Niladri Sekhar Chatterjee

Multiresidue analysis of multiclass pesticides and polyaromatic hydrocarbons in fatty fish by gas chromatography tandem mass spectrometry and evaluation of matrix effect

This paper reports a selective and sensitive method for multiresidue determination of 119 chemical residues including pesticides and polyaromatic hydrocarbons (PAH) in high fatty fish matrix. The novel sample preparation method involved extraction of the target analytes from homogenized fish meat (5 g) in acetonitrile (15 mL, 1% acetic acid) after three-phase partitioning with hexane (2 mL) and the remaining aqueous layer. An aliquot (1.5 mL) of the acetonitrile layer was aspirated and subjected to two-stage dispersive solid phase extraction (dSPE) cleanup and the residues were finally estimated by gas chromatography mass spectrometry with selected reaction monitoring (GC-MS/MS). The co-eluted matrix components were identified on the basis of their accurate mass by GC with quadrupole time of flight MS. Addition of hexane during extraction and optimized dSPE cleanup significantly minimized the matrix effects. Recoveries at 10, 25 and 50 μg/kg were within 60-120% with associated precision, RSD<11%.

Assessment of polyaromatic hydrocarbons and pesticide residues in domestic and imported pangasius (Pangasianodon hypophthalmus) fish in India

BACKGROUND The pangasius catfish (Pangasianodon hypophthalmus) is a ubiquitous item of seafood in global markets. However, pesticide residues in aquaculture fish, arising from agricultural run-off and/or direct application during pond preparation, pest control and harvest, are a potential food safety concern. This study assessed the level of chemical hazard in domestic and imported pangasius fish in India. RESULTS A total of 119 contaminants, including polyaromatic hydrocarbons (PAHs), organochlorines (OCs) and other groups of pesticides, were screened in 148 samples during 2014-2015 as the first endeavour of its kind in an Indian context, employing a validated gas chromatography/tandem mass spectrometry (GC/MS/MS) method. Apart from the low-level incidence of OCs, pesticides such as quinalphos, malathion, parathion-methyl, etc. were detected in 38% of the samples. In comparison with Indian pangasius, fewer contaminants at low residue level were detected in pangasius fillets imported from Vietnam. CONCLUSION The human dietary exposures of the residue concentrations detected were less than the maximum permissible intakes and hence appeared safe. However, detection of commonly used pesticides indicated their direct application in aquaculture and contamination from agricultural run-off. This emphasizes the need for continual residue monitoring in aquaculture fish.

Evaluation of chitosan as a wall material for microencapsulation of squalene by spray drying: Characterization and oxidative stability studies

The present study was aimed at investigating the efficacy of chitosan as a wall material for microencapsulation of squalene by spray drying for functional food applications. Based on different core to wall material ratio (1:1, 0.5:1 and 0.3:1 on w/w basis), emulsions were prepared and evaluated in terms of emulsion stability, particle size, zeta potential, polydispersity Index (PDI), rheology and microstructure. The optimized emulsion combination was spray dried and characterized, physically and chemically. The encapsulation efficiency of the powder was found to be 26±0.6% whereas other properties such as particle size, zeta potential, water activity, hygroscopicity, Carr Index, Hausner ratio have shown satisfactory results. SEM analysis showed that the squalene microcapsules were smooth spherical particles free from dents and fissures. FTIR data further confirmed the encapsulation of squalene with chitosan. However, TGA, oxidative stability and accelerated Rancimat results showed that chitosan was not able to protect squalene from oxidation during storage. The results suggest that chitosan is not an appropriate wall material for microencapsulation of squalene and hence a combination of wall materials could be attempted for the encapsulation of squalene.

Microencapsulation of sardine oil: application of vanillic acid grafted chitosan as a bio-functional wall material

Vanillic acid grafted chitosan (Va-g-Ch) was evaluated as a new antioxidant wall material for microencapsulation of polyunsaturated fatty acid rich sardine oil. A high grafting ratio of 305mg vanillic acid equivalent/g of polymer was achieved using a free radical mediated grafting reaction. Oil in water emulsion was prepared with an optimised combination of Va-g-Ch and Tween 20 (3.2:1). Sardine oil loaded microparticles (SO-M) were produced (∼75% yield) by spray drying. The average diameter and polydispersity Index (PDI) of the particles were found to be 2.3μ and 0.345. XRD spectra of SO-M showed reduction in crystallinity due to microencapsulation. After four weeks of storage, a moderate (∼12%) decrease in the EPA and DHA content and a low PV of 5.5±0.51meq/kg oil in SO-M demonstrated good oxidative stability. Satisfactory encapsulation efficiency (84±0.84%) and loading efficiency (67±0.51%) values, also demonstrated the suitability of Va-g-Ch for microencapsulation of sardine oil.

Biochemical Characterization of Liver Oil of Echinorhinus brucus (Bramble Shark) and Its Cytotoxic Evaluation on Neuroblastoma Cell Lines (SHSY-5Y)

The objective of the present study was to characterize the liver oil extracted from the deep sea shark, Echinorhinus brucus, caught from Central Indian Ocean and to evaluate its cytotoxic effect on neuroblastoma cell line (SHSY-5Y). Characterization of liver oil of Echinorhinus brucus revealed the presence of palmitic acid (15%), oleic acid (12%), stearic acid (8%), docosahexaenoic acid (DHA) (18%), and eicosapentaenoic acid (EPA) (16%). It was also found to be a good source of squalene (38.5%) and fat soluble vitamins such as A, D, and K (vitamin A: 17.08 mg/100 g of oil, vitamin D: 15.04 mg/100 g oil, and vitamin K: 11.45 mg/100 g oil). Since it was found to be rich in essential fatty acids, fat soluble vitamins, and squalene, it can be considered as better dietary supplement. The oil of Echinorhinus brucus also showed high in vitro cytotoxic effect against the human neuroblastoma cell line (SHSY-5Y) and the IC50 value laid between 35 and 45 ng.

Screening Natural Content of Water-Soluble B Vitamins in Fish: Enzymatic Extraction, HILIC Separation, and Tandem Mass Spectrometric Determination

Despite the potential of LC with tandem MS (MS/MS) in improving sensitivity and selectivity, analytical methods are scarce for the determination of protein-bound and phosphorylated forms of B vitamins in food. This prompted us to develop a method for LC-MS/MS determination of naturally occurring nicotinamide, nicotinic acid, thiamine, pyridoxine, riboflavin, pantothenic acid, biotin, folic acid, and cyanocobalamin in fish. Baseline separation of the vitamins was achieved in a hydrophilic interaction LC condition. An ultrasonication-assisted enzymatic extraction protocol for sample preparation was optimized and validated. The time required for extraction was significantly reduced (to 4 h), while maintaining good extraction efficiency. Acetonitrile content (80%, v/v) in the prepared sample was found to be optimum for excellent peak shape and sensitivity. The dynamic linear range of the vitamins ranged from 2.5 to 500 ng/g, and the regression coefficient values were greater than 0.99. LOQ values ranged from 0.4 to 50 ng/g for the different vitamins. The spike recovery values at 50 and 100 ng/g ranged from 87.5 to 97.5%. The intra- and interday precision values were satisfactory. Accuracy of the developed method was determined by analysis of a Certified Reference Material. The method could also be used for unambiguous determination of the natural content of the target vitamins in fish.

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.