吐温80增溶-紫外分光光度法测定辅酶Q_(10)脂质体的载量及包封率

在辅酶Q10脂质体的制备过程中,载量和包封率是评价辅酶Q10脂质体的2个重要质量指标。采用表面活性剂吐温80对辅酶Q10脂质体进行增溶,再结合紫外分光光度法测定其载量和包封率。研究结果表明,辅酶Q10浓度在2.5～50μg/mL范围内,吐温80增溶法与以乙醇为溶剂的反相高效液相色谱法以及紫外分光光度法有良好的相关性(R2>0.999);空白脂质体中,辅酶Q10的加样回收率在(98.26±0.63)%～(101.20±1.28)%之间,相对标准偏差RSD<2%(n=6);该法用于测定辅酶Q10脂质体中总辅酶Q10含量的RSD<5%(n=6);不同载量[(3.22±0.01)%～(13.62±0.31)%]的辅酶Q10脂质体的包封率均高于95%(RSD<1%,n=6)。与以乙醇为溶剂的反相高效液相色谱法以及紫外分光光度法相比,该法具有准确可靠、简单、重现性较好的优点。     

Tyrosinase inhibition by water and ethanol extracts of a far eastern sea cucumber, Stichopus japonicus

BACKGROUND: Tyrosinase plays a key role in hyperpigmentaion and enzymatic browning. The present study was aimed at investigating the inhibitory effects of water and 70% aqueous ethanol extracts of Stichopus japonicus, a sea cucumber long consumed as a tonic food and traditional medicine, on the diphenolase activity of tyrosinase. RESULTS: In the tyrosinase inhibition study, high‐performance liquid chromatography completely separated L ‐3,4‐dihydroxyphenylalanine and dopachrome from other compounds present in the extracts, and provided more reliable results than the commonly used spectrophotometry. The ethanol extract (IC₅₀ = 0.49–0.61 mg mL^?1) showed higher inhibitory activity than the water extract (IC₅₀ = 1.80–1.99 mg mL^?1). Enzyme inhibition by the extracts was reversible and of mixed type. For both extracts, the dissociation constants for binding to free enzyme were significantly smaller than those for binding to enzyme–substrate complex. Ethyl‐α‐D ‐glucopyranoside (IC₅₀ = 0.19 mg mL^?1), isolated for the first time from sea cucumber, and adenosine (IC₅₀ = 0.13 mg mL^?1), were identified as key tyrosinase inhibitors. CONCLUSION: The sea cucumber extracts were demonstrated to possess considerable inhibitory potency against the diphenolase activity of tyrosinase, suggesting that the sea cucumber may be a good source of safe and effective tyrosinase inhibitors. Copyright © 2011 Society of Chemical Industry     

Antioxidant activities of different Teucrium montanum L. Extracts

The sequential extraction of Teucrium montanum L. was realised with five solvents of different polarities (70% methanol, petroleum ether, chloroform, ethyl acetate, and n‐butanol) and HPLC method was used for identification of phenolic compounds. The total phenolic content of the extracts was determined spectrophotometrically according to the Folin–Ciocalteau procedure and range from 0 to 296 mg g^?1. The antioxidant activity of extracts was tested by measuring their ability to scavenge reactive hydroxyl radical during the Fenton reaction, using electron spin resonance (ESR) spectroscopy. Moreover, the influence of these extracts on lipid peroxyl radicals obtained during lipid peroxidation of: (1) sunflower oil (37 °C, 3 h) induced by 4,4′‐azobis(4‐cyanovaleric acid) (ACVA) and (2) liposomes induced by 2,2′‐azobis(2‐amidino‐propane)dihydrochloride (AAPH) was studied. n‐Butanol extract, because of the highest content of total phenolic compounds (296 mg g^?1) had the best antioxidant activity (100% at 0.16 mg mL^?1 in Fenton reaction system; 90.57% at 5 mg mL^?1 in system I; 100% at 5 mg mL^?1 in system II).     

