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Rapid and Quantitative Analysis of Histamine in Fish Using Surface Enhanced Raman Spectroscopy |
GUO Xiao-ying1, QIU Li1, ZHANG Jin-jie1*, YANG Dan-ting2*, TANG Chun-lan2, XU Da-lun1, LOU Qiao-ming1, YANG Wen-ge1, HU Qi-jie3 |
1. College of Food and Pharmaceutical Sciences, Ningbo University, Ningbo 315000, China
2. School of Medicine, Ningbo University, Ningbo 315000, China
3. Huzhou Institute for Food and Drug Control, Huzhou 313000, China |
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Abstract A rapid method for the detection of histamine in fish products was developed in this study based on Surface-enhanced Raman spectroscopy (SERS). Silver nanoparticles (Ag NPs) were used as the active substrate and sodium chloride solution as an aggregating agent to obtain histamine SERS peaks. The histamine content in fish extracts was determined by linear regression algorithm. In this research, the normal Raman spectra of histamine powder, the SERS characteristic peaks and vibrational assignment of histamine standard solutions as well as histamine in fish extracts were analyzed. The SERS reaction conditions for the concentration of silver nanoparticles and sodium chloride solution were optimized. Lastly, the optimization conditions were used to quantitatively analyze the histamine in fish extracts. The results showed that the silver nanoparticles had the maximum absorbance at 400 nm. The shape of the nanoparticles was mainly spherical and the average size was about 30 nm. Then using 4-MBA as a probe molecule to detect the stability and sensitivity of Ag NPs and obtained Raman peak has good repeatability and high Raman intensity. Therefore, not only is the synthesis method of the active substrate less time-consuming and easy to operate, but the synthesized Ag NPs can be used as a reliable reinforcing substrate in the SERS test. In addition, it was proved by UV-Vis spectrophotometer that the NaCl solution can arouse the agglomeration of Ag NPs to form hot spots, which can enhance SERS signal. The Raman spectrum of solid histamine reflects that the characteristic peak appeared at 1 167 cm-1 due to the in-plane bending of N-H, the in-plane bending of imidazole in C—H plane and ring breathing (1 236 cm-1), the ring stretching (1 291 cm-1), the in-plane bending vibration of imidazole N—H and ring stretching (1 474 cm-1). Then the optimized reaction conditions showed the highest enhancement effect when the concentration multiple of silver nanoparticle was 15× and the concentration of sodium chloride solution was 1 mol·L-1, and the optimal condition was detected at the of 5~250 mg·L-1 histamine aqueous solution, of which the lowest concentration was detected at 5 mg·L-1. At the same time, the SERS spectra of histamine in fish extracts ranging from 10 to 100 mg·L-1 were detected under the optimized conditions. It was revealed that 3 calibration curves established at the characteristic peaks at 1 180, 1 258, 1 425 cm-1 corresponding Raman peak intensities had a good linear relationship when the concentration of histamine in fish extracts ranged from 10 to 100 mg·L-1 with R2= 0.918 1~0.947 3, of which the R2 value of the prominent peak at 1 258 cm-1 was the largest. The lowest concentration detected in fish was 10 mg·L-1, much lower than maximum limit of detection of histamine in the national standard. The calibration curve had superior accuracy with the recovery between 100%~111%. And the applicability of the method was verified by high performance liquid chromatography (HPLC). The experimental results showed that surface-enhanced Raman spectroscopy using silver nanoparticles as an active substrate and sodium chloride solution as an aggregator coupled with linear regression to establish a standard curve for rapid detection of histamine in fish is feasible and accurate, which provides a reference basis for rapid quantitative analysis of histamine content in fish products.
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Received: 2018-06-20
Accepted: 2018-10-28
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Corresponding Authors:
ZHANG Jin-jie, YANG Dan-ting
E-mail: jackace@163.com;yangdanting@nbu.edu.cn
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