Chemical Synthesis and Analysis of Seaweed

Chemical Synthesis and Analysis of Seaweed Chapter 1 Literature review 2015 Introduction Marine-microalgae are normally termed as seaweed belonging to the primitive family of non-flowering plant known as thallophyta (Fig 1). The sea plant lacks the leaves, stem and roots of true plants. The body of the seaweed is called the thallus named after the thallophyta. The flattened leaf portion of the microalgae is called the blade and its function is to increase the photosynthesizing surface area. Seaweeds are attached to the bottom by a root-like structure called a holdfast. Seaweeds are autotrophic which grows in the intertidal and sub-tidal region of the sea. Seaweeds grow abundantly where rocks and coral are present as substratum (Marine Fisheries information Services October-November 1998; Kaliaperumal et al., 2004). Figure 1: Shows the general structure of seaweed ( Thallophyta)( Reproduced from http://dtc.pima.edu/blc/183/03_183/03_183answers.html). Seaweed may be classified into two categories namely phaephyceae (Brown) and Rhodophyceae (Red). Seaweeds contain different types of phycolloids such as alginate, carrageenan, agarose and agar agar. Seaweed is mainly used in Asia as food, fodders, fertilizer and also as drugs in the pharmaceutical industry for its therapeutics properties (Marine Fisheries information Services, October-November, 1998). 1.1 Brown seaweed (Phaephyceae) Brown seaweed is found in the family of seaweed which is called the phaephyceae. The brown seaweed composition varies according to the species, throughout the year and between different habitats. Brown seaweed is composed mainly of carbohydrate such as alginate, laminaran, mannitol, fucoidan and small traces of cellulose as shown in Table 1. (Fasahati et al., 2012) Table1. Brown seaweed composition Component Base design dry weight (%) Used in simulation dry weight (%) Ash 23.63 25.48 Algin 27.78 27.78 Laminaran 16.67 18.52 Mannitol 14.81 14.81 Fucoidan 3.7 Cellulose 6.6 Protein 7.41 7.41 1.2 Alginate Alginate is a major structural biopolymer found in cell wall and intercellular matrix which provides mechanical strength in brown seaweed. Seaweeds containing alginate are called alginophyte (Yabur et al., 2006). In the natural environment, alginate exists as a mixture of potassium, calcium and sodium salt (Gomez et al., 2009). 1.2.1 Structure of alginate The structure differs from species, age and from different part of seaweed. Alginate consists of two monomers and is linked to each other alternately in a linear form. The monomers are namely ÃŽ ²-D-Mannuronate and ÃŽ ±-L-Guluronate in Fig 2(B) and 2(D) respectively. The acid form of mannuronate and guluronate, is mannuronic acid and guluronic acid (Fig 2(C)) respectively. Figure 2(A) depicts the structure of sodium alginate in a linear form. Alginate is the salt of alginic acid and its simplest formula is NaC6H7O6 in Fig 2(A) (Mahmood et al., 2009). . 2(A) 2(B) 2(C) 2(D) Figure 2: Repeat unit of (A) Sodium alginate (B) guluronate, (C) guluronic acid and (D) mannuronate (Falkeborg et al., 2014) The monomers are linked through carbon 1 and 4 and have sequences forming blocks of MM(Fig 3(A)), GG(Fig 3(A)) or GM blocks in Fig 3(A) (Arzate-Và ¡zquez et al., 2012, Morais et al., 2013). Mannuronates and guluronates both have carboxylic group on the carbon 5 of their respective structures (Falkeborg et al., 2014). (A) G G M M (B) G G M M GMMMMGGGGGGGMGMGMGMGMMMMG (C) MM blocks GG blocks MG blocks MM blocks Figure 3: (A) Alginate structure with G and M repeat units, (B) showing the repeat of G and M joined together (C) block fractions of alginate polymer (Yuemei Lin et al., 2010) 1.3 Species of brown seaweeds Alginate are commercially manufactured mainly from Laminaria hyperborea, Macrocystis pyrifera, Laminaria digitata, Ascophyllumnodosum, Laminaria japonica, Eclonia maxima, Lessonia nigrescens, Durvillea Antarctica, Turbinaria conoides, Turbianria ornata and Turbinaria decurrens, Sargassum, Turbinaria ,Hormophysa, Cystoseira and Sargassum sp (Bertagnolli et al.,2014; Chennubhotla et al., 2013; Kaliaperumal et al., 1974). 1.4 Extraction of alginate Gomez et al., (2009) described the pre-treatment and the extraction of alginate from brown seaweed. The seaweed was crushed and was added to water to moisten the latter. 