A simple and efficient alginate hydrogel combined with surface-enhanced Raman spectroscopy for quantitative analysis of sodium nitrite in meat products.

Sodium nitrite is a commonly used preservative and color protectant in the food industry. Conventional analytical methods are highly susceptible to food matrix interference, time-consuming and costly. In this study, the ion cross-linking method was employed to prepare alginate hydrogel substrates, and phenosafranin was chosen as a single-molecule probe to analyze sodium nitrite. Our investigation centered on elucidating the effects of alginate and cross-linking ion concentrations on Raman signal characteristics. The optimal Raman response was observed in the precursor solution with 1% sodium alginate and 0.1 mol L-1 cross-linking ions. The relative standard deviations (RSDs) of the feature peaks from the three substrate batches ranged from 1.22% to 16.30%, attesting the robustness and consistency of the substrates. The signal reduction of the substrates after a four-week storage period remained below 10%, indicating that the substrates had good reproducibility and stability. The limits of detection (LODs) for sodium nitrite in extracts from cured meat, luncheon meat, and sliced ham were determined to range from 3.75 mg kg-1 to 8.11 mg kg-1, with low interference from the food matrix. The support vector machine algorithm was utilized to train and predict the data, which proved to be more accurate (98.6%-99.8% recovery) than the traditional linear regression model (81.9%-112.7% recovery) in predicting the spiked samples. The application of hydrogel-based surface-enhanced Raman spectroscopy (SERS) substrates for nitrite detection in food, combined with machine learning for regression prediction in data processing, collectively augmented the potential of SERS technology in the field of food analysis.

The Effect of Glucose on the Interaction of Bisphenol A and Bovine Hemoglobin Characterized by Spectroscopic and Molecular Docking Techniques.

The interaction mechanism of hemoglobin (Hb) with bisphenol A (BPA) in diabetic patients and the difference with healthy people have been studied using spectroscopic and molecular docking techniques at several glucose (Glc) concentration, with bovine hemoglobin (BHb) instead of Hb. It is found that Glc can interact with BHb-BPA and affect its molecular structure, resulting in an altered microenvironment for tyrosine (Tyr) and tryptophan (Trp) in BHb-BPA. It is also found that Glc can bind to BHb alone, and its effect on the molecular structure of BHb is weaker than that on the structure of BHb in BHb-BPA complex. The results of circular dichroism (CD) and Fourier transform infrared spectroscopy (FTIR) indicate that Glc causes an increase in the content of the α-helix and a decrease in that of the ß-sheet of BHb-BPA by 1.5-1.9% and 3.1%, respectively. The results of molecular docking show that Glc binds to BHb-BPA through hydrogen and hydrophobic bonds, and the position of binding differs from that of Glc binding to BHb alone, which may be attributed to the fact that BPA affects the protein molecular structure of BHb and has an effect on the binding of BHb to Glc. This study provides some theoretical basis for the mechanism of BPA toxicity in vivo for people with different blood glucose levels.

Kiwi-derived extracellular vesicles for oral delivery of sorafenib.

Sorafenib is an oral treatment for hepatocellular carcinoma (HCC). However, poor water solubility, harsh gastrointestinal environment and off-target effects contribute to the low bioavailability of oral sorafenib. Plant-derived extracellular vesicles (PDEVs) are biological nanovesicles with various bioactive functions that offer significant advantages in the field of oral drug delivery: protection from degradation by gastrointestinal fluids; crossing the intestinal epithelial barrier; specific targeting; safety; and abundant yield. However, there are fewer studies applying PDEVs for anti-tumor drug delivery to extra-digestive tissues. In this study, kiwifruit-derived extracellular vesicles (KEVs) were isolated and purified from kiwifruit, and their natural hepatic accumulation properties were exploited for targeted delivery of sorafenib (KEVs-SFB). Evidence showed that encapsulation of KEVs reduced the leakage of sorafenib in the gastrointestinal environment and enhanced the ability to cross the intestinal epithelium; KEVs-SFB was able to achieve liver accumulation and was predominantly taken up by HepG2 cells; KEVs-SFB was effective in inhibiting 4T1 cell proliferation; in the orthotopic liver cancer model, oral administration of KEVs-SFB inhibited tumor growth and improved the side effects of SFB. This PDEVs-based oral drug delivery platform is important for improving oral bioavailability and reducing drug side effects.

