Abstract
Combining surface-enhanced Raman spectroscopy (SERS) of aggregated graphene oxide/gold nanoparticle hybrids with immunomagnetic bead sample preparation method, a highly sensitive strategy to determine the clenbuterol content in animal urine was developed. Based on a linear calibration curve of the SERS characteristic peak intensity of clenbuterol at Δv = 1474 cm−1 versus the spiked clenbuterol concentration in the range of 0.5–20 ng·mL−1, the quantity of clenbuterol in real animal urine samples can be determined and matches well with those determined by LC-MS/MS, while the detection time is significantly reduced to 15 min/sample. The limits of detection and quantification in the urine are 0.5 ng·mL−1 and 1 ng·mL−1, respectively, and the recovery clenbuterol rates are 82.8–92.4% with coefficients of variation <9.4%. The day-to-day variation of the detection is less than 6.41%, and the shelving life of the SERS substrates is no less than 4 weeks. All these indicate that this proposed SERS detection protocol for clenbuterol is reproducible, reliable, and can be easily developed for the routine monitoring of the illicit use of clenbuterol in animal farming.
Highlights
Many rapid screening methods have been proposed for clenbuterol detection, such as enzyme-linked immunosorbent assay technique[10,11], immunochromatographic assay based on up-conversion phosphors[12], fluorescent multi-component immuno chromatography[13] and surface enhanced Raman spectroscopy (SERS)[14,15]
It was found that the adsorption of clenbuterol molecules was more effective on gold nanoparticles under acidic condition based on the direct interaction of the aromatic or aliphatic moieties through ionic or coordination bonds with the metal
We designed a unique hybrid graphene oxide/Au nanoparticle(GO/AuNPs) SERS probe for clenbuterol detection from urine sample.The hybrid probe was stable within four weeks under 4 °C storage temperature, and the limit of detection (LOD) and the limit of quantitation (LOQ) based on SERS for real urine sample were 0.5 ng·mL−1 and 1 ng·mL−1, respectively
Summary
Many rapid screening methods have been proposed for clenbuterol detection, such as enzyme-linked immunosorbent assay technique[10,11], immunochromatographic assay based on up-conversion phosphors[12], fluorescent multi-component immuno chromatography[13] and surface enhanced Raman spectroscopy (SERS)[14,15]. Zhu et al reported a SERS detection of clenbuterol based on a competitive SERS immunoassay with a LOD of 0.1 pg·mL−1 15. Xie et al prepared a modified AuMBA@Ag - Antibody probe (the notation “AuMBA@Ag” refers to the polyclonal antibody of clenbuterol labeled Au-Ag core-shell nanoparticles sandwiched with a Raman reporter 4-mercaptobenzoic acid (MBA)) and demonstrated a LOD of 0.24 pg·mL−1 for quantitative detection of clenbuterol from urine[14]. We designed a unique hybrid graphene oxide/Au nanoparticle(GO/AuNPs) SERS probe for clenbuterol detection from urine sample.The hybrid probe was stable within four weeks under 4 °C storage temperature, and the LOD and the limit of quantitation (LOQ) based on SERS for real urine sample were 0.5 ng·mL−1 and 1 ng·mL−1, respectively.
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