0.1 N Hydrochloric acid was added to the sample and the solution was stirred in order to reach a pH value of 4. This process was continued for 15 min at room temperature and the supernatant was eliminated. The pre-treatment was carried out three times for each sample using 0.1 N Hydrochloric acid. The extraction was followed by the addition of the pre-treated solution to a beaker with 1 N sodium carbonate (pH of solution 10). The mixture was stirred mechanically for a period of 2 hours at a temperature of 60 oC. To the extracted solution a silicon based rock was added and was stirred for 15 minutes. The mixture was then centrifuged to obtain the sodium alginate supernatant. Sodium alginate can be purified using three different routes namely (i) Ethanol route, (ii) Hydrochloric acid route and (iii) Calcium chloride route as summarised in scheme 1. (i) Ethanol pathway The ethanol pathway was performed by the direct precipitation of sodium alginate using ethanol. The precipitate was washed then with ethanol under soxhlet condition for 100 hours to obtain the pure biopolymer which was dried under vacuum. (ii) Alginic pathway The extract alginate was mixed up with 1 N of HCl at room temperature under constant stirring for 1 hour at a pH of 1. To obtain the alginic acid, the mixture was separated by centrifugation by adding water and 1 N sodium carbonate. The mixture was left at room temperature under stirring for 1 hour in order to obtain the soluble form of sodium alginate. The polymer was precipitated by the addition of 1:1 volume ratio of ethanol by the method described in the ethanol pathway. (iii) Calcium chloride pathway To the extract of sodium alginate, 1 M calcium chloride was added and the precipitate was thoroughly washed with distilled water using a soxhlet for 64 hours. The mixture was agitated at room temperature and distilled water was then added followed by the addition of 1 N Hydrochloric acid until a pH 2 was reached. The alginic acid which is insoluble was separated from the supernatant by centrifugation and the insoluble alginic acid was washed with 0.05 N HCl and the solution were stirred at room temperature. The precipitation of alginate was obtained by the addition of Na2CO3 to obtain sodium alginate. The purified alginate was collected by the soxhlet as discussed in the ethanol pathway. Scheme1. Extraction of alginate (Gomez et al., 2009) 1.5 Yield of alginate The yield of alginate is being affected by different factors such as the species, age, period of harvest, temperature of extraction, extracted with either alkali or acid and on the content of alginate in brown seaweed also the size of alginate also affects the yield.(Fertah et al., 2014) Seaweeds that are grown in summer show a lower content of alginate in the branches and stems (15.1 %) and no trace of alginate in the receptacles (Bertagnolli et al., 2014). Sample SS size is 1mm and BS for 1mmoC (51.8 for SS and 44.01for BS) but an increase in temperature proved to have a drastic decrease in the yield shown in table 2 (Fertah et al., 2014). Table 2. Yield of alginate at different size sample and temperature Size 1mm Temperature /o C 25 40 60 25 40 60 Yield/% 38.33 51.8 43.2 35.28 44.01 40.2 There are up to 40% of alginate content in brown seaweed and the content of alginate depending species of brown algae can vary from 10 to 25% (Yabur et al., 2006). The period of collected of seaweed Sargassum filipendula have different yield of alginate. During fall, spring and summer the yield are 17.0  ±0.1 %, 17.2 ±0.3 % and 15.1 ± 0.1 % respectively. The yield is lower in summer than spring and fall because summer algae have lower receptacle and branches which are characteristic in their reproductive period. (Bertagnolli et al., 2015). The yield of alginate is also affected to different species of Sargassum shown in table 3. The highest yield is from S.vulgare is 30.2 % and the lowest S.dentifulum is 3.3 %. Table 3: Yield % of alginate from different species of Sargassum Species of seaweed % Yield S.vulgare 30.2 S.polycystum 17.1-27.6 S.dentifilium 3.3 S.latifilium 17.7 S.asperifolium 12.4 S.oligocystum 16.3-20.5 S.fuiltans 21.1-24.5 S.filipendula 15.1-17.2 1.6 Biosynthesis of alginate The proposed biosynthesis of alginic acid in Fucus gardineri in the presence of bacteria Pseudomonas is shown in Scheme 2 (Lin and Hassid: 1966). Mannose 1-phosphate reacts with GTP/ H+ in the presence of mannose 1-phosphate guanyl transferase to produce GDP-ÃŽ ±-D-mannose and the by product is diphosphate. GDP-ÃŽ ±-D-mannose in the presence of enzyme GDP-mannose dehydrogenase react with water and NAD+ giving GDP-Mannuronate and finally reacting with GDP/H+ to give alginate. Scheme 2: Biosynthesis of alginate in brown seaweed Fucus gardineri and bacteria (Lin and Hassid; 1966) 1.7 Characterisation of alginate Extracted alginate may be characterised using a variety of techniques such as 1H-NMR, FT-IR, SEC, DSC, CHNS elemental analysis and viscosity. 1.7.1 1H-NMR 1H-NMR spectroscopy is used to determine the main structural pattern and composition of alginate. Structures of alginate are generally constituted of homopolymeric blocks i.e M and G blocks that can be separated by heteropolymeric of MG blocks. The M/G ratio has a strong effect on the physical properties of alginate. Figure 4:1H-NMR Spectra for solution of alginate form S. vulgare in D2O (a) SVLV (S.vulgare low viscosity) and (b) SVHV (S.vulgare High viscosity) (Torres et al., 2007) Figure 4 (a) and (b) show the 1H-NMR spectra of alginate extracted from SLHV and SLHV respectively. Peak I (5.06 ppm) was attributed to anomeric proton (G-1) guluronic acid, peak II (4.7 ppm) show the anomeric proton (M-1) of mannuronic acid and the C-5 alternating blocks (GM-5) which is overlapping each other at this peak and peak III (4.4 ppm) is for the anomeric peak of guluronic acid H-5 (G-5) (Torres et al., 2007). 