Characterisation of the phenolic and flavonoid fractions and antioxidant power of Italian honeys of different botanical origin

BACKGROUND: Twenty‐seven Italian honey samples of different floral origin were analysed for total phenolic and flavonoid contents by a spectrophotometric method and for antioxidant power and radical‐scavenging activity by the ferric‐reducing/antioxidant power (FRAP) and 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) assays respectively. In addition, the phenolic and flavonoid profiles were analysed using high‐performance liquid chromatography with UV detection (HPLC‐UV). RESULTS: The results of this study showed that honey contains copious amounts of phenolics and flavonoids. HPLC‐UV analysis showed a similar qualitative polyphenolic profile for all honey samples analysed. The main difference among samples was in the contribution of individual analytes, which was affected by floral origin. Total phenolic and flavonoid contents varied from 60.50 to 276.04 mg gallic acid equivalent kg^?1 and from 41.88 to 211.68 mg quercetin equivalent kg^?1 respectively. The antioxidant capacity was high and differed widely among samples. The FRAP value varied from 1.265 to 4.396 mmol Fe²⁺ kg^?1, while the radical‐scavenging activity expressed as DPPH‐IC₅₀ varied from 7.08 to 64.09 mg mL^?1. Correlations between the parameters analysed were found to be statistically significant (P < 0.05). CONCLUSION: The present study shows that honey contains high levels of phenolics and flavonoids and that the distribution of these compounds is influenced by the honey's floral origin. Copyright © 2009 Society of Chemical Industry     

Ferrous sulfate liposomes: preparation,stability and application in fluid milk

The effects of cholesterol and Tween 80 on the physical stability of empty liposomes were investigated. Results showed that the physical stability of liposomes, including electrostatic and steric stability, was improved by addition of cholesterol and Tween 80. Liposomes prepared by different methods, thin-film hydration, thin-film sonication, reverse-phase evaporation and freeze-thawing, were tested for their capacity to encapsulate ferrous sulfate. Technology parameters to microencapsulate ferrous sulfate, concentration of ferrous sulfate, hydrating media and sonication strength, were optimized. The concentration of ferrous sulfate and the hydrating medium had a significant effect on the amount of encapsulated ferrous ions. The encapsulation efficiency (EE) of 67% was obtained by using the reverse-phase evaporation method. In milk fortified with ferrous sulfate liposomes, the concentration of iron increased to 15 mg L⁻¹, and it was stable to heat sterilization (100 °C, 30 min) and storage at 4 °C during one week.     

Improved analysis of sinigrin in Ethiopian mustard

An ion‐pair reversed‐phase high‐performance liquid chromatography (HPLC) method was developed for the analysis of sinigrin in Ethiopian mustard (Brassica carinata A. Braun) seed and seed fractions. Separation was compared on several RP‐HPLC columns (Inertsil^® ODS‐4 C₁₈ and ZORBAX^® Eclipse XDB‐C₁₈) with an isocratic eluent containing 100% aqueous (aq.) tetramethylammonium bromide (10 mm , pH 5.0). Sinigrin retention was affected by HPLC variables including the type of ion‐pair reagent, buffer strength and pH, acetonitrile concentration, column temperature and eluent flow rate. Partial validation demonstrated this optimised chromatographic condition to be linear, accurate and precise. Multistage extraction using 70% (v/v) aq. methanol was more efficient than 50% (v/v) aq. acetonitrile. In addition, the matrix effect and recovery rate as well as processing efficiency of the analytical protocol were determined. This method is suitable for high throughput analysis of sinigrin in Ethiopian mustard seed and seed fractions.     

Determination of seven preservatives in cosmetic products by micellar electrokinetic chromatography

A micellar electrokinetic chromatography method using cetyltrimethylammonium bromide (CTAB) as a cationic surfactant, coupled with UV–Vis detection, was developed for the simultaneous determination of seven preservatives, including methyl‐, ethyl‐, propyl‐ and butyl‐paraben and phenol, phenoxyethanol and resorcinol. The method involved optimizing the pH of the phosphate buffer and concentrations of CTAB, ethanol and 2‐hydroxypropyl‐β‐cyclodextrin (HP‐β‐CD). The preservatives were well separated using optimum conditions and separated within 10 min at a separation voltage of ?12.5 kV with the 1.0 mM phosphate buffer (pH 7.0) containing 90 mM CTAB, 25 mM HP‐β‐CD and 10% (v/v) ethanol. Satisfactory recoveries (84.1–103.0%), migration time (RSD < 3.1%) and peak area (RSD < 4.5%) repeatabilities were achieved. Detection limits of the preservatives were between 0.31 and 1.52 μg mL^?1 (S/N = 3, n = 5). The optimized method was successfully applied to the simultaneous determination of these preservatives in 10 commercial cosmetic products.     

