Characteristics of collagen from the skin of clown featherback (Chitala ornata)

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) from the skin of clown featherback (Chitala ornata) were isolated and characterised. Yields of ASC and PSC were 27.64 and 44.63% (dry weight basis) with total collagen recovery of 82.08%. Both collagens contained glycine as the major amino acid with relatively high content of proline, hydroxyproline and glutamic acid/glutamine. Nevertheless, they had the low content of cysteine, histidine and tryrosine. The collagen was characterised as type I, comprising (α1)₂α2‐heterotrimer. Pepsin‐aided process did not affect triple‐helical structure of PSC as determined by FTIR spectra. Thermal transition temperature of ASC (36.28 °C) was slightly higher than that of PSC (35.23 °C). However, no differences in isoelectric point (5.54–5.68) between ASC and PSC were observed. Therefore, collagen from the skin of clown featherback could be successfully extracted for further applications.     

Characteristics of acid soluble collagen and pepsin soluble collagen from scale of spotted golden goatfish (Parupeneus heptacanthus)

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) were extracted from scale of spotted golden goatfish (Parupeneus heptacanthus) with the yields of 0.46% and 1.20% (based on dry weight basis), respectively. Both ASC and PSC were characterised as type I collagen, containing α₁ and α₂ chains. β and γ components were also found in both collagens. Based on FTIR spectra, the limited digestion by pepsin did not disrupt the triple helical structure of collagen. ASC and PSC contained glycine (336–340 residues/1000 residues) as the major amino acid and had imino acids of 186–189 residues/1000 residues. Maximal transition temperatures (T_max) were 41.58 and 41.01 °C for ASC and PSC, respectively. From zeta potential analysis, net charge of zero was found at pH 4.96 and 5.39 for ASC and PSC, respectively. Both collagens exhibited high solubility in acidic pH (2–4) and were soluble in the presence of NaCl at concentration up to 20 and 30 g/l for ASC and PSC, respectively.     

Isolation and characterization of collagen from bigeye snapper (Priacanthus macracanthus) skin

Acid‐solubilized collagen (ASC) and pepsin‐solubilized collagen (PSC) were isolated from the skin of bigeye snapper (Priacanthus macracanthus) with yields of 64 and 11 g kg^?1 wet weight, respectively. Both ASC and PSC were characterized as type I collagens with no disulfide bonds. Peptide maps of ASC and PSC digested by V8 protease and lysyl endopeptidase showed some differences in peptide patterns and were totally different from those of calf skin collagen. The maximum solubility was observed at pH 4 and 5 for ASC and PSC, respectively. A sharp decrease in solubility of both collagens in acetic acid was found with NaCl concentration above 30 g l^?1. Thermal transitions of ASC and PSC in deionized water were observed with T_max of 30.37 and 30.87 °C, respectively, and were lowered in the presence of acetic acid (0.05 mol kg^?1 solution). Therefore, ASC was a major fraction in bigeye snapper skin and it exhibited some different characteristics to PSC. Copyright © 2005 Society of Chemical Industry     

Isolation and characterization of acid and pepsin-solubilized collagens from the skin of balloon fish (Diodon holocanthus)

Acid-solubilized collagen (ASC) and pepsin-solubilized collagen (PSC) were successfully extracted from the skin of balloon fish (Diodon holocanthus) with yields of 4% and 19.5% respectively (based on dry weight). According to the electrophoretic patterns, both the ASC and PSC consisted of two different α chains (α1 and α2), were characterized to be type I, and contained imino acid of 179 and 175 residues/1000 residues, respectively. The PSC had a lower content of high-molecular weight cross-links than the ASC. The ultraviolet (UV) absorption spectrum of collagens showed that the distinct absorption was between 210 and 230 nm. A maximum solubility in 0.5 M acetic acid was observed at pH 1–5, and a sharp decrease in solubility above 4% (w/v) in both the ASC and PSC was observed in the presence of NaCl. The denaturation temperatures (T_d) of the ASC and PSC measured by viscometry were 29.01 °C and 30.01 °C, respectively. The maximum temperatures (T_max) of the ASC and PSC were 29.64 °C and 30.30 °C, respectively.     

