Search

Roast duck, a popular meat commodity in Asian countries especially Taiwan and China, can produce many types of toxic compounds such as cholesterol oxidation products (COPs) and heterocyclic amines (HAs). The objectives of this study were to evaluate analysis of COPs and HAs in duck by GC-MS and UPLC-MS/MS respectively, and study their formation as affected by roasting methods including traditional oven, fan oven and superheated steam oven. Results showed that both high precision and accuracy was attained by using the QuEChERS method with reduced separation time of 14 min for 7 COPs and 4.5 min for 21 HAs by GC-MS and UPLC-MS/MS, respectively. The duck meat roasted in both superheated steam oven and traditional oven generated significantly higher levels of total COPs than fan oven, while for total HAs, both traditional oven and fan oven produced significantly higher levels than superheated steam oven. Compared to roast duck skin, a much higher level of total COPs was shown in roast duck meat, while a reversed tendency was shown for total HAs. Furthermore, the correlation between formation of COPs and HAs in roast duck skin showed no significant correlation (p0.05) for traditional oven, fan oven and superheated steam oven, while in roast duck meat, only traditional oven showed significant correlation (p0.05). However, by taking both duck skin and meat into account, superheated steam oven produced the lowest level of total HAs, while fan oven generated the lowest level of total COPs. Thus, the developed method can form a basis for determination of COPs and HAs in duck as well as by choosing an appropriate roasting method their formation can be minimized.

Resveratrol, a phenolic compound possessing vital biological activities such as anti-cancer, is present abundantly in grape skin, a waste produced during the processing of grape juice. The objectives of this study were to prepare resveratrol-gold nanoparticles and a resveratrol nanoemulsion from grape skin and study their inhibition effects on pancreatic cancer cells BxPC-3. The spherical-shaped citrate gold nanoparticles (GNPs) and resveratrol-gold nanoparticles (R-GNPs) were, respectively, prepared with a surface plasmon resonance peak at 528 and 538 nm, mean particle size of 20.8 and 11.9 nm, and zeta-potential at −32.7 and −66.7 mV, by controlling an appropriate concentration of citrate/resveratrol and gold chloride as well as stirring time and temperature. The resveratrol nanoemulsion, composed of soybean oil, Tween 80, and sucrose fatty acid ester in glycerol and water, possessed a high storage stability with a mean particle size of 14.1 nm, zeta-potential of −49.7 mV, and encapsulation efficiency of 95.5%. An antiproliferation study revealed that both R-GNPs and resveratrol nanoemulsion could effectively inhibit the growth of pancreatic cancer cells BxPC-3, with the latter showing a higher inhibition effect. Western blot analysis implied that both can down-regulate expressions of cyclin A, cyclin B, CDK1, and CDK2 and up-regulate expressions of p53 and p21, accompanied by enhancing cytochrome C expression, decreasing BcL-2 expression, increasing Bax expression, and leading to the elevation of caspase-8, caspase-9, and caspase-3 activities for cell apoptosis execution. Future research is needed to study the inhibition of pancreatic tumors in vivo by R-GNPs and resveratrol nanoemulsions.

Organosulfur compounds, phenolic acids and flavonoids in raw and black garlic were determined, and followed by preparation of black garlic nanoemulsion for studying their effects on deoxycorticosterone acetate-salt-induced hypertension and associated mild cognitive impairment in rats. Three organosulfur compounds, including diallyl sulfide (87.8 μg/g), diallyl disulfide (203.9 μg/g) and diallyl trisulfide (282.6 μg/g) were detected in black garlic by GC-MS, while gallic acid (19.19 μg/g), p-coumaric acid (27.03 μg/g) and quercetin (22.77 μg/g) were detected by UPLC-MS/MS. High doses of both black garlic extract and nanoemulsion prepared using Tween-80, glycerol, grapeseed oil and water could decrease systolic blood pressure through the elevation of bradykinin and nitric oxide levels as well as diminish aldosterone and angiotensin II levels in rats. In Morris water maze test, they could significantly decrease escape latency and swimming distance and increase the time spent in the target quadrant, accompanied by a decline of acetylcholinesterase activity and malondialdehyde level in the hippocampus as well as a rise in glutathione level and activities of superoxide dismutase, catalase and glutathione peroxidase. In addition, the levels of tumor necrosis factor, interleukin-6 and interleukin-1β were reduced. Effects of lowering blood pressure and improving learning/memory ability in rats followed the order: lisinopril &gt, black garlic nanoemulsion &gt, black garlic extract.

Anti-cancer activity of catechin nanoemulsions prepared from Oolong tea leaf waste was studied on prostate cancer cells DU-145 and DU-145-induced tumors in mice. Catechin nanoemulsions composed of lecithin, Tween-80 and water in an appropriate proportion was prepared with high stability, particle size of 11.3 nm, zeta potential of −67.2 mV and encapsulation efficiency of 83.4%. Catechin nanoemulsions were more effective than extracts in inhibiting DU-145 cell growth, with the IC50 being 13.52 and 214.6 μg/mL, respectively, after 48 h incubation. Furthermore, both catechin nanoemulsions and extracts could raise caspase-8, caspase-9 and caspase-3 activities for DU-145 cell apoptosis, arresting the cell cycle at S and G2/M phases. Compared to control, catechin nanoemulsion at 20 μg/mL and paclitaxel at 10 μg/mL were the most effective in reducing tumor volume by 41.3% and 52.5% and tumor weight by 77.5% and 90.6% in mice, respectively, through a decrease in EGF and VEGF levels in serum.

This study aims to synthesize a magnetic activated carbon nanocomposite from green tea leaf waste (MNPs-GTAC) for evaluation of adsorption efficiency of 4 priority polycyclic aromatic hydrocarbons (PAHs). MNPs-GTAC contained spherically-shaped MNPs with cubic spinel structure, surface area at 118.8 m2/g, particle size at 8.6 nm and saturation magnetization at 34.2 emu/g. PAH adsorption reached a plateau at an MNPs-GTAC dose of 50 or 60 mg/L, pH of 2–4 and ionic strength of 0.1–10%, with PAH reduction in the presence of humic acid being compensated by addition of 0.1% sodium chloride. Kinetics was rapid attaining 80% removal within 5 min and the pseudo-second-order rate decreased in this order: Benzo[a]anthracene>Chrysene>Benzo[b]fluoranthene>Benzo[a]pyrene. Isotherm modeling revealed a Langmuir type-2 shape with the maximum adsorption capacity being 28.08, 22.75, 19.14 and 15.86 mg/g for Benzo[b]fluoranthene, Benzo[a]pyrene, Chrysene and Benzo[a]anthracene, respectively. Temperature study showed the PAH adsorption to be an endothermic and spontaneous process with increased randomness at solid-solution interface. Acetonitrile could completely recover the adsorbed PAH and MNPs-GTAC was successfully recycled 5 times with a minimum loss. Application to mineral water showed 86–98% and 72–89% removal for PAHs spiked respectively at 0.1 and 1 mg/L, while a complete removal was attained in tap and river waters.

A simple and rapid high performance liquid chromatographic (HPLC) method was developed to determine the major carotenoids in human serum. Lutein, b-carotene and lycopene were separated within 20 min using a C18 column and a mobile phase of methanol/methylene chloride (95:5, v/v) with sample solvent methanol/methylene chloride (45:55, v/v), detection at 476 nm and flow rate at 0.8 mL/min. A high recovery was achieved with this method, which amounted to 92, 90 and 87% for lutein, b-carotene and lycopene, respectively, while the detection limit was 5.0, 10.0 and 4.0 ng/mL. This method was applied to determine the major carotenoids in human serum samples collected from an elderly care center. A total of 62 samples were measured, and in most cases, bcarotene (17.5-1105.7 ng/mL) was present in the highest amount, followed by lutein (10.6-182.3 ng/mL) and lycopene (0-402 ng/mL).

