Microfluidic Distillation System for Separation of Propionic Acid in Foods.

A microfluidic distillation system is proposed to facilitate the separation and subsequent determination of propionic acid (PA) in foods. The system comprises two main components: (1) a polymethyl methacrylate (PMMA) micro-distillation chip incorporating a micro-evaporator chamber, a sample reservoir, and a serpentine micro-condensation channel; and (2) and a DC-powered distillation module with built-in heating and cooling functions. In the distillation process, homogenized PA sample and de-ionized water are injected into the sample reservoir and micro-evaporator chamber, respectively, and the chip is then mounted on a side of the distillation module. The de-ionized water is heated by the distillation module, and the steam flows from the evaporation chamber to the sample reservoir, where it prompts the formation of PA vapor. The vapor flows through the serpentine microchannel and is condensed under the cooling effects of the distillation module to produce a PA extract solution. A small quantity of the extract is transferred to a macroscale HPLC and photodiode array (PDA) detector system, where the PA concentration is determined using a chromatographic method. The experimental results show that the microfluidic distillation system achieves a distillation (separation) efficiency of around 97% after 15 min. Moreover, in tests performed using 10 commercial baked food samples, the system achieves a limit of detection of 50 mg/L and a limit of quantitation of 96 mg/L, respectively. The practical feasibility of the proposed system is thus confirmed.

Finger pump microfluidic detection system for methylparaben detection in foods.

A novel assay platform consisting of a finger pump microchip (FPM) and a WiFi-based analytical detection platform is presented for measuring the concentration of methylparaben (MP) in commercial foods. In the presented approach, a low quantity (5 µL) of distilled food sample is dripped onto the FPM and undergoes a modified Fenton reaction at a temperature of 40 °C to form a green-colored complex. The MP concentration is then determined by measuring the color intensity (RGB) of the reaction complex using APP software (self-written) installed on a smartphone. The color intensity Red(R) + Green(G) value of the reaction complex is found to be linearly related (R2 = 0.9944) to the MP concentration for standard samples with different MP concentrations ranging from 100 to 3000 ppm. The proposed method is used to detect the MP concentrations of 12 real-world commercial foods. The MP concentrations measurements are found to deviate by no more than 5.88% from the results obtained using a conventional benchtop method. The presented platform thus offers a feasible and low-cost alternative to existing macroscale techniques for measuring the MP concentration in commercial foods.

Light-shading reaction microfluidic PMMA/paper detection system for detection of cyclamate concentration in foods.

Cyclamate is an artificial sweetener with high sweetness and low calories, and is a common sugar substitute for weight control and diabetic patients. However, excessive cyclamate consumption is associated with various health disorders, and hence it is prohibited as a food additive in many countries around the world. The current research proposes a light-shading reaction microfluidic PMMA/paper detection (MPD) system for determining the cyclamate concentration in food. In the current system, inject 10 µL of the extracted sodium cyclamate sample into the sample chamber of the MPD device, perform the diazotization reaction under shading conditions, and then suck it into the detection area through a paper strip, which consists of a paper chip embedded with modified Bratton-Marshall reagent. Once the paper chip is thoroughly wetted, the MPD device is inserted into a microanalysis box, where a fuchsia azo reaction compound is produced through heating at 40 °C for 3 min. The reaction complex is observed by a camera and the reaction image is wirelessly transmitted to a smartphone, and the concentration of sodium cyclamate is measured through the self-developed grayscale software. The results obtained for the sodium cyclamate samples with a concentration in the range of 50-1000 ppm show that the measured gray value changes linearly with the sodium cyclamate concentration, and the correlation coefficient (R2) is 0.9898. By analyzing the concentration of sodium cyclamate in 10 real-world samples, the practical feasibility of the current MPD system is proved. The results showed that the concentration measurement value did not deviate by more than 4.8 % from the value obtained using the conventional liquid chromatography/tandem mass spectrometry (LC-MS/MS) method.

Antioxidant Activity and Inhibitory Effects of Black Rice Leaf on the Proliferation of Human Carcinoma Cells.

The leaves of black rice, well-known as postharvest agricultural waste, contain a rich source of antioxidants with multiple benefits for human health. In the present study, the ethyl acetate fraction obtained from black rice leaf was separated into five subfractions using Sephadex LH-20 column chromatography, and their antioxidant and anticancer activities were investigated. The results revealed that among all the subfractions, subfraction 5 (Sub5) showed the highest total phenolic and flavonoid values. The antioxidant activity was also superior in Sub5 (the IC50 values are 3.23, 31.95, and 72.74 µg/mL, in the DPPH, ABTS, and reducing power assays, respectively) compared to the other subfractions. All subfractions, in a time-dependent manner, inhibited the proliferation of hepatoma (HepG2), breast (MCF-7), and colorectal (Caco-2) cancer cells, especially the Sub5. Thus, Sub5 was employed to conduct the cell cycle and cell apoptosis by flow cytometry. Sub5 significantly increased the accumulation of cells at the Sub-G1 phase in HepG2 cells (44.5%, at 48 h). Furthermore, it could trigger annexin V-detected apoptosis through mitochondrial and death receptor pathways accompanied by the suppression of PI3K/Akt and Erk signaling pathways. In addition, HPLC-DAD-MS/MS was conducted to characterize the bioactive constituents in the most potent antioxidant, cytotoxic, and apoptosis-inducing subfraction. Conclusively, Sub5 may have high potential as functional dietary supplements to inhibit the development of HepG2 liver cancer.