Isolation and characterization of acid soluble collagens and pepsin soluble collagens from the skin and bone of Spanish mackerel (Scomberomorous niphonius)

The objective of this study was to extract and characterize acid soluble collagens (ASCs) and pepsin soluble collagens (PSCs) from the skin and bone of Spanish mackerel (Scomberomorous niphonius) and to provide a simultaneous comparison of the four collagens. The yields of ASC-S (ASC from skin), PSC-S (PSC from skin), ASC-B (ASC from bone) and PSC-B (PSC from bone) were 13.68 ± 0.35, 3.49 ± 0.24, 12.54 ± 0.83 and 14.27 ± 0.66% (on wet weight basis), respectively. The four collagens contained glycine (341.6–352.6 residues/1000 residues) as the major amino acid and the contents of imino acids were between 177.1 and 184.3 residues/1000 residues. Amino acid composition, SDS-PAGE and FTIR confirmed that ASC-S, ASC-B and PSC-B were mainly composed of type I collagen with slight molecular structure differences, and PSCs had lower content of high-molecular weight cross-links than that of ASCs. The denaturation temperatures of ASC-S, PSC-S, ASC-B and PSC-B were 15.12, 14.66, 18.02 and 16.85 °C, respectively, which were much lower than those of collagens from the mammalian and tropical fish species due to low imino acid contents. All the collagens were soluble at acidic pH (1–4) and lost their solubility when the NaCl concentrations were above 2% (w/v). The four lyophilized collagens displayed a uniform and regular network ultrastructure based on the ultrastructural analysis. The isolated collagens from Spanish mackerel could serve as an alternative source of collagens for further application in food and neutraceutical industries.     

Isolation and characterisation of collagen extracted from the skin of striped catfish (Pangasianodon hypophthalmus)

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) from the skin of striped catfish (Pangasianodon hypophthalmus) were isolated and characterised. The yields of ASC and PSC were 5.1% and 7.7%, based on the wet weight of skin, respectively, with the accumulated yield of 12.8%. Both ASC and PSC comprising two different α-chains (α1 and α2) were characterised as type I and contained imino acid of 206 and 211 imino acid residues/1000 residues, respectively. Peptide maps of ASC and PSC hydrolysed by either lysyl endopeptidase or V8 protease were slightly different and totally differed from those of type I calf skin collagen, suggesting some differences in amino acid sequences and collagen structure. Fourier transform infrared (FTIR) spectra of both ASC and PSC were almost similar and pepsin hydrolysis had no marked effect on the triple-helical structure of collagen. Both ASC and PSC had the highest solubility at acidic pH. A loss in solubility was observed at a pH greater than 4 or when NaCl concentration was higher than 2% (w/v). T_max of ASC and PSC were 39.3 and 39.6 °C, respectively, and shifted to a lower temperature when rehydrated with 0.05 M acetic acid. Zeta potential studies indicated that ASC and PSC exhibited a net zero charge at pH 4.72 and 5.43, respectively. Thus, ASC and PSC were slightly different in terms of composition and structure, leading to somewhat different properties.     

Physicochemical properties of acid-soluble collagens from different tissues of large yellow croaker (Larimichthys crocea)

Acid-soluble collagens (ASCs) were extracted and characterised from different tissues of large yellow croaker (Larimichthys crocea). The yields of ASCs in bones, skins, scales and muscles were 3.22 ± 0.47%, 42.30 ± 1.15%, 2.82 ± 0.31% and 0.89 ± 0.12%, respectively. SDS-PAGE and ATR-FTIR analysis showed that all ASCs were type I collagen with intact triple helical structure and consisted of α1,3-chain, α2-chain, β-chain and γ-chain. Quantitative analysis of SDS-PAGE revealed that the ratio of β-chains in ASC from scales was lower than other tissues. The imino acid contents of ASC from bones, skins, scales and muscles were 169, 167, 162 and 173 residues/1000 residues, respectively. The maximum transition temperature (T_m) of ASC from scales was 30.15 °C, lower than the other three ASCs, indicated that the thermal stability of collagen was affected by both the content of imino acid and β-chains. The results also implied that thermal stability of ASC might exist the tissue specificity among different fish species.     