Soy isoflavones are phytoestrogens mainly present in soybean and soybean products. Isoflavones have been reported to possess physiological activities such as inhibition of cancer cell proliferation, reduction of cardiovascular risk, prevention of osteoporosis and alleviation of postmenopausal syndrome. This paper gives an overview of soy isoflavone analysis in foods and biological fluids. Extraction of isoflavones is often carried out by polar solvents such as methanol, ethanol, acetone or acetonitrile, or in combination with acid-containing water. The separation, identification and quantification of isoflavones are usually conducted by gas chromatography coupled with flame ionization detector or mass spectrometer (GC/FID or GC/MS), high-performance liquid chromatography with UV or mass spectrometer (HPLC/UV or HPLC/MS) and immunoassay. GC methods provide high sensitivity and adequate separation of only few soy isoflavones; however, a time-consuming derivatization step is needed before analysis. HPLC with a gradient solvent system with or without acid has been applied to analyze soy isoflavones, but most HPLC methods failed to separate all 12 isoflavones in a single run or the separation time is lengthy. Nevertheless, an appropriate choice of mobile phase and gradient condition in a C18 column with UV detection could provide a simultaneous separation of 12 soy isoflavones within short time. For immunoassay, it is convenient, fast, highly sensitive and can analyze a large number of samples at the same time. Yet, the drawback is that only a few soy isoflavones can be determined.

The objectives of this study were to determine the variety and amount of various functional components in Scutellaria barbata D. Don as well as study their anti-inflammatory activity on RAW 264.7 cells. Both ethanol and ethyl acetate extracts were shown to contain the functional components including phenolics, flavonoids, chlorophylls, and carotenoids, with the former mainly composed of phenolics and flavonoids, and the latter of carotenoids and chlorophylls. Both extracts could significantly inhibit (p

The objectives of this study were to extract curcuminoid from Curcuma longa L. (C. longa), a vital medicinal plant demonstrated to possess many biological activities, and prepare the curcuminoid extract and microemulsion for studying the inhibition mechanism of HT-29 colon cancer cells. Results showed that a total of 3 curcuminoids including curcumin, demethoxycurcumin (DMC) and bisdemethoxycurcumin (BDMC), were separated within 10 min by using an Eclipse XDB-18 column and a gradient mobile phase of 0.1% formic acid solution (A) and acetonitrile (B). The curcuminoid microemulsion composed of soybean oil, Tween 80, ethanol and water was prepared with a high stability and mean particle size of 10.9 nm, zeta-potential of −65.2 mV and encapsulation efficiency of 85.7%. Both curcuminoid extract and microemulsion were effective in inhibiting HT-29 cell growth with the IC50 being 3.83 and 2.51 μg mL−1 after 24 h incubation, respectively, but further reduced to 2.23 and 1.94 μg mL−1, after 48 h incubation. Both treatments could decrease the proportion of both viable and necrosis cells and increase the proportion of both early and late apoptosis cells in a dose-dependent manner, with the cell cycle arrested at the S phase. Also, both treatments could up-regulate p53 expression and down-regulate cyclin A and CDK2 expressions through a p21-independent pathway. In addition, the expressions of Bax and cytochrome C as well as the activities of caspase-8, caspase-9 and caspase-3 increased for the curcuminoid extract, while the curcuminoid microemulsion showed the same trend with the exception that an insignificant change (p > 0.05) in Bax expression was observed. Collectively, this study demonstrated that the curcuminoid microemulsion prepared from C. longa may possess great potential for the treatment of colon cancer in the future.

Pomelo (Citrus grandis), an important fruit crop grown in tropical and subtropical areas, is cultivated mainly in Asian countries. The dominant pigment in pomelo leaves, chlorophyll, has been reported to possess many biological activities such as antioxidant, anti-inflammation and anticancer. The objectives of this study were to determine chlorophylls in Pomelo leaves by high-performance liquid chromatography-mass spectrometry (HPLC-MS) and to encapsulate the isolated chlorophylls from preparative column chromatography into a nanoemulsion system for elucidating the inhibition mechanism on the growth of melanoma cells A375. The results showed that chlorophyll a and chlorophyll b could be separated within 25 min by using a C18 column and a gradient ternary mobile phase of acetone, acetonitrile and methanol. Pomelo leaves mainly contained chlorophyll a (2278.3 μg/g) and chlorophyll b (785.8 μg/g). A highly stable chlorophyll nanoemulsion was prepared with the mean particle size being 13.2 nm as determined by a dynamic light scattering (DLS) method. The encapsulation efficiency of chlorophyll nanoemulsion was 99%, while the zeta potential was −64.4 mV. In addition, the chlorophyll nanoemulsion possessed high thermal stability up to 100 °C and remained stable over a 90-day storage period at 4 °C. Western blot analysis revealed that chlorophyll nanoemulsion and extract could upregulate p53, p21, cyclin B and cyclin A as well as downregulate CDK1 and CDK2 in a concentration-dependent manner for inhibition of melanoma cells A375. Furthermore, chlorophyll nanoemulsion and extract could upregulate Bax and cytochrome C and downregulate Bcl-2, leading to activation of caspase-9, caspase-8 and caspase-3 for the induction of cell apoptosis. Compared to chlorophyll extract, chlorophyll nanoemulsion was more effective in inhibiting the growth of melanoma cells A375.

This study aims to determine toxic compounds polycyclic aromatic hydrocarbons (PAHs) and cholesterol oxidation products (COPs) in thin slices of dried pork as affected by different flavorings and roasting temperature treatments through employing a QuEChERS method coupled with gas chromatograph–tandem mass spectrometer (GC-MS/MS) and gas chromatograph–mass spectrometer (GC-MS), respectively. By employing this method, high accuracy and precision was attained for freeze-dried pork hind leg sample. Following addition of 8 different flavorings with roasting temperature at 120, 160, and 200 °C, the levels of total COPs and PAHs in thin slices of dried pork followed a temperature-dependent increase during roasting, which was further confirmed by principle component analysis. High level of soy sauce or sugar inhibited COP formation, while the low-level minimized PAH formation in thin slices of dried pork during roasting. Sugar was more effective in inhibiting COP formation while soy sauce was more efficient in reducing PAH formation.

Background: Curcuminoid from Curcuma longa Linnaeus has been demonstrated to be effective in anti-cancer and anti-inflammation. The objectives of the present study were to prepare curcuminoid dispersion and nanoemulsion from C. longa and determine their oral bioavailabilities in rats.; Results: After curcuminoid extraction using 99.5% ethanol, bisdemethoxycurcumin (BDMC), demethoxycurcumin (DMC) and curcumin were separated within 10 min by high-performance liquid chromatography using an Eclipse XDB-C18 column (Agilent, Palo Alto, CA, USA) and a gradient mobile phase of 0.1% aqueous formic acid and acetonitrile, with a flow rate of 1 mL min-1 , column temperature of 35 °C and detection wavelength of 425 nm. Curcuminoid nanoemulsion at a particle size of 12.1 nm and encapsulation efficiency 98.8% was prepared using lecithin, Tween 80 and water. A pharmacokinetic study in rats revealed that the parameters including Tmax , Cmax , t1/2 and the area under the curve were higher for curcuminoid nanoemulsions than for curcuminoid dispersion at the same dose employed for gavage administration, whereas, for intravenous injection, an opposite trend was shown. The oral bioavailabilities of BDMC, DMC, curcumin and total curcuminoids in nanoemulsion and dispersion were 34.39 and 4.65%, 39.93 and 5.49%, 47.82 and 9.38%, and 46 and 8.7%, respectively.; Conclusion: The results of the present study demonstrate a higher oral bioavailability after incorporation of curcuminoid into nanoemulsion, facilitating its application as a botanic drug. © 2017 Society of Chemical Industry.; © 2017 Society of Chemical Industry.

