Compositional characteristics and antioxidant properties of fresh and processed sea cucumber (Cucumaria frondosa)

The antioxidant activity of fresh and rehydrated sea cucumber (Cucumaria frondosa) samples with/without internal organs was evaluated for the first time. In addition, their proximate, amino acid, and fatty acid compositions were examined. Rehydrated sea cucumber samples in distilled water were prepared from oven-dried products. All samples contained 83-90% moisture, but showed a significant difference among groups in their protein and lipid contents. Glutamic acid was the predominant amino acid in sea cucumber, followed by glycine and aspartic acid. Essential amino acids such as leucine and lysine were also present at high levels. The trend for free amino acid was different from that of total amino acids and varied among groups. Lipids in sea cucumber were dominated by eicosapentaenoic acid (EPA, C20:5n-3), ranging from 43.2 to 56.7% of the total fatty acids. Docosahexaenoic acid (DHA, C22:6n-3) was present at a much lower concentration of 2.0-5.8%. All sea cucumber samples exhibited radical scavenging property against 2,2'-azobis(2-aminopropane) dihydrochloride (AAPH) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, with rehydrated samples, especially those with internal organs, possessing higher antioxidant activity than their fresh counterparts. No correlation existed between radical scavenging capacity and total phenolics content, suggesting that other components, in addition to phenolic compounds, contribute to the antioxidant activity of sea cucumber.     

蚕豆发芽过程中蛋白质和抗氧化能力的变化

为更好地开发利用蚕豆资源,研究蚕豆发芽过程中蛋白质及蛋白抗氧化能力的变化规律,采用凯氏定氮法测定发芽蚕豆的蛋白质含量,采用氨基酸分析仪测定氨基酸组分含量,运用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)技术观察发芽蚕豆蛋白组成的变化,通过测定发芽蚕豆蛋白的1,1-二苯基-2-三硝基苯肼(DPPH)自由基和2,2'-联氮双-(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)自由基的清除能力分析其抗氧化能力。结果表明,蚕豆发芽过程中蛋白和氨基酸含量升高,促进部分蚕豆蛋白由大分子量转化分解成小分子量,提高了蚕豆蛋白的抗氧化能力,在蚕豆发芽第9天时蛋白质含量达到34.1 g·100g^-1、氨基酸含量为29.16 g·100g^-1,均达到最大值,蚕豆蛋白的自由基清除能力达到最强。综上,蚕豆的发芽过程可以增加蚕豆的营养物质成分含量,增强蚕豆的抗氧化能力。本研究结果为蚕豆蛋白、蚕豆芽等功能食品的开发利用提供了参考依据。     

Evolution of phenolic compounds and antioxidant activity during malting

Two barley varieties, Gan4 and Hamelin, were malted to investigate the evolution of phenolic compounds and antioxidant activity during malting. The antioxidant activity was evaluated with DPPH radical scavenging activity, ABTS radical cation scavenging activity, reducing power, and metal chelating activity. Results showed that malting had significant influences on individual and total phenolic contents as well as antioxidant activities of two barley varieties. The contents of some phenolic compounds and the antioxidant activities decreased significantly during steeping and the early stages of germination and then increased remarkably during the later stages of germination and subsequent kilning. The most phenolic compounds identified in barley were (+)-catechin and ferulic acid, which both changed significantly during malting. Moreover, results from the Pearson correlation analysis showed that there were good correlations among DPPH radical scavenging activity, ABTS radical cation scavenging activity, reducing power, total phenolic content and sum of individual phenolic contents during malting.     

Botanical origin causes changes in nutritional profile and antioxidant activity of fermented products obtained from honey

Honey as rich source of enzymatic and nonenzymatic antioxidants serves as health-promoting nutrient in the human body. Here, we present the first time a comparative study of nutritional profiles (e.g., acidities, sugar, organic acid profile, total polyphenolic, flavonoid content) for different unifloral, multifloral honeys and their fermented products, in correlation with their antioxidant activity. Additionally, an optimized method for HPLC separation of organic acids from honey was established. The total phenolic content of honey samples varied widely among the honey types compared to fermented products. High amounts of total flavonoids were quantified in heather honey, followed by raspberry, multifloral, black locust, and linden honey. A positive correlation between the content of polyphenols, flavonoids, and antioxidant activity was observed in honey samples. After fermentation, the flavonoid content of dark honey fermented products decreased significantly. Black locust and linden honeys are more suitable for fermentation because the decrease in antioxidant substances is less pronounced.     

