JFS E: Food Engineering and Physical Properties

Extraction and Physicochemical Characterization of Greater Lizardsh (Saurida tumbil) Skin and Bone Gelatin
A. TAHERI, A.M. ABEDIAN KENARI, A. GILDBERG, AND S. BEHNAM
ABSTRACT: Type A gelatins were extracted from skins and bones of lizardfish and analyzed to determine their functional and chemical properties. Lizardfish skin gelatin had ash content of 2.2 0.3% while bone gelatin had ash content of 12.2 0.2%. Gel strength was 159.1 14 and 135 7.9 g, respectively, for skin and bone gelatins compared to 224.3 7.7 g for porcine gelatin. Gelatin from skin exhibited higher viscosity and lower setting time than bone. Skin gelatin had higher imino acid content than bone gelatin. The total imino acid content was 21.71% and 19.83% for skin and bone, respectively. Both skin and bone gelatins contained more chains than and components. Both bone and skin gelatins also contained low molecular weight (< ) peptides. The differences in functional properties between the skin and bone gelatins appeared to be related to differences in amino acid composition and molecular weight distribution of the gelatins. Keywords: gelatin, gel strength, imino acids, lizardfish, viscosity

E: Food Engineering & Physical Properties

Introduction
elatin is a polypeptide obtained by extraction of collagen, which is the major constituent of animal connective tissue. It has found widespread use in the photographic, pharmaceutical and, food industries over the years. Gim enez and others (2005) reported that in the food industry, gelatin is used as an ingredient to enhance the elasticity, consistency, and stability of food products, and therefore good rheological properties are required. Processing of fish leads to enormous amounts of waste. It is estimated that waste from fish processing after filleting accounts for approximately 75% of the total fish weight (Shahidi 1994) and Gom ez-Guill en and others (2002) reported that 30% of the waste is in the form of bones and skins. The fish skins and bones can be processed into gelatin, thus contribute to solve the problem of waste disposal and in addition creating a value-added product. With regard to the occurrence of bovine spongiform encephalopathy (BSE) and the fact that Muslims and Jews advocate abstinence from pork, recently the use of fish skin and bone to process gelatin has gained advocacy. Recently, several studies of gelatins from the skin of various fish species have been published (Gudmundsson 2002; Simon and others 2002; Sadowska and others 2003; Terao and others 2003; Haug and others 2004; Kolodziejska and others 2004; Muyonga and others 2004). The quality of gelatin depends on its physicochemical properties, which are greatly influenced, not only by the species or tissue from which it is extracted, but also by the severity of the manufacturing method (Johnston-Banks 1990). Good rheological properties are required for many applications, such as thickening of sauces and gelling of pate.

In the South East Asian region, most lizardfish is used for dried fish products and as low-quality minced meat for low-priced fishcake products (Morrissey and Tan 2000). Also lizardfish (Saurida tumbil) is one of the major fish used for filleting in Iran by deskinning and deboning. The aim of this study was to extract the gelatin from the skins and bones of the Greater lizardfish (S. tumbil) and to compare their physicochemical characteristics with commercial porcine skin gelatin (Sigma Chemical Co. 2500, St. Louis, Mo., U.S.A.) with high gel strength (300 g).

Materials and Methods

Raw materials
Greater lizardfish (S. tumbil) skin and bone was prepared by Marin Food Industries after filleting in Qazvin, Iran, and stored at 20 C for maximum 10 d until usage. Porcine gelatin from Sigma Co. was used for comparison. The length of fishes was ranged between 40 and 50 cm. Preparation of raw material was done by filleting the fish first by hand, and then the skin of the fillet was separated by a skinner machine and cut in pieces of size 20 to 30 cm. The bone of the fillet was deboned with a deboner machine (TRIO FTC Sweden AB, Arlandastad, Sweden) to segments of size 3 to 10 cm.