Colloidal SERS measurement of enrofloxacin with petaloid nanostructure clusters formed by terminal deoxynucleotidyl transferase catalyzed cytosine-constituted ssDNA.

We developed petal-like plasmonic nanoparticle (PLNP) clusters-based colloidal SERS method for enrofloxacin (EnFX) detection. PLNPs were synthesized by the regulation of single-stranded DNA composed of homo-cytosine deoxynucleotides (hC) catalyzed by terminal deoxynucleotidyl transferase. SERS hot spots were created via the agglomeration process of PLNPs by adding an inorganic salt potassium iodide solution, in which EnFX molecules were attached to the negatively charged PLNPs surface by electrostatic interactions. This approach enabled direct in situ detection of antibiotic residues, achieving a limit of detection (LOD) of 1.15 µg/kg for EnFX. The spiked recoveries of the SERS method were approximately 92.7% to 107.2% and the RSDs ranged from 1.05% to 7.8%, indicating that the method can be applied to actual sample detection. This colloidal SERS measurement platform would be very promising in various applications, especially in real-time and on-site food safety screening owing to its rapidness, simplicity, and sensitivity.

Rapid detection of thiabendazole residues in apple juice by surface-enhanced Raman scattering coupled with silver coated gold nanoparticles.

In recent years, the excessive use of pesticides has posed significant hazards to the ecological environment and human health in the pursuit of high crop yields. In this work, we developed a simple, sensitive, and eco-friendly approach for rapid detection of thiabendazole in apple juice using surface-enhanced Raman scattering (SERS) coupled with silver-coated gold nanoparticles (Au@Ag NPs). The developed Au@Ag NPs exhibited excellent sensitivity, allowing for the detection of thiabendazole in standard solutions at a minimum concentration of 50 ng/mL. Furthermore, two sample preparation methods were compared for detecting thiabendazole in apple juice. As the direct detection method for SERS analysis failed to detect thiabendazole at levels below the maximum residue limit based on the Chinese standard (3000 ng/mL), the effects of main matrix components in apple juice on the detection of thiabendazole were further investigated. The results revealed that both sugars and organic acids in apple juice interfered with the SERS measurement to varying degrees. Consequently, we optimized the QuEChERS method for sample preparation and achieved a higher sensitivity with a minimum detectable concentration of 250 ng/mL, a limit of detection of 0.06 mg/L and the recoveries of spiked samples were ranged from 80.2 % to 108.6 %. This study demonstrated the feasibility of proposed SERS method for pesticide residue analysis, addressing the need for food safety monitoring.

Formation of advanced glycation end-products in minced pork during frozen-then-chilled storage and subsequent heating.

The influences of frozen-then-chilled storage of minced pork on the formation of advanced glycation end-products (AGEs) including Nε-carboxymethyllysine and Nε-carboxyethyllysine, and their corresponding α-dicarbonyl precursors (α-DPs; glyoxal and methylglyoxal) during storage and subsequent heating were investigated in comparison with chilled storage. During cold storage, the levels of AGEs, trichloroacetic acid-soluble peptides, and Schiff bases in minced pork continuously increased while α-DPs decreased. The 30 min heating (100 °C) resulted in 64-560% increase of AGEs in pork, corresponding with an increase of Schiff bases and decreases of α-DPs. Compared to the chilled storage, the frozen-then-chilled storage led to no significant difference (P > 0.05) on the levels of AGEs and α-DPs in raw or heat-treated pork, implying that the formation and thawing of ice crystals in pork during the frozen-then-chilled storage had minor to no effects on the formation of AGEs and their α-DPs.

Modified paper-based substrates fabricated via electrostatic attraction of gold nanospheres for non-destructive detection of pesticides based on surface-enhanced Raman spectroscopy.