Roasting of chicken generates many toxic compounds such as heterocyclic amines (HAs) and cholesterol oxidation products (COPs). The objectives of this study were to evaluate analysis of HAs and COPs in raw, boiled and roasted chicken by QuEChERS coupled with UPLC-MS/MS and GC-MS, respectively, and study their formation as affected by different flavorings (red pepper, black pepper, rosemary and soy sauce) and roasting methods (fan oven and superheated steam oven). Results showed that the average recovery respectively ranged from 54.1 to 109.3% and 65.1 to 116.6% for 20 HAs and from 87.9 to 102.8% and 87.5 to 101.3% for 7 COPs in the skin and meat portions of boiled chicken breast, while the coefficient of variation (CV) of the intra-day and inter-day variability for HAs respectively ranged from 5.27 to 12.09% and 4.23 to 12.68% in chicken skin, as well as 7.34-18.32% and 6.13-15.54% in chicken meat. For COPs, the CV of the intra-day and inter-day variability respectively ranged from 0.82 to 4.14% and 1.80 to 6.30% in chicken skin, as well as 1.40-9.04% and 3.02-9.81% in chicken meat. A total of 8 HAs were formed in roasted chicken, with a higher level of total HAs being shown in the skin portion of roasted chicken flavored with soy sauce and cooked in a fan oven. Also, for both roasting methods, the skin portion contained a higher level of total HAs than the meat portion, but a reversed trend was shown for total COPs with the exception of black pepper flavoring. Furthermore, in roasted chicken with 4 different flavorings, all of which showed no significant correlation (p 0.05) between formation of HAs and COPs in the skin portion, while in the meat portion, only rosemary showed significant correlation (p 0.05). By taking both skin and meat into account, the incorporation of rosemary flavoring into chicken during roasting was the most effective in inhibiting the formation of total HAs and COPs, while soy sauce flavoring was the least effective.

Background: Anthocyanins, a flavonoid class of water-soluble pigments, are reported to possess several biological activities, including antioxidant, anti-inflammatory, and anti-cancer. However, anthocyanins are highly susceptible to degradation in high pH, light, heat, and oxygen during processing and storage. Conventional microencapsulation techniques fail to provide stability to anthocyanins under physiological environments mainly because of their large particle size as well as low zeta potential and encapsulation efficiency. Methods: Nanotechnology provides novel strategies for preparing nanoformulations to enhance the physicochemical stability of anthocyanins. Nanoemulsion and nanoliposome are the two most commonly used nanosystems in pharmaceutical and food-related fields. In this review, an overview of various nanoemulsion and nanoliposome systems reported recently for enhancing stability, bioavailability, and bioactivity of anthocyanins is presented. Results: Anthocyanin nanoemulsions with different oil, water, surfactant, and cosurfactant ratios were prepared from extracts of mangosteen peel, purple sweet potato, cranberry, red cabbage, blueberry, jaboticaba peel, and acai berry and evaluated for their antioxidant activity, enhancement of physicochemical stability, topical skin application, and urinary tract infection. Likewise, unilamellar and multilamellar nanoliposomes were prepared using different types and levels of lecithin without or with cholesterol from anthocyanin standards and extracts of Hibiscus sabdariffa, mulberry, elderberry, black carrot, and pistachio green hull for the evaluation of physicochemical and oxidative stability, in vitro bioaccessibility, and melanogenic activity, as well as protective effects against diabetes mellitus and cataract. Conclusion: This review provides an insight into the current nanotechnology updates on enhancement of anthocyanin stability and biological activity.

Yin-Jieh Tsai, Bing-Huei ChenDepartment of Food Science, Fu Jen Catholic University, New Taipei City, Taiwan, Republic of China Abstract: Green tea is one of the most commonly consumed natural health beverages in Taiwan’s market, with the major functional component catechin being shown to possess several biological activities such as antioxidation, anticancer, and prevention of cardiovascular disease. The objectives of this study were to develop a high-performance liquid chromatography–mass spectrometry method to determine the variety and content of catechins in green tea leaf waste, a by-product obtained during processing of tea beverage. In addition, catechin nanoemulsion was prepared to study its inhibition effect on prostate cancer cell PC-3. Results showed that a total of eight catechin standards were separated within 25minutes by using a Gemini C18 column and a gradient mobile phase of 0.1% formic acid (A) and acetonitrile (B) with flow rate at 1mL/min, column temperature at 30°C, and detection wavelength at 280nm. Among various extraction solvents, 50% ethanol generated the highest yield of total catechins from tea leaf waste, of which five catechins were identified and quantified. The catechin nanoemulsion was composed of catechin extract, lecithin, Tween 80, and deionized water in an appropriate proportion, with the mean particle size being 11.45nm, encapsulation efficiency 88.1%, and zeta potential -66.3mV. A high stability of catechin nanoemulsion was shown over a storage period of 120days at 4°C. Both catechin extract and nanoemulsion could inhibit growth of PC-3 tumor cells, with the half maximal inhibitory concentration being 15.4µg/mL and 8.5µg/mL, respectively. The PC-3 cell cycle was arrested at S phase through elevation of P27 expression and decline of cyclin A, cyclin B, cyclin-dependent kinase 2, and cyclin-dependent kinase 1 expression. In addition, both catechin extract and nanoemulsion could induce apoptosis of PC-3 cells through decrease in B-cell lymphoma 2 (bcl-2) expression and increase in cytochrome c expression for activation of caspase-3, caspase-8, and caspase-9. Taken together, both caspase-dependent and caspase-independent pathways may be involved in apoptosis of PC-3 cells. Keywords: green tea leaf waste, HPLC-MS, catechin nanoemulsion, prostate cancer cell PC-3, apoptosis

This study aims to determine the formation of heterocyclic amines (HAs) and cholesterol oxidation products (COPs) in fried chicken fiber (FCF) processed under different traditional conditions. An ultra-performance liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry technique was used to analyze various HAs and COPs respectively in chicken meat and FCF. Five HAs were produced in FCF during processing with the amount ranging from 0.052 to 106.005 ng/g and decreasing in the following order: 1-methyl-9H-pyrido[3,4-b]indole(Harman)>9H-pyrido[3,4-b]indole(Norharman)>2-amino-5-phenylpyridine(Phe-P-1)>2-amino-3-methyl-imidazo[4,5-f]-quinoxaline(IQx)>3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole(Trp-P-1). Chicken fiber fried in lard at 180 °C/20 min generated a higher level of HAs (106.005 ng/g) than in soybean oil (77.823 ng/g), but with frying at 150 °C/40 min, the total HAs contents were reduced by 26.4% and 6.2% respectively. Seven COPs were produced in FCF with the maximum level being 35.220 μg/g and both cholesterol-5α-6α-epoxide (5,6α-EP) and cholesterol-5β-6β-epoxide (5,6β-EP) dominating. A lower level of COPs was formed in chicken fiber fried in lard (12.999 or 11.182 μg/g) than in soybean oil (35.220 or 28.553 μg/g) during frying at 150 °C/40 min or 180 °C/20 min, with the former reducing COPs by 77.9% without soy sauce. Comparatively, the formation of both HAs and COPs could be substantially reduced by processing FCF in lard at 150 °C/40 min without soy sauce flavoring.

Allyl isothiocyanate (AITC), a volatile and water-insoluble compound present in several cruciferous vegetables, has been shown to possess several biological qualities such as anti-bacterial, anti-fungal, and anti-cancer activity. In this study, water-soluble allyl isothiocyanate nanoparticles (AITC-NPs) were prepared by oil dispersed in water (O/W) microemulsion and complex coacervation techniques and evaluated for their anti-inflammatory activity towards macrophage cell RAW 264.7 and anti-cancer effect on human bladder cancer cell HT1376. RESULTS The AITC-NPs with a particle size of 9.4?nm were stable during heating up to 110?°C or three freeze-thawing cycles. No significant cytotoxicity was shown on Caco-2 and intestine epithelial IEC-6 cells at AITC-NP doses ranging from 0.25 to 2?g?L?1 (8.75?70?mg?L?1 AITC). However, at 2?g?L?1 dosage, AITC-NPs could inhibit the growth of human bladder cancer cells HT1376 by 90%, while their low dosage at 0.25?g?L?1 could inhibit migration ability by 83.7, 71.3, 58.4 and 31.4% after 4, 8, 12, and 24?h of incubation, respectively. Compared to AITC and NPs, AITC-NPs showed a better inhibition on lipopolysaccharide (LPS)-induced TNF-α, IL-6, NO and iNOS production in RAW 264.7 macrophage cells. CONCLUSION The results demonstrate the potential of AITC-NPs as therapeutic agents for the treatment of bladder cancer and the enhancement of immune function. ? 2018 Society of Chemical Industry