Evaluation of Phytochemical Contents and In Vitro Antioxidant, Anti-Inflammatory, and Anticancer Activities of Black Rice Leaf (Oryza sativa L.) Extract and Its Fractions.

Black rice leaves (Oryza sativa L.) are a major part of rice straw left in open fields after rice harvest as agricultural waste. In this study, crude ethanolic extract (CEE) and various solvent fractions (hexane (Hex), ethyl acetate (EtOAc), n-butanol (n-BuOH), and aqueous fractions) of black rice leaves were investigated for their bioactive compound contents as well as antioxidant, anti-inflammatory, and anticancer activities. The results demonstrated that among all the fractions, the n-BuOH fraction presented the greatest contents of total phenolics and flavonoids, while anthocyanins were found to be abundant in the n-BuOH and aqueous fractions, which also exhibited powerful antioxidant abilities according to DPPH and ABTS radical-scavenging assays and a reducing power assay. Regarding anti-inflammatory activity, CEE and EtOAc reduced the production of NO and cytokine secretion (PGE2, IL-6, and IL-1ß) but displayed less effect on tumor necrosis factor α (TNF-α) release in lipopolysaccharide (LPS)-induced RAW 264.7 cells. They also significantly decreased iNOS and COX-2 protein expression. Additionally, the phenolics-rich ethyl acetate fraction showed the greatest activity against HepG2 liver carcinoma cells, inhibited cell growth, increased the Sub-G1 population, and induced apoptosis via mitochondrion-dependent mechanisms. In conclusion, black rice leaves, a byproduct of rice, exhibited strong antioxidant, anti-inflammatory, and anticancer capacities and might be useful for application in functional foods and the pharmaceutical industry.

Microfluidic colorimetric analysis system for sodium benzoate detection in foods.

Sodium benzoate (SBA) is a widely-used additive for preventing food spoilage and deterioration and extending the shelf life. However, the concentration of SBA must be controlled under safe regulations to avoid damaging human health. Accordingly, this study proposes a microfluidic colorimetric analysis (MCA) system composing of a wax-printed paper-microchip and a self-made smart analysis equipment for the concentration detection of SBA in common foods and beverages. In the presented method, the distilled SBA sample is mixed with NaOH to obtain a nitro compound and the compound is then dripped onto the reaction area of the paper-microchip, which is embedded with two layers of reagents (namely acetophenone and acetone). The paper-microchip is heated at 120 °C for 20 min to cause a colorimetric reaction and the reaction image is then obtained through a CMOS (complementary metal oxide semiconductor) device and transmitted to a cell-phone over a WiFi connection. Finally, use the self-developed RGB analysis software installed on the cell-phone to obtain the SBA concentration. A calibration curve is constructed using SBA samples with known concentrations ranging from 50 ppm (0.35 mM) to 5000 ppm (35 mM). It is shown that the R + G + B value (Y) of the reaction image and SBA concentration (X) are related via Y = -0.034 X +737.40, with a determination coefficient of R2 = 0.9970. By measuring the SBA concentration of 15 commercially available food and beverage products, the actual feasibility of the current MCA system can be demonstrated. The results show that the difference from the measurement results obtained using the macroscale HPLC method does not exceed 6.0%. Overall, the current system provides a reliable and low-cost technique for quantifying the SBA concentration in food and drink products.

Multifunctional microchip-based distillation apparatus II - Aerated distillation for sulfur dioxide detection.

A multifunctional microchip-based distillation apparatus is presented for the distilled of sulfur dioxide (SO2) in food products. The microchip is fabricated on poly(methyl methacrylate) (PMMA) substrates, and comprises a sample zone, a buffer zone, a serpentine distillation column, and a collection zone. In the process, the sample is introduced into the sample zone and is heated under carefully controlled temperature and time conditions. The resulting SO2 and water vapor are carried by nitrogen (N2) gas to the distillation column, where the SO2 is separated from the water vapor via the condensing effects of a continuous cold water flow. Finally, the SO2 is transported to the collection zone, where it is collected with hydrogen peroxide (H2O2) and its concentration determined using an alkali-based titration and paper-based detection method. A distillation efficiency of 90.5% is obtained under the optimal distillation conditions at concentrations of 20-4000 ppm. Moreover, a linear correlation (R2 = 0.9997) is observed between the experimental measurements of the SO2 concentration and the known concentration. The validity of the presented microchip-based distillation apparatus is further investigated by distilling the SO2 concentrations of 25 commodity samples. The detection results show that the deviation does not exceed 5.4% compared with the traditional official method.

