Authentication of Two Bio-Active Fish Oils by Qualitative Lipid Profiling Using Semi-Targeted Approach: An Exploratory Study.

BACKGROUND: Fish oils, which are rich in health-promoting polyunsaturated fatty acids (PUFA), have emerged as promising functional foods in the global health and wellness food market. Their source regarding the fish type, season, and location of harvesting might influence the nutritional value of such bioactive oils and determine their market price. The differences in price among such oils often lead to economically motivated mislabeling and adulteration. OBJECTIVE: In this study, our objective was to demonstrate how a qualitative targeted shotgun lipid profile workflow using an electrospray ionization-quadrupole-linear ion trap MS (QTrap) could differentiate fish oils originating from two different species. METHODS: Five samples each of sardine (Sardinella longiceps) oil and shark (Echinorhinus brucus) liver oil were diluted to a concentration of 80 µg/mL in chloroform-methanol (1 + 2, v/v) with 5 mM ammonium acetate. These samples were directly infused into a QTrap MS. The data were acquired for 23 precursor ion and 4 neutral loss scan experiments in the positive ionization mode and compared. RESULTS: We identified the following major lipid classes: cholesteryl ester, diacyl glycerol, triacylglycerol, monoalkyldiacylglycerol, and phophatydyl choline. The relative peak areas of the identified lipid species, when subjected to supervised multivariate analysis, could effectively distinguish the sardine oil and shark liver oil. CONCLUSIONS: The approach will be useful in establishing authenticity of fish oil and to support the regulatory agencies in dispute resolution. It can also be extended to establish authenticity in other agricultural and food commodities. HIGHLIGHTS: This paper reports a proof of concept for authenticating PUFA-rich fish supplements. A shotgun targeted lipidomics profile and chemometrics modeling successfully discriminated sardine oil and shark liver oil.

Dietary supplementation of thiamine and pyridoxine-loaded vanillic acid-grafted chitosan microspheres enhances growth performance, metabolic and immune responses in experimental rats.

In the present investigation, the effect of dietary supplementation of thiamine and pyridoxine loaded vanillic acid-grafted chitosan microspheres (TPVGC) on growth, metabolic and immune responses in Wistar strain albino rats was studied. Eight experimental groups, namely four groups each for male and female rats were fed with 0, 0.4, 0.8 and 1.6% of TPVGC in the diet. At the end of 45days feeding trials, both male and female rats supplemented with TPVGC had higher weight gain% and specific growth rate than the control groups. Significantly (p<0.05) lower blood glucose level and higher respiratory burst activity were recorded in the treatment groups than the control groups of both male and female rats. Activity of metabolic enzymes (aspartate amino transferase, alanine aminotransferase, alkaline phosphatase and acid phosphatase) and antioxidant enzymes (superoxide dismutase, catalase and glutathione S-transferase) were significantly higher (p<0.05) in the control groups and a decreasing trend in the same was observed with a gradual increase in the inclusion level of TPVGC in the diet of the treatment groups. However, a reverse trend was observed for acetylcholine esterase. It was inferred that dietary supplementation of thiamine and pyridoxine loaded vanillic acid-grafted chitosan enhanced the growth performance, metabolic and immune responses in the animal-model.

Antiaging effect of dietary chitosan supplementation on glutathione-dependent antioxidant system in young and aged rats.

Aging has been defined as the changes that occur in living organisms with the passage of time that lead to functional impairment and ultimately to death. Free radical-induced oxidative damage has long been thought to be the most important consequence of the aging process. In the present study, an attempt has been made to study the salubrious effects of dietary supplementation of chitosan on glutathione-dependent antioxidant defense system in young and aged rats. The dietary supplementation of chitosan significantly reduced the age-associated dyslipidemic abnormalities noted in the levels of total cholesterol, HDL-cholesterol, and LDL-cholesterol in plasma and heart tissue. Its administration significantly (P < 0.05) attenuated the oxidative stress in the heart tissue of aged rats through the counteraction of free radical formation by maintaining the enzymatic [glutathione peroxidase (GPx) and glutathione reductase (GR)] and non-enzymatic [reduced glutathione (GSH)] status at levels comparable to that of normal young rats. Our results conclude that dietary intake of chitosan restores the depleted myocardial antioxidant status and suggest that it could be an effective therapeutic agent in treatment of age-associated disorders where hypercholesterolemia and oxidative stress are the major causative factors.