1.7.2 FT-IR analysis FT-IR spectroscopy is used to determine the functional groups present in the structure of alginate. The FT-IR spectra of the extracted alginate from brown seaweed Turbinaria ornata is given in (figure 5). The bands which is around 800 cm-1 corresponds to the C-H which is ‘out plane’ of the aromatic ring. The peak near 1040 cm-1 is due to the stretching vibrations of C-O bond. The bands at 1230 and 1600 cm-1 corresponding to C-N stretching and N-H bending vibration respectively show the presence of aliphatic amine. The O-H alcoholic groups shows stretching vibrations band at around 3400 cm-1 and C=O stretching vibrations due to the carboxylic acids group present in the structure (Gowtham Sriram et al., 2014). Figure 5: FT-IR spectra of Turbinaria ornata seaweed (Gowtham Sriram et al., 2014) 1.7.3 CHNS elemental analysis Table 4 shows the result for elemental analysis for SVHV and SVLV and both sample proved to have similar results. The nitrogen content is due to the protein found in the seaweed. The % C are 27.7 and 28.1, %H are 5.15 and 4.4 for SVLV and SVHV respectively. The %N (nitrogen) is due to the presence of protein which contains amine groups in seaweed. % S are zero for both SVLV and SVHV (Torres at al., 2007) Table 4 : Analytical data of Sargassum Vulgare alginate Analytical parameter (m/m %) SVLV(S.Vulgare low viscosity SVHV(S.Vulgare high viscosity H 5.15 4.49 C 27.7 28.1 N 0.178 0.164 S 0 0 1.7.4. Viscosity Intrinsic viscosity is the measurement of hydrodynamic volume occupied by complex molecule at unlimited dilution in specific solvents at a given temperature. The intrinsic viscosity depends on the molar mass (Mv), composition and the sequences of M and G residues. Intrinsic viscosity is represented as equation 1 [ÆÅ ¾] = sp/C Equation (1) Where ÆÅ ¾sp is specific viscosity and ÆÅ ¾sp/C is reduced viscosity Another equation of reduced viscosity is defined as ÆÅ ¾red = Equation (2) the intrinsic viscosity was found by the classical method of a plotting straight line graph according to the Huggin’s Equation (3) to which [ÆÅ ¾] y-intercept is obtained. ÆÅ ¾red = [ÆÅ ¾] + kH[ÆÅ ¾]2 C Equation (3) Graph of reduced viscosity versus alginate concentration for SVLV and SVHV sample shown in figure 6 (Torres et al., 2007). Figure 6: Specific viscosity of SVLV (S.Vulgare low viscosity) and SVHV(S.vulgare high viscosity) The viscosities were performed in 0.1 N NaCl at 25 oC for both SVLV and SVHV. Intrinsic viscosites are 6.9 and 4.14 g/dL for SVLV and SVHV respectively (Torres et al., 2007). Table 5 shows different [ÆÅ ¾] for different species of brown seaweed. High vis The intrinsic viscosities of S.Vulgare algiante ranges from (2.5-15.4) g/dL cosity was found SVLV due to high content of MM blocks. Table 5 :Intrinsic viscosity and average molar masses of alginates from different sources(Torres et al., 2007) Alginate source [ÆÅ ¾] (dL/g) Mwa 105 (g/mol) L. hyperborean 6.4 3.05 F. vesiculosus 2.5 1.17 A. nodosum 2.8 1.32 L. japonica 15.4 7.44 S. fluitans 6.30 3.00 S. dentifolium 12.6 6.06 S. asperifolium 15.2 7.34 S. latifolium 8.7 4.16 S. vulgare This study 4.1 1.94 LV (low viscosity ) HV (high viscosity) 6.9 3.30 1.7.4.1. Molar mass Molar mass (Mv) is determined by using the Mark-Houwink equation (4), [ÆÅ ¾] = k Mva - Equation (4) where k and a are empirical coefficients that are dependent on the polymer and the solvent-temperature system used. As for alginate, a value ranges from 0.73 to 1.31 can be acquired depending on the composition of alginate and ionic strength. G-rich alginate has higher a values while lower a values are observed in M-rich alginate which is flexible in nature Proposed empirical relations for [ņ¹] and the weight-average molar mass (Mw) for SVHV and SVLV was computed using equation 4 and the result shown in table 3 [ÆÅ ¾] = 0.023 Mw 0.984 - Equation 4 where [ÆÅ ¾] is given in dL/g and M in kDaltons. The a value used is 0.984 which is different on at specific solvent used. Average molar mass for SVHV was higher than SVLV and similar values to S.fluitans. SVLV has a lower value than other Sargassum species reported in table 5 (Torres et al., 2007). 