The objectives of this study are to investigate antiproliferative effect and mechanisms of bioactive compounds from Gynostemma pentaphyllum (G. pentaphyllum) on lung carcinoma cell A549. Saponins, carotenoids and chlorophylls were extracted and fractionated by column chromatography, and were subjected to high-performance liquid chromatography-mass spectrometry analyses. The saponin fraction, which consisted mainly of gypenoside (Gyp) XXII and XXIII, rather than the carotenoid and chlorophyll ones, was effective in inhibiting A549 cell growth in a concentration- and a time-dependent manner as evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The estimated half maximal inhibitory concentration (IC50 ) of Gyp on A549 cells was 30.6 μg/ml. Gyp was further demonstrated to induce an apparent arrest of the A549 cell cycle at both the S phase and the G2/M phase, accompanied by a concentration- and a time-dependent increase in the proportions of both the early and late apoptotic cells. Furthermore, Gyp down-regulated cellular expression of cyclin A and B as well as BCL-2, while up-regulated the expression of BAX, DNA degradation factor 35 KD, poly [ADP-ribose] polymerase 1, p53, p21 and caspase-3. Nevertheless, both the treatment of a p53 inhibitor, pifithrin-α, and the small hairpin RNA-mediated p53 knockdown in the A549 cells did not alter the growth inhibition effect induced by Gyp. As a result, the cell cycle arrest and apoptosis of A549 cells induced by Gyp would most likely proceed through p53-independent pathway(s).

Metal oxide nanoparticles, especially alumina nanoparticles (AN) have drawn considerable attention in biotechnological and biomedical applications. Interaction of nanoparticles with protein plays a crucial role in several medical applications. Also, nanoparticles may exert toxic effects on cancer cells that can be beneficial for application in cancer therapy. The objectives of this study were to synthesize and characterize poly(γ-glutamic acid) (γ-PGA)-functionalized alumina nanoparticles (γ-PAN) for evaluation of protein adsorption ability using bovine serum albumin (BSA) and lysozyme (LSZ) as well as cytotoxicity towards human prostate cancer cell PC-3. Characterization of both AN and γ-PAN revealed a spinel lattice structure belonging to γ-Al2O3 with the mean particle size as determined by TEM being 5.4 nm for AN and 6.7 nm for γ-PAN. The zeta potential at different pH changed from positive to negative upon coating γ-PGA onto AN with a shift in point-of-zero-charge from 9.1 to 3.2 mV. Positively-charged AN at pH 7 could preferentially adsorb BSA compared to LSZ, while an opposite trend was observed for negatively-charged γ-PAN. The Langmuir adsorption capacity for AN and γ-PAN was 224.5 and 36.1 mg g−1 for BSA, respectively, and 16.2 and 110.1 mg g−1 for LSZ. Both AN and γ-PAN could lower cell viability of PC-3 cells in a dose-dependent manner with the latter more efficient than the former. The ROS production also increased with dose, which may lead to cytotoxicity towards PC-3 cells through oxidative stress and mitochondrial dysfunction. Thus, the γ-PGA functionalized alumina nanoparticles may be used as a promising material for future biomedical applications.

The variety and content of phenolic acids and flavonoids in a Chinese herb Rhinacanthus nasutus were determined by developing an HPLC method coupled with photodiode-array-detection and tandem-mass-spectrometry. A high yield of phenolic acids and flavonoids was obtained using 30% ethanol-in-water and shaking in 60 °C water-bath for 3 h. By employing a C18 column and gradient mobile phase of 0.1% formic acid-in-water and acetonitrile, a total of 20 phenolic acids and 3 flavonoids were separated within 45 min with detection at 280 nm, flow rate at 0.8 mL/min and column temperature at 35 °C. Internal standards vanillic acid and quercetin were used for quantitation of phenolic acids and flavonoids in R. nasutus, respectively, which included caffeic acid derivatives (6.40 mg/g), quercetin derivatives (4.43 mg/g), ferulic acid derivatives (3.20 mg/g), p-coumaric acid derivatives (1.63 mg/g), sinapic acid hexoside (1.11 mg/g), kaempferol-3-O-rutinoside (1.02 mg/g), hydroxycinnamic acid derivatives (0.44 mg/g) and protocatechuic acid (0.39 mg/g).

Objectives The aim of this study was to examine the effects of several vegetable oils and blended oil composed of soybean and camellia oils on blood lipid reduction and antioxidative activity. Methods Forty male hamsters were fed an AIN-93 G diet for 1 wk, followed by dividing into five groups: control group-1 was fed a low-fat diet containing 5% oil for 6 wk, and the other four groups were fed high-fat diets with group-2 containing 14% palm oil, group-3 containing 14% camellia oil, group-4 containing 14% soybean oil, and group-5 containing 14% blended oil (8.4% soybean oil and 5.6% camellia oil) along with 0.2% cholesterol and 0.1% bile acid. Results High-fat diets raised serum triacylglycerol, total cholesterol, and aspartate aminotransferase in hamsters without affecting alanine aminotransferase. Compared with palm oil-containing diet, the other three high-fat diets reduced serum total cholesterol, low-density lipoprotein cholesterol, and the ratio of low-density lipoprotein to high-density lipoprotein cholesterol with an opposite trend for liver total cholesterol. However, compared with the control group, the serum high-density lipoprotein cholesterol level was raised for all four high-fat diets. The higher the degree of oil unsaturation, the higher the serum thiobarbituric acid reactive substances and the lower the liver triacylglycerol level and activities of fatty acid synthase, glucose 6-phosphate dehydrogenase, and malic enzymes. Both soybean and blended oils lowered the antioxidative activity of liver. Conclusion Camellia and blended oils were more efficient than soybean oil in elevating serum high-density lipoprotein cholesterol and decreasing the ratio of low-density lipoprotein to high-density lipoprotein cholesterol in hamsters.

The traditional way to analyze heterocyclic amines (HAs) is time-consuming and uses large amounts of solvents. The objective of this study is to develop a quick and simultaneous analysis method for multiple types of HAs contained in meat products. Results showed that 20 HAs and 1 internal standard (4,7,8-TriMeIQx) can be separated within 30 min using an Inspire C18 column and a gradient solvent system containing 10 mM ammonium acetate (pH 2.9) and acetonitrile. This process resulted in a high degree of separation. Using acetonitrile with 1% acetic acid as an extraction solvent, followed by primary and secondary amine, MgSO4, and C18EC as purified reagent, is highly suitable for extracting HAs using the quick, easy, cheap, effective, rugged, and safe method (QuEChERS). Tandem mass spectrometry with selected reaction monitoring mode were used for analysis, which indicated reasonable recovery (58.9–117.4%) for all 20 types of HAs along with limits of detection and quantification in the range of 0.003–0.05 an...

Chu-Ching Yang,1,* Chi-Feng Hung,2,* Bing-Huei Chen1 1Department of Food Science, 2School of Medicine, Fu Jen Catholic University, Taipei, Taiwan *These authors contributed equally to this work Abstract: Coffee grounds, a waste by-product generated after making coffee, contains approximately 15% coffee oil which can be used as a raw material in cosmetics. Algae oil rich in docosahexaenoic acid (DHA) has been demonstrated to possess anticancer and anti-inflammation functions. Theobjectives of this study were to develop a gas chromatography-mass spectrometry (GC-MS) method for the determination of fatty acids in coffee oil and algae oil and prepare a nanoemulsion for studying its inhibition effect on ultraviolet A-induced skin damage in mice and growth of melanoma cells B16-F10. A total of 8 and 5 fatty acids were separated and quantified in coffee oil and algae oil by GC-MS, respectively, with linoleic acid (39.8%) dominating in the former and DHA (33.9%) in the latter. A nanoemulsion with a particle size of 30 nm, zeta potential -72.72 mV, and DHA encapsulation efficiency 100% was prepared by using coffee oil, algae oil, surfactant (20% Span 80 and 80% Tween 80), and deionized water. Differential scanning calorimetry (DSC) analysis revealed a high stability of nanoemulsion when heated up to 110°C at a pH 6, whereas no significant changes in particle size distribution and pH occurred over a 90-day storage period at 4°C. Animal experiments showed that a dose of 0.1% coffee oil-algae oil nanoemulsion was effective in mitigating trans-epidermal water loss, skin erythema, melanin formation, and subcutaneous blood flow. Cytotoxicity test implied effective inhibition of melanoma cell growth by nanoemulsion with an IC50 value of 26.5 µg/mL and the cell cycle arrested at G2/M phase. A dose-dependent upregulation of p53, p21, cyclin B, and cyclin A expressions and downregulation of CDK1 and CDK2 occurred. Also, both Bax and cytochrome c expressions were upregulated and bcl-2 expression downregulated, accompanied by a rise in caspase-3, caspase-8, and caspase-9 activities for apoptosis execution. Collectively, the apoptosis pathway of melanoma cells B16-F10 may involve both mitochondria and death receptor. Keywords: coffee ground, coffee oil-algae oil nanoemulsion, DHA, melanoma cell apoptosis, mouse skin protection, GC-MS