Solvent effects on antioxidant properties of persimmon (Diospyros kaki L. cv. Daebong) seeds

The aim of this study was to investigate the antioxidant activities of persimmon seed extracts (PSE) using different solvents such as ethanol, methanol, acetone, and their aqueous 80% solvents. The EC₅₀ values of the extracts from absolute ethanol (EE) and methanol (ME) in 2, 2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical–scavenging assay were 49.71 and 51.15 μg mL^?1, respectively, while the EC₅₀ of butylated hydroxyanisole (BHA) was 70.82 μg mL^?1. However, the EC₅₀ value of reducing power for the absolute acetone extract (AE) was higher (210.06 μg mL^?1) than that of BHA (212.67 μg mL^?1). Although the absolute ME had the highest antioxidant activity, it exhibited the lowest total phenolics and flavonoids. In contrast, the antioxidant activities of the aqueous solvent extracts showed a good correlation with total phenolics and flavonoids when compared to the absolute solvent extracts. The results showed that PSE could potentially be used as an inexpensive source of natural antioxidant in food and pharmaceutical industries.     

Influence of extraction methods on functional properties of protein concentrates prepared from South African bambara groundnut landraces

Functional properties of protein concentrates prepared from three bambara groundnut landraces using acid precipitation and salt solubilisation methods were evaluated. The protein content of bambara grains (26–27%) was similar for the three landraces. The acid precipitation gave a much higher yield of protein concentrates (52%), which were also high in protein (79%) compared to the salt solubilisation method (yield: 25%, protein content: 57%). Functional properties of proteins were more influenced by the methods of preparation rather than the landraces. Protein concentrate prepared by salt solubilisation method showed higher emulsifying (63–66%), foaming (53–57%), water (1.4–2.0 mg mL^?1) and oil absorption properties (2.2–2.6 mg mL^?1) than the acid‐precipitated concentrates (53–57%, 63–66%, 2.0–2.7 mg mL^?1, 1.4–1.7 mg mL^?1). The foaming capacity and stability of all the protein concentrates decreased with increasing pH from 3 to 8. Salt solubilisation may be the most appropriate method for the enhanced functionality and utilisation of bambara groundnuts’ protein concentrates.     

Coenzyme Q10 and Q9 contents in 6 commercial vegetable oils and their average daily intakes in Korea

This study investigated the concentration of coenzyme Q₁₀ (CoQ₁₀) and Q₉ (CoQ₉) in 6 commercial vegetable oils commonly consumed in Korea and estimated the average daily intake of CoQ₉ and CoQ₁₀ from oils selected. The analytical method employed saponification before solvent extraction and quantification using high performance liquid chromatography (HPLC) with a mass spectrometer (LC-MS). Contents of CoQ₉ and CoQ₁₀ in 6 cooking oils varied from 233.7 to 1.4 and from 84.9 to 1.3 μg/g oil, respectively. Maize germ oil was the richest sources of CoQ₉ (233.7±8.2 μg/g), while CoQ₁₀ was found the highest contents in perilla oil (84.9±7.6 μg/g). However, the major oil source of CoQ intake in the Korean population was soybean oil (63.0%). The estimated daily intake of total CoQ (Q₉+Q₁₀) was 2.92 mg/day/person.     