BACKGROUND: Flexible surface-enhanced Raman spectroscopy (SERS) substrates such as paper-based substrates show great potential for rapid detection of residual chemicals on food surfaces. However, controlling the density and distribution of metallic nanoparticles adsorbed on the paper is still challenging. RESULTS: The amount of gold (Au) nanospheres (51 ± 4 nm) attached on the filter paper modified with 3-aminopropyltriethoxysilane (APTES) was tunable, increasing as the level of APTES (2.5-15.0 g kg-1 ) applied for paper modification increased. Moreover, the Au nanospheres were relative evenly distributed on the filter paper modified with 2.5-10.0 g kg-1 of APTES, which resulted in excellent intra- and inter-reproducibility of SERS signals for pesticides including thiram, diquat dibromide, and paraquat dichloride (relative standard deviation = 2.2-10.1%). The modified paper-based substrate could be used to detect as low as 0.05-0.2 mg L-1 of pesticides in standard solutions, and as low as 5-20 ng cm-2 of residual pesticides on apple skins with minimum sample pretreatment. CONCLUSION: This paper-based substrate with tunable feature for the density and distribution of nanoparticles is applicable for rapid SERS detection of residual pesticides in fruits and vegetables. © 2023 Society of Chemical Industry.

Effects of acetic, malic, and citric acids on the large deformation behaviors of fish gelatin gels.

This research was aimed to quantify the effects of acetic acid, malic acid, and citric acid (0, 0.5, 1.0, and 2.0 g/100 g H2 O) on the stress-strain responses of fish gelatin (FG) gels (2, 4, and 6.67 g/100 g H2 O) under uniaxial compression up to 68% of deformation. The first-order Ogden model fitted quite well for the compression responses of FG gels (R2 = 0.9909-0.9997). Protons from the acids played a key role on weakening the FG gel structures (gel rigidity, µ, decreased 11%-27%), as the µ values and pH values of FG gels were linearly correlated (R2 = 0.8240-0.9748), regardless of the acid type. The addition of an acid also resulted in a significant increase (p < .002) in the strain hardening capacity (α) of gels with 2 g FG/100 g H2 O. Both µ and α values of FG gels with higher gelatin concentrations were less affected by an acid partly due to their stronger buffering effects. The µ and α values of FG gels as affected by acids could not be fully explained based upon the pH changes, implying that the effects of acetate, malate, and citrate ions on the gel structure could not be ignored.

Selection and quantification of volatile indicators for quality deterioration of reheated pork based on simultaneously extracting volatiles and reheating precooked pork.

This study was to screen and quantify characteristic volatiles tied to the quality deterioration of reheated pork via simultaneously reheating (75 °C, 30 min) and collecting headspace volatiles of precooked pork (100 °C, 10 min; stored: 0 °C, 0-14 d) for GC-MS analysis. The concentrations of hexanal (6.05 ± 0.86-12.05 ± 0.44 mg/kg), (E)-2-octenal (1.54 ± 0.16-3.07 ± 0.08 mg/kg), (E,E)-2,4-heptadienal (1.52 ± 0.44-2.58 ± 0.31 mg/kg) and 8 other selected volatiles in reheated pork increased as the storage time of the precooked counterparts increased. The increase rate of hexanal was 2.9-199 times faster than that of other volatiles based on zero-order reaction fitting (R2 = 0.876-0.997). Results from clustering analysis of these volatiles were consistent with their formation pathways tied to lipid autooxidation. This simple approach, reheating and collecting volatiles of precooked meat concurrently, introduces a new possibility for standardizing volatile analysis of precooked meats required being reheated before consumption.

Effect of Sucrose on the Formation of Advanced Glycation End-Products of Ground Pork during Freeze-Thaw Cycles and Subsequent Heat Treatment.

The changes in protein degradation (TCA-soluble peptides), Schiff bases, dicarbonyl compounds (glyoxal-GO, methylglyoxal-MGO) and two typical advanced glycation end-products (AGEs) including Nε-carboxymethyllysine (CML), Nε-carboxyethyllysine (CEL) levels in ground pork supplemented with sucrose (4.0%) were investigated under nine freeze-thaw cycles and subsequent heating (100 °C/30 min). It was found that increase in freeze-thaw cycles promoted protein degradation and oxidation. The addition of sucrose further promoted the production of TCA-soluble peptides, Schiff bases and CEL, but not significantly, ultimately leading to higher levels of TCA-soluble peptides, Schiff bases, GO, MGO, CML, and CEL in the ground pork with the addition of sucrose than in the blank groups by 4%, 9%, 214%, 180%, 3%, and 56%, respectively. Subsequent heating resulted in severe increase of Schiff bases but not TCA-soluble peptides. Contents of GO and MGO all decreased after heating, while contents of CML and CEL increased.

Stimulus-Responsive DNA Hydrogel Biosensors for Food Safety Detection.

