Chemical composition, vitamins, and minerals of family farming biribiri (Averrhoa bilimbi L.) in the Middle Doce River region, Minas Gerais, BrazilTítulo 12

ABSTRACT: This study evaluated physical characteristics, chemical composition, content of vitamin C, vitamin E, carotenoids, and minerals in biribiri fruits (Averrhoa bilimbi) from the Middle Doce River region (Minas Gerais, Brazil). Titratable acidity was determined by volumetric neutralization, pH by direct potentiometry, soluble solids by refractometry, humidity by gravimetry, ash by calcination in muffle, proteins by the micro-Kjeldahl method, dietary fiber by non-enzymatic gravimetric method and lipids using a Soxhlet extractor. Carotenoids and vitamin C were analyzed by high-performance liquid chromatography (HPLC) and vitamin E by HPLC with fluorescence detector. Fourteen minerals were analyzed by inductively coupled plasma atomic emission spectrometry. Biribiri showed high yield of edible portion (100%), low lipid, protein and carbohydrate content, and; consequently, low total energy value (25.36 kcal 100 g-1). The fruit also showed low dietary fiber content (0.62 g 100 g-1), total vitamin E (17.62 µg 100 g-1), total carotenoids (0.32 g 100 g-1), and high vitamin C, zinc, copper, iron content, manganese, molybdenum and chrome content. Regarding the heavy metals, the fruit showed no cadmium, and traces of aluminum and nickel. In conclusion, biribiri presented low energy value and expressive contents of dietary fibers, vitamin C, iron, manganese, molybdenum, chromium, zinc, and copper.

Rational Programming of Cas12a for Early-Stage Detection of COVID-19 by Lateral Flow Assay and Portable Real-Time Fluorescence Readout Facilities

Coronavirus disease 2019 (COVID-19) caused by the SARS-CoV-2 virus has led to a global pandemic with a high spread rate and pathogenicity. Thus, with limited testing solutions, it is imperative to develop early-stage diagnostics for rapid and accurate detection of SARS-CoV-2 to contain the rapid transmission of the ongoing COVID-19 pandemic. In this regard, there remains little knowledge about the integration of the CRISPR collateral cleavage mechanism in the lateral flow assay and fluorophotometer. In the current study, we demonstrate a CRISPR/Cas12a-based collateral cleavage method for COVID-19 diagnosis using the Cas12a/crRNA complex for target recognition, reverse transcription loop-mediated isothermal amplification (RT-LAMP) for sensitivity enhancement, and a novel DNA capture probe-based lateral flow strip (LFS) or real-time fluorescence detector as the parallel system readout facility, termed CRICOLAP. Our novel approach uses a customized reporter that hybridizes an optimized complementary capture probe fixed at the test line for naked-eye result readout. The CRICOLAP system achieved ultra-sensitivity of 1 copy/µL in ~32 min by portable real-time fluorescence detection and ~60 min by LFS. Furthermore, CRICOLAP validation using 60 clinical nasopharyngeal samples previously verified with a commercial RT-PCR kit showed 97.5% and 100% sensitivity for S and N genes, respectively, and 100% specificity for both genes of SARS-CoV-2. CRICOLAP advances the CRISPR/Cas12a collateral cleavage result readout in the lateral flow assay and fluorophotometer, and it can be an alternative method for the decentralized field-deployable diagnosis of COVID-19 in remote and limited-resource locations.

Histamine Detection in Mackerel (Scomberomorus Sp.) and its Products Derivatized with 9-Flourenilmethylchloroformate

Histamine is commonly present in food containing proteins, like in mackerel. Consuming fish is imperative for the improvement of human muscles. Nevertheless, some studies reported ingesting fish containing histamine more than 50 mg·kg-1 can cause toxicity. This study analyzed and determined the composition of histamine in mackerel and its products commonly consumed in Malaysia, especially on the East Coast of Malaysia. These included processed mackerel such as canned products, satay (skewed fish) and keropok lekor (fish cake/ cracker). Histamine analysis was performed using High Performance Liquid Chromatography (HPLC) equipped with a fluorescence detector. A derivatizing reaction was applied to increase the sensitivity of HPLC to histamine using 9-flourenilmethylchloroformate (FMOC-Cl). The chromatographic separation was achieved in 15 min. Method validation was in accordance to Commission Decision 657/2002/CE. The linear range was at 0.16 – 5.00 µg·mL-1 (histamine) with the LOD at 0.10 µg·mL-1 and LOQ at 0.30 µg·mL-1 . Method applicability was checked on seven real samples involving raw, cooked, and dry products, yielding acceptable recovery.

