	125	125.8 ± 14.5	11.5	0.7	128.8 ± 17.9	13.9	2.9
	500	514.7 ± 26.2	5.1	2.9	510.9 ± 26.1	5.1	2.1
Urine	78.1	71.4 ± 11	15.6	−9.4	72.6 ± 1.4	19.1	−4.7
	156.3	143.7 ± 16.5	11.5	−8.7	157.2 ± 23	14.6	0.7
	625	585.9 ± 61.5	10.5	−6.7	568.9 ± 33	5.8	−9.9
	2500	2637.5 ± 308.6	11.7	5.2	2537.3 ± 294.3	11.6	1.5
Methotrexate
Plasma	15.6	13.9 ± 1.5	11.0	−12.6	14.0 ± 1.6	11.6	−11.9
	31.3	36.7 ± 5.5	15.1	11.5	33.8 ± 4.7	13.8	7.6
	125	124.1 ± 14.5	11.7	−0.8	119.2 ± 14.2	11.9	−4.9
	500	505.2 ± 51.5	10.2	1.0	474.4 ± 39	8.2	−5.4
Brain	15.6	15.4 ± 1.0	6.7	1.9	16.9 ± 2.5	14.6	7.6
	31.3	29.5 ± 2.7	9.4	−10.6	30.1 ± 1.4	4.6	−3.9
	125	123.1 ± 8.0	6.5	−1.5	130.0 ± 7.5	5.8	3.8
	500	512.2 ± 6.1	1.2	2.4	503.3 ± 40.3	8.0	0.7
Urine	78.1	71.6 ± 10.7	14.9	−9.0	78.7 ± 14.2	18.1	0.8
	156.3	167.8 ± 22.7	13.5	6.9	142.7 ± 20	14.0	−9.4
	625	565.3 ± 55.4	9.8	−10.1	629.6 ± 98.2	15.6	0.7
	2500	2513.3 ± 83	3.3	0.51	2446.4 ± 203.1	8.3	−2.1

^a

Precision (CV %) = (standard deviation/mean) × 100.

^b

Bias (RE %) = [(mean observed − theoretical value)/theoretical value] × 100.

Concentrations measured are reported as means ± SD; n = 5 for interday and n = 6 for intraday.