In vitro inhibitory effects of cranberry bean (Phaseolus vulgaris L.) extracts on aldose reductase, α‐glucosidase and α‐amylase

The in vitro inhibitory activities of different seed extracts prepared from cranberry bean mutant SA‐05 and its wild‐type variety Hwachia against aldose reductase, α‐glucosidase and α‐amylase were examined. The results indicated that the polyphenolics‐rich extracts obtained using 800 g kg^?1 methanol and 500 g kg^?1 ethanol demonstrated inhibitory activities against aldose reductase (IC₅₀ of 0.36–0.46 mg mL^?1) and α‐glucosidase (IC₅₀ of 1.32–1.94 mg mL^?1). The 500 g kg^?1 ethanol extracts also showed α‐amylase inhibitory activities (IC₅₀ of 70.11–80.22 μg mL^?1). Subsequent extracts, prepared further with NaCl and H₂O from precipitates of 800 g kg^?1 methanol or 500 g kg^?1 ethanol extracts, exhibited potent α‐amylase inhibitory activities (IC₅₀ of 17.68–38.68 μg mL^?1). A combination of 500 g kg^?1 ethanol extraction plus a subsequent H₂O extraction produced highest polyphenolics and α‐amylase inhibitors. The SA‐05 α‐amylase inhibitor extracts showed greater inhibitory activities than that of Hwachia. Thus, cranberry bean mutant SA‐05 is an advantageous choice for producing anti‐hyperglycaemic compounds.     

Determination of melamine in milk powder,milk and fish feed by capillary electrophoresis: a good alternative to HPLC

BACKGROUND: Since September 2008, an increased incidence of kidney stones and renal failure in infants, associated with the ingestion of infant formula contaminated with melamine has been reported in China. Furthermore, melamine was not only found in many protein‐based food commodities, but also in the feeds for cattle and poultry. So it is necessary to develop a suitable method to determine melamine. RESULTS: A capillary zone electrophoresis (CZE) method for analysis of melamine was developed by use of running electrolyte containing 35 mmol L^?1 sodium dihydrogen phosphate at pH 3.5, with UV detection at 210 nm. Regression equation revealed linear relationships (r = 0.9999) between the peak‐area and the content of melamine from 0.8 to 80 µg mL^?1. The detection limit was 0.08 µg mL^?1. The method was successfully applied to the determination of melamine in milk powder, milk and fish feed, with the recoveries from 94.5% to 103.7%. CONCLUSION: The performance of the CZE method evaluated in terms of precision, limits of detection, accuracy and quantification were comparable and in good agreement with those obtained by the HPLC method, with the advantage of shorter analysis time and lower cost. Copyright © 2010 Society of Chemical Industry     

Microbial inhibitory and radical scavenging activities of cold‐pressed terpeneless Valencia orange (Citrus sinensis) oil in different dispersing agents

BACKGROUND: Due to their low solubility in water, oil‐based bioactive compounds require dispersion in a surface‐active agent or appropriate solvents to ensure maximum contact with microorganisms. These combinations, however, may change their physical and/or chemical characteristics and consequently alter the desired functionality. The objective of this study was to determine the impact of selected dispersing agents, ethanol, dimethyl sulfoxide (DMSO), and Tween‐80, on cold‐pressed terpeneless (CPT) Valencia orange oil to function as a free radical scavenger and an antimicrobial food additive. RESULTS: When dissolved in ethanol or DMSO, the orange oil fraction had similar minimum inhibitory concentrations (MIC) for Listeria monocytogenes ATCC 19 115 (0.3% and 0.25% v/v respectively), which were significantly lower (P ≤ 0.5) than the MIC for Salmonella typhimurium ATCC 14 028 (1% v/v). Both ethanol and DMSO oil dispersion systems exhibited an intermediate MIC (0.75% v/v) for Lactobacillus plantarum WCFS1. The orange oil (up to 3%) in an aqueous solution of 0.1% Tween‐80 yielded no inhibitory activities against any of the test bacteria. However, the 1% natural orange oil dispersed in Tween‐80 exhibited 56.86% 2,2‐diphenyl‐1‐picryl hydrazyl (DPPH) radical inhibition versus 18.37% and 16.60% when the same level of orange oil was dissolved in DMSO or ethanol, respectively. At the same orange oil concentration, the oil/Tween‐80 suspension yielded 57.92% neutralization of hydroxyl radicals. This represents 71.37% of the mannitol antioxidant activity, which was used as a positive control. CONCLUSIONS: These findings suggest that Tween‐80 is an appropriate dispersing agent only if the antioxidant functionality is desired. If both antimicrobial and antioxidant properties are needed, the CPT Valencia orange oil should be dispersed in either DMSO or ethanol. Copyright © 2010 Society of Chemical Industry     