Collagens from the skin of arabesque greenling (Pleurogrammus azonus) solubilized with the aid of acetic acid and pepsin from albacore tuna (Thunnus alalunga) stomach

BACKGROUND: Due to the low extraction efficiency of collagen from fish skin by the typical acid solubilization process, pepsin has been widely used to aid further extraction of collagen from the residue. The aim of this study was to characterize collagen from the skin of arabesque greenling extracted with the aid of albacore tuna pepsin, in comparison with collagen obtained from the acid solubilization process. RESULTS: Acid‐solubilized collagen (ASC) from the skin of arabesque greenling was extracted with acetic acid. Pepsin‐solubilized collagen (PSC) was further extracted from the skin residue with the aid of pepsin from albacore tuna. The yields of ASC and PSC were 303 and 140 g kg^?1 (dry weight), respectively. Both collagens contained α‐ and β‐chains as their major components and were characterized as type I collagen. Both collagens contained glycine as a major amino acid and had imino acid content of 157–159 residues per 1000 residues. The degradation induced by lysyl endopeptidase and V8‐protease was more pronounced in PSC compared with ASC. Maximal transition temperatures of both collagens were in the range of 15.4–15.7 °C. Fourier transform infrared spectra revealed some differences in molecular order between ASC and PSC. Nevertheless, the triple‐helical structure of PSC was still predominant. Based on ζ‐potential, pI of ASC and PSC was estimated to be 6.31 and 6.38, respectively. CONCLUSION: Isolation of collagens from the skin of arabesque greenling could be achieved by acid or albacore tuna pepsin solubilization. However, there was a slight difference in properties between ASC and PSC. Copyright © 2010 Society of Chemical Industry     

Isolation and characterization of collagen from the cartilages of brownbanded bamboo shark (Chiloscyllium punctatum) and blacktip shark (Carcharhinus limbatus)

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) from the cartilages of brownbanded bamboo shark (BBS; Chiloscyllium punctatum) and blacktip shark (BTS; Carcharhinus limbatus) were isolated and characterized. ASC and PSC extracted from BBS cartilage showed the yields of 1.27 and 9.59 g/100 g (Based on dry weight), respectively, while yields of ASC and PSC from BTS cartilage were 1.04 and 10.30 g/100 g (based on dry weight), respectively. All collagens had protein as a major constituent with the trace amount of ash and fat. They contained glycine as the major amino acid with high contents of alanine, proline and hydroxyproline. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoretic patterns and subunit compositions, all collagens more likely comprised 2 types of collagen, type I and II, and contained α- and β-chains as the major components. Peptide maps of those collagens from both species digested by V8-protease and lysyl endopeptidase were different and were completely different from those of type I collagen from calf skin. Thermal transition temperature of ASC from those collagens (36.28-36.73 °C) was slightly higher than their corresponding PSC (34.56-35.98 °C). From zeta potential analysis, isoelectric points (pI) of collagen from the cartilages of BBS and BTS were estimated to be from 6.53 to 7.03 and from 6.96 to 7.26, respectively. Fourier transform infrared (FTIR) spectra of both ASC and PSC were quite similar, suggesting that pepsin hydrolysis did not affect the secondary structure of collagen, especially triple-helical structure.     

Isolation and Characterisation of collagen from the skin of brownbanded bamboo shark (Chiloscyllium punctatum)

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) from the skin of brownbanded bamboo shark (Chiloscyllium punctatum) were isolated and characterised. The yield of ASC and PSC were 9.38% and 8.86% (wet weight basis), respectively. Based on protein patterns and TOYOPEARL^® CM-650M column chromatography, both collagens contained α- and β-chains as their major components. These were characterised as type I collagen with the cross-link of α2-chain. As digested by V8-protease and lysyl endopeptidase, peptide maps of both ASC and PSC were similar, but differed from that of type I collagen from calf skin. Fourier transform infrared (FTIR) spectra of both collagens were similar and pepsin hydrolysis had no effect on triple-helical structure of collagen. Transition temperature (T_max) of ASC and PSC were 34.45 and 34.52 °C, respectively, as determined by differential scanning colorimetry (DSC). From zeta potential study, the isoelectric points of ASC and PSC were estimated to be 6.21 and 6.56, respectively. Therefore, the skin of brownbanded bamboo shark could serve as an alternative source of collagen for different applications.     

