Caffeine ELISA Kit Wako Data

This kit is based on a competitive assay, and absorbance intensity decreases as the amount of caffeine in the sample increases.

Calibration curve range	0.244 - 1,000 ng/mL
Assay target	Caffeine
Analysis sample	Human saliva/serum/plasma (EDTA/heparin) Mouse serum/plasma Rat serum/plasma
Sample volume	55 μL (2-fold dilution, duplicate)
Measurement duration	Approx. 2 hours 20 minutes
Detection method	Chromogenic (Primary wavelength 450 nm / reference wavelength 620 nm)

Human

	Amount spiked (ng/mL)	Measured value (ng/mL)	Recovery volume (ng/mL)	Recovery rate (%)
Saliva 1	0	11.3	-	-
	10	21.1	9.80	98.0
	50	64.7	53.4	107
	250	268	257	103
	Average			103
Saliva 2	0	3.15	-	-
	10	13.8	10.7	107
	50	53.0	49.9	99.8
	250	276	273	109
	Average			105
Saliva 3	0	7.79	-	-
	10	18.2	10.4	104
	50	58.3	50.5	101
	250	267	259	104
	Average			103
Serum	0.00	1.31	－	－
	1.00	2.33	1.02	102
	50.0	49.9	48.6	97.2
	300	295	294	98.0
	Average			99.1
Plasma (EDTA)	0.00	2.11	－	－
	1.00	3.15	1.04	104
	50.0	45.5	43.4	86.8
	300	289	287	95.7
	Average			95.5
Plasma (Heparin)	0.00	3.21	－	－
	1.00	4.32	1.11	111
	50.0	54.9	51.7	103
	300	312	309	103
	Average			106

Mouse

	Amount spiked (ng/mL)	Measured value (ng/mL)	Recovery volume (ng/mL)	Recovery rate (%)
Serum	0.00	9.72	-	-
	1.00	10.7	0.980	98.0
	50.0	62.6	52.9	106
	300	314	304	101
	Average			102
Plasma	0.00	8.88	-	-
	1.00	9.87	0.990	99.0
	50.0	58.3	49.4	98.8
	300	317	308	103
	Average			100

Rat

	Amount spiked (ng/mL)	Measured value (ng/mL)	Recovery volume (ng/mL)	Recovery rate (%)
Serum	0.00	17.1	-	-
	1.00	18.1	1.00	100
	50.0	66.1	49.0	98.0
	300	322	305	102
	Average			100
Plasma	0.00	4.23	-	-
	1.00	5.22	0.990	99.0
	50.0	49.9	45.7	91.4
	300	322	318	106
	Average			98.8

Human

Mouse

Rat