Profile of antioxidant and antibacterial activities of different solvent extracts from Rabdosia rubescens

The objective of this work was to investigate the antioxidant and antibacterial activities of different extracts from Rabdosia rubescens and to further evaluate the antibacterial mechanism of extracts. The results showed that 80% acetone extracts had the highest contents of total polyphenols (8.09 mg GAE g^?1) and flavonoids (5.69 mg RE g^?1) and exhibited the strongest antioxidant activities, followed by 80% methanol and 80% ethanol, and the lowest for hexane extracts. Others except for hexane extracts showed different antibacterial activities against Gram‐positive strains, while no inhibitory effects were found on tested Gram‐negative bacterial strains. Among these extracts, 80% acetone and ethanol extracts had relatively higher antibacterial activities with the lowest minimum inhibitory concentration and minimum bactericidal concentration values of 5 and 10 mg mL^?1. The antibacterial mechanism of ethanol extracts against Staphylococcus aureus might be described as it disrupts cell wall, increases cell membrane permeability and then leads to the leakage of cell constituents.     

Preparative separation and purification of phenolic compounds from Canarium album L. by macroporous resins

BACKGROUND: Canarium album L. (also called Chinese olive) is a traditional medicine material in China, and phenolic compounds from C. album possess great pharmacological activities. To obtain high‐purity phenolics from C. album, a crude extract of C. album phenolics was prepared by ethanol extraction. The use of macroporous resins for further separation and purification of phenolics in the extract was studied. RESULTS: Through static adsorption and desorption tests, AB‐8 resin was chosen for the separation of phenolics because of its higher adsorption capacity and desorption ratio than other resins. Then, dynamic adsorption and desorption experiments were carried out on an AB‐8 resin packed column to obtain optimal separation parameters. The highest adsorption capacity of AB‐8 was achieved when variables including initial concentration (C₀), feed flow rate and feed volume were 10 mg mL^?1, 2 mL min^?1 and 9 bed volumes (BV), respectively, and saturated resin was first washed with 5 BV of water to remove impurities, then a purified product containing more than 85% of C. album phenolics was obtained by desorbing the resin with 2.5 BV of 70% (v/v) aqueous ethanol at flow rate of 1 mL min^?1, and the recovery of phenolics was up to 75%. In addition, five phenolic compounds in the product were identified as gallic acid, ellagic acid, corilagin, hyperin and kaempferol‐3‐glucopyranoside by UV and LC–ESI–MS analysis. CONCLUSION: The results in this study could provide scientific references for the large‐scale production of phenolics from C. album. Copyright © 2007 Society of Chemical Industry     

A Miniaturized Preconcentration Method Based on Dispersive Liquid–Liquid Microextraction for the Spectrophotometric Determination of Aziridine in Food Simulants

In this work a simple, rapid and sensitive method using dispersive liquid–liquid microextraction (DLLME) combined with UV–Vis spectrophotometry has been developed for the preconcentration and determination of trace amounts of aziridine in food simulants. The method is based on derivatization of aziridine with Folin's reagent (1,2-naphthoquione-4-sulphonic acid) and extraction of color product using DLLME technique. Some important parameters, such as reaction conditions, and type and volume of extraction solvent and disperser solvent were studied and optimized. Under optimum conditions, a linear calibration curve in the range of 2.0–350 ng mL^?1 of aziridine was obtained. Detection limit based on 3S_b was 1.0 ng mL^?1, and the relative standard deviation for 50 ng mL^?1 of aziridine was 2.49c (n?=?7). The proposed method was applied for the determination of aziridine in food simulants.     