海马水解蛋白的氨基酸组成与抗氧化能力的关系

为了探讨大海马蛋白质的高效利用方法,本试验将脱脂大海马粉经碱性蛋白酶和胰蛋白酶联合消解后,酶解液(PH)经超滤膜分离为5组分(PH-Ⅰ、PH-Ⅱ、PH-Ⅲ、PH-Ⅳ、PH-Ⅴ),结合各组分的氨基酸组成,回归分析了各组分的自由基清除能力与氨基酸组成间的相关关系。结果表明,酶解液经超滤膜分离的5组分中PH-Ⅴ(<2 500 Da)抗氧化能力较强,清除DPPH自由基能力为34.2%±0.1%,对超氧阴离子的清除能力为29.2%±0.1%,Fe³⁺还原力为0.28%±0.1%。海马酶解多肽各组成中疏水性氨基酸含量低于亲水性氨基酸,但疏水性氨基酸对自由基的清除能力起着决定性作用,极性氨基酸和非极性氨基酸可通过协同作用来增强海马多肽清除自由基的能力,且每个单位极性氨基酸的抗氧能力是非极性氨基酸的1.2倍,每个单位芳香族氨基酸的抗氧化能力是脂肪族氨基酸的2倍。本研究不仅制备了一类具有开发价值的抗氧化多肽,并且分析了多肽抗氧化能力与氨基酸组成的相关关系,为大海马的高值化利用提供了理论依据。     

高氨基酸和高茶氨酸茶树资源筛选

为筛选高氨基酸和高茶氨酸茶树资源及探究茶树中茶氨酸与氨基酸含量之间的关系,本研究利用茚三酮显色法结合酶标仪检测技术测定218份茶树资源春季一芽二叶蒸青样的氨基酸含量,利用高效液相色谱法测定了其茶氨酸含量。结果表明,218份资源的氨基酸含量介于1.50%~8.98%之间,其中高氨基酸茶树资源(氨基酸含量≥5%)55份;茶氨酸含量介于0%~4.03%之间,其中高茶氨酸茶树资源(茶氨酸含量≥3%)13份,不含茶氨酸的茶树资源4份;茶氨酸与氨基酸含量的比值介于0~0.78之间,且集中分布于0.2~0.5之间,二者的相关性系数达0.752,呈极显著正相关。本研究筛选获得了一批氨基酸特异的茶树资源,为高氨基酸和高茶氨酸茶树新品种选育及其形成的分子机理研究奠定了基础。     

Quantification of polyphenolics and their antioxidant capacity in fresh plums

Total phenolics, total flavonoids, and antioxidant capacity of 11 cultivars of fresh plums were determined using spectrophotometric methods. Identification and quantification of individual polyphenolics were performed using reversed-phase high-performance liquid chromatography equipped with a diode array detector. The total phenolic contents of various cultivars widely varied from 125.0 to 372.6 mg/100 g expressed as gallic acid equivalents. The level of total flavonoids in fresh plums ranged between 64.8 and 257.5 mg/100 g expressed as catechin equivalents. Antioxidant capacity, expressed as vitamin C equivalent antioxidant capacity (VCEAC), ranged from 204.9 to 567.0 mg/100 g with an average of 290.9 mg/100 g of fresh weight. Cv. Beltsville Elite B70197 showed the highest amounts of total phenolics and total flavonoids and the highest VCEAC. A positive relationship (correlation coefficient r (2)() = 0.977) was presented between total phenolics and VCEAC, suggesting polyphenolics would play an important role in free radical scavenging. The level of IC(50) value of superoxide radical anion scavenging activity of the plum cultivars ranged from 13.4 to 45.7 mg of VCEAC/100 g. Neochlorogenic acid was the predominant polyphenolic among fresh plums tested. Flavonols found in plum were commonly quercetin derivatives. Rutin was the most predominant flavonol in plums. Various anthocyanins containing cyanidin aglycon and peonidin aglycon were commonly found in all plums except for cv. Mirabellier and NY 101.     

