含功能性油的水飞蓟宾超饱和自纳米乳的制备与体外评价

目的制备含功能性油的水飞蓟宾超饱和自纳米乳(SLB-S-SNEDDS),并对其进行表征及体外评价研究,以提高难溶性药物水飞蓟宾的生物利用度。方法铁氢化钾还原力与1,1-二苯基-2-苦肼基(DPPH)自由基清除实验筛选功能性油脂;伪三元相图考察乳化剂乳化能力;测定粒径、多分散指数(PDI)、Zeta电位等考察混合油相比例与载药量;相容性与溶出度实验筛选促过饱和物质并考察其质量浓度;从外观、粒径分布、自乳化效率、形态学等方面表征SLB-S-SNEDDS,并进行溶出度、抗氧化能力、细胞毒性等体外评价。结果所得SLB-S-SNEDDS处方为(1)小麦胚芽油/Capryol 90-Cremophor ELP-Transcutol HP与(2)沙棘籽油/Capryol 90-Cremophor ELP-Transcutol HP,1 g基质(包含0.043 g小麦胚芽油或沙棘籽油、0.387 g Capryol 90、0.380 g Cremophor ELP、0.190 g Transcutol HP),水飞蓟宾的添加量为各组分平衡溶解度之和的20%,Soluplus的添加量为上述总质量的0.1%。小麦胚芽油、沙棘籽油体系分别为淡黄色、亮黄色透明状均一液体,2种体系自乳化分散后均呈近球形白色扁平乳滴,粒径约为50 nm,乳化时间均为65 s。与药物原料及SLB-SNEDDS相比,SLB-S-SNEDDS中水飞蓟宾的累积溶出率8h内均维持在85%～110%,表明该体系能够显著提高药物的溶出度。SLB-S-SNEDDS与铁氰化钾反应后的吸光度(A值0.452～0.782,0.488～0.765)以及DPPH自由基清除率(39.09%～96.02%,30.54%～89.20%)均高于相应质量浓度下水飞蓟宾原料的A值与清除率(0.411～0.760,22.89%～63.21%),表明2种处方体系均能提高水飞蓟宾的抗氧化能力。细胞毒性实验结果显示,在5、10μmol/L药物浓度下,水飞蓟宾原料组、水飞蓟宾S-SNEDDS组及其相应的空白S-SNEDDS组细胞生存率均90%,说明SLB-S-SNEDDS及其所用辅料对人克隆结肠腺癌细胞(Caco-2)毒性较小、安全性较好。结论制备的含功能性油SLB-S-SNEDDS在提高水飞蓟宾累积溶出率的同时,增强了其抗氧化能力,为将超饱和自纳米乳(S-SNEDDS)用于改善难溶性药物水溶性及其生物活性提供有益参考。     

The effect of complexation with cyclodextrins on the antioxidant and antimicrobial activity of ellagic acid

Purpose: The aim of the paper was to develop the simple procedures for preparation of inclusion complexes of ellagic acid (EA) with cyclodextrins (CDs) and to investigate their antioxidant and antimicrobial activity.

Methods: The structural characterization was carried out using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and nuclear magnetic resonance (NMR) methods. The phase solubility technique was used to investigate the interactions between ‘host’ and ‘guest’ molecules and to estimate the molar ratio between them. The antioxidant and antimicrobial activity of EA and inclusion complexes were determined.

Results: The apparent stability constants were found to be 117?dm³ mol^?1 for the complex with β-CD and 161?dm³ mol^?1 for the complex with (2-hydroxypropyl)-β-cyclodextrin (HP-β-CD). The results of phase-solubility studies showed that EA formed the inclusion complexes with CDs in the molar ratio of 1:1. The calculated half-maximal inhibitory concentration was 41.18?μg cm^?3 for butyl hydroxy toluene, 1.96?μg cm^?3 for EA, 0.88?μg cm^?3 for inclusion complex with HP-β-CD, and 1.27?μg cm^?3 for inclusion complex with β-CD.