Dietary chitosan supplementation attenuates isoprenaline-induced oxidative stress in rat myocardium.

Despite considerable advances in diagnosis and management over the last three decades, acute myocardial infarction continues to be a major public health problem. It is predicted that ischemic heart diseases will constitute the major disease-burden worldwide in the year 2020. In the present study, an attempt has been made to examine the effects of dietary chitosan supplementation on lipid peroxidation and cardiac antioxidant defense system in isoprenaline-induced myocardial infarction in rats, an animal model of myocardial infarction in man. Dietary chitosan intake significantly attenuated the isoprenaline-induced lipid peroxidation and maintained the level of reduced glutathione at near normal. Its administration demonstrated an antioxidant effect by maintaining the activities of myocardial glutathione dependent antioxidant enzymes (glutathione peroxidase and glutathione-S-transferase) and antiperoxidative enzymes (superoxide dismutase and catalase) at levels comparable to that of controls. The results of the present study indicate that the salubrious effects of dietary supplementation of chitosan is probably related to a counteraction of free radicals and/or to normal maintenance of the activities of free radical enzymes and the level of GSH, which protect myocardial membrane against oxidative damage by decreasing lipid peroxidation.

Evaluation of Azadirachta indica leaf fractions for in vitro antioxidant potential and protective effects against H2O2-induced oxidative damage to pBR322 DNA and red blood cells.

We evaluated the protective effects of subfractions of the ethyl acetate fraction (EAF) and the methanolic fraction (MF) from the crude ethanolic extract (CEE) of Azadirachta indica A. Juss (neem) leaves against various free radicals and hydrogen peroxide (H2O2)-induced oxidative damage to red blood cells (RBCs) and pBR322 DNA. Neem leaf fractions reduced DPPH(*), ABTS(*+), superoxide (O(*-)), hydroxyl (OH(*)), and nitric oxide radicals to nonradical forms in a concentration-dependent manner. Treatment with the benzene insoluble fraction from EAF (EBIF), the chloroform insoluble fraction from EAF (ECIF), the chloroform insoluble fraction from MF (MCIF), and the ethyl acetate insoluble fraction from MF (MEIF) significantly mitigated H2O2-induced oxidative damage to RBCs and pBR322 DNA. Although we found low in vitro free radical scavenging activity for the benzene insoluble fraction from EAF (EBSF), the chloroform soluble fraction from EAF (ECSF), the chloroform soluble fraction from MF (MCSF), and the ethyl acetate soluble fraction from MF (MESF), these fractions showed no effect on H2O2-induced lipid peroxidation and pBR322 DNA damage. High-performance liquid chromatography (HPLC) and TLC-Iatroscan analysis revealed that the greater efficacy of EBIF, ECIF, MCIF, and MEIF may be due to the presence of more polar compounds such as nimbolide and quercetin. Our studies suggest that the antioxidant and protective effects of active neem leaf fractions against H2O2-induced lipid peroxidation and pBR322 DNA damage can be attributed to their ability to inhibit various free radicals.

Antinociceptive and anti-pyretic activity of Benincasa hispida (thunb.) cogn. in Wistar albino rats.

The seeds of Benincasa hispida (Thunb) COGN. (Family: Cucurbitaceae) was extracted with ethanol and was used to study acute toxicity, antinociceptive and anti-pyretic effects. Brewer's yeast (15%) was used to induce pyrexia in rats. The extract was non lethal to the rats up to the dose of 5000 mg/kg b.w. At doses of 250 and 500 mg/kg b.w, the extract significantly (P<0.05) increased the antinociceptive effective in a dose dependent manner in rats. Similarly, at doses of 250 and 500 mg/kg b.w the extract significantly (P<0.05) decreased yeast-induced pyrexia in rats. These results indicate that ethanolic extract of Benincasa hispida possesses potent antinociceptive and antipyretic effects and thus pharmacologically justifying its folkloric use in the management of fever and pain conditions.

Anti-bacterial activity of ethanolic extract of Indoneesiella echioides (L) nees. evaluated by the filter paper disc method.