Phytochemical profile of main antioxidants in different fractions of purple and blue wheat, and black barley

Two pigmented wheat genotypes (blue and purple) and two black barley genotypes were fractionated in bran and flour fractions, examined, and compared for their free radical scavenging properties against 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical cation (Trolox equivalent antioxidant capacity, TEAC), ferric reducing antioxidant power (FRAP), total phenolic content (TPC), phenolic acid composition, carotenoid composition, and total anthocyanin content. The results showed that fractionation has a significant influence on the antioxidant properties, TPC, anthocyanin and carotenoid contents, and phenolic acid composition. Bran fractions had the greatest antioxidant activities (1.9-2.3 mmol TEAC/100 g) in all four grain genotypes and were 3-5-fold higher than the respective flour fractions (0.4-0.7 mmol TEAC/100 g). Ferulic acid was the predominant phenolic acid in wheat genotypes (bran fractions) while p-coumaric acid was the predominant phenolic acid in the bran fractions of barley genotypes. High-performance liquid chromatography analysis detected the presence of lutein and zeaxanthin in all fractions with different distribution patterns within the genotypes. The highest contents of anthocyanins were found in the middlings of black barley genotypes or in the shorts of blue and purple wheat. These data suggest the possibility to improve the antioxidant release from cereal-based food through selection of postharvest treatments.     

Free radical scavenging properties and phenolic content of Chinese black-grained wheat

Free radical scavenging properties and phenolic content of extracts from a novel Chinese black-grained wheat were evaluated for comparison with selected wheat controls. Extracts of bran and whole meal were compared for their scavenging activities against the 2,2-diphenyl-1-picryhydrazyl (DPPH) free radical. The total phenolic content and phenolic acid levels were determined using colorimetric and high-performance liquid chromatography (HPLC) methods, respectively. There were significant differences in radical scavenging activities and phenolic contents among bran or whole meal samples of Chinese black-grained wheat and selected wheat controls. Chinese black-grained wheat had the strongest scavenging activity and the highest total phenolic content among the wheat samples. The scavenging activity and total phenolic content of wheat bran was generally twice as high as that of whole meal. A positive correlation was found between DPPH radical scavenging activity and total phenolic content of bran (R = 0.86) and whole meal (R = 0.96). In addition, HPLC analysis detected the presence of gallic, p-hydroxybenzoic, caffeic, syringic, p-coumaric, vanillic, gentisic, o-coumaric acid, and ferulic acids in wheat bran. Ferulic acid content was highest among the phenolic acids. Chinese black-grained wheat may be considered as a potential source of natural antioxidants given its high free radical scavenging ability and phenolic content. Additional research is needed to further investigate other phenolic compounds and evaluate their contribution to the antioxidant activity in order to understand the nutraceutical value of the novel black-grained wheat genotype.     

不同种类及加工方式对杂粮酸奶体外抗氧化活性的比较

为探究杂粮品种及加工方式对杂粮酸奶体外抗氧化活性的影响,选择小米、黄米、燕麦、藜麦、糙米、荞麦、高粱米7种杂粮为原料,经过蒸制、煮制、打浆3种常见加工方式,比较其多酚含量及抗氧化活性,对特性较好的杂粮经恰当处理后与牛奶共发酵制备酸奶,研究该杂粮酸奶的多酚含量及抗氧化活性,开发具备抗氧化活性的杂粮酸奶。结果表明,7种杂粮之间的抗氧化能力存在显著（P<0.05）差异,采用抗氧化综合（antioxidant potency composite,APC）指数法评定杂粮的抗氧化活性,发现抗氧化活性最高的杂粮为荞麦。进一步对荞麦进行加工处理,发现蒸制处理后其抗氧化活性优于煮制和打浆。将蒸制的荞麦与牛奶混合制备酸奶,制成的荞麦酸奶总酚含量为52.85 mg/100g,是普通酸奶的5.16倍;且其抗氧化能力显著高于普通酸奶（P<0.05）。该研究为功能性杂粮酸奶的开发提供借鉴。     

