Research Article Extraction, Characterization, and ...

7
Research Article Extraction, Characterization, and Molecular Weight Determination of Senna tora (L.) Seed Polysaccharide Harshal A. Pawar 1 and K. G. Lalitha 2 1 Department of Quality Assurance, Dr. L. H. Hiranandani College of Pharmacy, Ulhasnagar, Maharashtra 421003, India 2 Department of Pharmaceutical Chemistry, Ultra College of Pharmacy, No. 4/235, College Road, asildar Nagar, Madurai, Tamil Nadu 625020, India Correspondence should be addressed to Harshal A. Pawar; [email protected] Received 1 September 2015; Revised 22 October 2015; Accepted 29 October 2015 Academic Editor: Vijaya K. Rangari Copyright © 2015 H. A. Pawar and K. G. Lalitha. is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. e objective of the present work was extraction of polysaccharide from Senna tora L. seed and its characterization as a pharmaceutical excipient. Polysaccharide extraction was based on mechanical separation of the endosperm of seeds of Senna tora, water dissolution, centrifugation, and precipitation with acetone. Standard procedures were used to study the viscosity, micromeritic properties, and microbial bioburden. Accelerated stability study was carried out on isolated polysaccharide for six months at 40 C/75 RH as per ICH guidelines. e gum obtained from S. tora seeds was an amorphous free flowing odourless powder with dull brown colour (yield = 35% w/w). e bulk density, tapped density, and angle of repose data reveal that S. tora gum possesses good flow property. e intrinsic viscosity obtained was 1.568dL/g. e average molecular weight of purified S. tora gum was found to be 198 kDa by intrinsic viscosity method. e results indicated that viscosity of gum solution increases with increase in temperature. FTIR study revealed the absence of degradation or decomposition of polysaccharide at accelerated stability conditions for six months. It has been concluded that extracted polysaccharide can be used as pharmaceutical excipient in terms of flow behavior, microbial properties, and stability. 1. Introduction e pharmaceutical applications of galactomannans obtained from different commercial and noncommercial sources have been extensively studied over the past decade. Galactoman- nans show potential in the international trend towards the use of more plant-based products for ecological motives, and their production and application do not cause pollution or disturbance to the ecosystem. Galactomannans are used in a variety of pharmaceutical dosage forms such as tablets, suspensions, hydrogels, and films as an excipient. Besides the simple use as inert excipient, these polysaccharides play role in the modification of drug release, especially in colonic environment, as a matrix or coating material. Senna tora (L.) Roxb. (S. tora) belonging to the family Fabaceae is an annual undershrub which grows all over the tropical countries (India, Pakistan, Bangladesh, and west China). It grows well in wasteland as a rainy season weed. It is also known as “Chakramard” in Ayurveda [1, 2]. e S. tora seed is composed of hull (27%), endosperm (32%), and germ (41%). Seeds of S. tora contain about 23.2% of proteins, rich in all essential amino acids, par- ticularly, methionine and tryptophan. Several compounds belonging to anthraquinone and naphthopyrone groups have been isolated from seeds of this plant. It also contains phenolic glycosides, namely, rubrofusarine triglucoside, nor- rubrofusarin gentiobioside, demethylflavasperone gentiobio- side, torachrysone gentiobioside, torachrysone tetragluco- side, and torachrysone apioglucoside. Seed oil contains dif- ferent percentage of oleic, linoleic, palmitic, stearic, and lignoceric acids [3–5]. Several studies have been conducted throughout the last decade to investigate the biological properties of S. tora. Different parts of the plant (Leaves, seed, and root) are reputed for their medicinal value. e leaves of S. tora are reported to have antirheumatic activity in folklore practice. Decoction of the leaves is used as laxative. e seeds of S. tora have been used in Chinese medicine as aperients, Hindawi Publishing Corporation International Journal of Biomaterials Volume 2015, Article ID 928679, 6 pages http://dx.doi.org/10.1155/2015/928679

Transcript of Research Article Extraction, Characterization, and ...