Gelatin extraction
Gelatin from fish skins was prepared following the procedure described by Gudmundsson and Hafsteinsson (1997) with slight modifications based on practice of Gim enez and others (2005). Frozen fish skin and bone were thawed at 4 C for about 20 h. The thawed skin was first thoroughly cleaned and rinsed with plenty of water to remove superfluous material and then soaked thrice in 0.2% (w/v) sodium hydroxide solution, each time for 40 min. After washing off NaOH with running tap water, 2 successive acidic incubations were performed, each time 40 min, first in a 0.2% (w/v) sulfuric and then in a 1% (w/v) citric acid solution, each with 3 repetitions. The acidic solutions were drained off and then the samples were washed with
C

MS 20080686 Submitted 9/7/2008, Accepted 1/26/2009. Author Taheri, Abedian Kenari, and Behnam are with Fisheries Dept., Marine Sciences Faculty, Tarbiat Modares Univ., Noor, P .O. Box 64414-356, Noor, Mazandaran, Iran. Author Gildberg is with Nofima Marin, Troms, P .O. Box 6122, 9291 Troms, Norway. Direct inquiries to author Abedian Kenari (E-mail: aabedian@ modares.ac.ir).

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Further reproduction without permission is prohibited

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running tap water until they had about pH 7. The ratio of skin to washing liquid used for each treatment was 1 kg skin (wet weight) to 7 L of acid or alkali solution. The skins were then subjected to a final wash with distilled water to remove any residual matter. The final extraction was carried out in distilled water for 12 h at controlled temperature of 40 to 50 C. The ratio used was 1 kg (weight of wet skin) to 3 L of distilled water. Based on practice of Muyonga and others (2004), the bones used for gelatin extraction were cleaned by scraping with a knife to eliminate some of the flesh and then degreased by tumbling in warm (35 C) water. The degreased bones were then demineralized using 3% HCl at ambient temperature (20 to 25 C) until the bones did not have any hard cores. The acidification solution was changed at 3 d intervals. The time required for complete demineralization was 12 d. The leached bones (ossein) were washed with water until the wash water pH was greater than 4 and reached near 6. This required 6 to 7 washes (ossein to water ratio of 1:2). The pretreated materials were transferred to beakers, covered with warm distilled (60 C) water, and the gelatin was extracted in water baths of 60 C for 12 h. The ratio used was 1 kg (weight of wet bone) to 3 L of distilled water. The clear extract obtained from skin and bone was filtered in a Buchner funnel with an MN filter paper (nr 1; MACHEREY-NAGEL GmbH & Co. KG, Duren, Germany) followed by evaporation in a hot air dryer and then subjected freeze-drying. Protein samples were heat-denatured for 5 min at 90 C and analyzed by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDSPAGE) according to Laemmli (1970) using 3% stacking gels and 5% resolving gels in a Mini Protean II unit (Bio-Rad Laboratories, Hercules, Calif., U.S.A.) at 25 mA/gel. The loading volume was 15 L in all lines. Protein bands were stained with Coomassie brilliant Blue R250. Wide range molecular protein size markers (Fermentas SM0661) were used as a ladder.

Viscoelastic properties
Dynamic viscoelastic studies were performed on a Bohlin CSR-10 rheometer rotary viscometer (Bohlin Instruments Ltd., Gloucestershire, U.K.) using cone-plate geometry (cone angle 4 , gap = 0.15 mm). Cooling and heating, from 40 to 5 C and back to 40 C, was performed at a scan rate of 0.2 C/min, frequency 1 Hz, and oscillating stress of 3 Pa was applied. Dry gelatin powder was dissolved in water to 6.67% dilution (w/v) by heating at 40 C and shaking gently just before starting the test. The elasticity modulus (G ; Pa), viscosity modulus (G ; Pa), and the phase angle ( ; ), were represented as a function of temperature.

