Improved preparation of a chitosan-based immunoaffinity column using antibody against methandrostenolone as ligand

Figures & data

Figure 1. Schematic illustration of the preparation of mAb-chitosan CL.

Table 1. The colour and stability of the cross-linked chitosan when using different glutaraldehyde concentration.

Glutaraldehyde concentration (%)	Colour	Stability in pH 1.5 solution
0.5	Light yellow	Partial solubility
1.0	Light yellow	Insolubility
1.5	Deep yellow	Insolubility
2.0	Brown	Insolubility

Figure 2. The amount of antibody coupled to chitosan CL when using different epichlorohydrin concentrations. Data and error bars represent mean±SD (n=3).

Figure 3. Scanning electron micrograph (magnification 60×) of cross-linked chitosan.

Figure 4. IR spectra of chitosan, chitosan CL and mAb-chitosan CL. (a) chitosan, (b) chitosan CL and (c) mAb-chitosan CL.

Figure 5. Recovery of MA from the IAC column with different methanol concentrations. Data and error bars represent mean±SD (n=3).

Table 2. MA immunoaffinity column-to-column variability.

Column ID	Measured amount (ng)	Recovery (%)
100 ng of MA standard applied
1	96.57	96.57
2	97.68	97.68
200 ng of MA standard applied
3	187.48	96.74
4	191.92	98.96
300 ng of MA standard applied
5	283.83	94.61
6	290.55	96.85
Average recovery (%)	96.9±1.48

Figure 6. IAC column capacity and recovery of MA for 10 cycles of usage.

Table 3. Extraction recoveries of MA from spiked animal tissue and feed samples by IAC column cleanup (n=3).

Sample	Spiked level (ng/g)	Recovery rate (%)	RSD (%)
Animal tissue	1.5	87.1	4.1
	6.0	85.0	4.9
Animal feed	1.5	85.3	7.5
	6.0	84.9	5.4