Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 10

HPLC Related Publications

10 Vitamin Content of Swordfish (Xhiphias gladius) Affected by Salting and Frying

Authors: L. Gómez-Limia, I. Franco, S. Martínez, L. Piñeiro, N. Cobas

Abstract:

The swordfish (Xiphias gladius) is a large oceanic fish of high commercial value, which is widely distributed in waters of the world’s oceans. They are considered to be an important source of high quality proteins, vitamins and essential fatty acids, although only half of the population follows the recommendation of nutritionists to consume fish at least twice a week. Swordfish is consumed worldwide because of its low fat content and high protein content. It is generally sold as fresh, frozen, and as pieces or slices. The aim of this study was to evaluate the effect of salting and frying on the composition of the water-soluble vitamins (B2, B3, B9 and B12) and fat-soluble vitamins (A, D, and E) of swordfish. Three loins of swordfish from Pacific Ocean were analyzed. All the fishes had a weight between 50 and 70 kg and were transported to the laboratory frozen (-18 ºC). Before the processing, they were defrosted at 4 ºC. Each loin was sliced and salted in brine. After cleaning the slices, they were divided into portions (10×2 cm) and fried in olive oil. The identification and quantification of vitamins were carried out by high-performance liquid chromatography (HPLC), using methanol and 0.010% trifluoroacetic acid as mobile phases at a flow-rate of 0.7 mL min-1. The UV-Vis detector was used for the detection of the water- and fat-soluble vitamins (A and D), as well as the fluorescence detector for the detection of the vitamin E. During salting, water and fat-soluble vitamin contents remained constant, observing an evident decrease in the values of vitamin B2. The diffusion of salt into the interior of the pieces and the loss of constitution water that occur during this stage would be related to this significant decrease. In general, after frying water-soluble and fat-soluble vitamins showed a great thermolability with high percentages of retention with values among 50–100%. Vitamin B3 is the one that exhibited higher percentages of retention with values close to 100%. However, vitamin B9 presented the highest losses with a percentage of retention of less than 20%.

Keywords: HPLC, Vitamins, frying, salting, swordfish

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9 Changes in Amino Acids Content in Muscle of European Eel (Anguilla anguilla) in Relation to Body Size

Authors: L. Gómez-Limia, I. Franco, T. Blanco, S. Martínez

Abstract:

European eels (Anguilla anguilla) belong to Anguilliformes order and Anguillidae family. They are generally classified as warm-water fish. Eels have a great commercial value in Europe and Asian countries. Eels can reach high weights, although their commercial size is relatively low in some countries. The capture of larger eels would facilitate the recovery of the species, as well as having a greater number of either glass eels or elvers for aquaculture. In the last years, the demand and the price of eels have increased significantly. However, European eel is considered critically endangered by the International Union for the Conservation of Nature (IUCN) Red List. The biochemical composition of fishes is an important aspect of quality and affects the nutritional value and consumption quality of fish. In addition, knowing this composition can help predict an individual’s condition for their recovery. Fish is known to be important source of protein rich in essential amino acids. However, there is very little information about changes in amino acids composition of European eels with increase in size. The aim of this study was to evaluate the effect of two different weight categories on the amino acids content in muscle tissue of wild European eels. European eels were caught in River Ulla (Galicia, NW Spain), during winter. The eels were slaughtered in ice water immersion. Then, they were purchased and transferred to the laboratory. The eels were subdivided into two groups, according to the weight. The samples were kept frozen (-20 °C) until their analysis. Frozen eels were defrosted and the white muscle between the head and the anal hole. was extracted, in order to obtain amino acids composition. Thirty eels for each group were used. Liquid chromatography was used for separation and quantification of amino a cids. The results conclude that the eels are rich in glutamic acid, leucine, lysine, threonine, valine, isoleucine and phenylalanine. The analysis showed that there are significant differences (p < 0.05) among the eels with different sizes. Histidine, threonine, lysine, hydroxyproline, serine, glycine, arginine, alanine and proline were higher in small eels. European eels muscle presents between 45 and 46% of essential amino acids in the total amino acids. European eels have a well-balanced and high quality protein source in the respect of E/NE ratio. However, eels with higher weight showed a better ratio of essential and non-essential amino acid.

Keywords: Amino Acids, HPLC, body size, European eels

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8 Fatty Acid and Amino Acid Composition in Mene maculata in The Sea of Maluku

Authors: Semuel Unwakoly, Reinner Puppela, Maresthy Rumalean, Healthy Kainama

Abstract:

Fish is a kind of food that contains many nutritions, one of those is the long chain of unsaturated fatty acids as omega-3 and omega-6 fatty acids and essential amino acid in enough amount for the necessity of our body. Like pelagic fish that found in the sea of Maluku. This research was done to identify fatty acids and amino acids composition in Moonfish (M. maculata) using transesterification reaction steps and Gas Chromatograph-Mass Spectrophotometer (GC-MS) and High-Performance Liquid Chromatography (HPLC). The result showed that fatty acids composition in Moonfish (M. maculata) contained tridecanoic acid (2.84%); palmitoleic acid (2.65%); palmitic acid (35.24%); oleic acid (6.2%); stearic acid (14.20%); and 5,8,11,14-eicosatetraenoic acid (1.29%) and 12 amino acids composition that consist of 7 essential amino acids, were leucine, isoleucine, valine, phenylalanine, methionine, lysine, and histidine, and also 5 non-essential amino acid, were tyrosine, glycine, alanine, glutamic acid, and arginine.Thus, these fishes can be used by the people to complete the necessity of essential fatty acid and amino acid.