Gel strength
The gel strength was determined by the British Standard 757: 1975 method (BSI 1975) on a 6.67% gel (w/v), which was prepared by dissolving the dry gelatin in distilled water at 60 C, and cooling the solution in a refrigerator of 7 to 8 C (maturation temperature) for 16 18 h. Gel strength at 8 to 9 C was determined on a TXA with a load cell of 500 kN, crosshead speed of 0.5 mm/s, and equipped with a 1.27-cm-dia flat-faced cylindrical Teflon plunger. The maximum force in grams (called the Bloom strength) was determined when the plunger had penetrated 4 mm into the gelatin gel.

Determination of pH of raw materials

The pH level of raw fish skin and bone was measured using the British Standard Institution method BSI 757 (1975). Fish skins and bones were chopped and blended in distilled water to form 1% (w/v) suspension. The pH was measured with a glass electrode after standardizing the pH meter (Cyberscan Euteoh.Pc 300, Singapore) with 4 and 7 pH buffers.

Determination of amino acid composition

The amino acid composition was determined following hydrolysis with 6 N HCl for 16 h at 110 C. The amino acids were analyzed following online derivatization and reversed phase high-performance liquid chromatography (RP-HPLC) analysis in an Agilent 1100 (Agilent Technologies, Palo Alto, Calif., U.S.A.) assembly system using a Zorbax 80A C18 column (4.6 i.d. 180 mm; Agilent Technologies) running at 0.5 mL/min.

Determination of pH of gelatin solution

The pH of gelatin solution was measured using the British Standard Institution method, BSI 757, (1975). A 1% (w/v) gelatin solution was prepared in distilled water and cooled to 25 C in a water bath. The pH was measured with a glass electrode after standardizing the pH meter with 4 and 7 pH buffers.

Determination of gelatin recovery

Statistical analysis

Independent t -test analysis was carried out for proximate comThe recovery percentage of gelatin was calculated from the folposition and pHs of lizardfishes, using of computer program SPSSlowing formula: 11.5 for Windows (SPSS Co. Chicago, Ill., U.S.A.). Confidence level Recovery (% ) = protein content of gelatin (g/g) was set at P 0.05. weight of gelatin (g) Results and Discussion 100/protein content of raw material (g/g) weight of raw material used (g)

Approximate composition of raw materials

Approximate compositions of gelatins

The protein content of the fish skins and bones was found to be 26.6 3.3% and 21.3 2.17%, respectively (Table 1). The protein