Food safety has always been a major global challenge to human health and the effective detection of harmful substances in food can reduce the risk to human health. However, the food industry has been plagued by a lack of effective and sensitive safety monitoring methods due to the tension between the cost and effectiveness of monitoring. DNA-based hydrogels combine the advantages of biocompatibility, programmability, the molecular recognition of DNA molecules, and the hydrophilicity of hydrogels, making them a hotspot in the research field of new nanomaterials. The stimulus response property greatly broadens the function and application range of DNA hydrogel. In recent years, DNA hydrogels based on stimulus-responsive mechanisms have been widely applied in the field of biosensing for the detection of a variety of target substances, including various food contaminants. In this review, we describe the recent advances in the preparation of stimuli-responsive DNA hydrogels, highlighting the progress of its application in food safety detection. Finally, we also discuss the challenges and future application of stimulus-responsive DNA hydrogels.

Study on Maillard reaction mechanism by quantum chemistry calculation.

CONTEXT: The Maillard reaction is a high-temperature reaction of amino acids and carbohydrates to produce macromolecular substances such as melanoidins and intermediate reducing ketones, aldehydes, and volatile compounds. At present, only very limited researches involved the reaction mechanisms of Maillard reaction, which causes a lot of confusion in understanding numerous food processes. The detailed calculations of Maillard reaction are urgently needed. METHODS: The density functional theory (DFT) method (M06-2X/6-311G*) was used to deeply explore the specific mechanism of the primary and intermediate stages of Maillard reaction for a selected model system. RESULTS: The results show that the basic reaction processes in primary stage are the formation of Schiff-base by the condensation of amino and carbonyl groups, and then, Schiff-base tautomerization twice through proton transfer to generate Amadori rearrangement products. In the intermediate stage, two main reaction paths, 1-2 and 2-3 enolization, were comprehensively investigated. The first route finally generates 5-hydroxymethylfurfural through isomerization, dehydration, hydrolysis, elimination, and condensation, and the second route products dicarbonyl compounds through isomerization and elimination and then Strecker degradation forms aldehydes through condensation, decarboxylation, hydrolysis, and elimination. The results show that both paths are involved in complex reactions, some are lower barrier reactions, and some higher barrier reactions. An important aspect is that water catalysis is critical in all of these reactions; it is present in most processes. Our study deepens the understanding of the Maillard reaction from molecular level and facilitate the regulation of some harmful products.

A highly sensitive surface-enhanced Raman scattering sensor with MIL-100(Fe)/Au composites for detection of malachite green in fish pond water.

Concerns about food safety have been arisen due to the improper use of chemicals in aquaculture. Malachite green (MG) has attracted attention because of its illegal usage and its potential negative impacts on the environment and public health. Surface-enhanced Raman scattering (SERS) platforms coupled with different SERS substrates have been employed for rapid analysis of MG residues in food. However, the most commonly used SERS substrates were non-reusable and showed limited detection sensitivity. In this study, a novel SERS substrate with a good recyclability and a high sensitivity was prepared by electrostatically assembling together a metal-organic framework material called materials of institute lavoisie-100(Fe) (MIL-100(Fe)) and Au NPs. The lowest detectable concentration of MG was 10-13 M based on the optimal substrate. The SERS sensor was applied for the detection of the trace MG in fish pond water, which was accomplished with the correlation coefficients R2 = 0.991-0.996 in a concentration range of 10-6-10-13 M. Moreover, MIL-100(Fe)/Au was recycled at least five times, realizing a "detection to degradation", showing great potential for food contamination monitoring due to its distinguished performance.

Investigation of Advanced Glycation End-Products, α-Dicarbonyl Compounds, and Their Correlations with Chemical Composition and Salt Levels in Commercial Fish Products.

