LabAssay NEFA Data

Performance

Sample	Serum, plasma(Mouse, Rat, dog, cat)
Standard curve range	0.4～1.97 mEq/L
Measurement time	approx. 20 min.
Amount of sample	4 μL
Measurement wavelength	550 nm

Standard curve

Procedure

Assay in a microplate
Perform the assay in the wells according to the following table scheme.

No.	Standard Solution	Distilled or deionized water	Sample volume	Concentration
1	80 µL	120 µL	4 μL	0.40 mEq/L
2	120 µL	80 µL	4 μL	0.60 mEq/L
3	160 µL	40 µL	4 μL	0.80 mEq/L
4	Undiluted	-	4 μL	1.00 mEq/L
5	Undiluted	-	8 μL	1.97 mEq/L^*1

*1: The test sample volume is usually 4 μL, but 8 μL is taken in this case. The NEFA concentration must be corrected accordingly as indicated in the table above.

Data

Spike-and-Recovery Test

Spike and Recovery Test of NEFA, a known amount of standard is spiked into in mouse and rat sample n=2.

Rat Sample

	Spike (mEq/L)	Mean (mEq/L)	Recovery (mEq/L)	Recovery (%)
Serum	-	0.57
	0.40	1.00	0.43	106.4%
	0.61	1.21	0.65	106.2%
	0.78	1.40	0.83	106.4%
	1.01	1.57	1.01	99.7%
Plasma (EDTA)	-	0.41
	0.40	0.86	0.45	111.5%
	0.61	1.07	0.66	108.5%
	0.78	1.27	0.86	109.5%
	1.01	1.44	1.02	101.4%

Mouse Sample

	Spike (mEq/L)	Mean (mEq/L)	Recovery (mEq/L)	Recovery (%)
Serum	-	0.70
	0.40	1.07	0.37	91.7%
	0.61	1.31	0.61	100.1%
	0.78	1.44	0.74	94.5%
	1.01	1.62	0.92	91.3%
Plasma (EDTA)	-	0.62
	0.40	0.99	0.37	92.5%
	0.61	1.29	0.68	111.5%
	0.78	1.38	0.77	97.8%
	1.01	1.62	1.00	99.1%

Linearity-of-Dilution Test

Rat sample

Mouse sample

D-MEM/10% FBS

Examples of measurement

20% Diluted Nomal Mouse serum, CD-1(ICR), male
No.	Mean (mEq/L)	SD	CV (%)
1	1.118	0.001	0.1
2	0.788	0.007	0.9
3	0.715	0.022	3.1
4	0.794	0.002	0.2
5	0.829	0.005	0.6

Nomal Mouse plasma K2EDTA, CD-1(ICR), male
No.	Mean	SD	CV (%)
1	*0.309	0.002	0.6
2	*0.320	0.002	0.6
3	*0.205	0.004	2.0
4	*0.179	0.008	4.6
5	*0.305	0.017	5.6

10% Diluted Nomal Rat serum, Sprague Dawley, male
No.	Mean	SD	CV (%)
1	*0.300	0	0.0
2	*0.371	0.004	1.1
3	*0.289	0.017	5.8
4	*0.365	0	0.1
5	*0.382	0.029	7.6

Nomal Rat plasma K2EDTA, Sprague Dawley, male
No.	Mean	SD	CV (%)
1	*0.333	0.004	1.2
2	*0.356	0.003	0.8
3	*0.325	0.020	6.0
4	*0.218	0.005	2.6
5	0.515	0.010	1.9

Nomal Canine serum, Beagle, male
No.	Mean (mEq/L)	SD	CV (%)	Age (years)
1	0.656	0.0027	0.4	4
2	0.665	0.0320	4.8	6
3	0.425	0.0090	2.0	6
4	*0.38	0.0057	1.5	11
5	0.882	0.0070	0.8	12
6	0.599	0.0060	1.1	6
7	1.006	0.0267	2.7	10
8	0.506	0.0060	1.2	4

Nomal Canine plasma EDTA, Beagle, male
No.	Mean (mEq/L)	SD	CV (%)	Age (years)
1	0.442	0.0033	0.7	4
2	0.427	0.0110	2.6	6
3	*0.308	0.0037	1.2	10
4	0.478	0.0123	2.6	12
5	0.473	0.0050	1.0	6
6	*0.367	0.0047	1.2	11
7	0.780	0.0133	1.7	10
8	0.411	0.0387	9.4	12

Nomal Feline serum, Short hair, male
No.	Mean (mEq/L)	SD	CV (%)	Age (years)
1	0.717	0.0117	1.6	3
2	0.635	0.0100	1.6	3
3	1.004	0.0053	0.5	6
4	0.981	0.0093	0.9	4
5	0.636	0.0170	2.7	6
6	1.193	0.0267	2.3	6
7	1.022	0.0893	8.8	1
8	*0.382	0.0157	4.1	1

Nomal Feline plasma EDTA, Short hair, male
No.	Mean (mEq/L)	SD	CV (%)	Age (years)
1	1.099	0.0103	0.9	0.5
2	*0.174	0.0033	2.0	1
3	*0.241	0.0043	1.7	3
4	*0.320	0.0067	2.2	0.5
5	*0.359	0.0060	1.7	3
6	*0.197	0.0120	6.2	3
7	*0.215	0.0240	11.1	0.5
8	*0.131	0.0160	12.3	1