Approximate analysis of the raw lizardfish was performed acTable 1 --- Proximate composition and pHs of Greater cording to the procedures of the Assn. of Official Analytical lizardsh raw material.A Chemists for moisture (method 24003), ash (method 1821), and Raw material protein (method 24024) (AOAC 1995). Crude fat was determined acComposition (%) Skin Bone cording to the method of Bligh and Dyer (1959). Moisture 69.23 2.9a 43.23 4.8b Protein 26.63 3.3a 21.28 2.17a Electrophoretic analysis a Fat 2.41 0.21 6.57 0.95b Dry gelatin was dissolved in distilled water at 60 C and then 2- Ash 1.44 0.37a 25.06 3.08b fold concentration of loading buffer containing -mercaptoethanol pH 6.28 0.1a 6.05 0.42b was added until a final concentration of 2 mg/mL of gelatin was A Values with standard deviations of triplicate samples are presented; values in reached, as described earlier by Gom ez-Guill en and others (1997). the same row followed by same letter are not signicantly different at = 0.05.
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content of the collagenous material represents the maximum possible yield of gelatin that can be expected (Table 2). There was a significant difference between the skin contents and the gelatin extracted from it (P < 0.05) except in the ash content (Table 3). Also the bone contents and the gelatin extracted from it show a significant difference (Table 4). The ash content of the bone gelatin is very high (11.17 0.19%) which can be due to the low concentration of acid used or a short period of acidification of the bone. Accordingly for forthcoming studies, we recommend that the bone ash content be measured before extraction of gelatin. Other differences are due to the reduction of moisture and lipid during the process of gelatin powder production. The lizardfish skin and bone contained 30.5% and 52.9% dry matter, respectively. The extraction of gelatine from lizardfish skin and bone gave yields of 10.7% and 5.1%, respectively (dry gelatin/wet skin). However, the total yields of skin and bone gelatin on a dry matter basis were 34.9% and 8.9%, respectively. According to Arnesen and Gildberg (2007), reporting gelatin yield as dry gelatin weight compared to the weight of wet skin is common, but not very reliable. Water content may vary because of different treatment of the skin (freezing, salting, scraping, draining, and so on). Therefore, the gelatin yield should be reported rather as the amount of dry gelatin compared to the amount of dry matter in the skin. As reported by Muyonga and others (2004), the yield of gelatin in a 4-stage extraction process of backbones from young and adult Nile perch (Lates niloticus) was 1.3% and 2.4%, respectively. Also, Kolodziejska and others (2008) reported the yield of gelatin as ranging between 43% and 71% in 45 C from backbones of Baltic cod (Gadus morhua) the gelatin was extracted directly from the raw material without preliminary treatment. In our study, a long leaching process was used in preparing gelatin from lizardfish backbones, which process could lead to loss of collagen. Gelatin recoveries of protein from lizardfish skin and bone were 33.85% and 19.55%, respectively (Table 5). Arnesen and Gildberg (2007) notified that the difference in gelatin recovery can be attributed to differences in the extent of intermolecular cross-linking of the collagens. Lizardfish bones generally gave a lower yield of gelatin than skins. The difference may be attributed to 2 factors: a high proportion of flesh attached to the bones compared to the skins and to a higher loss of collagen due to the long leaching process. The flesh mainly consists of noncollagenous material and this was dissolved during the acidification process. Sims and others (2000) proved that 2 kinds of tissue differ in the type and quantities of cross-links. Extractability instance at low temperature is expected to be higher if the collagen is less crosslinked. The bone gelatin had a higher content of ash in comparison to that of skin gelatin (Table 2). Muyonga and others (2004) reported that Nile perch bone gelatin had much higher ash content (mostly in the range 3% to 10%) than the skin gelatin, indicating that the leaching process was inadequate. They are suggesting, therefore, that the manufacture of fish bone gelatin may require improvement of the leaching process, by adding an ion exchange step to remove the salts, for example, by application of a counter-current process.

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Table 2 --- Proximate composition and pHs of gelatine obtained from Greater lizardsh.A Gelatine Composition (%) Moisture Protein Fat Ash pH Skin 10.07 0.42 83.94 0.15a 0.03 0.01a 1.98 0.25a 3.68 0.12a
a

pH determination
The pH of the lizardfish skin and bone gelatin was 3.7 0.1 and 4.1 0.1, respectively, and that of raw material was 6.3 0.1 for skin and 6.1 0.4 for bone. The pH of lizardfish skin gelatin is significantly different (P < 0.05) from the pH of the gelatin from porcine (4.8 0.1), but the pH of the lizardfish bone gelatin was similar to that of porcine gelatin. This difference in pH of the gelatins may also be due to the type and strength of acids employed during the extraction procedures (Table 1).

Bone 8.27 0.1a 81.89 1.01a 0.01 0.005a 11.17 0.19b 4.15 0.006a

A Values with standard deviations of triplicate samples are presented; values in the same row followed by same letter are not signicantly different at = 0.05.