1.8 Uses of alginate Alginate has many areas of application. For example in food industry and is use to stabilise mixture dispersion and emulsion which increase the viscosity of the gel and forms gel such as jam and jellies. Alginate gel films are also utilized in the food industry as edible films on fruits and vegetables and as coatings designed to improve stored meat quality, industrial and pharmaceutics because the water in alginate has the ability to hold and form gel, to form and stabilise emulsion, textile industry, rubber adhesive paper products, cosmetics and in dental mould (Chennubhotla et al., 2013; Crossingham et al., 2014). Alginate also protects against carcinogenic substances, it clears the digestive system and protects the membrane of the stomach and intestine. Alginate is used in the manufacturing of soft capsules and is consumed as beverage for lowering blood sugar level. Alginate is used in the textile industry as an additive for textile and has also been investigated recently in the composition in vaginal film dosage in pharmaceutical industries and alginate is also used in wound dressings (Paul et al., 2004).

The Signalman Analysis

The story begins with the narrator calling â€Å"Halloa! Below there! † into a railway cutting. The signalman standing on the line below does not look up, as the narrator expects, but rather turns about and stares into the railway tunnel it is his responsibility to monitor. The narrator calls down again and asks permission to descend. The signalman seems reluctant, but assents and waits with an air of ‘expectation and watchfulness’. The railway cutting is a damp, gloomy and lonely place. The signalman seems still to be in fear of the narrator, who tries to put him at ease. The signalman appears to have seen the narrator before. The narrator assures him that this is impossible. Reassured, the signalman welcomes the newcomer into his little cabin and the two men speak of the signalman's work. His labour consists of a dull, monotonous routine, but the signalman feels he deserves nothing better, as he misused his youthful academic opportunities. The narrator remarks that the signalman seems a sane and dutiful employee at all times but when he looks to his signal bell at two moments when it is not ringing. The visitor leaves with a promise to return on the following night. Before he makes his exit, the signalman asks him why he used the words â€Å"Halloa! Below! † on his arrival; were they not suggested to him â€Å"†¦in any supernatural way†? The narrator says no. The signalman implores him to by no means call out so again. He says that he is â€Å"troubled†. The following day the narrator returns and does not call. The signalman tells him that he will reveal to him the nature of his trouble, which is that he is haunted by a recurring apparition: he has seen a spectre at the entrance to the tunnel on two separate occasions and that each appearance has foreshadowed a tragedy. In the first instance, the signalman heard the shouted words that the narrator spoke and saw a figure with its arm across its face, waving the other in desperate warning. He questions it but it vanishes. He then runs into the tunnel but finds nobody. Within a few hours there occurs a terrible train crash with many casualties. On its second appearance, the figure is silent, with both hands before the face in an attitude of mourning. Soon afterward, a beautiful young woman dies in a passing train. Finally the signalman admits that he has seen the spectre several times within the past week. It seems to the narrator that the signalman is suffering from hallucinations. During the conversation the signalman sees the spectre, and hears his bell toll out a phantom ring, but the narrator sees and hears nothing of these events. The signalman is convinced these supernatural incidents are portents of a third tragic event yet to happen, and is sick with fear and frustration: he does not understand why he should be burdened with knowledge of an incipient tragedy when he, as a lowly railway functionary, has neither the authority nor the ability to prevent it. The sceptical narrator believes that his new friend's imagination has been overtaxed and suggests taking him to see a doctor. The following day the narrator visits the railway cutting for a third time, and is struck to see a figure at the mouth of the tunnel. This figure is no ghost, however. It is a man, one of a group of officials investigating an incident on the line. The narrator discovers his friend the signalman is dead, having been struck by an oncoming train. He had been standing on the line, looking intently at something, and failed to get out of the way. The driver of the train explains that he did attempt to warn the signalman of his danger: as the train bore down on the signalman the driver called out to him â€Å"Below there! Look out! Look out! For God’s sake, clear the way! † Moreover, the driver waved his arm in warning even as he covered his face to avoid seeing the train strike the hapless signalman. The narrator notes the significance of the similarity between the driver's actions and the actions of the spectre as the signalman had earlier described them, but leaves the nature of that significance to the reader.

