Development of a rapid and reliable high-performance liquid chromatography method for determination of water-soluble vitamins in veterinary feed premix

Background and Aim: Determination of trace amounts of vitamins in multi-component feed premix is a troublesome analytical procedure. In this study, a simple and rapid high-performance liquid chromatography (HPLC) method was developed and validated for the concurrent detection and quantitation of four water-soluble vitamins such as thiamine, riboflavin, pyridoxine, and cyanocobalamin in veterinary feed premixes. Materials and Methods: The chromatographic separation of the vitamins was carried out at 35°C temperature on a reversed-phase C18 column using a gradient pump mode. Mobile phase constituents were solvent (a): 25 mM Potassium dihydrogen phosphate and 5 mM sodium hexanesulfonate in deionized water having pH-4.0 and solvent and (b) 5 mM sodium hexanesulfonate in methanol. Detection was performed with HPLC ultraviolet/visible detection set at 278 and 361 nm wavelength in two different channels. The flow rate was 1.2 mL/min and the total run time was 25 min. Results: The method was validated according to the International Conference on Harmonization and Food and Drug Administration guidelines and acceptance criteria for system suitability, precision, linearity, and recovery were met in all cases. The relative standard deviation for system suitability and precision was <2% for all vitamins. The linearity of the calibration curves was excellent (R2>0.999) at concentration of 5, 10, 15, 20, 25, and 30 μg/mL for all vitamins. The limits of detection values were 0.0125, 0.0017, 0.0064, and 0.0065 μg/mL for thiamine, riboflavin, pyridoxine, and cyanocobalamin, respectively, and the limits of quantification values were 0.0378, 0.0051, 0.0213, and 0.0198 μg/mL for thiamine, riboflavin, pyridoxine, and cyanocobalamin, respectively. The recovery percentages ranged from 88% to 115%. Conclusion: The overall parameters of the proposed method met the validation criteria and this method could be a highly desirable technique for routine analysis of water-soluble vitamins in veterinary feed premix.

APPLICATION OF FERROCIN-CONTAINING SORBENT AND PREMIX TO REDUCE THE TRANSITION OF 137CS TO MEAT CALVES

The studies were carried out in order to study the effectiveness of the use of a ferrocinecontaining sorbent and premix to reduce the intake of 137Cs in calf meat under the condition of a significant intake of mineral substances in the diet of animals in the form of a feed premix.

Effect of nutrients enriched feed premix on the growth performance of major carps Rohu, Catla and Mrigal in a polyculture system

To evaluate the effect of nutrients enriched feed premix on the growth performance of major carps Rohu (Labeo rohita), Catla (Catla catla) and Mrigal (Cirrhinus mrigala) in a polyculture system, the present experiment was carried out in six earthen ponds located at Khulna University of Bangladesh for a period of 90 days. Two treatments each with three replications were stocked with 40% Rohu, 30% Catla and 30% Mrigal with a total stocking density of 10000/ha. T-2 was treated with feed premix at the rate of 4g/1 kg feed while T-1 was not treated with any feed premix. Both treatments were applied normal feed twice a day at the rate of 2% to 4% of the body weight of the fingerlings. The net fish production was found to be 2238.06 kg/ha and 3607.33 kg/ha in T-1 and T-2 respectively. The results showed that fish production was significantly higher in T-2 than T-1 (P<0.05). T-2 showed 0.60 times greater fish production than that of T-1.Asian J. Med. Biol. Res. September 2018, 4(3): 298-306

Use of n-alkanes for determination of Kikuyu grass (Pennisetum clandestinum) intake in free-range broilers