Multifunctional microchip-based distillation apparatus I - Steam distillation for formaldehyde detection.

A multifunctional microchip-based distillation apparatus for distilling and detecting formaldehyde (CH2O) in food products is developed. The presented apparatus comprises a disposable microchip, a steam supply system, and a recirculating cooling water supply. The microchip is formed on PMMA substrates by laser ablation and includes a sample zone, a flash distillation zone, a cooling zone, a condensation zone, and a collection zone. In the presented method, the CH2O sample is placed in the microchip and is vaporized by the high-throughput vapor supply and driven through the condensed zone. The condensed CH2O liquid is guided into the collection zone of the microchip. Finally, the distilled CH2O solution is determined using an AHMT spectrometry method and a paper-based RGB (red, green and blue) intensity analysis method. A distilled efficiency is as high as 98%, when a vapor stream rate is 0.4 ml/min and a distilled time is 10 min. Moreover, both detection methods show linear relationships of the corresponding CH2O concentrations. The actual sample suitability of the presented multifunctional microchip-based distillation apparatus is confirmed by analyzing the CH2O concentrations of 21 commodities.

A PET/paper chip platform for high resolution sulphur dioxide detection in foods.

A convenient assay platform comprising a PET/paper chip (PP-chip) and a smart analytical device is developed for detection of sulphur dioxide (SO2) concentration. In the presented approach, the distilled SO2 solution is dropped onto the detection region of the PP-chip and undergoes a reaction with an acid-based reagent. The resulting color variation is analyzed through a high-resolution camera (CMOS) and the reacted image is processed by a RGB (red, green and blue) analytical app installed on a smartphone. Results show that the known SO2 concentrations ranging from 10 to 300 ppm indicate that the high linear relationship (R2 = 0.9981) between the (R (red) + G (green) - B (blue)) value and SO2 concentration. Moreover, a high measurement resolution is equal to 1.45 ppm/a.u. The presented assay platform was proved to detect the SO2 concentrations of twenty-five practical food samples. Compared with the developed assay platform and certified inspection technique, the deviation of SO2 measurement does not exceed 3.82%. It was satisfactory to apply this developed assay platform to analyze the SO2 concentration in the practical samples.

Microfluidic paper-based chip platform for benzoic acid detection in food.

An integrated microfluidic platform comprising a microfluidic paper-based analytical device (µPAD) and a portable detection system is proposed for the concentration detection of benzoic acid via Janovsky reaction theory. In the proposed approach, the reaction zone of the µPAD is implanted with 5 N sodium hydroxide and dried at 30 °C for 20 min. The benzoic acid sample is derived to 3,5-Dinitrobenzoic acid using KNO3 and H2SO4 at 40 °C for 40 min and is then dripped on the reaction zone of the µPAD. Finally, the µPAD is transferred to the portable detection system and heated at a temperature of 45 °C for 20 min on a hot plate to prompt a Janovsky reaction. The resulting color change of the detection zone is observed using a CMOS camera. The reaction color image is delivered to a smartphone via a connector and the benzoic acid concentration is determined using self-written RGB analysis software. The experimental results obtained using control samples with known benzoic acid concentrations in the range of 500-4000 ppm show that the R(ed) + B(lue) intensity (Y) and benzoic acid concentration (X) are related as Y =  -0.0264 X + 408.79. Moreover, the correlation coefficient is equal to R2 = 0.9953. The proposed detection platform is used to measure the benzoic acid concentrations of twenty-one commercial food samples. It is shown that the concentration measurements deviate by no more than 6.6% from those obtained using a standard HPLC macroscale method. Overall, the results presented in this study show that the proposed integrated microfluidic paper-based chip platform provides a compact and reliable tool for benzoic acid concentration measurement purposes.

Microfluidic distillation chip for methanol concentration detection.

An integrated microfluidic distillation system is proposed for separating a mixed ethanol-methanol-water solution into its constituent components. The microfluidic chip is fabricated using a CO2 laser system and comprises a serpentine channel, a boiling zone, a heating zone, and a cooled collection chamber filled with de-ionized (DI) water. In the proposed device, the ethanol-methanol-water solution is injected into the microfluidic chip and driven through the serpentine channel and into the collection chamber by means of a nitrogen carrier gas. Following the distillation process, the ethanol-methanol vapor flows into the collection chamber and condenses into the DI water. The resulting solution is removed from the collection tank and reacted with a mixed indicator. Finally, the methanol concentration is inversely derived from the absorbance measurements obtained using a spectrophotometer. The experimental results show the proposed microfluidic system achieves an average methanol distillation efficiency of 97%. The practicality of the proposed device is demonstrated by detecting the methanol concentrations of two commercial fruit wines. It is shown that the measured concentration values deviate by no more than 3% from those obtained using a conventional bench top system.