Page 1: Research Article Extraction, Characterization, and ...

Research ArticleExtraction Characterization and Molecular WeightDetermination of Senna tora (L) Seed Polysaccharide

Harshal A Pawar1 and K G Lalitha2

1Department of Quality Assurance Dr L H Hiranandani College of Pharmacy Ulhasnagar Maharashtra 421003 India2Department of Pharmaceutical Chemistry Ultra College of Pharmacy No 4235 College Road Thasildar NagarMadurai Tamil Nadu 625020 India

Correspondence should be addressed to Harshal A Pawar harshaldlhhcopgmailcom

Received 1 September 2015 Revised 22 October 2015 Accepted 29 October 2015

Academic Editor Vijaya K Rangari

Copyright copy 2015 H A Pawar and K G LalithaThis is an open access article distributed under theCreativeCommonsAttributionLicense which permits unrestricted use distribution and reproduction in anymedium provided the originalwork is properly cited

The objective of the present work was extraction of polysaccharide from Senna tora L seed and its characterization as apharmaceutical excipient Polysaccharide extraction was based on mechanical separation of the endosperm of seeds of Senna torawater dissolution centrifugation andprecipitationwith acetone Standard procedureswere used to study the viscositymicromeriticproperties andmicrobial bioburdenAccelerated stability studywas carried out on isolated polysaccharide for sixmonths at 40∘C75RH as per ICH guidelines The gum obtained from S tora seeds was an amorphous free flowing odourless powder with dull browncolour (yield = 35 ww) The bulk density tapped density and angle of repose data reveal that S tora gum possesses good flowproperty The intrinsic viscosity obtained was 1568 dLg The average molecular weight of purified S tora gum was found to be198 kDa by intrinsic viscosity method The results indicated that viscosity of gum solution increases with increase in temperatureFTIR study revealed the absence of degradation or decomposition of polysaccharide at accelerated stability conditions for sixmonths It has been concluded that extracted polysaccharide can be used as pharmaceutical excipient in terms of flow behaviormicrobial properties and stability

1 Introduction

Thepharmaceutical applications of galactomannans obtainedfrom different commercial and noncommercial sources havebeen extensively studied over the past decade Galactoman-nans show potential in the international trend towards theuse of more plant-based products for ecological motives andtheir production and application do not cause pollution ordisturbance to the ecosystem Galactomannans are used ina variety of pharmaceutical dosage forms such as tabletssuspensions hydrogels and films as an excipient Besidesthe simple use as inert excipient these polysaccharides playrole in the modification of drug release especially in colonicenvironment as a matrix or coating material

Senna tora (L) Roxb (S tora) belonging to the familyFabaceae is an annual undershrub which grows all over thetropical countries (India Pakistan Bangladesh and westChina) It grows well in wasteland as a rainy season weed Itis also known as ldquoChakramardrdquo in Ayurveda [1 2]

The S tora seed is composed of hull (27) endosperm(32) and germ (41) Seeds of S tora contain about232 of proteins rich in all essential amino acids par-ticularly methionine and tryptophan Several compoundsbelonging to anthraquinone and naphthopyrone groups havebeen isolated from seeds of this plant It also containsphenolic glycosides namely rubrofusarine triglucoside nor-rubrofusarin gentiobioside demethylflavasperone gentiobio-side torachrysone gentiobioside torachrysone tetragluco-side and torachrysone apioglucoside Seed oil contains dif-ferent percentage of oleic linoleic palmitic stearic andlignoceric acids [3ndash5]

Several studies have been conducted throughout the lastdecade to investigate the biological properties of S toraDifferent parts of the plant (Leaves seed and root) arereputed for their medicinal value The leaves of S tora arereported to have antirheumatic activity in folklore practiceDecoction of the leaves is used as laxative The seeds ofS tora have been used in Chinese medicine as aperients