The contents of free and protein-bound advanced glycation end-products (AGEs) including Nε-carboxymethyllysine (CML) and Nε-carboxyethyllysine (CEL), along with glyoxal (GO), methylglyoxal (MGO), chemical components, and salt in commercially prepared and prefabricated fish products were analyzed. Snack food classified as commercially prepared products exhibited higher levels of GO (25.00 ± 3.34-137.12 ± 25.87 mg/kg of dry matter) and MGO (11.47 ± 1.39-43.23 ± 7.91 mg/kg of dry matter). Variations in the contents of free CML and CEL increased 29.9- and 73.0-fold, respectively. Protein-bound CML and CEL in commercially prepared samples were higher than those in raw prefabricated ones due to the impact of heat treatment. Levels of GO and MGO demonstrated negative correlations with fat (R = -0.720 and -0.751, p < 0.05) in commercially prepared samples, whereas positive correlations were observed (R = 0.526 and 0.521, p < 0.05) in raw prefabricated ones. The heat-induced formation of protein-bound CML and CEL showed a negative correlation with the variations of GO and MGO but was positively related to protein levels in prefabricated products, suggesting that GO and MGO may interact with proteins to generate AGEs during heating. The influence of NaCl on the formation of GO and MGO exhibited variations across different fish products, necessitating further investigation.

Heat-induced formation of advanced glycation end-products in ground pork as affected by the addition of acetic acid or citric acid and the storage duration prior to the heat treatments.

The heat-induced (121 °C, 10 or 30 min) formation of two potentially hazardous advanced glycation end-products (AGEs), protein-bound Nɛ -carboxymethyllysine (CML) and Nɛ -carboxyethyllysine (CEL), in pork as affected by citric or acetic acid (0.5, 1 g/100 pork) and the storage duration (0 °C, 0 - 8 d) prior to the heating was investigated. A longer storage time of raw pork resulted in higher levels of AGEs produced during the later heating, likely due to the accumulation of some AGE precursors during the storage. Depending on the acid level and heating time, adding acid in pork led to 30 - 54% (citric acid) or 14 - 48% (acetic acid) average reduction of heat-induced production of CML/CEL, which corresponded to the reduction of thiobarbituric acid reactive substances and Schiff bases. The marinating time of raw pork with an acid did not significantly affect (P = 0.959 - 0.998) the acid's inhibition effect on heat-induced formation of CML/CEL.

Effects of Acidulants on the Rheological Properties of Gelatin Extracted from the Skin of Tilapia (Oreochromis mossambicus).

This study aimed to evaluate the effects of lactic acid (LA), citric acid (CA), and malic acid (MA) varying in concentration (0.5−2.0% w/w) on the rheological properties of fish gelatin (1.5−6.67% w/w) obtained from the skin of tilapia (Oreochromis mossambicus). The addition of LA, CA, or MA in gelatin dispersions significantly (p < 0.05) weakened their gel strengths, leading to a 14.3−62.2 reduction in gel strength. The gel strength, elastic (G'), and viscous (G″) moduli, as well as the gelling (TG) and melting (TM) temperatures of gelatin dispersions decreased with an increased level of acid added, implying the weakening effects of these acids on junction zones of the gelatin network in aqueous media. The addition of LA had less effect on these rheological properties of gelatin dispersions as compared to that of MA and CA, which were consistent with their effects on the pH of gelatin dispersions. Moreover, the reductions of TG and TM for gelatin dispersions with a higher gelatin concentration (e.g., 6.67% gelatin with 0.5% LA, TG dropped 0.4 °C) due to the addition of LA, CA, or MA were less pronounced compared to those with a lower gelatin content (e.g., 2% gelatin with 0.5% LA, TG dropped 7.1 °C), likely attributing to the stronger buffering effect of the high gelatin dispersion and less percentage reduction in the junction zones in the dispersion due to the addition of an acid. Incorporation of the effects of acids on the linear relationships (R2 = 0.9959−0.9999) between the square of gelatin concentrations and G' or G″ could make it possible to develop a model to predict G', G″, phase transition temperatures of gelatin dispersions containing different amounts of gelatin and acid (within the tested range) in the future.

Detection of thiram on fruit surfaces and in juices with minimum sample pretreatment via a bendable and reusable substrate for surface-enhanced Raman scattering.