Amino acid composition

Table 6 shows the amino acid (a.a) composition of the gelatins from lizardfish skin and bone and from porcine. The gelatin from Table 3 --- Proximate composition and pHs of Greater lizardfish skin and bone had a 4.9% and 6.8% lower molar conlizardsh skin and gelatine. tent of imino acids, respectively, than porcine gelatin. Apart from Skin this, the only significant difference was that lizardfish gelatin had Composition (%) Raw material Gelatine a higher content of glysine and serine. Probably the hydroxyl side Moisture 69.23 2.9a 10.07 0.42b chains of serine and other hydroxylated amino acids play an imProtein 26.63 3.3b 83.94 0.15a portant role in the generation of hydrogen bonds and helical strucFat 2.41 0.21a 0.03 0.01b tures during the storage for gel strengthening. Skin and bone gelatin Ash 1.44 0.37a 1.98 0.25a pH 6.28 0.1a 3.68 0.12b showed a slightly lower imino acid content (21.3% and 19.8%, respectively) compared to porcine gelatin (26.7%). Hyp is believed to A Values with standard deviations of triplicate samples are presented; values in play a singular role in the stabilization of the triple-stranded colthe same row followed by same letter are not signicantly different at = 0.05. lagen helix due to its hydrogen bonding ability through its OH Table 4 --- Proximate composition and pHs of Greater group (Burjandze 1979; Ledward 1986). Johnston-Banks (1990) reported that the imino acids (proline and hydroxyproline) impart lizardsh bone and gelatine. considerable rigidity to the collagen structure and that a relatively Bone limited imino acid content should result in a less sterically hindered Composition (%) Raw material Gelatine
Moisture Protein Fat Ash pH
A

43.23 4.8a 21.28 2.17b 6.57 0.95b 25.06 3.08b 6.05 0.42a

8.27 0.1b 81.89 1.01a 0.01 0.005a 11.17 0.19a 4.15 0.006b

Table 5 --- Yield and extraction recovery of lizardsh skin and bone gelatine (%). Based on wet weight Skin Bone 10.74 5.08 Based on dry weight 34.92 8.95 Recovery 33.85 19.55

Values with standard deviations of triplicate samples are presented; values in the same row followed by same letter are not signicantly different at = 0.05.

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helix and may affect the dynamic properties of the gelatin. Moreover, lizardfish gelatin showed a lower content of Ala. This amino acid, together with Pro and Hyp, is found in nonpolar regions where sequences of the type Gly-Pro-Y predominate (Ledward 1986). A higher content of these amino acids, that is, Pro, Hyp, and Ala, in commercial gelatin from porcine is one of the major causes responsible for its higher viscoelastic properties. The a.a profile obtained was from an acid hydrolysate. A proportion of the acidic amino acids occur as the sidechain amides glutamine and asparagine in collagen (Ward and Courts 1977). During acid hydrolysis of gelatin, some of the glutamine and asparagine will be converted into the acidic forms, that is, glutamic acid and aspartic acid, respectively. decrease of -component and an almost disappearance of higher molecular weight polymers was observed. The proportion of low molecular weight (< ) fractions (peptides) was higher in the molecular weight profile of bone which proved that bone gelatins have a higher content of low molecular weight peptides than skin gelatins. This difference of molecular weight distribution in bone and skin gelatin may be responsible for the lower viscosity, setting, and melting temperature for the lizardfish skin and bone gelatin as compared to porcine gelatin. According to Tavernier (1989), the high incidence of low molecular weight peptides is associated with low viscosity, melting point, setting point and high setting time. These experiments were done with freeze-stored lizardfish skin and bone. Fern andez-D az and others (2003) explained that tissue damage occurred by ice-crystal formation during the freezing process. Therefore, the more covalently cross-linked collagen makes the extraction and solubility of -chain dimers and trimers and high molecular weight polymers more difficult, and on the contrary, smaller collagen fractions could be more easily extracted. This is of great importance since, as reviewed by Johnston-Banks (1990), the average molecular weight of a gelatin is largely responsible for its gelling behavior. Also, in this study molecular weights of skin and bone gelatin have a wide distribution. According to Yau and others (1979), wide molecular weight distribution negatively affects some functional properties of macromolecules. The difference between the properties of the fish gelatins and porcine gelatin may be attributed partly to differences in molecular weight distribution. Muyonga and others (2004) reported that Nile perch skin gelatin was generally found to contain higher proportion of the fractions than the Nile perch bone gelatin. They concluded that the bone gelatin had a consistently higher incidence and/or stability of cross-links than found in the skin collagen, resulting in more cleavage of peptide bonds during the manufacture of bone gelatins.