Conclusion: The stability constants indicated the rapid release of EA from the inclusion complexes in the aqueous medium at 25?°C. The antioxidant activity of EA was increased, while the antimicrobial activity was preserved after complexation with CDs.     

白芍总苷体外抗氧化活性研究

目的研究白芍总苷体外对不同化学体系和生物体系的抗氧化活性。方法通过Fenton反应生成·OH自由基,采用DPPH·自由基,观察不同质量浓度的白芍总苷溶液对化学体系自由基的清除作用。利用邻二氮菲-Fe2+-H2O2产生的·OH自由基造成红细胞膜破裂,以及肝、脑细胞脂质的过氧化,观察不同质量浓度的白芍总苷溶液对生物体系自由基的清除作用。结果白芍总苷对·OH有一定的清除作用,其IC50为0.62 g/L。白芍总苷对DPPH·自由基有较强的清除作用,IC50为6.6 mg/L;对·OH自由基引发的红细胞膜破裂有一定的抑制作用,其IC50为30.17 mg/L,对肝、脑匀浆脂质过氧化有一定的抑制作用,其IC50分别为6.9、16.3 mg/L。结论白芍总苷具有清除自由基作用,在不同体外实验体系中抗氧化量效关系存在差异。     

Extract from Ceratonia siliqua Exhibits Depigmentation Properties

Skin hyper‐pigmentation is a condition initiated by the overproduction of melanin existing in the melanocytes. Melanin pigment is responsible for the colour of skin in humans. It is formed through a series of oxidative reactions involving the amino acid tyrosine in the presence of the key enzyme tyrosinase. In continuation with our efforts to identify tyrosinase inhibitors from plants sources, the methanol extract from leaf, bark and fruit of Ceratonia siliqua were screened for tyrosinase inhibition and diphenolase activity. The bark extract exhibited significant inhibition on mushroom tyrosinase using L‐tyrosine as a substrate and showed diphenolase activity. The extract further significantly lowered tyrosinase mRNA levels in B16‐F10 mouse melanocytes. Bioassay‐guided fractionation led to the isolation of six compounds. Compounds (?)‐epicatechin‐3‐O‐gallate, 1,2,3,6‐tetra‐O‐galloyl‐ß‐D‐glucose and gallocatechin‐3‐O‐gallate showed tyrosinase inhibitions with the IC₅₀ values of 27.52, 83.30 and 28.30 µg/mL, respectively. These compounds also exhibited L‐DOPA activities with IC₅₀ values of >200, 150 and 200 µg/mL, respectively. A clinical study was conducted using 20 volunteers in a patch testing trial for irritancy potential and skin depigmentation. The clinical results showed the sample to be non‐irritant with irritancy potential of ?34.21 and depigmentation trial showed an improvement in the even skin tone of UV induced pigmentation at 3% after 28 days of application. Copyright © 2015 John Wiley & Sons, Ltd.     

Polyphenols,methylxanthines, and antioxidant capacity of chocolates produced in Serbia

Different kinds of chocolates produced in Serbia were analyzed regarding total polyphenol, flavonoid and proanthocyanidin content using spectrophotometric methods. Flavan-3ols and methylxanthines in all samples were determined with RP-HPLC. DPPH, FRAP, ABTS and ORAC assays were applied for measuring antioxidant capacity. The average of all four antioxidant tests for each cocoa product was used for calculating antioxidant potency composite index (ACI). Obtained results for all four assays have shown that antioxidant capacity of analyzed chocolate/cocoa extracts followed cocoa, polyphenol, flavonoid, and proanthocyanidin contents. Although the addition of raspberries to dark chocolates had no significant influence on their total polyphenol, flavonoid and proanthocyanidin contents, statistical analysis showed that there was significant increase in the antioxidant capacity of dark chocolates with raspberry compared to plain dark chocolates (p = 0.007). Overall range for theobromine content varied from 5.5 to 22.3 mg/g depending on the product type, while the content of caffeine was 13–30 times lower in all analyzed cocoa products. In addition, correlation between antioxidant potency composite index and declared percentage of cocoa was high (R² = 0.798, p < 0.05) and indicated that declared cocoa content was a reliable indication for antioxidant capacity of chocolates produced in Serbia.     