Flavonoids, containing mainly kaempferol rhamnohexoside derivatives, were extracted from Gynostemma pentaphyllum (G. pentaphyllum) and their potential growth inhibition effects against H460 non-small cell lung cancer cells was explored and compared to that on A549 cells. The extracted flavonoids were found to exhibit antiproliferation effects against H460 cells (IC50 = 50.2 μg/mL), although the IC50 of H460 is 2.5-fold that of A549 cells (IC50 = 19.8 μg/mL). Further investigation revealed that H460 cells are more susceptible to kaempferol than A549, whereas A549 cell growth is better inhibited by kaempferol rhamnohexoside derivatives as compared with H460. In addition, flavonoids from G. pentaphyllum induced cell cycle arrest at both S and G2/M phases with concurrent modulated expression of the cellular proteins cyclin A, B, p53 and p21 in A549 cells, but not H460. On the contrary, apoptosis and concomitant alteration in balance of BCL-2 and BAX expression as well as activation of caspase-3 were equally affected between both cells by flavonoid treatment. These observations strongly suggest the growth inhibition discrepancy between H460 and A549 following flavonoid treatment can be attributed to the lack of cell cycle arrest in H460 cells and the differences between H460 and A549 cells may serve as contrasting models for further mechanistic investigations.

Accumulation of aluminum in human has been reported to be associated with dementia, Parkinson's disease, and Alzheimer's disease. The objectives of this study were to evaluate an edible biopolymer poly(γ-glutamic acid) (γ-PGA) for aluminum removal efficiency under in vitro conditions as affected by pH, contact time, aluminum concentration, temperature, ionic strength, and essential metals in both aqueous aluminum solution and simulated gastrointestinal fluid (GIF). A low aluminum adsorption occurred at pH 1.5-2.5, followed by a maximum adsorption at pH 3.0-4.0 and precipitating thereafter as aluminum hydroxide at pH > 4. Adsorption was extremely fast with 81-96% of total adsorption being attained within 1 min, reaching equilibrium in 5-10 min. Kinetic data at low (10 mg/L) and high (50 mg/L) concentrations were well described by pseudo-first-order and pseudo-second-order models, respectively. Equilibrium adsorption isotherms at different temperatures were precisely fitted by both Langmuir and Redlich-Peterson models with the maximum adsorption capacities at 25, 37, and 50 °C being 35.85, 38.68, and 44.23 mg/g, respectively. Thermodynamic calculations suggested endothermic and spontaneous nature of aluminum adsorption by γ-PGA with increased randomness at the solid/solution interface. Variation in ionic strengths did not alter the adsorption capacity, however, the incorporation of essential metals significantly reduced the aluminum adsorption by following the order copper > iron > zinc > calcium > potassium. Compared to aqueous solution, the aluminum adsorption from simulated GIF was high at all studied pH (1-4) with Langmuir monolayer adsorption capacity being 49.43 mg/g at 37 °C and pH 4. The outcome of this study suggests that γ-PGA could be used as a safe detoxifying agent for aluminum.

A high performance liquid chromatography-diode array detection-mass spectrometry method with atmospheric pressure chemical ionization mode (HPLC-DAD-MS-APCI) was developed to determine carotenoids in Scutellaria barbata D. Don (Banzhilian). Results showed that a total of 19 carotenoids including internal standard were separated within 37 min by employing a YMC C30 column and a gradient mobile phase composed of methanol–acetonitrile (86:14, v/v) and methylene chloride. The various carotenoids in S. barbata were identified and quantified using internal standard β-apo-8′-carotenal, in which all-trans-lutein and its cis isomers constituted the largest portion, followed by all-trans-β-carotene and its cis isomers. A high recovery of 93.0–100.9% and a high reproducibility were obtained with this method. This would allow determination of carotenoids in herbal samples, and provide a basis for possible production of functional foods with S. barbata D. Don as raw material.

The present study investigated the anti-oxidative and hepatoprotective effects of Glossogyne tenuifolia (GT) Cassini, against acetaminophen-induced acute liver injury in BALB/c mice. The extracts of GT by various solvents (hot water, 50% ethanol and 95% ethanol) were compared for their 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, reducing power, total phenolic content, and total anti-oxidant capacity. The results showed that hot water (HW) extracts of GT contained high levels of phenolics and exerted an excellent anti-oxidative capacity; thus, these were used in the animal experiment. The male BALB/c mice were randomly divided into control group, acetaminophen (APAP) group, positive control group and two GT groups at low (GT-L) and high (GT-H) dosages. The results showed that mice treated with GT had significantly decreased serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). GT-H increased glutathione levels and the ratios of reduced glutathione and oxidized glutathione (GSH/GSSG) in the liver, and inhibited serum and lipid peroxidation. This experiment was the first to determine phenolic compounds, chlorogenic acid and luteolin-7-glucoside in HW extract of GT. In conclusion, HW extract of GT may have potential anti-oxidant capacity and show hepatoprotective capacities in APAP-induced liver damaged mice.

Polycyclic aromatic hydrocarbons (PAHs) represent an important pollutant in foods and/or the environment. This study aimed to determine the PAH contents in sugar-smoked meat by employing a quick, easy, cheap, effective, rugged, safe (QuEChERS) method combined with a GC-MS technique and assess the dietary exposure of PAHs in Taiwan. Results showed that the longer the sugar-smoking duration, the more the total PAH formation. By sugar-smoking for 6 min, the total PAH contents generated in red meat (33.9 ± 3.1-125.5 ± 9.2 ppb) were higher than in poultry meat (19.1 ± 2.0-28.2 ± 1.2 ppb) and seafood (9.1 ± 1.4-31.8 ± 1.8 ppb), with lamb steak containing the largest amount of total PAHs. Most importantly, the highly carcinogenic benzo[a]pyrene remained undetected in all of the sugar-smoked meat samples. In addition, the cancer risk due to dietary PAH exposure based on total intake of meat in Taiwan was

Carotenoids have been known to reduce the risk of several diseases including cancer and cardiovascular. However, carotenoids are unstable and susceptible to degradation. Rhinacanthus nasutus (L.) Kurz (R. nasutus), a Chinese medicinal herb rich in carotenoids, was reported to possess vital biological activities such as anti-cancer. This study intends to isolate carotenoids from R. nasutus by column chromatography, identify and quantify by HPLC-MS, and prepare carotenoid microemulsions for determination of absolute bioavailability in rats. Initially, carotenoid fraction was isolated using 250 mL ethyl acetate poured into an open-column packed with magnesium oxide-diatomaceous earth (1:3, w/w). Fourteen carotenoids including internal standard β-apo-8′-carotenal were resolved within 62 min by a YMC C30 column and gradient mobile phase of methanol-acetonitrile-water (82:14:4, v/v/v) and methylene chloride. Highly stable carotenoid microemulsions were prepared using a mixture of CapryolTM90, Transcutol®HP, Tween 80 and deionized water, with the mean particle being 10.4 nm for oral administration and 10.7 nm for intravenous injection. Pharmacokinetic study revealed that the absolute bioavailability of carotenoids in microemulsions and dispersion was 0.45% and 0.11%, respectively, while a much higher value of 6.25% and 1.57% were shown for lutein, demonstrating 4-fold enhancement in bioavailability upon incorporation of R. nasutus carotenoids into a microemulsion system.