Optimisation of hydrolysis of purple sea urchin (Strongylocentrotus nudus) gonad by response surface methodology and evaluation of in vitro antioxidant activity of the hydrolysate

BACKGROUND: Hydrolysates prepared from sea urchin (Strongylocentrotus nudus) gonad by enzymatic treatment showed strong 1,1‐diphenyl‐2‐picrylhydrazyl radical scavenging activity and reducing power. RESULTS: Hydrolysis of S. nudus gonad by the commercial protease papain was optimised for maximum degree of hydrolysis (DH) and trichloroacetic acid‐soluble peptide index (TCA‐SPI) using response surface methodology. Results showed that the optimal conditions were the following: temperature of 48.83 °C, pH of 6.92, enzyme‐to‐substrate ratio of 3143 U g^?1, and substrate concentration of 83.5 g L^?1. Under these conditions, a DH of 27.96 ± 0.54% and a TCA‐SPI of 57.32 ± 0.63% were obtained. The hydrolysate prepared in the optimal conditions was fractionated by an ultra‐filtration system and the resultant fraction below 10 kDa was found to effectively scavenge hydroxyl radical (EC₅₀ = 13.29 ± 0.33 mg mL^?1) and hydrogen peroxide (EC₅₀ = 16.40 ± 0.37 mg mL^?1), inhibit lipid peroxidation (EC₅₀ = 11.05 ± 0.62 mg mL^?1), chelate Fe²⁺ (EC₅₀ = 7.26 ± 0.44 mg mL^?1), and protect mice macrophages against death induced by tert‐butyl hydroperoxide. CONCLUSION: Hydrolysates prepared from S. nudus gonad have the potential to be applied as natural antioxidant agents. Copyright © 2012 Society of Chemical Industry     

Determination of the veterinary drug maduramicin in food by fluorescence polarisation immunoassay

A fluorescence polarisation immunoassay (FPIA) using a specific polyclonal antiserum for the detection of maduramicin (MD) was developed and optimised. The polyclonal antiserum was produced against MD linked to bovine serum albumin. Fluorescein‐labelled MD (tracer) was synthesised by N‐hydroxysuccinimide active ester method and purified using thin layer chromatography. The developed FPIA for MD had a dynamic range from 0.01 to 5.6 μg mL^?1 with an IC₅₀ value of 0.16 μg mL^?1 and a limit of detection of 0.002 μg mL^?1. Recoveries from chicken muscle, fat and egg samples spiked at 0.25, 5 and 10 μg g^?1 levels were 82–130%. The FPIA results from analysis of incurred residues in chicken muscle samples showed that the simple procedure is viable.     

Isolation of peptides from a novel brewers spent grain protein isolate with potential to modulate glycaemic response

The in vitro dipeptidyl peptidase‐IV (DPP‐IV) inhibitory activity of a Brewers’ spent grain protein‐enriched isolate (BSG‐PI) Alcalase? hydrolysate (AlcH), which had previously been identified as a relatively potent angiotensin‐converting enzyme (ACE) inhibitor, was determined. The half maximal DPP‐IV inhibitory concentration (IC₅₀) value of AlcH following 240‐min digestion was 3.57 ± 0.19 mg mL^?1. Ultrafiltration fractionation did not significantly increase the DPP‐IV inhibitory activity of the AlcH fractions. Subjection of AlcH to simulated gastrointestinal digestion (SGID), which yielded SAlcH, resulted in a significant increase in DPP‐IV inhibitory activity (P < 0.05), particularly after the intestinal phase of digestion. Following semi‐preparative reverse phase high performance liquid chromatography (RP‐HPLC) fractionation of SAlcH, fraction 28 was identified as having highest mean DPP‐IV inhibitory activity. Two novel DPP‐IV inhibitory peptides, ILDL and ILLPGAQDGL, with IC₅₀ values of 1121.1 and 145.5 μm , respectively, were identified within fraction 28 of SAlcH following ultra‐performance liquid chromatography (UPLC)‐tandem mass spectrometry (MS/MS). BSG protein‐derived peptides were confirmed as having dual ACE and DPP‐IV inhibitory activities.