Keywords: HPLC, Amino Acid, GC-MS, fatty acid, Moonfish (M. maculata)

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7 Characterization of Penicillin V Acid and Its Related Compounds by HPLC

Authors: Bahdja Guerfi, N. Hadhoum, I. Azouz, M. Bendoumia, S. Bouafia, F. Z. Hadjadj Aoul

Abstract:

Background: 'Penicillin V' is a narrow, bactericidal antibiotic of the beta-lactam family of the naturally occurring penicillin group. It is limited to infections due to the germs defined as sensitive. The objective of this work was to identify and to characterize Penicillin V acid and its related compounds by High-performance liquid chromatography (HPLC). Methods: Firstly phenoxymethylpenicillin was identified by an infrared absorption. The organoleptic characteristics, pH, and determination of water content were also studied. The dosage of Penicillin V acid active substance and the determination of its related compounds were carried on waters HPLC, equipped with a UV detector at 254 nm and Discovery HS C18 column (250 mm X 4.6 mm X 5 µm) which is maintained at room temperature. The flow rate was about 1 ml per min. A mixture of water, acetonitrile and acetic acid (65:35:01) was used as mobile phase for phenoxyacetic acid ‘impurity B' and a mixture of water, acetonitrile and acetic acid (650:150:5.75) for the assay and 4-hydroxypenicillin V 'impurity D'. Results: The identification of Penicillin V acid active substance and the evaluation of its chemical quality showed conformity with USP 35th edition. The Penicillin V acid content in the raw material is equal to 1692.22 UI/mg. The percentage content of phenoxyacetic acid and 4-hydroxypenicillin V was respectively: 0.035% and 0.323%. Conclusion: Through these results, we can conclude that the Penicillin V acid active substance tested is of good physicochemical quality.

Keywords: Characterization, HPLC, related substances, Penicillin V acid

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6 Development and Validation of a HPLC Method for 6-Gingerol and 6-Shogaol in Joint Pain Relief Gel Containing Ginger (Zingiber officinale)

Authors: Tanwarat Kajsongkram, Saowalux Rotamporn, Sirinat Limbunruang, Sirinan Thubthimthed

Abstract:

High Performance Liquid Chromatography (HPLC) method was developed and validated for simultaneous estimation of 6-Gingerol(6G) and 6-Shogaol(6S) in joint pain relief gel containing ginger extract. The chromatographic separation was achieved by using C18 column, 150 x 4.6mm i.d., 5μ Luna, mobile phase containing acetonitrile and water (gradient elution). The flow rate was 1.0 ml/min and the absorbance was monitored at 282 nm. The proposed method was validated in terms of the analytical parameters such as specificity, accuracy, precision, linearity, range, limit of detection (LOD), limit of quantification (LOQ), and determined based on the International Conference on Harmonization (ICH) guidelines. The linearity ranges of 6G and 6S were obtained over 20- 60 and 6-18 μg/ml respectively. Good linearity was observed over the above-mentioned range with linear regression equation Y= 11016x- 23778 for 6G and Y = 19276x-19604 for 6S (x is concentration of analytes in μg/ml and Y is peak area). The value of correlation coefficient was found to be 0.9994 for both markers. The limit of detection (LOD) and limit of quantification (LOQ) for 6G were 0.8567 and 2.8555 μg/ml and for 6S were 0.3672 and 1.2238 μg/ml respectively. The recovery range for 6G and 6S were found to be 91.57 to 102.36 % and 84.73 to 92.85 % for all three spiked levels. The RSD values from repeated extractions for 6G and 6S were 3.43 and 3.09% respectively. The validation of developed method on precision, accuracy, specificity, linearity, and range were also performed with well-accepted results.

Keywords: HPLC, ginger

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5 Identification of Phenolic Contents in Malaysian Variety of Pummelo (Citrus Grandis L. Osbeck) Fruit Juice Using HPLC-DAD

Authors: N. N. A. K. Shah, R. A. Rahman, R. Shamsuddin, N. M. Adzahan

Abstract:

Pummelo is known to be the largest of all citrus fruits, with expected ratio of 2:1 (w/v) of producing juice, is an attractive opportunity for Malaysia to expand pummelo-s influence and marketability over the international market of juices. The purpose of this study is to identify and quantify the phenolic compounds in two Malaysian varieties of pummelo fruit juice: Ledang (PO55) and Tambun (PO52). Identifications of polyphenols composition were done using High Performance Liquid Chromatography Diode Array Detection (HPLC-DAD). The phenolic compounds that were found in both varieties were hydroxycinnamic acids and flavonones. This study proved that Tambun variety has the highest antioxidant and phenolic compounds in comparison to Ledang variety. However, considerations have to be made to suit consumer-s taste bud and the amount of enzyme needed to clarify the juice for its marketability.