This study examines the influence of the temperature on the low-frequency resonant spectrum of piezoelectric ceramics based on lead zirconate titanate solid solution Pb(Zr,Ti)O 3 -based piezoelectric ceramics with heat variations ranging from 25 to 150 °C, concluding that temperature increases cause the resonant spectrum to shift to higher frequencies. Furthermore, the variation in fundamental series resonant frequency f s (with temperature variation) is larger than the variation of fundamental parallel resonant frequency f p (with temperature variation). Another result of increased temperature is the decrease in effective electromechanical coupling factor k eff . Finally, the effect of temperature on the overtone mode is more obvious than that of the fundamental mode.

An improved high performance liquid chromatography-diode array detection-mass spectrometry method was developed for determination of various carotenoids and their precursors phytoene and phytofluene in human serum. A polymeric C30 column and mobile phase of (A) methanol/acetonitrile/water (84:14:4, v/v/v) and (B) dichloromethane (100%) were employed with the gradient condition of 100% A and 0% B initially, raised to 10% B at 4 min, 18% B at 12 min, 21% B at 17 min, 30% B at 20 min and maintained until 25 min and increased further to 39% B at 28 min, 60% B at 40 min and returned to 100% A and 0% B at 45 min. A total of 30 carotenoids, including 6 all-trans forms, 20 cis-isomers, 2 β-carotene epoxides, phytoene and phytofluene, were resolved within 45 min at a flow-rate of 1 mL/min, column temperature 25 °C and detection wavelengths 450, 348 and 286 nm. Identification of carotenoids was carried out by comparing retention behavior, absorption and mass spectral data with those of reference standards, isomerized standards and reported values. An internal standard parared was found appropriate for quantitation of all the carotenoids. The developed method provided high sensitivity with low detection and quantitation limits (2-14 and 6-43 ng/mL), high recovery (91-99%), and small intra-day and inter-day variations (0.14-6.01% and 0.31-7.28%). Application of the developed method to Taiwan subjects supplemented with carotenoid-rich capsules revealed β-carotene plus its cis isomers as well as epoxide derivatives to be present in largest amount (1069.8-2783.1 ng/mL) in serum, followed by lutein plus its cis isomers (511.6-2009.5 ng/mL), phytofluene plus its cis isomer (515.0-1765.0 ng/mL), lycopene plus its cis isomers (551.1-1455.1 ng/mL), β-cryptoxanthin plus its cis isomers (458.0-965.0 ng/mL), all-trans-zeaxanthin (110.0-177.0 ng/mL), phytoene (41.8-165.0 ng/mL) and all-trans-α-carotene (37.5-95.9 ng/mL).

The objectives of this research were to develop a method for the determination of 16 polycyclic aromatic hydrocarbons (PAHs) in poultry meat by combining the quick, easy, cheap, effective, rugged, and safe (QuEChERS) method with gas chromatography-mass spectrometry (GC-MS) and study their formation during marinating and frying. The recoveries of 16 PAHs ranged from 94.5 to 104% in blank samples and from 71.2 to 104% in poultry meat samples. The quantitation limits of 16 PAHs were from 0.02 to 1 ng/mL, with the intraday variability being from 2.4 to 6.6% [percent relative standard deviation (RSD%)] and interday variability being from 3.3 to 7.1% (RSD%). Most PAHs followed a time-dependent increase over a 24 h marinating period, with naphthalene being generated in the largest amount. Among the various poultry meat, chicken gizzard produced the highest level of total PAHs after 24 h of marinating. A similar tendency was observed for most PAHs during frying of poultry meat, but a high amount of total PAHs was shown in duck drumstick after 15 min of frying.

Rhinacanthus nasutus (L.) Kurz, a traditional Chinese herb possessing antioxidant and anti-cancer activities, has been reported to contain functional components like carotenoids and chlorophylls. However, the variety and amount of chlorophylls remain uncertain. The objectives of this study were to develop a high performance liquid chromatography-photodiode array detection-atmospheric pressure chemical ionization-mass spectrometry (HPLC-DAD-APCI-MS) method for determination of chlorophylls and their derivatives in hot-air-dried and freeze-dried R. nasutus. An Agilent Eclipse XDB-C18 column and a gradient mobile phase composed of methanol/N,N-dimethylformamide (97:3, v/v), acetonitrile and acetone were employed to separate internal standard zinc-phthalocyanine plus 12 cholorophylls and their derivatives within 21 min, including chlorophyll a, chlorophyll a', hydroxychlorophyll a, 15-OH-lactone chlorophyll a, chlorophyll b, chlorophyll b', hydroxychlorophyll b, pheophytin a, pheophytin a', hydroxypheophytin a, hydroxypheophytin a' and pheophytin b in hot-air-dried R. nasutus with flow rate at 1 mL/min and detection at 660 nm. But, in freeze-dried R. nasutus, only 4 chlorophylls and their derivatives, including chlorophyll a, chlorophyll a', chlorophyll b and pheophytin a were detected. Zinc-phthalocyanine was found to be an appropriate internal standard to quantify all the chlorophyll compounds. After quantification by HPLC-DAD, both chlorophyll a and pheophytin a were the most abundant in hot-air-dried R. nasutus, while in freeze-dried R. nasutus, chlorophyll a and chlorophyll b dominated.

Magnetite nanoparticles coated with an anionic biopolymer poly(γ-glutamic acid) (PGA-MNPs) were synthesized and characterized for their methylene blue dye adsorption capability. Both bare- and dye-loaded PGA-MNPs were characterized by FTIR, TEM and VSM measurements, revealing the PGA-MNPs to be superparamagnetic with average particle diameter being 12.4 nm and magnetization value 59.2 emu/g. The synthesized PGA-MNPs were stable in deionized, tap and river waters as well as in acidic and basic media. Redlich-Peterson and Langmuir models precisely described the isotherm and the maximum adsorption capacity was 78.67 mg/g. A pseudo-second-order equation best predicted the kinetics with a maximum adsorption attained within 5 min. Incorporation of sodium or calcium ions reduced the dye adsorption, while a raise in pH enhanced adsorption and a complete desorption occurred at pH 1.0. Dye removal mechanism by PGA-MNPs was probably due to electrostatic interaction through exchange of protons from side-chain α-carboxyl groups on PGA-MNPs surface.

The objectives of this study were to determine the effect of two CLAs , 9-cis, 11-trans CLA and 10-trans, 12-cis on cholesterol degradation and cholesterol oxidation products (COPs) formation in lard during heating for varied length of time. No CLAs and approximately 1770.1 µg/mL of cholesterol were detected in lard. Additionally, there was no significant change in the level of cholesterol and COPs in lard during heating at 100°C over a period of 240 min. But, at 150 or 200°C, the degradation of cholesterol was prominent with substantial amount of COPs being formed. Formation profiles of 7-OOH, 7-OH, and 7-keto at 150°C as well as 5,6-epoxides at both 150 and 200°C were fitted by a first-order equation, while a pseudo-second-order model described the kinetic pathway of 7-OH and 7-keto formation at 200°C. The formation of 7-OOH at 200°C were fitted as multiple first-order formation and first-order degradation curves as its level reached a peak at 60 min and declined to zero. Incorporation of 100 µg/mL CLA showed antioxidant activity, whereas a prooxidant activity was observed for CLA at 500 µg/mL. The outcome of this study demonstrated the potential of CLA to be an antioxidant in oil system.

A preparative column chromatographic method was developed to isolate flavonoids and saponins from Gynostemma pentaphyllum, a Chinese Medicinal herb, and evaluate their antiproliferation effect on hepatoma cell Hep3B, with the standards rutin and ginsenoside Rb(3) being used for comparison. Initially the powdered G. pentaphyllum was extracted with ethanol, followed by eluting flavonoids and saponins with ethanol-water (30:70, v/v) and 100% ethanol, respectively, in an open-column containing 5 g of Cosmosil 75C(18)-OPN, and then subjected to HPLC-MS analysis. The flavonoid fraction was mainly composed of quercetin- and kaempferol-glycosides, while in saponin fraction, both ginsenoside Rb(3) and ginsenoside Rd dominated. Both fractions were more effective against Hep3B cells than the standards rutin and ginsenoside Rb(3), with the cell cycle being arrested at G0/G1 phase for all the treatments. Additionally, the inhibition effect followed a dose-dependent increase for all the sample treatments. The result of this study may be used as a basis for possible phytopreparations in the future with G. pentaphyllum as raw material.