Hindawi Publishing CorporationInternational Journal of BiomaterialsVolume 2015 Article ID 928679 6 pageshttpdxdoiorg1011552015928679

2 International Journal of Biomaterials

antiasthenic and diuretic agent It is also given to improvevisual activity (eye diseases) and to treat liver disordersIn Korea the hot extract of seeds is taken orally for liverprotection Leaves and seeds are used in the treatment of skindisorders (ringworm and itch) Stem bark extract is used forvarious skin ailments and rheumatic diseases and as laxativeIn Ayurveda the plant is used in ldquoDadrughani Vatirdquo andldquoPamari Tailardquo [6ndash10]

The objective of the present research work was extractionof polysaccharide from Senna tora L seeds and furthercharacterization of it as pharmaceutical excipient Studyincludes evaluation of micromeritic properties viscosity andeffect of temperature on viscosity microbial bioburden andstability and determination of molecular weight of extractedpolysaccharide

2 Materials and Methods

21 Collection of Plant Material The pods of S tora werecollected fromThane district ofMaharashtra India Plantwasauthenticated by Dr Rajendra D Shinde Associate ProfessorBlatter Herbarium St Xavierrsquos College Mumbai and wasidentified as Senna tora (L) Roxb (herbarium specimennumber 8361) The herbarium specimen of Senna tora wasstored in Ultra College of Pharmacy Madurai for futurereference The seeds were separated from pods and stored incool place till further use

22 Isolation andPurification ofGum Thegumwas extractedfrom endosperm using solvent precipitation method asreported in literature [11] The dried seeds were dehuskedand degermed by mechanical treatment followed by millingand screening of the endosperm The powder was soakedin benzene-ethanol solution (1 1) overnight to remove lipidsand then it was dried in vacuum oven The endosperm pow-der of S tora seeds (10 g) was soaked in 200mL of distilledwater and stirred under overhead stirrer for 3-4 hours Theviscous solution obtainedwas passed through themuslinThemarc obtainedwas pressed to remove themucilage and boiledwith 200mL of water for one hour Viscous solution obtainedwas filtered through muslin cloth The marc obtained wasnot discarded but it was sent for multiple extractions withdecreasing quantity of extracting solvent that is water withthe increase of number of extractions The isolation wascontinued until the material becomes free frommucilage Allthe viscous solutions obtained weremixed together An equalquantity of 10 trichloroacetic acid was added to the mixtureto precipitate protein The solution was centrifuged and thesupernatant was precipitated out by addition of acetone inthe ratio 1 05 with continuous stirring The coagulatedmucilage which formed as a white mass was transferredto an evaporating dish and dried in vacuum oven at 40∘Cpowdered and stored in airtight containers

23 Characterization of Gum The yield of isolated polysac-charide was calculated and organoleptic features were notedThe polysaccharide was evaluated further for the followingproperties

24Micromeritic Properties Powder fluidity or flowpropertyis one of the important requirements of pharmaceuticalexcipient which decides its utility and application in thedevelopment of different dosage formsThe flowability of theextracted gumpowderwas evaluated by determining its angleof repose bulk and tapped density using methods reportedin the literature [12 13]

241 Angle of Repose The flow characteristics were mea-sured by angle of repose Improper flow of powder is due tofrictional forces between the particles These frictional forcesare quantified by angle of repose It can be calculated by thefollowing formula

tan 120579 = ℎ119903

or 120579 = tanminus1 ℎ119903

(1)

where ℎ is the height of pile 119903 is the radius of the base of thepile and 120579 is the angle of repose

A dry and clean funnel was fixed on to a burette standat particular height (2-3 cm) A graph paper was placed onthe flat surface and a sufficient quantity of the powder (10 g)was allowed to flow slowly through the funnel until theheap touched the tip of the funnel The circumference ofthe heap was drawn and the midpoint was located and itsradius was measured The experiment was repeated thriceand the average height and radius were calculated Usingthese readings and the above formula the angle of repose wascalculated