The study was carried out to investigate the antibacterial activity of the ethanolic extracts of Indoneesiella echioides (L) Nees. was evaluated by the filter paper disc method. This method is based on the diffusion of an antibiotic from a filter paper disc through the solidified culture media of a Petri dish used for study. The growth of inoculated is inhibited entirely in a circular area "Zone around the filter" paper disc containing a solution of antibiotic and the plant extract. The microorganisms used were: 1. Staphylococcus aureus (Gram positive 2. Escherichia coli (Gram negative). The organisms were maintained on nutrient agar slants. These were tested using nutrient broth. One loop full of the respective cultures was taken in slants which were maintained below 40 degrees C were taken and inoculated in the broth and incubated at 37 degrees C for 24 hrs and were observed for the growth of the organism with naked eye for their turbid nature. It was compared with that of sterile broth. The presence of turbidity indicated growth and suitability of the culture for further work.

Protective effect of dietary squalene supplementation on mitochondrial function in liver of aged rats.

Mitochondria are an important intracellular source and target of reactive oxygen species. The life span of a species is thought to be determined, in part, by the rate of mitochondrial damage inflicted by oxygen free radicals during the course of normal cellular metabolism. In the present study, we have investigated the protective effect of squalene supplementation for 15 days and 30 days on energy status and antioxidant defense system in liver mitochondria of 18 young and 18 aged rats. The dietary supplementation of 2% squalene significantly minimized aging associated alterations in mitochondrial energy status by maintaining the activities of TCA cycle enzymes (isocitrate dehydrogenase, alpha-ketoglutarate dehydrogenase, succinate dehydrogenase and malate dehydrogenase) and respiratory marker enzymes (NADH dehydrogenase and cytochrome-c-oxidase) at higher level in the liver mitochondria of aged rats compared with unsupplemented controls. It exerted an antioxidant effect by inhibiting mitochondrial lipid peroxidation (malondialdehyde) in liver of young and aged rats. Supplementation with squalene also maintained the mitochondrial antioxidant defense system at higher rate by increasing the level of reduced glutathione and the activities of glutathione-dependent antioxidant enzymes (glutathione peroxidase and glutathione-S-transferase) and antiperoxidative enzymes (superoxide dismutase and catalase) in the liver of young and aged rats. The results of this study provide evidence that dietary supplementation with squalene can improve liver mitochondrial function during aging and minimize the age-associated disorders in which reactive oxygen species are a major cause.

Cardioprotective effect of squalene on lipid profile in isoprenaline-induced myocardial infarction in rats.

We studied the cardioprotective effect of squalene on isoprenaline-induced myocardial infarction in male albino rats with respect to changes in the levels of lipid components in plasma and heart tissue. Prior administration of 2% squalene in feed for 45 days significantly reduced the isoprenaline-induced elevation in the levels of cholesterol, triglycerides, and free fatty acids in plasma and heart tissue of rats following myocardial infarction. It exerted an antilipidemic effect by reducing the level of low-density lipoprotein cholesterol with a parallel rise in the level of high-density lipoprotein cholesterol in plasma of experimental rats. A tendency to prevent the isoprenaline-induced depletion of phospholipids in the myocardium of experimental rats was also observed. In the present study, the pretreatment with squalene significantly counteracted the isoprenaline-induced lipid peroxidation and maintained the rats at near normal status. The results of the present study indicate that the overall cardioprotective effect of squalene is probably related to an inhibition of lipid accumulation by its hypolipidemic properties and/or its antioxidant properties.

Cardioprotective effects of Picrorrhiza kurroa against isoproterenol-induced myocardial stress in rats.

The cardioprotective effect of the ethanol extract of Picrorrhiza kurroa rhizomes and roots (PK) on isoproterenol-induced myocardial infarction in rats with respect to lipid metabolism in serum and heart tissue has been investigated. Oral pre-treatment with PK (80 mg kg(-1) day(-1) for 15 days) significantly prevented the isoproterenol-induced myocardial infarction and maintained the rats at near normal status.

Protective effect of Hemidesmus indicus against rifampicin and isoniazid-induced hepatotoxicity in rats.

Oral treatment with the ethanol extract of Hemidesmus indicus roots (100 mg/kg, for 15 days) significantly prevented rifampicin and isoniazid-induced hepatotoxicity in rats.