Extraction and Quantitation of Phytosterols from Edible Brown Seaweeds: Optimization, Validation, and Application.

Brown seaweeds are known as important marine food sources, from which phytosterols have been recognized as functional food components with multiple health-beneficial effects. However, studies on phytosterol extraction and quantitation from edible brown seaweeds are limited. In the present work, extraction methods for seaweed phytosterols were compared and optimized by one-factor-at-one-time method and response surface methodology. Moreover, the quantitation method of total sterols and major sterol components, including fucosterol, saringosterol, and ostreasterol, was established and validated using 1H NMR. Furthermore, the developed extraction and determination methods were applied to investigate three common edible seaweeds from Japan (Hijiki, Wakame, and Kombu). As a result, the finally optimized conditions were ultrasound-assisted extraction with CHCl3-MeOH 2:3 for 15 min followed by saponification with 1.65 mL of 1.85 M KOH for 14.5 h. Based on the developed methods, phytosterols in three seaweeds were compared, and Hijiki showed an abundant total sterol amount (2.601 ± 0.171 mg/g DW), significantly higher than Wakame (1.845 ± 0.137 mg/g DW) and Kombu (1.171 ± 0.243 mg/g DW). Notably, the composition of the sterol components varied in different seaweeds. These findings might help the nutritional investigation and functional food development concerning phytosterols from seaweeds.

Hizikia fusiformis: Pharmacological and Nutritional Properties.

The brown seaweed Hizikia fusiformis (syn. Sargassum fusiforme), commonly known as "Hijiki", has been utilized in traditional cuisine and medicine in East Asian countries for several centuries. H. fusiformis has attracted much attention owing to its rich nutritional and pharmacological properties. However, there has been no comprehensive review of the nutritional and pharmacological properties of H. fusiformis. The aim of this systematic review was to provide detailed information from the published literature on the nutritional and pharmacological properties of H. fusiformis. A comprehensive online search of the literature was conducted by accessing databases, such as PubMed, SpringerLink, ScienceDirect, and Google Scholar, for published studies on the nutritional and pharmacological properties of H. fusiformis between 2010 and 2021. A total of 916 articles were screened from all the databases using the preferred reporting items for systematic reviews and meta-analyses method. Screening based on the setdown criteria resulted in 59 articles, which were used for this review. In this review, we found that there has been an increase in the number of publications on the pharmacological and nutritional properties of H. fusiformis over the last 10 years. In the last 10 years, studies have focused on the proximate, mineral, polysaccharide, and bioactive compound composition, and pharmacological properties, such as antioxidant, anticancer, antitumor, anti-inflammatory, photoprotective, neuroprotective, antidiabetic, immunomodulatory, osteoprotective, and gastroprotective properties of H. fusiformis extracts. Overall, further studies and strategies are required to develop H. fusiformis as a promising resource for the nutrition and pharmacological industries.

A Simplified Questionnaire for the Assessment of Inorganic Arsenic Intake in a Japanese Population.

A simplified questionnaire was developed to assess inorganic arsenic (iAs) intake level in a Japanese population. The two page questionnaire included photographs of single serving sizes of rice and cooked hijiki (Hizikia fusiforme: brown algae), and asked subjects about the number of servings of rice and cooked hijiki, two predominant dietary sources of iAs in Japan, they consume in a day. Daily intake of iAs was estimated for 72 Japanese subjects using the questionnaire together with data of iAs content in rice and hijiki seaweed, and the estimated intakes were compared with actual iAs intakes of the subjects as measured for a duplicate diet using liquid chromatography-inductively coupled plasma mass spectrometry. A highly significant correlation was found between the estimated and measured intakes (r = 0.65, p < 0.001); however, the slope of regression indicated a systematic error in the intake estimation. Possible sources of error are discussed herein. It was concluded that this approach is promising if minor improvements are made to the questionnaire.

A New Candidate Reference Material for Inorganic Arsenic and Arsenosugars in Hijiki Seaweed: First Results from an Inter-laboratory Study.

An inter-laboratory study was carried out to characterize a candidate hijiki seaweed for its concentrations of total arsenic and water-soluble arsenic compounds, particularly arsenosugar compounds. The candidate material, a dried hijiki seaweed powder, was analyzed by individual techniques in two laboratories. The water-soluble arsenic compounds were separated by anion exchange, and reversed-phase columns, and As(V), DMA and four kinds of arsenosugars, namely glycerol (-OH), phosphate (-PO4), sulfonate (-SO3), and sulfate (-SO4) types were detected by HPLC-ICP-MS. The methods applied were validated by analyzing a second sample, the NMIJ CRM 7405-a hijiki seaweed, which is certified for both total arsenic and As(V). Techniques for the inter-laboratory study, extraction efficiencies under different extraction conditions, some chromatographic techniques and sequential extraction were investigated. The results from the two laboratories for the candidate hijiki material showed good agreement within the measurement uncertainties for total and water-soluble arsenic compounds.

Hijiki seaweed consumption elevates levels of inorganic arsenic intake in Japanese children and pregnant women.