Antioxidant Capacity of Newly Developed Pigmented Rice Cultivars in Korea

The antioxidant capacity of newly developed and highly popular pigmented rice cultivars (black rice, Galsaekchalmi, Jeoktomi, Hongchalmi, and Nogwonmi) in South Korea was analyzed. The rice grains were ground into powder, extracted with 70% ethanol, filtered, and concentrated with a rotary evaporator. The samples were analyzed for phenolic, flavonoid, and phytic acid contents, free radical scavenging activity, reducing power, ferrous ion chelating ability, lipid peroxidation inhibition, and superoxide dismutase‐like activity. The ethanolic extracts from pigmented rice cultivars showed greater antioxidant activity than that of the normal white rice. The black rice exhibited the highest free radical scavenging activity, ferrous chelating ability, and total phenolic and flavonoid contents. The reducing power and phytic acid content were found to be highest in Hongchalmi cultivar. The inhibition of lipid peroxidation was markedly higher in Jeoktomi compared with the other rice samples. The Nogwonmi rice showed the lowest antioxidant activity among the pigmented varieties analyzed. These findings provide valuable information on the antioxidant potential of newly developed pigmented rice varieties and may assist plant breeders in the selection of cultivars for the development of new lines of rice with enhanced functional quality.     

The Effect of Bioprocessing on the Phenolic Acid Composition and Antioxidant Activity of Wheat Bran

In the present study, bioprocessing with eight microbial strains including Bacillus species, yeasts, and filamentous fungi was evaluated for its potential to improve the phenolic acid composition and antioxidant activity of wheat bran. The soluble free and soluble conjugated fractions of ethanolic extracts of the treated bran samples were compared for their total phenolic contents, phenolic acid composition, and in vitro antioxidant activities. In general, total phenolic content in the soluble free fraction increased significantly, accounting for 241.11 ± 1.25 μg of gallic acid equivalents (GE)/g (Rhizopus oryzae), 230.50 ± 1.05 μg of GE/g (Mucor circinelloides), and 230.19 ± 1.02 μg of GE/g (Saccharomycopsis fibuligera). The phenolic acid composition, especially of the soluble free fraction, was improved most by S. fibuligera (hydroxybenzoic, vanillic, syringic, and trans‐ferulic acids), M. circinelloides (chlorogenic acid), and R. oryzae (protocatechuic, trans‐coumaric, and benzoic acids). Comparatively, bioprocessing exhibited less effectiveness on conjugated phenolic acid composition. Fermented wheat bran displayed enhanced reducing capacity, superoxide anion radical scavenging activity, and 1,1‐diphenyl‐2‐picrylhydrazyl radical scavenging activity in comparison with the nonfermented sample. The antioxidant activity was significantly correlated to the total phenolic content.     

Buckwheat honey increases serum antioxidant capacity in humans

Honey has been known to exert significant in vitro antioxidant activity, in part due to its phenolic content. However, conclusions that the antioxidants in honey are or are not efficacious in the human body cannot be reached if its antioxidant action is not assessed as part of a human study. In the present study, the acute effect of consumption of 500 mL of water, water with buckwheat honey, black tea, black tea with sugar, or black tea with buckwheat honey on serum oxidative reactions was examined in 25 healthy men. Antioxidant capacity of human serum samples was measured using different methods: the oxygen radical absorbance capacity (ORAC) assay, ex vivo susceptibility of serum lipoprotein to Cu(2+)-induced oxidation, and the thiobarbituric acid reactive substances (TBARS) assay. The results showed that the serum antioxidant capacity determined by ORAC increased significantly (p < 0.05) by 7% following consumption of buckwheat honey in water. No significant changes in serum antioxidant capacity could be established after the consumption of any of the other beverages. Ex vivo serum lipoprotein oxidation and TBARS values were not significantly altered after consumption of any of the five beverages. This study provides primary evidence of the in vivo antioxidant activity of buckwheat honey. However, long-term studies and epidemiological data are necessary to investigate whether honey consumption can exert overall antioxidant-related health benefits.     

