Table 1 Chemiluminescence Signal Calculated by Averaging the Ten Brightest Pixels Or Integration

Supplementary Material

Simultaneous determination of four different antibiotic residues in honey by chemiluminescence multianalyte chip immunoassays

Klaus Wutz, Reinhard Niessner and Michael Seidel*

Institute of Hydrochemistry and Chair for Analytical Chemistry, Technische Universität München, Marchioninistraße 17, 81377 München, Germany

*To whom correspondence should be addressed. Phone: +49-89-2180-78238. Fax +49-89- 2180-78255. e-mail:

Supp Fig.1 Chemiluminescence signals of five spots of the same analyte with inhomogeneous spot sizes

Supp Table 1 Chemiluminescence signal calculated by averaging the ten brightest pixels or integration of all pixels (grid cell size: 25x25 pixels)

Spot / Mean value [a.u.] / Integral [a.u.]
1 / 34716 / 2672517
2 / 36008 / 2818014
3 / 35495 / 2381813
4 / 33190 / 2505971
5 / 33179 / 1531150
M.V. [a.u.] / 34518 / 2381893
S.D. [a.u.] / 1301 / 503444
R.S.D. [%] / 3.8 / 21.1

Supp Fig.2 Chemiluminescence signals of five spots of the same analyte. Spot 5 shows significantly decreased signal intensity

Supp Table 2 Outlier control by calculation of the standard deviations of all 4-spot combinations (n=5, k=4)

Spot / Mean value [a.u.] / Mean value [a.u.] / Mean value [a.u.] / Mean value [a.u.] / Mean value [a.u.] / Mean value [a.u.]
1 / 12528 / 12528 / 12528 / 12528 / 12528
2 / 13667 / 13667 / 13667 / 13667 / 13667
3 / 11786 / 11786 / 11786 / 11786 / 11786
4 / 10575 / 10575 / 10575 / 10575 / 10575
5 / 3929 / 3929 / 3929 / 3929 / 3929
M.V. [a.u.] / 10497 / 9989 / 9705 / 10175 / 10478 / 12139
S.D. [a.u.] / 3840 / 4236 / 3933 / 4355 / 4434 / 1298
R.S.D. [%] / 36.6 / 42.4 / 40.5 / 42.8 / 42.3 / 10.7

SuppTable3 Spot sizes and signal-to-noise ratios (SNR) of the immobilized antibiotic derivatives in dependency on the spotting concentration. SNR were calculated as ratio of the MVanalyte and the MVnegative control

antibiotic derivative [mgmL-1] / Spot size [µm] / SNR
SMZ 0.1 / 500 / 158 : 1
SMZ 1.0 / 500 / 173 : 1
SMZ 10 / 500 / 205 : 1
SDZ 0.1 / 370 / 158 : 1
SDZ 1.0 / 410 / 167 : 1
SDZ 10 / 450 / 183 : 1
Streptomycin 0.1 / 290 / 16 : 1
Streptomycin 1.0 / 410 / 73 : 1
Streptomycin 10 / 410 / 112 : 1
Clinafloxacin 0.01 / 250 / 73 : 1
Clinafloxacin 0.1 / 370 / 117 : 1
Clinafloxacin 1.0 / 450 / 199 : 1