Study on the self‐assembly property of type I collagen prepared from tilapia (Oreochromis niloticus) skin by different extraction methods

Type I collagen was prepared from tilapia (Oreochromis niloticus) skin by acetic acid and pepsin process at 4 °C, respectively (ASC and PSC), and hot‐water method separately at 25, 35 and 45 °C (C‐25, C‐35 and C‐45). Their structure and self‐assembly property were discussed. SDS‐PAGE patterns suggested that pepsin hydrolysis and the 35 and 45 °C extraction produced collagen with much reduced proportions of α‐ and β‐chains. Fourier transform infrared spectroscopy spectra revealed that pepsin hydrolysis did not change the conformation of collagen, but higher extraction temperature did. Self‐assembly curves and atomic force microscopy (AFM) observations showed that only ASC, PSC and C‐25 could self‐assemble into fibrils with D‐periodicity, but the reconstruction rate of C‐25 was lower. Besides, PSC had relatively higher resolution ratio compared with others. Overall, pepsin‐extracted collagen displayed higher solubility and better fibril‐forming capacity, having the potential of applying in biomaterials and food‐packaging materials.     

Isolation and characterisation of collagens from the skin of largefin longbarbel catfish (Mystus macropterus)

Acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) were extracted from the skin of largefin longbarbel catfish (Mystus macropterus) with yields of 16.8% and 28.0%, respectively, on the basis of dry weight. Both ASC and PSC contained α1 and α2 chains and the amino acid composition of collagen was close to that of calf skin type ? collagen. The intrinsic viscosities of ASC and PSC were 14.9 dl/g and 14.5 dl/g, respectively. Similar ultraviolet and FTIR spectra of ASC and PSC were observed. However, peptide maps of ASC and PSC, hydrolysed by trypsin, revealed some differences in primary structures between the two fractions. Denaturation temperatures of ASC and PSC were 32.1 °C and 31.6 °C, respectively. The higher T_m showed that it is possible to use largefin longbarbel catfish skin collagen as an alternative source of vertebrate collagens for industrial purposes.     

Detection of five novel GMO maize events by qualitative, multiplex PCR and fluorescence capillary gel electrophoresis

Acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) from the skin of blacktip shark (Carcharhinus limbatus) were isolated and characterized. The yield of ASC (20.01%) was much higher than that of PSC isolated from the residue of ASC extraction (0.86%). Both collagens had protein as their major constituent with the trace amounts of ash and fat. Based on protein patterns and TOYOPEARL^® CM-650M column chromatography, both collagens contained α- and β-chains as their main components and were characterized as type I collagen with the cross-link of α2-chain. Similar peptide maps of both collagens, digested by either V8 protease or lysyl endopeptidase, were observed but they were totally different from those of type I collagen from calf skin hydrolyzed by the same enzyme. Thermal transition temperature (T _max) of ASC and PSC were 34.23 and 34.37 °C, respectively. Fourier-transform infrared spectra suggested that both collagens were in triple-helical structure. From zeta potential analysis, isoelectric points (pI) of ASC and PSC were estimated to be 6.78 and 7.02, respectively. Thus, blacktip shark skin may serve as an alternative source of collagen and acid solubilization process could be implemented with ease and high yield.     

Distinctive characteristics of collagen and gelatin extracted from Dosidicus gigas skin

Squid skin, often discarded as processing by-product, is a good resource of collagen/gelatin. In this study, acid soluble collagen (ASC), pepsin soluble collagen (PSC) and water soluble gelatin (WSG) were extracted from squid (Dosidicus gigas) skin and physicochemically examined. The lowest yield of 33.5% was obtained for ASC extracted at 4 °C, and the addition of pepsin increased the collagen yield by around 35.0% (PSC). The highest yield of 81.9% (WSG) was achieved by thermal extraction at 60 °C. A low temperature can largely retain the native helix structures of ASC and PSC, contrariwise, thermal treatment converted collagen into gelatin with unordered and renatured structures. The proline and hydroxyproline contents of ASC, PSC and WSG were 183/1000 residues, 194/1000 residues and 175/1000 residues, respectively. In addition, WSG showed a denaturation temperature at 80.7 °C which was much higher than that of ASC (24.2 °C) and PSC (26.2 °C), while a significant lower resistance towards enzymatic digestion.     