Keywords: HPLC, antioxidant, phenolic contents and pummelo fruit juice

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4 High Performance Liquid Chromatographic Method for Determination of Colistin Sulfate and its Application in Medicated Premixand Animal Feed

Authors: S.Choosakoonkriang, S. Supaluknari, P. Puangkaew

Abstract:

The aim of the present study was to develop and validate an inexpensive and simple high performance liquid chromatographic (HPLC) method for the determination of colistin sulfate. Separation of colistin sulfate was achieved on a ZORBAX Eclipse XDB-C18 column using UV detection at λ=215 nm. The mobile phase was 30 mM sulfate buffer (pH 2.5):acetonitrile(76:24). An excellent linearity (r2=0.998) was found in the concentration range of 25 - 400 μg/mL. Intra- day and inter-day precisions of method (%RSD, n=3) were less than 7.9%.The developed and validated method was applied to determination of the content of colistin sulfate in medicated premix and animal feed sample.The recovery of colistin from animal feed was satisfactorily ranged from 90.92 to 93.77%. The results demonstrated that the HPLC method developed in this work is appropriate for direct determination of colistin sulfate in commercial medicated premixes and animal feed.

Keywords: Animal Feed, HPLC, Colistin sulfate, medicated premix

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3 Metabolic Analysis of Fibroblast Conditioned Media and Comparison with Theoretical Modeling

Authors: Priyanka Gupta, Paul Verma, Kerry Hourigan, Jayesh Bellare, Sameer Jadhav

Abstract:

Understanding the consumption and production of various metabolites of fibroblast conditioned media is needed for its proper and optimized use in expansion of pluripotent stem cells. For this purpose, we have used the HPLC method to analyse the consumption of glucose and the production of lactate over time by mouse embryonic fibroblasts. The experimental data have also been compared with mathematical model fits. 0.025 moles of lactate was produced after 72 hrs while the glucose concentration decreased from 0.017 moles to 0.011 moles. The mathematical model was able to predict the trends of glucose consumption and lactate production.

Keywords: HPLC, Conditioned media, metabolite analysis, mouse embryonic fibroblast

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2 Pervasiveness of Aflatoxin in Peanuts Growing in the Area of Pothohar, Pakistan

Authors: Mateen Abbas, Abdul Muqeet Khan, Muhammad Rafique Asi, Javed Akhtar

Abstract:

Mycotoxin (aflatoxins) contamination of peanuts is a great concern for human health. A total of 72 samples of unripe, roasted, and salty peanuts were collected randomly from Pothohar plateau of Pakistan for the assessment of aflatoxin. Samples were dried, ground and extracted by acetonitrile (84%). The filtered extracts were cleaned up by MycoSep-226 and analyzed by high performance liquid chromatography with flourescence detector. Quantification limit of Aflatoxin was 1 μg/kg and 70% Recovery was observed in spiked samples in the range 1–10 μg/kg. The screening of mycotoxins indicated that aflatoxins were present in most of the samples being detected in 82%, in concentrations from 14.25 μg/kg to 98.80 μg/kg. Optimal conditions for mycotoxin production and fungal growth are frequently found in the crop fields as well as in store houses. Human exposure of such toxin can be controlled by pointed out such awareness and implemented the regulations.

Keywords: HPLC, Pakistan, peanuts, aflatoxin, Punjab

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1 Determination of EDTA in Dairy Wastewater and Adjacent Surface Water

Authors: Congmin Z. Xie, Terry Healy, Peter Robinson, Kevin Stewart

Abstract:

An HPLC-UV analytical method was developed to determine ethylenediaminetetraacetic acid (EDTA) in dairy wastewater and surface water. The optimizing separation was achieved by reversed–phase ion-pair liquid chromatography on a C18 column using methanol as mobile phase solvent, tetrabutylammonium bromide as the ion-pair reagent in pH 3.3 formate buffer solution at a flow rate of 0.9 mL min-1 with a UV detector at 265 nm. No interference of Ca, Mg or NO3 - was detected. Method performance was evaluated in terms of linearity, repeatability and reproducibility. The method detection limit was 5 μg L-1. The contents of EDTA in dairy effluents were 72 ~ 261 μg L-1 at a large dairy site. A change of EDTA concentration was observed downstream of the dairy effluent discharge, but this was well under the predicted no effect concentration for aquatic ecosystem.

Keywords: HPLC, Surface Water, EDTA, dairy wastewater

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