Comparison of Swiss red wheat grain and fractions for their antioxidant properties

Swiss red wheat grain, bran, aleurone, and micronized aleurone were examined and compared for their free radical scavenging properties against 2,2-diphenyl-1-picrylhydrazyl radical (DPPH*), radical cation ABTS*+ and peroxide radical anion O(2)*-, oxygen radical absorbance capacity (ORAC), chelating capacity, total phenolic content (TPC), and phenolic acid composition. The results showed that micronized aleurone, aleurone, bran, and grain may significantly differ in their antioxidant properties, TPC, and phenolic acid composition. Micronized aleurone had the greatest antioxidant activities, TPC, and concentrations of all identified phenolic acids, suggesting the potential of postharvesting treatment on antioxidant activities and availability of TPC and phenolic acids. Ferulic acid was the predominant phenolic acid in Swiss red wheat and accounted for approximately 57-77% of total phenolic acids on a weight basis. Ferulic acid concentration was well correlated with scavenging activities against radical cation and superoxide anion, TPC, and other phenolic acid concentrations, suggesting the potential use of ferulic acid as a marker of wheat antioxidants. In addition, 50% acetone and ethanol were compared for their effects on wheat ORAC values. The ORAC value of 50% acetone extracts was 3-20-fold greater than that of the ethanol extracts, indicating that 50% acetone may be a better solvent system for monitoring antioxidant properties of wheat. These data suggest the possibility to improve the antioxidant release from wheat-based food ingredients through postharvesting treatment or processing.     

Comparison of free amino acids profile in honey from three Argentinian regions

The physical chemistry characteristics of honey are directly related to floral origin and, as a result, to the production region. There are some available methods that can determine the botanical or geographical origin of honey such as the free amino acids profile analysis. This paper reports data on the free amino acid composition, determined by reversed-phase high-performance liquid chromatography UV detection on 56 honey samples from three different Argentine regions, with characteristic apiarian flora. To evaluate if the quantified amino acid could be used to verify the geographical or botanical origin of honey, statistical analyses were performed. The cluster analysis showed that samples were grouped in clusters related to sampling regions and more strictly to apiarian flora around apiaries. Each cluster appears associated, in accordance with the principal component analysis, to high or low concentrations of different amino acids.     

Effect of principal polyphenolic components in relation to antioxidant characteristics of aged red wines.

Aged red wines possess significantly different polyphenolic composition compared with young ones, mainly due not only to formation of polymeric compounds but also because of oxidation, hydrolysis, and other transformations that may occur in native grape phenolics during aging. Representative Greek, single-variety, aged red wines were examined for total phenol, total flavanol, and total anthocyanin content using spectrophotometry, and attempts were made to establish correlations with the antiradical, reducing, and hydroxyl free radical scavenging activity. In addition, HPLC analyses were carried out, to ascertain whether individual polyphenols are actually responsible for the antioxidant effects of aged red wines. It was found that total flavanols are the class of polyphenols that account for hydroxyl free radical scavenging efficacy and to a lesser extent for antiradical and reducing ability, whereas there was a less significant link between the antioxidant properties and the total phenolics and only a weak relationship to total anthocyanin content. The correlation of the antioxidant properties with the principal polyphenols showed that individual compounds are weakly associated with all the antioxidant parameters, suggesting that the expression of antioxidant activity in aged red wines is rather a consequence of synergism between various phenolics, and it is not simply attributed to specific constituents.     

Free and Bound Phenolic Acids and Total Phenolics in Black,Blue, and Yellow Barley and Their Contribution to Free Radical Scavenging Capacity

Barley is considered a healthy food because of its high content of β‐glucan and phenolic antioxidants. In the current study, 28 black, blue, and yellow barleys were investigated in terms of their composition of free and bound phenolic acids and 2,2‐diphenyl‐1‐picrylhydrazyl radical scavenging capacity. Free phenolics were based on aqueous methanol extraction, whereas bound phenolics were extracted following alkaline hydrolysis. Phenolics were then separated and quantified by liquid chromatography and the Folin–Ciocalteu method. Significant differences were observed between the three barley color groups, and within each color group a wide range of phenolics concentrations existed. Ferulic acid was the predominant phenolic acid in free and bound extracts, followed by p‐coumaric acid in all the barleys investigated. Total phenols content and individual phenolic acids strongly correlated with free radical scavenging capacity of barley. Black and blue barley were found to be related and distinct from yellow barley. The results showed significant variations in phenolics among barleys, with a potential for the development of barley grains with high content of phenolic compounds as antioxidant potential.     