Molecular weight distribution

To characterize differences in molecular weight, gelatin preparations from lizardfish skins and bones were analyzed by polyacrylamide gel electrophoresis in the presence of SDS (Figure 1). Lizardfish skin and bone gelatins demonstrated by sharp appearance in electrophoresis a notable amount of -chains, but there was less content of and -components, while a strong
Table 6 --- Amino acid composition of gelatine from skin and bone of lizardsh and porcine gelatine.a Amino acid Ala Arg Asp Glu Gly His Hyp Ile Leu Lys Met Phe Pro Ser Tyr Thr Val
a

Skin 9.58 5.34 5.36 7.3 30.34 0.9 9.9 0.85 1.92 2.13 1.35 1.4 11.81 3.46 0.3 1.53 1.8

Bone 9.1 5.29 5.36 7.23 30.34 0.9 8.13 0.85 1.9 2.1 1.35 1.4 11.7 3.4 0.3 1.53 1.84

Porcine 11.1 7.27 5.04 10.16 20.98 0.64 13.96 1.13 2.66 3.36 0.77 2.15 12.7 2.91 0.58 1.71 2.35

Viscoelastic properties
Dry gelatins were dissolved in distilled water at 40 C and their viscoelastic properties upon gelling and subsequent melting were compared. Two independent parameters were obtained from dynamic measurements: the storage modulus (G ) describing the amount of energy that is stored elastically in the structure and the loss of modulus (G ) indicating amount of energy loss or the viscous response. Phase angle ( ) is a measure of how much the stress and strain are out of phase. Figure 2 shows thermal transitions by changes in the phase angle ( ) of dissolved gelatins during cooling (40 to 7 C) and subsequent melting (7 to 40 C). The onset of gelling in the dissolved skin and bone gelatin took place at around 14 C. The subsequent melting curves revealed that gelatins started melting at around 17 C. Porcine gelatins gel at a high temperature of 26 C and melt at 33 C. The minimal presence of dimers and trimers of -chains in the lizardfish skin and bone gelatin, as noted earlier in the electrophoretic analysis, suggests that refolding could be more the result of a random association of individual -chains, rather than a partial reformation of collagen native structure (Ledward 1986). The thermal stability of a gelatin gel has been shown to be directly correlated to the number and stability of Pro-rich regions in the collagen or gelatin molecules (Ledward 1986). The imino acid content, as well as the pro-hydroxylation degree, has been found to be significantly lower in lizardfish skin and bone than in the better gelling porcine gelatins resulting in poorer viscoelastic properties. However, Gudmundsson and Hafsteinsson (1997) reported that the viscosity of the gel may be mainly due to the molecular weight distribution rather than the amino acid composition of the gelatin. On the other hand, the extraction procedure should
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Figures show percent of amino acids in hydrolyzed gelatine samples.

components
200 150 120 100 85 kd 70 kd 60 kd 50 kd

components -chains

Low molecular weight components

Figure 1 --- Electrophoretic analysis of the gelatine reparations. Dry gelatine preparations extracted from lizardsh bone (b), skin (c), porcine (d) were analyzed by SDS-PAGE on 5% gels in the presence of -mercaptoethanol. Fermentase wide range molecular weight marker was used as ladder.

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not be ignored. Gelatin is subjected to chemical hydrolysis during extraction at acidic conditions. Hence, the average molecular weight will be reduced by increasing extraction time, temperature, and acidity (Arnesen and Gildberg 2007). It should also be realized that only a partial extraction of denatured collagen molecules is achieved and thus, the material obtained after the gelatin extraction procedure may not present the same properties regarding aggregation or even amino acid composition as the material present in the intact skin. In the present study, the electrophoretic analysis of the extracted gelatins (Figure 1) reveals differences regarding the molecular weight distribution, mainly affecting the collagen chains which were notably higher than the - and -components.