Effects of cooking on in vitro sinigrin bioaccessibility,total phenols,antioxidant and antimutagenic activity of cauliflower (Brassica oleraceae L. var. Botrytis)

Cauliflower (Brassica oleraceae L. var. Botrytis) is a good source of bioactive compounds, such as glucosinolates, phenolic compounds and vitamins. In this study, the effects of some processes (i.e. boiling, steaming) on the sinigrin bioaccessibility as a major glucosinolate found in cruciferous vegetables after in vitro digestion, also in vitro antimutagenic activities, total phenols and total antioxidant capacities of cauliflower were determined. The sinigrin content was reduced by approximately 9.6% and 29.1% in steamed and boiled cauliflower (p > 0.05), respectively. After in vitro simulated digestion, sinigrin content was decreased by 26.4% in raw samples, increased by 29.5% and 114.7% in steamed and boiled samples, respectively. In all samples, mutagenic effect to Salmonella typhimurium TA 100 was not seen. When samples were steamed, phenol content was increased by 14.83%. After boiling total phenol content of cauliflower was decreased by 1.8%. Total antioxidant capacities (TAC) measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3 ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods were increased by 47% and 39%, respectively (p < 0.05) in steamed samples and decreased by 8% and 7% with boiling, respectively (p > 0.05). TAC in raw sample of cauliflower, which was investigated in phosphomolybdenum assays, was determined as 18.7 mg ascorbic acid equivalents (AAE)/100 g. In all cases, the highest antioxidant activity was determined in the steamed samples, while the lowest antioxidant activity was in boiled samples.     

Evaluation of yield,quality and antioxidant activity of essential oil of in vitro propagated Kaempferia galanga Linn.

ObjectiveTo determine chemical constituents and antioxidant properties of essential oil from rhizome of the medicinal plant, Kaempferia galanga (K. galanga) Linn. (Zingiberaceae) in conventionally propagated (CP) and in vitro propagated (IVP) plants.MethodsIn vitro (micro) propagation of K. galanga was done by inoculating explants on to Murashige and Skoog agar medium, supplemented with suitable combinations of phytohormones; the regenerants were transferred to soil for further growth. Essential oil preparations of both CP and IVP rhizomes grown in soil, obtained by the hydro-distillation method were analyzed by gas chromatography-mass spectrometry. Antioxidant activities of essential oil samples were monitored.ResultsMaximum numbers of regenerated shoots were found in the medium supplemented with 1 mg/L benzyl adenine and 0.5 mg/L indole-3-acetic acid. A total of 6 compounds were identified from rhizomes from CP and IVP plants that yielded 96.9% and 97.81% of the total oil contents, respectively. The major compound of rhizome oil identified from CP and IVP rhizomes was ethyl p-methoxy cinnamate in quantities, 82.01% and 71.77%, respectively, without any compositional variation. Antioxidant properties of essential oil preparations were assessed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydrogen peroxide radical scavenging assays. Moreover, antioxidant activities of rhizome-oil from IVP plants were better than that of CP oil samples.ConclusionsAs IVP rhizomes had better oil yield, those could be used for a large scale commercial propagation for sustainable use of essential oil. The principal chemical in the essential oil, ethyl p-methoxy cinnamate could help apothecary, for several ailments.     

Investigation of Phytochemical and Antioxidant Properties of Methanol Extract and Fractions of Ballota limbata (Lamiaceae)