Free radical scavenging properties of wheat extracts

Three hard winter wheat varieties (Akron, Trego, and Platte) were examined and compared for their free radical scavenging properties and total phenolic contents (TPC). Free radical scavenging properties of wheat grain extracts were evaluated by spectrophotometric and electron spin resonance (ESR) spectrometry methods against stable 2,2-diphenyl-1-picryhydrazyl radical (DPPH*) and radical cation ABTS*+ (2,2'-azino-di[3-ethylbenzthiazoline sulfonate]). The results showed that the three wheat extracts differed in their capacities to quench or inhibit DPPH* and ABTS*+. Akron showed the greatest activity to quench DPPH radicals, while Platte had the highest capacity against ABTS*+. The ED50 values of wheat extracts against DPPH radicals were 0.60 mg/mL for Akron, 7.1 mg/mL for Trego, and 0.95 mg/mL for Platte under the experimental conditions. The trolox equivalents against ABTS*+ were 1.31 +/- 0.44, 1.08 +/- 0.05, and 1.91 +/- 0.06 micromol/g of grain for Akron, Trego, and Platte wheat, respectively. ESR results confirmed that wheat extracts directly reacted with and quenched free radicals. The TPC were 487.9 +/- 927.8 microg gallic acid equivalents/g of grain. No correlation was observed between TPC and radical scavenging capacities for DPPH* and ABTS*+ (p = 0.15 and p > 0.5, respectively).     

红外热处理对油茶籽油活性成分及综合抗氧化水平的影响

为探究热处理后油茶籽油活性成分的变化规律与其氧化稳定性内在联系,该研究以油茶籽为原料,经热处理后对所得油茶籽油的7个指标进行测定分析（氧化诱导时间、脂肪酸组成、多酚、美拉德产物、β-谷甾醇、角鲨烯、生育酚）,并对整体抗氧化水平构建综合评价体系。结果表明：热处理后油茶籽油各指标均有显著性变化（P<0.05）：氧化诱导时间最大和最小值与未处理对照相比分别增加47.05%和降低36.02%;不饱和脂肪酸在脂肪酸组成中占比最高且易被氧化;总酚、5-羟甲基糠醛(5-HMF)、丙酮醛(MGO)、3-脱氧葡萄糖醛酮(3-DG)、β-谷甾醇、角鲨烯与α-生育酚含量最高分别是对照的7.12、1.69、7.19、4.27、1.13、1.03和1.25倍;总酚与抗氧化能力呈极显著相关（P<0.01）;美拉德产物5-HMF、MGO、3-DG含量与抗氧化能力呈显著相关（P<0.05）,β-谷甾醇、角鲨烯与α-生育酚与温度分别呈正相关、显著负相关、显著正相关;综合评价模型表明极性组分（总酚、美拉德产物）抗氧化能力权重大于非极性组分,热处理110 ℃前非极性组分对油茶籽油的抗氧化水平起主要作用,110 ℃后极性组分起主要作用。与未处理对照相比,适度热处理可以改变油茶籽油中活性物质含量及抗氧化能力并以此调控油脂的氧化稳定性。研究结果为探究油脂氧化稳定机理提供了数据支持,为油茶籽油加工工艺参数选择提供参考。     

Antioxidative properties of cardoon (Cynara cardunculus L.) infusion against superoxide radical,hydroxyl radical,and hypochlorous acid

Polyphenols are able to act as antioxidants by virtue of their hydrogen-donating and metal-chelating capacities. Cardoon (Cynara cardunculus L.) is a species containing considerable amounts of polyphenolic compounds, namely flavonoids and phenolic acids. This study examined the antioxidant activity of cardoon lyophilized infusion against superoxide radical, hydroxyl radical, and hypochlorous acid. Superoxide radical was generated either in an enzymatic system or nonenzymatically, and the scavenging ability was assessed by the inhibition of superoxide radical-induced reduction of nitroblue tetrazolium. Hydroxyl radical was generated by the Fe3+-EDTA/ascorbate Fenton system, and scavenging capacity was estimated by evaluating the inhibition of hydroxyl radical-induced deoxyribose degradation into thiobarbituric acid-reactive substances. Inhibition of hypochlorous acid-induced 5-thio-2-nitrobenzoic acid oxidation to 5,5'-dithiobis(2-nitrobenzoic acid) was used in order to test the hypochlorous acid scavenging activity.