Although exposure to inorganic arsenic is a health concern, especially in developing foetuses and children, dietary exposure levels among pregnant women and children have not been extensively studied in Japan. To address this shortcoming, we completed a 3-day duplicate diet study for 104 children and 101 pregnant women in two cities, Shimotsuke, Tochigi and Asahikawa, Hokkaido. The levels of intake of total and inorganic arsenic were estimated using the concentrations of total and inorganic arsenic in food and drinking water measured by inductively coupled plasma-mass spectrometry. Estimated intakes of total and inorganic arsenic were 8.46 ± 3.02 [µg/kg BW/week] and 1.74 ± 1.07 [µg/kg BW/week] in pregnant women and 20.07 ± 3.53 [µg/kg BW/week] and 8.46 ± 3.02 [µg/kg BW/week] in children, respectively. Weekly arsenic exposure per kg body weight was significantly higher in children than in pregnant women. Concentrations of total arsenic were generally very low in collected drinking water samples with a small number of exceptions, and drinking water was not considered as a major source of inorganic arsenic exposure in Japan. We found that total and inorganic arsenic intake were higher among frequent consumers of hijiki seaweed, in both pregnant women and children. Although rice and rice products that are staple foods of the Japanese have been reported to be major sources of inorganic arsenic exposure in Japan, our results indicate that hijiki consumption elevates levels of inorganic arsenic in Japanese children and pregnant women. More efforts are necessary to reduce the risk of exposure to inorganic arsenic in populations highly sensitive to environmental pollutants.

Hijiki and sodium arsenite stimulate growth of human colorectal adenocarcinoma cells through ERK1/2 activation.

Hijiki seaweed (Hijikia fusiformes) contains high levels of inorganic arsenic, a known carcinogen. However, scientific reports on carcinogenic risks associated with the consumption of this seaweed are limited. This study investigated the effects of seaweed extracts contaminated with arsenic on two colorectal cancer cell lines. Two seaweed extracts, including Hijiki and red seaweed, induced H508 but not HT29 cell proliferation. Growth induction of H508 cells after treatments with Hijiki and sodium arsenite at concentrations equivalent to arsenic found in Hijiki was observed by both MTT and BrdU assays. Hijiki and sodium arsenite induced epidermal growth factor receptor (EGFR) and ERK1/2 activations. AG1478, an EGFR inhibitor, decreased the activation of EGFR and ERK1/2 induced by Hijiki and sodium arsenite. U0126, an ERK1/2 upstream inhibitor, and atropine, a muscarinic acetylcholine receptor (mAChR) antagonist, but not AG1478 completely inhibited the proliferative effect of Hijiki. Altogether, the results suggest that the presence of arsenic in seaweed may partly contribute to the proliferation of colorectal cancer cells. EGFR-dependent, and -independent ERK1/2 signaling pathways, and mAChR may be involved in the growth stimulation by Hijiki. These results raise concern regarding the potential colorectal cancer risks from regular consumption of Hijiki containing high contents of inorganic arsenic.

Effects of Edible Seaweed on Physicochemical and Sensory Characteristics of Reduced-salt Frankfurters.

The effects of sea tangle, sea mustard, hijiki, and glasswort were investigated based on the proximate composition, salinity, cooking loss, emulsion stability, pH, color, texture profile analysis, apparent viscosity, and sensory characteristics of reduced-salt (NaCl) meat batter and frankfurters. The moisture content, salinity, lightness of the meat batter and frankfurter, hardness, gumminess, and chewiness of the reduced-salt frankfurters with sea weeds were lower than the control without seaweed (p<0.05). The protein content, springiness, and cohesiveness of the reduced-salt frankfurters were not significantly different among the treatments (p>0.05). The moisture content, salinity, cooking loss, lightness, redness, hardness, gumminess, and chewiness of treatments with sea tangle and with sea mustard were lower than the control (p<0.05). Among the sensory traits, color was highest in the control (p<0.05). The flavor was also highest in the control. The treatments with sea tangle and with sea mustard samples had high tenderness, juiciness, and overall acceptability scores similar to the control (p<0.05). The results of this study show that the combination of low-salt and seaweed in the formulation successfully improved reduced-salt frankfurters, improving sensory characteristics to levels similar to the regular salt control (1.5%).

Stability of toxic arsenic species and arsenosugars found in the dry alga Hijiki and its water extracts.

The achievement of reliable results in speciation analysis requires not only sensitive techniques but also sureness of species stability. Therefore, it is necessary to carry out stability studies because it is important to know with absolute certainty that there is not any species transformation during sample treatment and/or storage. Although several procedures have been recommended for the preservation of species integrity, there is no general agreement, as arsenic species stability depends on the sample matrix, the concentration level and the sample treatment procedure, so it is necessary to assess the arsenic species stability for each case. Thus, the present paper reports the stability tests of arsenic species carried out on the commercially available edible alga Hijiki (Hizikia fusiformis), from Japan, in both the dry sample and its water extracts, which were stored in amber glass and polystyrene containers at -18 and +4°C in the dark. Extractions were carried out with deionized water by microwave-assisted extraction, at a temperature of 90°C and three extraction steps of 5 min each, whereas arsenic speciation analysis was performed by anion exchange high performance liquid chromatography-photo-oxidation-hydride generation-atomic fluorescence spectrometry. The results obtained for the dry alga showed that the arsenic species present in it (arsenate (As(V)), dimethylarsinic acid (DMA) and the arsenosugars glycerol (Gly-sug), phosphate (PO4-sug), sulfonate (SO3-sug) and sulfate (SO4-sug)) were stable for at least 12 months when the sample was stored in polystyrene containers at +20°C in the dark. Regarding water extracts, the best storage conditions consisted of the use of polystyrene containers and a temperature of +4°C, for a maximum storage time of seven days. Therefore, the immediate analysis of Hijiki water extracts would not be necessary, and they could be stored for one week before analysis, ensuring arsenic species stability. This information about species integrity in extracts is especially useful when the sample treatment for arsenic species extraction is time-consuming.