Surface-modified magnetite nanoparticles (MNPs) were synthesized by co-precipitation of aqueous solution of ferrous and ferric salts (molar ratio 1:2) upon adding a base followed by calcium salt of poly(γ-glutamic acid) (Ca-γ-PGA) for uniform coating on the surface of MNPs. Both uncoated and Ca-γ-PGA-coated MNPs were characterized using various techniques including Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), thermogravimetric analysis (TGA) and vibrating sample magnetometric (VSM) studies. Compared with bare MNPs, the IR spectra of coated MNPs showed characteristic peaks of γ-PGA, implying the γ-PGA coating on MNPs did occur. The TEM images depicted an average size of 8–10 nm for bare MNPs and 14 nm for coated MNPs, with their shape being spherical in nature. In the presence of applied magnetic field, a superparamagnetic behavior was observed at room temperature for both bare and Ca-γ-PGA-coated MNPs, with no magnetism left upon magnetic-field removal.

Fruit of Zizyphus jujuba Mill, a traditional Chinese herb widely consumed in Asian countries, has been reported to possess several vital biological activities. This study intends to develop an appropriate analytical method for isolation of polysaccharides from Z. jujuba fruits and evaluate their antioxidant activity. Initially, powdered Z. jujuba fruits were subjected to hot water extraction, followed by ethanol precipitation, deproteination, dialysis and fractionation in a DEAE-Sepharose CL-6B column. One neutral polysaccharide fraction (ZJPN) and 3 acidic polysaccharide fractions (ZJPa1, ZJPa2 and ZJPa3) were isolated with the average MW ranging from 40,566 to 129,518 Da. GC analysis revealed that 6 monosaccharides, namely, rhamnose, arabinose, xylose, mannose, glucose and galactose were present in polysaccharide fractions. The galacturonic acid content in polysaccharide fractions followed the order: ZJPa3 > ZJPa2 > ZJPa1 > ZJPN. All the 4 polysaccharide fractions were found to be more effective in scavenging superoxide anions than hydroxyl radicals, while acidic polysaccharides showed a more pronounced effect in chelating ferrous ion.

A preparative column chromatography method was developed to isolate carotenoids, flavonoids and polysaccharides, from Lycium barbarum L., possessing vital biological activity, and their antioxidant activity was evaluated. Carotenoids were isolated by a column containing magnesium oxide and diatomaceous earth (1.5:1, w/w), and β-carotene was eluted with hexane, β-cryptoxanthin and neoxanthin with ethyl acetate and zeaxanthin with ethyl acetate–ethanol (80:20, v/v). Flavonoids and phenolic acids were separated using a Cosmosil 140 C18-OPN column, with phenolic acids being eluted with deionized water and neutral flavonoids with methanol. Polysaccharides were fractionated using a DEAE-Sepharose CL-6B column; neutral polysaccharides were eluted with water and acidic polysaccharides with different concentrations of NaCl. For antioxidant activity, the flavonoid fraction was the most effective in scavenging DPPH· and ABTS + free radicals, chelating metal ions and reducing power, while the zeaxanthin fraction and polysaccharides showed the most pronounced effect in scavenging hydroxy free radicals and superoxide anions, respectively.

Polysaccharides in Lycium barbarum Linnaeus have been shown to be effective in preventing cancer. The objectives of this study were to develop an appropriate method for molecular weight determination of polysaccharides in L. barbarum . The most suitable analytical condition was: a volume-ratio of L. barbarum sample to deionized water at 1:10, followed by shaking in a 100 °C water bath for 30 min, concentrating to 50 mL and adding 250 mL of 95% ethanol for precipitation at −20 °C for 8 h, hydrolysing protein with 2.5 U/mL of proteinase at pH 8 and 60 °C for 4 h and separating polysaccharide into five fractions by high-performance size exclusion chromatography (HPSEC) with the molecular weight of two major fractions being 79,250 and 24,468 Da. Analysis of monosaccharides by gas chromatography (GC) indicated the presence of rhamnose, arabinose, xylose, mannose, glucose and galactose, with the molar ratio at 0.3:2.7:0.3:0.2:2.7:0.9, respectively.

Antioxidant activity of polysaccharide fractions isolated from Lycium barbarum Linnaeus was evaluated. Polysaccharides were extracted with boiling water, followed by precipitating with ethanol, protein hydrolysis, dialysis, and fractionation with a DEAE–Sepharose CL-6B column. A total of 4 fractions, including 1 neutral polysaccharide (LBPN) and 3 acidic polysaccharides were obtained, and compared with crude polysaccharide (CP), crude extract of polysaccharide (CE), deproteinated polysaccharide (DP), and deproteinated and dialyzed polysaccharide (DDP) for antioxidative activity. With the exception of CE and DDP, most polysaccharides were effective in scavenging DPPH and ABTS + free radicals, superoxide anion and hydroxyl radical at 1000 μg/mL.

Adsorption of mercury(II) by an extracellular biopolymer, poly(γ-glutamic acid) (γ-PGA), was studied as a function of pH, temperature, agitation time, ionic strength, light and heavy metal ions. An appreciable adsorption occurred at pH > 3 and reached a maximum at pH 6. Isotherms were well predicted by Redlich–Peterson model with a dominating Freundlich behavior, implying the heterogeneous nature of mercury(II) adsorption. The adsorption followed an exothermic and spontaneous process with increased orderliness at solid/solution interface. The adsorption was rapid with 90% being attained within 5 min for a 80 mg/L mercury(II) solution, and the kinetic data were precisely described by pseudo second order model. Ionic strength due to added sodium salts reduced the mercury(II) binding with the coordinating ligands following the order: Cl− > SO 4 2 - ≫ NO 3 - . Both light and heavy metal ions decreased mercury(II) binding by γ-PGA, with calcium(II) ions showing a more pronounced effect than monovalent sodium and potassium ions, while the interfering heavy metal ions followed the order: Cu2+ ≫ Cd2+ > Zn2+. Distilled water adjusted to pH 2 using hydrochloric acid recovered 98.8% of mercury(II), and γ-PGA reuse for five cycles of operation showed a loss of only 6.5%. IR spectra of γ-PGA and Hg(II)-γ-PGA revealed binding of mercury(II) with carboxylate and amide groups on γ-PGA.

The objectives of this study were to develop a high performance liquid chromatography-mass spectrometry (HPLC-MS) method for determination of chlorophylls and their derivatives in Gynostemma pentaphyllum Makino, a traditional Chinese herb possessing vital biological activities. Chlorophylls were extracted with a quaternary solvent system of hexane-acetone-ethanol-toluene (10:7:6:7, v/v/v/v), followed by separation of a total of 15 chlorophylls and their derivatives within 32 min using a gradient mobile phase of acetone, acetonitrile and methanol and a HyPURITY C18 column, with detection at 660 nm and flow rate at 1 mL/min. Identification was carried out on the basis of retention behavior, absorption spectra and mass spectra using atmospheric pressure chemical ionization (APCI) in positive ion mode for detection. Of the 15 analytes, chlorophyll a, chlorophyll b, pheophytin a and pheophytin b were quantified by using standard calibration curves, with the other 11 being quantified with an internal standard Fast Green FCF. Chlorophyll extracts in G. pentaphyllum were found to contain pheophytin a (2508.3 microg/g), pheophytin a' (111.2 microg/g), chlorophyll a (113.8 microg/g), chlorophyll a' (11.0 microg/g), hydroxypheophytin a (88.6 microg/g), hydroxypheophytin a' (66.5 microg/g), pyropheophytin a (76.0 microg/g), hydroxychlorophyll a (23.8 microg/g), pheophytin b (319.6 microg/g), pheophytin b' (13.2 microg/g), chlorophyll b (287.9 microg/g), chlorophyll b' (11.1 microg/g), hydroxychlorophyll b (15.0 microg/g), hydroxypheophytin b (11.2 microg/g) and hydroxypheophytin b' (8.5 microg/g).