Exposure and risk characterizations of ochratoxins A and aflatoxins through maize (Zea mays) consumed in different agro-ecological zones of Ghana

Mycotoxin contamination of foodstuffs is a serious food safety concern globally as the prolonged ingestion of these toxins has the tendency to worsen the risk of hepatocellular carcinoma. This study aimed at estimating ochratoxin A (OTA) and aflatoxin (AF) levels above international (European Food Safety Authority, EFSA) and local (Ghana Standards Authority, GSA) standards as well as the health risks associated with the consumption of maize (n = 180) sampled from six (6) regions representing the agro-ecological zones of Ghana. OTA and AF were measured with High-Performance Liquid Chromatography with a Fluorescence detector. Out of the 180 samples analyzed for total aflatoxins (AFtotal), 131/180 tested positive and 127 (70.50%) exceeded the limits of EFSA and ranged 4.27–441.02 µg/kg. While for GSA, 116 (64.44%) of samples exceeded this limit and ranged between 10.18 and 441.02 µg/kg. For OTA, 103/180 tested positive and 94 (52.22%) of samples between the range 4.00–97.51 µg/kg exceeded the tolerable limit of EFSA, whereas 89 (49.44%) and were in the range of 3.30–97.51 µg/kg exceeded the limits of GSA. Risk assessment values for total aflatoxins (AFtotal) ranged between 50 and 1150 ng/kg bw/day, 0.4–6.67, 0–0.0323 aflatoxins ng/kg bw/day and 1.62–37.15 cases/100,000 person/year for Estimated Daily Intake (EDI), Margin of Exposure (MOE), Average Potency, and Cancer Risks respectively. Likewise, ochratoxin (OTA) values were in the ranges of 8.6 × 10–3–450 ng/kg bw/day, 0.05–2059.97, 0–0.0323 ochratoxins ng/kg bw/day and 2.78 × 10–4–14.54 cases/100,000 person/year. Consumption of maize posed adverse health effects in all age categories of the locations studied since the calculated MOE values were less than 10,000.

Fluorescence Based Comparative Sensing Behavior of the Nano-Composites of SiO2 and TiO2 towards Toxic Hg2+ Ions

We have synthesized sulfonamide based nano-composites of SiO2 and TiO2 for selective and sensitive determination of toxic metal ion Hg2+ in aqueous medium. Nano-composites (11) and (12) were morphologically characterized with FT-IR, solid state NMR, UV-vis, FE SEM, TEM, EDX, BET, pXRD and elemental analysis. The comparative sensing behavior, pH effect and sensor concentrations were carried out with fluorescence signaling on spectrofluorometer and nano-composites (11) and (12), both were evaluated as “turn-on” fluorescence detector for the toxic Hg2+ ions. The LODs were calculated to be 41.2 and 18.8 nM, respectively of nano-composites (11) and (12). The detection limit of TiO2 based nano-composites was found comparatively lower than the SiO2 based nano-composites.

Natural Occurrence of Deoxynivalenol in Cereal-Based Baby Foods for Infants from Western Poland

The study examined 110 samples of baby products based on rice, wheat, maize and multi-grains available on the western Polish market in order to detect the level of deoxynivalenol (DON) by means of HPLC (high-performance liquid chromatography) with a fluorescence detector (HPLC-FLD). DON was detected in 9.09% of the infant food samples, with an average and maximum level of 107.8 ± 30 and 148 μg/kg, respectively. The highest concentration of DON was detected in food for infants: wheat-based (mean 121 ± 7.07, 4.8%), multi-grain (mean 118 ± 5.65, 4.25%) and maize-based (mean 100 ± 37.96; 35.30%). No high DON content and high estimated daily intake were observed in the analyzed products. However, in order to minimize the harmfulness associated with the presence of DON in food for infants and young children, a risk assessment should be performed based on the monitoring results.

Multiple Compact Camera Fluorescence Detector for Real-Time PCR Devices

The polymerase chain reaction is an important technique in biological research because it tests for diseases with a small amount of DNA. However, this process is time consuming and can lead to sample contamination. Recently, real-time PCR techniques have emerged which make it possible to monitor the amplification process for each cycle in real time. Existing camera-based systems that measure fluorescence after DNA amplification simultaneously process fluorescence excitation and emission for dozens of tubes. Therefore, there is a limit to the size, cost, and assembly of the optical element. In recent years, imaging devices for high-performance, open platforms have benefitted from significant innovations. In this paper, we propose a fluorescence detector for real-time PCR devices using an open platform camera. This system can reduce the cost, and can be miniaturized. To simplify the optical system, four low-cost, compact cameras were used. In addition, the field of view of the entire tube was minimized by dividing it into quadrants. An effective image processing method was used to compensate for the reduction in the signal-to-noise ratio. Using a reference fluorescence material, it was confirmed that the proposed system enables stable fluorescence detection according to the amount of DNA.

Trace-element XAFS sensitivity: a stress test for a new XRF multi-detector

X-ray absorption fine-structure (XAFS) spectroscopy can assess the chemical speciation of the elements providing their coordination and oxidation state, information generally hidden to other techniques. In the case of trace elements, achieving a good quality XAFS signal poses several challenges, as it requires high photon flux, counting statistics and detector linearity. Here, a new multi-element X-ray fluorescence detector is presented, specifically designed to probe the chemical speciation of trace 3d elements down to the p.p.m. range. The potentialities of the detector are presented through a case study: the speciation of ultra-diluted elements (Fe, Mn and Cr) in geological rocks from a calcareous formation related to the dispersal processes from Ontong (Java) volcanism (mid-Cretaceous). Trace-elements speciation is crucial in evaluating the impact of geogenic and anthropogenic harmful metals on the environment, and to evaluate the risks to human health and ecosystems. These results show that the new detector is suitable for collecting spectra of 3d elements in trace amounts in a calcareous matrix. The data quality is high enough that quantitative data analysis could be performed to determine their chemical speciation.