Cell Phone Effects Essay

Introduction: Introduction about using mobile phones in the present Thesis: Using mobile phones in the present is a popular for all ages and there are many advantages to user. I. Argument or reason to support – Mobile phones is convenient communication tool. – Good feature of mobile phone – Mobile phones no limited about time II. Argument against – Negative healthy effects – The effect on children – Mobile phone and driving – The effect on the social Refutation: Mobile phones are a necessity in daily life. Human must be use in communicate. Conclusion: Using mobile phone is popular in the present and there are important to human as if it’s past of life. Human must used it for communication. From a good feature, human can use many advantages from mobile phone. At present, mobile phones are popular for everyone in our society. Mobile phones are common facilities for all ages. They can be used to communication between two persons or more than two persons everywhere and every time. So, mobile phones are important equipment for people nowadays. And mobile phones have many disadvantages. To have good health and live happily in our society, we should not use mobile phones. The most important advantage of using mobile phones is that mobile phones are convenient communication tools that we can use to communicate with our friends. Besides, you can carry mobile phones everywhere by keeping them in shopping bags and school bags because mobile phones are small and easy to carry. Furthermore, mobile phones have a lot of good functions such as cameras, music players, radio, the Internet, games, dictionaries, sound recorders, video players, calculators, map, bluetooth devices, notes, calendars and etc. In contrast, mobile phones have many disadvantages. The first one is they have effects to our†¦

Essay about Animal Farm, by George Orwell - 1648 Words

nbsp;nbsp;nbsp;nbsp;nbsp;This novel takes place at a farm, which its name is Manor Farm, who’s owned by Mr. Jones. He a drunk that never really took care of his animals. There was a Boar that lived on the farm his name was Old Major. He is twelve years old, had lately grown rather stout, majestic- looking pig, tushes never been cut, and wise and benevolent appearance. He sets up a meeting to discuss an important matter that happened to him the night before. nbsp;nbsp;nbsp;nbsp;nbsp;The discussion to place at night after Mr. Jones was asleep. In a barn is where all the animals had gather to listen to Old Major speech. Three dogs, a cat, pigeons, hens, pigs, cows, sheep’s, two horses named Clover, mother that never got her figure†¦show more content†¦nbsp;nbsp;nbsp;nbsp;nbsp;In the past years Mr. Jones lost money in a lawsuit. Afterwards he drank more, the field was full of weeds, and the animals were getting underfed. When June came they just started to forget feeding the animals. They got so hungry they went to the store-shed a broke the doors open. Mr. Jones and four men went there and started whipping the animals. Their reaction was different this time, and they ended chasing Mr. Jones and his men off of their property. They’ve been learning how to read and write and with the sign that said Manor Farm, they changed to Animal Farm. Napoleon took puppies from the mother because he was going to teach them how to read and write. He took them and stuck them on the second floor, which you needed a ladder to get up there. Eventually everyone forgot about the puppies. They also wrote on a wall were the Seven Commandments, which are 1) Whatever goes upon two legs is an enemy. 2) Whatever goes upon four legs, or has wings, is a friend. 3) No animal shall wear clothes. 4) No animal shall sleep in bed. 5) No animal shall drink alcohol. 6) No animal shall kill any other animal. 7) All animals are equal. nbsp;nbsp;nbsp;nbsp;nbsp; They went to the hayfield, to collect food for the harvest. It was the biggest harvest ever. The pigs were the ones who directed the animals all of the work. During the summer they all worked. Every Sunday they had a meeting and ended the meeting with the song Beast of England.Show MoreRelatedAnimal Farm And George Orwell By George Orwell1034 Words   |  5 Pages Eric Arthur Blair, under the pseudonym of George Orwell, composed many novels in his lifetime that were considered both politically rebellious and socially incorrect. 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