Biochemical properties of bone and scale collagens isolated from the subtropical fish black drum (Pogonia cromis) and sheepshead seabream (Archosargus probatocephalus)

Acid-soluble collagen (ASC) and pepsin-solubilized collagen (PSC) were isolated from the bones and scales of black drum (Pogonia cromis) and sheepshead seabream (Archosargus probatocephalus) caught in the Gulf of Mexico. ASC and PSC were analyzed for molecular weight by SDS–PAGE, amino acid composition, secondary structure, and denaturation temperature. The molecular masses of the collagen subunits were about 130 kDa for α₁ and 110 kDa for α₂, respectively. The amino acid composition of the PSCs was closer to that of calf skin ASC than to that of cod skin ASC. The melting temperatures of ASC and PSC were >34 °C. Intrinsic viscosity of the PSCs was similar to the intrinsic viscosity of collagen from fish species such as hake, cod, and catfish. Black drum and sheepshead bone and scale collagens were typical type-I collagens and may find applications in the functional food, cosmetic, biomedical, and pharmaceutical industries.     

Temperature effects on type I pepsin‐solubilised collagen extraction from silver‐line grunt skin and its in vitro fibril self‐assembly

BACKGROUND: Fish skin is a potential source of collagen. Increasing the extraction temperature increases the yield of collagen. However, it may also result in degradation of the peptide chains, thus damaging the 3D structure of collagen that is vital for its application as a biomaterial. This work investigated the effects of extraction temperature on the yield and characteristics, including fibril self‐assembly, of type I pepsin‐solubilised fish skin collagen. RESULTS: Pepsin‐solubilised collagens were extracted from fresh skin of silver‐line grunt at 4, 10, 20 and 28 °C for 6 h. Extraction at 10 °C gave the highest yield of collagens (439.32 ± 96.43 mg g^?1 fresh skin, dry basis), which were identified as type I and comprised β, α1 and α2 subunits. Extraction at higher temperatures (20 and 28 °C) resulted in the formation of low‐molecular‐weight peptide fragments, thus reducing the yield of the resultant type I collagen. The denaturation temperatures of collagens extracted at 4 and 10 °C, as determined by thermal analysis using differential scanning calorimetry, were 39.5 and 37.5 °C respectively. In vitro fibril self‐assembly of 1 mg mL^?1 collagen solution (pH 6) incubated at 25 °C was only observed with collagens extracted at 4 and 10 °C. The 10 °C collagen not only showed a higher rate of self‐assembly, but its matrix also had a larger fibril diameter of 0.50 ± 0.07 µm (compared with 0.41 ± 0.07 µm for the 4 °C collagen) after 4 h of incubation. CONCLUSION: The results indicated strong effects of extraction temperature on the yield and characteristics of the collagen obtained. Extraction of pepsin‐solubilised collagen from silver‐line grunt skin at 4–10 °C gave a high yield of type I collagen with molecular integrity suitable for tissue‐engineering applications. Copyright © 2010 Society of Chemical Industry     

Isolation and Characterization of Collagen from the Body Wall of Sea Cucumber Stichopus monotuberculatus

To exploit a new collagen resource from the body wall of tropical sea cucumber, pepsin‐solubilized collagen of Stichopus monotuberculatus (PSC‐Sm) was isolated and characterized with UV‐vis spectra, sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE), amino acid composition, enzyme‐digested peptide maps, Fourier transform infrared spectroscopy (FTIR), maximum transition temperature (T_m), and solubilities. The maximum absorbance of PSC‐Sm was exhibited at 218 nm in UV‐vis spectra. The triple helical structure and activity of PSC‐Sm could be indicated by FTIR. SDS‐PAGE showed that the triple helix of PSC‐Sm was formed as (α₁)₃ by 3 α₁ chain homologous with molecular weight of 137 kDa. The T_m of PSC‐Sm and calf skin collagen (CSC) were 30.2 and 35.0 ºC, respectively, which consistent with the result of FTIR that CSC contained more stable triple‐helix than PSC‐Sm. Peptide maps were different between PSC‐Sm and CSC, indicating the differences in their amino acid compositions and sequences. The maximum and minimum solubilities of PSC‐Sm were observed at pH 2.0 and 4.0, respectively. A sharp decrease in solubility appeared when NaCl concentration was between 3% and 5%. These results showed that collagen from S. monotuberculatus had the type I collagen characteristics and good thermal stability, and therefore, it could be used as an alternative resource of collagen.     