BACKGROUND: Surface-enhanced Raman scattering (SERS) substrates based on metallic nanoparticles locked in some flexible materials have great potential for rapid detection of pesticide residues in foods, but these substrates are generally not reusable. RESULTS: A bendable and reusable sponge based on polydimethylsiloxane (PDMS) and Au nanospheres was synthesized and employed as SERS substrate to analyze thiram on the surfaces of apples and grapes (20-1000 ng cm-2 ) and in their juices (0.5-5.0 mg L-1 ) with minimum sample pretreatments. The lowest detectible concentrations for thiram in fruit juices and on fruit skins were 0.5 mg L-1 and 20 ng cm-2 , respectively. The Au-PDMS substrate had acceptable intra-reproducibility for SERS analysis of thiram in fruit juices and on fruit skins, resulting in 3.6-16.9% relative standard deviation (RSD) for the SERS signal of the primary peak of thiram. Moreover, the Au-PDMS substrate exhibited distinguished reusability and stability, which could provide a reproducible SERS signal of thiram in apple juice even after the substrate being reused ten times (RSDs for the three major characteristic peaks of thiram were 2.7-10.5% during the ten reused cycles). CONCLUSION: This flexible and reusable Au-PDMS SERS substrate for thiram detection could be readily extended to the analysis of other trace chemicals in a broad range of foods, providing a new possibility for SERS application. © 2022 Society of Chemical Industry.

Fishing unfunctionalized SERS tags with DNA hydrogel network generated by ligation-rolling circle amplification for simple and ultrasensitive detection of kanamycin.

Simple assay format-based SERS methods for sensitive target substance analysis is of great significance for the development of on-site monitoring biosensors. Herein, taking the typical antibacterial kanamycin (KANA) as a subject, a simple, highly sensitive and specific SERS aptasensor was developed by manipulating DNA hydrogel network to fish plasmonic core-shell nanoparticles. A competitive binding mode of aptamer, ligation-rolling circle amplification (L-RCA), gap-containing Au@Au nanoparticles (GCNPs) with embedded Raman reporters were integrated into the sensor. In the presence of KANA, the double stranded DNA (dsDNA) structure of the aptamer was disrupted, and the released primers were used to construct two kinds of circularized padlock probes (CPPs) which were partially complementary. DNA hydrogel network was formed through the intertwining and self-assembly of two RCA-generated single stranded DNA (ssDNA) chains, during which GCNPs and magnetic beads (MBs) were entangled and incorporated. Finally, KANA quantification was successfully achieved through the quantification of the DNA hydrogel. Overall, this novel SERS aptasensor realized a simple and ultrasensitive quantification of KANA down to 2.3 fM, plus excellent selectivity, and precision even for real food samples. In view of innovative fusion across L-RCA-based DNA hydrogel and SERS technique, the proposed method has promising potential for application in on-site detection and quantification of trace food contaminants.

Formation of protein-bound Nε-carboxymethyllysine and Nε-carboxyethyllysine in ground pork during commercial sterilization as affected by the type and concentration of sugars.

This research was aimed to investigate the formation of protein-bound Nε-carboxymethyllysine (CML) and Nε-carboxyethyllysine (CEL) in ground pork at 121 °C (5-30 min) as affected by sugars (1-9% w/w, glucose, fructose, lactose, and sucrose).The addition of reducing sugar significantly (P < 0.05) increased the levels of CML and CEL in heat treated pork samples. Even adding 1% of glucose in pork could lead to 3.8 and 4.0 times increase in the formation rate constant (zero-order) of CML and CEL, respectively. In a typical commercial sterilization process (121 °C, 30 min), adding glucose, fructose or lactose in pork resulted in an average increase of 224-581%, 26-276%, and 8-189% CML, and 217-720%, 213%-15.8 times, and 20-150% CEL, respectively, depending on the sugar concentration. Sucrose did not promote the formation of CML and CEL in pork during heating.

Trace analysis of organic compounds in foods with surface-enhanced Raman spectroscopy: Methodology, progress, and challenges.

Surface-enhanced Raman spectroscopy (SERS) provides a potential solution for rapid analysis of trace compounds such as residual pesticides, naturally occurred toxicants, banned or restricted drugs, and food additives in complex food matrices. In this review, the basic principles of SERS and general approaches to successfully apply SERS in food analysis are covered from an applications perspective. The key steps including substrate selection and evaluation, sample preparation and simplification, spectral collection, and data analysis during the development of SERS methods for food analysis are summarized, together with the discussion of typical underlying technical barriers or major challenges of these methods and their applications. Future directions in successfully applying SERS technology as a routine analytical approach to solve real-world food problems are analyzed. This comprehensive review summarizes the recent progress on theory, application, and scope of SERS for food analysis, providing a basic understanding of the technology; more importantly, key methodology, potential pitfalls, and possible solutions during the development of rapid SERS methods based on authors' years of SERS experience are shared with researchers in the field.