Gel strength
Figure 3 shows the gel strength of skin and bone gelatin and porcine gelatin after maturation overnight at 7 to 8 C. Gel strength

90 80 70 Phase angle(rad) 60 50 40 30 20 10 0 5 10 15 20 25 30 35 40 Temperature(C) Porcine gelatine Lizardfish skin gelatine Lizardfish bone gelatine

Figure 2 --- Viscoelastic properties upon cooling and heating of gelatine preparations. Dry gelatines were dissolved at 6.67% (w/v) at 40 C. Changes in phase angle were monitored during cooling from 40 to 5 C and subsequent heating from 5 to 40 C.

Phase angle (rad)

Gel strenght (g)

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90 80 70 60 50 40 30 20 10 0 5 10 15 20 25 30 35 40 Temperature(C) Lizardfish bone gelatine Lizardfish skin gelatine Porcine gelatine

250 200 150 100 50 0 skin gelatine

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Figure 3 --- Gel strength of gels made from lizardsh skin and bone and porcine gelatines at 10 C.

bone gelatine

porcin gelatine

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is the most important attribute of gelatin and determines the quality of produced gelatin. Skin gelatin showed higher gel strength (159.1 g) than the bone gelatin (135 g) and both of them are significantly different (P < 0.05) from porcine gelatin gel strength (224.3 g). This may be due to the higher content of hydroxyproline in porcine gelatin (see Table 3) According to Arnesen and Gildberg (2002), the low hydroxyproline content in fish gelatin was a major reason for the low gel strength of this gelatins. Also lower gel strength in this study can be explained based on the findings of Gom ez-Guill en and others (1997) study to be due to the lower percentage of - and -components as compared to individual chains in lizardfish gelatin, a longer maturation time may be required to allow for the growth of the existing nucleation sites. Also, the presence of protein degradation fragments in this preparation, according to Ledward (1986) and Normand and others (2000) may reduce the ability of -chains to anneal correctly during stabilization overnight and thus hindering the growth of the existing nucleation sites. In a resembling study of Muyonga and others (2004), it was reported that there was no significant difference (P > 0.05) between the viscosity of young Nile perch skin and the Nile perch bone gelatins. But Gel hardness was found to be higher for Nile perch skin than for bone gelatins.
Arnesen JA, Gildberg A. 2007. Extraction and characterisation of gelatine from Atlantic salmon (Salmo salar ) skin. Bioresour Technol 98:537. Bligh EG, Dyer WJ. 1959. A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37(8):9117. [BSI] British Standard Institution. 1975. Methods for sampling and testing gelatine (physical and chemical methods). London, U.K.: BSI. Burjandze TV. 1979. Hydroxy-proline content and location in relation to collagen thermal stability. Biopolymers 18:9316. Fern andez-D az MD, Montero P, G omez-Guill en MC. 2003. Effect of freezing fish skins on molecular and rheological properties of extracted gelatine. Food Hydrocoll 17:2816. Gim enez B, Gom ez-Guill en MC, Montero P. 2005. Storage of dried fish skins on quality characteristics of extracted gelatine. Food Hydrocoll 19:958 63. Gom ez-Guill en MC, Turnay TF, Fernandez-Diaz MD, Ulmo N, Lizarbe MA, Montero P. 2002. Structural and physical properties of gelatine extracted from different marine species: a comparative study. Food Hydrocoll 16:2534. Gudmundsson M. 2002. Rheological properties of fish gelatine. J Food Sci 67:2172 6. Gudmundsson M, Hafsteinsson H. 1997. Gelatine from cod skins as affected by chemical treatments. J Food Sci 62:379. Haug IJ, Draget KI, Smidsrd O. 2004. Physical and rheological properties of fish gelatine compared to mammalian gelatine. Food Hydrocoll 18:20313. Johnston-Banks FA. 1990. Gelatine. In: Harris, P, editor. Food gels. London, U.K.: Elsevier Applied Science. p 23389. Kolodziejska I, Kaczorowski K, Piotrowska B, Sadowska M. 2004. Modification of the properties of gelatine from skins of Baltic cod (Gadus morhua) with transglutaminase. Food Chem 86:2039. Koodziejska I, Skierka EZ, Sadowska M, Koodziejski W, Niecikowska C. 2008. Effect of extracting time and temperature on yield of gelatine from different fish offal. Food Chem 107:7006. Laemmli UK. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:6805. Ledward DA. 1986.Gelation of gelatine. In: Mitchell JR, Ledward DA, editors. Functional properties of food macromolecules. London, U.K.: Elsevier Applied Science. p 171201. Morrissey MT, Tan SM. 2000. World resources for surimi. In: Park JW, editor. Surimi and surimi seafood New York: Marcel Dekker Inc. p 121. Normand V, Muller S, Ravey JC, Parker A. 2000. Gelation kinetics of gelatine: a master curve and network modelling. Macromolecules 33:10631071. Muyonga JH, Cole CGB, Duodu KG. 2004. Extraction and physico-chemical characterisation of Nile perch (Lates niloticus) skin and bone gelatine. Food Hydrocoll 18:58192. Sadowska M, Kolodziejska I, Niecikowska C. 2003. Isolation of collagen from the skins of Baltic cod (Gadus morhua). Food Chem 81:25762. Shahidi F. 1994. Seafood processing by-products. In: Shahidi F, Botta JR, editors. Seafoods chemistry, processing, technology and quality. Glasgow: Blackie Academic & Professional. p 32034. Simon A, Vandanjon L, Levesque G, Bourseau P. 2002. Concentration and desalination of fish gelatine by ultrafiltration and continuous diafiltration processes. Desalination 144:3138. Sims JT, Avery NC, Bailey AJ. 2000. Quantitative determination of collagen crosslinks. In Streuli C, Grant M, editors. Methods in molecular biology. Extracellular matrix protocols, Totowa, N.J.: Humana Press. Vol. 139. p 1126. Tavernier BH. 1989. Molecular mass distribution of gelatine and physical properties. Photogr Gelatin Proc 1:21728. Terao K, Nagasawa N, Nishida H, Furusawa K, Mori Y, Yoshii F, Dobashi T. 2003. Reagent-free cross-linking of aqueous gelatine: manufacture and characteristics of gelatine irradiated with gamma- ray and electron beam. J Biomater Sci, Polym Edn 14:1197208. Ward AG, Courts A. 1977. The science and technology of gelatine. New York, London, U.K.: Academic Press. p 10936. Yau WW, Kirkland JJ, Bly DD. 1979. Modern size-exclusion liquid chromatography practice of gel permeation and gel filtration chromatography. New York: Wiley. 479 p.

Conclusions

here are considerable differences between extractability and yield of gelatin from lizardfish skins and bones. Lizardfish skin and bone gelatins also differ in their functional properties and molecular weight distribution. As a result, lizardfish bone gelatin consists of a high proportion of low molecular weight fractions, which are associated with poor gelling properties. There is a potential for exploitation of lizardfish processing waste for gelatin extraction. The potential is higher for lizardfish skins than for bones because lizardfish skins give higher gelatin yield and the skin gelatin exhibits better functional properties than lizardfish bone gelatin. Additional pretreatment such as liming may be required to obtain bone gelatin with better functional properties.

Acknowledgments
Financial support from The Tarbiat Modarres Univ. is acknowledged. The authors thank Ellen V. Tavakoli for editing the English text.

References
[AOAC] Assn. of Official Analytical Chemists. 1995. Official methods of analysis. 16th ed. Washington, D.C.: AOAC. Arnesen JA, Gildberg A. 2002. Preparation and characterisation of gelatine from the skin of harp seal (Phoca groendlandica). Bioresour Technol 82:1914.

Vol. 74, Nr. 3, 2009JOURNAL OF FOOD SCIENCE

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E: Food Engineering & Physical Properties