N-alkanes, long-chained saturated hydrocarbons occurring in plant cuticles, were used for estimating grass intake in fast-growing, free-range broilers. A total of 1440 as hatched mixed sex Cobb 500 broilers were equally divided between one of four experimental treatments in a 2 by 2 factorial arrangement involving conventional (barn) or free-range (barn and range access) production systems and diets (crude protein: 21%, metabolisable energy: 13 MJ/K) with or without in-feed antibiotic (Zinc Bacitracin: Albac® G 150 antibiotic feed premix, Pfizer Australia Pty Ltd, Sydney, NSW, Australia). Day-old chicks were randomly allocated to 48 pens (12 replicates/treatment) with 30 birds in each pen. Chicks were assigned to treatment diets on Day 1 while free-range access was available to birds from Day 21 onwards. Alkane concentrations in litter were measured and compared with alkane profiles of the intake components (grass, diet pellets and woodchip). Correction for incomplete recovery followed by estimation using a non-negative least square procedure resulted in calculation of total grass intake from the range area. Kikuyu grass consumption was estimated to be 13.5–14.7% of total ‘as-fed’ intake, equating to 6.34–6.78 g of grass per bird per hour of range access in this study. Taking into account grass consumption, this resulted in a significant increase in feed intake by 8.7–8.9% (P < 0.01) and feed conversion ratio from 2.30 to 2.54 points (P < 0.01). It can be concluded that broilers reared under free-range conditions eat a substantial quantity of grass. However, the nutrient profile of grass is not complementary to the formulated ration and its consumption is likely to lead to an array of nutritional changes for the bird, thus affecting performance.

Amelioration of oxidative stress in broilers during summer

A comparative study on antistressor and antioxidative effects of synthetic vitamin C and polyherbal feed premix Ayucee supplementation in broilers was conducted during the summer months of June-July when the mean temperature-humidity index was 84.74?2.51. Day old broiler chicks (n =60) were randomly divided into three groups. Control group I was given basal diet and treatment groups (II&III) were supplemented with Synthetic vitamin C (100g/tonne of feed) and Ayucee (100g/tonne of feed) from day 0 to six weeks of age. Biochemical parameters were analysed after 3rd & 5th week & erythrocytic antioxidant enzymes were analysed after 3rd & 6th week of experiment. Hormonal & immunological parameters were analysed after 6th of experimental study. After 3rd week, mean plasma glucose, cholesterol & antioxidant enzyme glutathione reductase (GSSG) were significantly (P? 0.01) lower in treated groups (II & III) than control (I); however total protein, albumin to globulin ratio & antioxidant enzyme superoxide dismutase (SOD) were significantly (P? 0.05) different in group II & III compared to group I. After 5th week, mean plasma glucose, total protein, albumin globulin ratio were significantly (P? 0.05) different in both the treatments compared to control. Erythrocytic GSSG were significantly (P? 0.05) different in both the treatments than control, as observed after 6th week. Stress hormones namely cortisol & thyroxine (T4) were observed to be significantly (P?0.05) higher in untreated control than the treated groups. However, the two treatments did not differ significantly. Mean total immunoglobulin (Ig) level was significantly (P?0.01) higher in AYUCEE & vit-C treated birds than control after 6th week of study. It can be concluded from the results that oxidative stress in broilers during summer could be ameliorated using antioxidant synthetic vitamin C & the polyherbal antistressor, immunomodulator & adaptogenic feed premix AYUCEE.

Rapid Determination of Supplemental Vitamin E Acetate in Feed Premixes by Capillary Gas Chromatography

A rapid capillary gas chromatographic method was developed for determination of supplemental cc-tocopherol acetate (containing and verified at 50–5000 mg/kg feed) in various kinds of feed premixes. The method is based on simple n-hexane extraction of feed samples pretreated with a solution of acetic acid in ether for matrix breakdown followed by column cleanup on deactivated alumina to eliminate background interference. Fused silica chromatography is completed in 3 min. The withinday precision expressed as the relative standard deviation for recovery examples was 3.4,2.9, and 4.0% for high, medium, and low concentrated feed premixes, respectively; extraction recovery was 99–110%. The storage stabilities of standards and feed premix preparations were found to be excellent up to 3 months and up to 60° C.

Fractional Factorial Design Approach for Optimizing Analytical Methods

Experimental designs based on procedures of Taguchi are described for optimizing analytical methods. Methods were efficiently developed, with improvements in precision and accuracy, for the determination of cholesterol in foods, magnesium in feed premix, and moisture in mayonnaise. Significant factors and interactions were identified by using 2-level designs, analysis of variance techniques, and commercially available computer software. The designs were prepared by using Taguchi linear diagrams. The methods developed as a result of these experiments have proven to be rugged and reliable.