Compositional and physicochemical characteristics of acid solubilized collagen extracted from the skin of unicorn leatherjacket (Aluterus monoceros)

Acid solubilized collagen (ASC) was extracted from the skin of unicorn leatherjacket (Aluterus monoceros) using 0.5 M acetic acid, followed by precipitation with 2.6 M NaCl. ASC with the yield of 4.19% (wet weight basis) was identified as type I collagen, which was composed of two α₁ chains and one α₂ chain. Different peptide maps were observed between ASC hydrolyzed by V8 protease and lysyl endopeptidase. The maps were also different from those of type I collagen from calf skin, suggesting the differences in amino acid sequences between both collagens. Glycine was the most predominant amino acid. ASC contained the relatively higher content of alanine, but lower contents of proline and hydroxyproline, compared with calf skin collagen. FTIR analysis showed that ASC was in triple helix structure. T_max of ASC dispersed in 0.05 M acetic acid and deionized water were 27.7 and 35.8 °C, respectively. Relative viscosity of 0.03% (w/v) ASC dissolved in 0.1 M acetic acid decreased continuously as the temperature increased from 4 to 52 °C, indicating thermal destabilization or denaturation of ASC molecules. ASC had the solubility greater than 90% in very acidic pH range (pH 1–4) and the solubility decreased continuously with increasing NaCl concentrations (0–6%). Net charge of ASC and calf skin collagen became zero at pHs of 5.58 and 5.68, respectively as determined by zeta potential titration. Therefore, skin of unicorn leatherjacket can be used as an alternative collagenous source.     

草鱼鱼鳞胶原蛋白的流变学性能研究

以草鱼鱼鳞为原料,在低于蛋白变性温度的条件下提取酸溶性胶原蛋白(acid-soluble collagens,ASC)和酶溶性胶原蛋白(pepsin-soluble collagens,PSC),并对其溶液流变学性能进行系统测定。实验结果表明,在实验选择的剪切速率范围内,ASC 和PSC 溶液均表现为典型非牛顿流体的假塑性流动行为,即溶液黏度随剪切速率的增加而减小;在相同浓度条件下,溶液黏度的大小依次为ASC ＞ PSC ＞明胶;剪切速率、温度、pH 值等因素对ASC 和PSC 溶液流体性能均有显著影响;在0.6% 浓度条件下,ASC 和PSC 溶液主要表现为流体行为,基本不具备凝胶的弹性行为;胶原蛋白发生热变性后,其流体的弹性行为增强而黏性行为下降。     

Isolation and characterization of pepsin‐solubilized collagen from the integument of sea cucumber (Stichopus vastus)

BACKGROUND: Sea cucumber (Stichopus vastus) is considered an underutilized resource, since only its stomach and intestines are eaten raw as salad in a few countries and the remaining parts, especially the integument rich in collagen, is discarded. Hence a valuable by‐product having potential nutraceutical and pharmaceutical applications is wasted. In the present investigation, pepsin‐solubilized collagen (PSC) from the integument of S. vastus was isolated, purified and characterized. RESULTS: Sodium dodecyl sulfate–polyacrylamide gel electrophoretic analysis showed that the purified collagen was of type I, consisting of three α1 chains of approximately 122 kDa each. The peptide map of PSC digested by V8 protease was different from that of calf skin type I collagen. Fourier transform infrared spectroscopy revealed that the triple helical structure was well preserved in isolated collagen. The denaturation temperature of PSC was 21.23 °C and showed good gel‐forming capability at pH 6.5 and 300 mmol L^?1 NaCl. CONCLUSION: It is inferred that the collagen isolated from S. vastus integument has potential for use as an alternative to land‐based mammalian collagen in food, nutraceuticals and pharmaceutical industries. © 2012 Society of Chemical Industry