Ballota limbata (Lamiaceae) has been used for its antispasmodic, antiulcer, diuretic, vermifuge and sedative effects in folk medicine. However, little is known about how does it work to produce these therapeutic actions. Present research investigated phytochemical components and antioxidant properties of methanol extract and different fractions of Ballota limbata. In this study, phytochemical investigation was done by performing different chemical tests. Here, antioxidant property of the extract and fractions was investigated by using 1,1-diphenyl-2-picryl hydrazyl radical scavenging activity, total antioxidant activity by the phosphomolybdenum method, linoleic acid peroxidation, ferric thiocyanate analysis and ferric-reducing antioxidant power. Methanol extract and fractions showed presence of numerous chemical principles including alkaloids, cardiac glycosides, tannins and flavonoids. The ethyl acetate fraction exhibited higher scavenging activity compared to the other fractions under investigation. This fraction displayed 84.16±1.02% 1,1-diphenyl-2-picryl hydrazyl radical inhibition at a dose of 60 μg/ml. IC₅₀ for 1,1-diphenyl-2-picryl hydrazylradical-scavenging activity was 13.53±0.22 μg/ml, relative to the standard, butylatedhydroxytoluene, having IC₅₀ of 12.33±0.88 μg/ml. Thus, Ballota limbata showed significant antioxidant activity, which may contribute in the mechanism of above pharmacological actions.     

Antioxidant effect of 4-nerolidylcatechol and α-tocopherol in erythrocyte ghost membranes and phospholipid bilayers

4-Nerolidylcatechol (4-NC) is found in Pothomorphe umbellata root extracts and is reported to have a topical protective effect against UVB radiation-induced skin damage, toxicity in melanoma cell lines, and antimalarial activity. We report a comparative study of the antioxidant activity of 4-NC and α-tocopherol against lipid peroxidation initiated by two free radical-generating systems: 2,2′-azobis(2-aminopropane) hydrochloride (AAPH) and FeSO₄/H₂O₂, in red blood cell ghost membranes and in egg phosphatidylcholine (PC) vesicles. Lipid peroxidation was monitored by membrane fluidity changes assessed by electron paramagnetic resonance spectroscopy of a spin-labeled lipid and by the formation of thiobarbituric acid-reactive substances. When lipoperoxidation was initiated by the hydroxyl radical in erythrocyte ghost membranes, both 4-NC and α-tocopherol acted in a very efficient manner. However, lower activities were observed when lipoperoxidation was initiated by the peroxyl radical; and, in this case, the protective effect of α-tocopherol was lower than that of 4-NC. In egg PC vesicles, malondialdehyde formation indicated that 4-NC was effective against lipoperoxidation initiated by both AAPH and FeSO₄/H₂O₂, whereas α-tocopherol was less efficient in protecting against lipoperoxidation by AAPH, and behaved as a pro-oxidant for FeSO₄/H₂O₂. The DPPH (2,2-diphenyl-1-picrylhydrazyl) free-radical assay indicated that two free radicals were scavenged per 4-NC molecule, and one free radical was scavenged per α-tocopherol molecule. These data provide new insights into the antioxidant capacity of 4-NC, which may have therapeutic applications for formulations designed to protect the skin from sunlight irradiation.     

苦菊乙酸乙酯提取物的体外抗氧化活性研究

目的:研究苦菊乙酸乙酯提取物(CEE)的体外抗氧化活性。方法:苦菊经95%乙醇加热回流、石油醚脱脂和乙酸乙酯萃取,挥干乙酸乙酯后得到CEE。通过二苯代苦味酰自由基(DPPH.)清除能力、还原能力和2,2′-偶氮-双(-2-脒基丙烷)氯化二氢(AAPH)诱导的红细胞溶血模型,检测CEE的抗氧化活性。结果:在DPPH.清除能力试验中,CEE和阳性对照药的清除率为50%时所需待测液的浓度(EC50)分别为(59.76±6.11)mg·L-1和(3.89±0.23)mg·L-1,两者相差1.54个数量级。CEE和阳性对照药(抗坏血酸)皆能使还原能力测试体系的吸光度增加,并呈浓度-效应关系(P<0.01),但CEE的还原能力不如抗坏血酸(P<0.01)。在AAPH诱导的红细胞溶血模型中,CEE能有效降低兔血红细胞的溶血率(P<0.01),且高剂量(20 mg·L-1)CEE可使溶血率恢复到空白对照的水平。结论:CEE能有效清除自由基,具有一定的还原能力,并对AAPH诱导的兔红细胞溶血具有很好的保护作用。提示CEE具有一定的抗氧化活性,可对其抗氧化活性及其有效成分作进一步的研究。