Gynostemma pentaphyllum (Thunb.) Makino, a traditional Chinese herb possessing antitumor and antioxidant activities, has been shown to contain several functional components like saponins and flavonoids. However, their identities remain uncertain. The objectives of this study were to develop an appropriate extraction, purification and HPLC-MS method to determine saponins and flavonoids in G. pentaphyllum. Both flavonoids and saponins were extracted with methanol, followed by purification with a C18 cartridge to elute the former with 50% methanol and the latter with 100% methanol. A total of 34 saponins were separated within 40 min by a Gemini C18 column and a gradient mobile phase of acetonitrile and 0.1% formic acid in water, in which 18 saponins were identified by LC-MS with ESI mode and Q-TOF (LC/MS/MS). Similarly, a total of eight flavonoids were separated within 45 min by the same column and a gradient solvent system of methanol and 0.1% formic acid in water, with identification being carried out by a post-column derivatization method and LC-MS with ESI mode. The amounts of flavonoids in G. pentaphyllum ranged from 170.7 to 2416.5 mug g(-1), whereas saponins were from 491.0 to 89,888.9 mug g(-1).

The fruit of Lycium barbarum Linnaeus, a traditional Chinese herb containing functional components such as carotenoids, flavonoids and polysaccharides, has been widely used in the health food industry because of its possible role in the prevention of chronic disease like age-related macular degeneration. The objectives of this study were to develop a high performance liquid chromatography-photo diode array detection-mass spectrometry (HPLC-DAD-MS) method with atmospheric pressure chemical ionization (APCI) mode for qualitative and quantitative analyses of carotenoids in fruits of L. barbarum. Dried samples of L. barbarum were subjected to extraction without saponification or extraction followed by saponification. A C30 column with a gradient mobile phase of methylene chloride (100%) and methanol-acetonitrile-water (81:14:5, v/v/v) was used to separate carotenoids, with a total of 11 free carotenoids and 7 carotenoid esters being resolved from unsaponified and saponified L. barbarum extracts within 51 and 41 min, respectively. The fatty acid composition of carotenoid esters was confirmed by gas chromatography. Zeaxanthin dipalmitate (1143.7 microg/g) was present in the largest amount, followed by beta-cryptoxanthin monopalmitate and its two isomers (32.9-68.5 microg/g), zeaxanthin monopalmitate and its two isomers (11.3-62.8 microg/g), all-trans-beta-carotene (23.7 microg/g) and all-trans-zeaxanthin (1.4 microg/g).

Inflammatory mediators such as TNF-alpha and interleukin (IL)-1beta, and IL-8, which can enhance binding of low-density lipoprotein (LDL) to endothelium and upregulate expression of leukocyte adhesion molecules on endothelium during atherogenesis. Lycopene, a natural carotenoid from tomato and other sources, has been shown to prevent cardiovascular diseases in epidemiological studies. However, its anti-inflammatory action mechanism remains unclear. In the present study, we studied the effect of lycopene on TNF-alpha-induced signaling in human umbilical endothelial cells (HUVECs). We found that TNF-alpha-induced intercellular adhesion molecule-1 (ICAM-1) expression in HUVECs was inhibited by lycopene, whereas cyclooxygenase-2 (COX-2) and platelet-endothelial cell adhesion molecule (PECAM-1) expression were not affected. A further analysis indicated that lycopene attenuated TNF-alpha-induced IkappaB phosphorylation, NF-kappaB expression, and NF-kappaB p65 translocation from cytosol to nucleus. In line with this, TNF-alpha-induced NF-kappaB-DNA but not AP1-DNA complexes formation was inhibited by lycopene, as determined by the electrophoretic mobility shift assay (EMSA). On the other hand, lycopene did not affect TNF-alpha-induced p38 and extracellular matrix-regulated kinase1/2 (ERK1/2) phosphorylation and interferon-gamma (IFN-gamma)-induced signaling, suggesting that lycopene primarily affects TNF-alpha-induced NF-kappaB signaling pathway. In a functional study, lycopene dose-dependently attenuated monocyte adhesion to endothelial monolayer but not that adhesion to extracellular matrix. Taken together, we provided here the first evidence showing that lycopene is able to inhibit TNF-alpha-induced NF-kappaB activation, ICAM-1 expression, and monocyte-endothelial interaction, suggesting an anti-inflammatory role of lycopene and possibly explaining in part why lycopene can prevent cardiovascular diseases.

Soybean cake has been shown to be a rich source of isoflavone and can be produced during processing of soybean oil as byproduct. The objectives of this study were to compare the extraction yield of isoflavone from soybean cake by solvent and supercritical carbon dioxide, and study the conversion of isoflavone glucosides to the biologically active aglycone by employing β-glucosidase. Results showed that with supercritical carbon dioxide extraction, a maximum yield of malonylglucoside and glucoside was generated at 60 °C and 350 bar, while a high level of acetylglucoside and aglycone was produced at 80 °C and 350 bar. Supercritical carbon dioxide extraction resulted in a lower yield of total isoflavone than solvent extraction, but the former was more applicable to extraction of acetylglucoside and aglycone, and the latter to malonylglucoside and glucoside. A peak level of aglycone was attained from conversion of isoflavone glucoside by β-glucosidase at 55 °C, pH 5, concentration 50 U/ml and incubation time of 2 h.

Mycotoxins are low-molecular weight natural products produced as secondary metabolites by filamentous fungi and more than 500 mycotoxins with different physicochemical properties are currently known. Contamination by mycotoxins can result in liver and kidney diseases, nervous system damage, immunosuppression, and carcinogenicity. Consequently, the food safety authorities have set strict maximum permissible limits for mycotoxins. Over the past two decades numerous analytical techniques have been developed for mycotoxin detection in food. Recently, the application of nanotechnology in biosensors can range from transducer device to recognition ligand, label, and running systems. The application of nanomaterials in sensor development provides excellent advantages such as miniaturization of devices and signal enhancements. Such nanomaterial-based sensor devices can be sensitive, cost effective, and highly beneficial for the food industry ensuring on-site safety and preservation of food quality. This chapter reviews some recent developments on nanomaterial-based sensors developed for analysis of mycotoxin in food.

The d-galactose (d-gal)-injected animal model, which is typically established by administering consecutive subcutaneous d-gal injections to animals for approximately six or eight weeks, has been frequently used for aging research. In addition, this animal model has been demonstrated to accelerate aging in the brain, kidneys, liver and blood cells. However, studies on aging in male reproductive organs that have used this animal model remain few. Therefore, the current study aimed to optimize a model of male reproductive aging by administering d-gal injections to male mice and to determine the possible mechanism expediting senescence processes during spermatogenesis. In this study, C57Bl/6 mice were randomized into five groups (each containing 8–10 mice according to the daily intraperitoneal injection of vehicle control or 100 or 200 mg/kg dosages of d-gal for a period of six or eight weeks). First, mice subjected to d-gal injections for six or eight weeks demonstrated considerably decreased superoxide dismutase activity in the serum and testis lysates compared to those in the control group. The lipid peroxidation in testis also increased in the d-gal-injected groups. Furthermore, the d-gal-injected groups exhibited a decreased ratio of testis weight/body weight and sperm count compared to the control group. The percentages of both immotile sperm and abnormal sperm increased considerably in the d-gal-injected groups compared to those of the control group. To determine the genes influenced by the d-gal injection during murine spermatogenesis, a c-DNA microarray was conducted to compare testicular RNA samples between the treated groups and the control group. The d-gal-injected groups exhibited RNA transcripts of nine spermatogenesis-related genes (Cycl2, Hk1, Pltp, Utp3, Cabyr, Zpbp2, Speer2, Csnka2ip and Katnb1) that were up- or down-regulated by at least two-fold compared to the control group. Several of these genes are critical for forming sperm-head morphologies or maintaining nuclear integration (e.g., cylicin, basic protein of sperm head cytoskeleton 2 (Cylc2), casein kinase 2, alpha prime interacting protein (Csnka2ip) and katanin p80 (WD40-containing) subunit B1 (Katnb1)). These results indicate that d-gal-injected mice are suitable for investigating male reproductive aging.