242 Bulk Density and Tapped Density A 10 g quantity ofgum powder was placed in a 50mL measuring cylinder andthe volume occupied by gum powder without tapping wasnoted After 100 taps the occupied volumewas readThe bulkand tap densities were calculated as the ratio of weight tovolume

243 Hausnerrsquos Ratio and Carr Index Hausnerrsquos ratio wascalculated as the ratio of tapped density to bulk density of thesamples

Carr index (compressibility index) was calculated usingthe following formula

Compressibility

=Tapped density minus bulk density

Tappeddensity times 100

(2)

25 Viscosity Measurement The viscosity of 1wv solutionof S tora gum was measured by Brookfield viscometer at50 RPM using spindle number 5

26 Molecular Weight Determination by Intrinsic ViscosityMethod Viscosity measurements were carried out with Ost-wald viscometer tubes after 24 hours of hydration of the gumFifty milligrams of the gum was dissolved in 100mL of waterto get concentration of 005wv The prepared solutionwas agitated vigorously for approximately 15min until thesolution become viscous and homogeneous

International Journal of Biomaterials 3

Table 1 Accelerated stability protocol of S tora gum

Product S tora seed polysaccharideQuantity loaded 20 g approx in each Petri dish

Quantity Sample 2 g approxSampling interval Storage conditions TestInitialFirst monthSecond monthThird monthSixth month

Temperature = 40∘CHumidity = 75 RH

(1) Organoleptic evaluation(2) pH of 1 wv solution(3) Moisture content(4) Microbial count

(5) FTIR

The relative viscosity of S tora gum dissolved in dis-tilled water (001ndash005 gdL) was measured using Ostwaldviscometer at room temperature Average molecular weight119872V was calculated using the Mark-Houwink relationship[Equation (3)] given by Doublier and Launay (1981) for guargum asmodified by Gaisford Harding Mitchell and Bradley(1986) to take into account the different values of mannose(M)galactose (G) of the galactomannans [14] Consider

120578 = 1155 times 10minus6

[(1 minus 120572) times119872V]098

(3)where 120572 = 1[(MG) + 1] and [120578] is expressed in dLg

27 Effect of Temperature on Viscosity of S tora Gum Effectof temperature on the viscosity of dilute solution (01wvin water) of S tora gum was studied experimentally withincrease in temperature from 36∘C to 100∘C The viscositymeasurements were performed using Ostwald viscometer

28 Total Microbial Load The total microbial load is animportant parameter which decides the suitability of asubstance for use as excipient in pharmaceutical dosageforms According to many Pharmacopoeias for syntheticand semisynthetic substances the total aerobic count shouldnot be more than 100 colony forming units (cfu) per gramand the total fungal count (including yeasts and molds)should not exceed 50 cfug In the case of excipients fromnatural origin the total aerobic count should not be morethan 1000 cfug and total fungal count should not exceed100 cfug The total microbial load of the isolated gumsample was determined by plate count method as per IndianPharmacopoeia [15]

29 Tests for Presence of Specific Microorganisms Materialsto be used as excipients in dosage forms need to be free fromEscherichia coli Salmonella typhi Pseudomonas aeruginosaand Staphylococcus aureus according to PharmacopoeiasInternational Conference for Harmonization (ICH) has alsosupported this criterion The tests for these microorganismswere carried out as described in Indian Pharmacopoeia

210 Accelerated Stability Study S tora seed polysaccharidewas subjected to accelerated stability studies for six monthsaccording to ICH guidelines to predict the stability of Stora seed polysaccharide [16] The samples were analyzed atregular intervals as per the stability protocol (Table 1)

Table 2 Results of micromeritic properties of S tora seed gum

Parameters Resultslowast

Bulk density (gmL) 051 plusmn 0047Tapped density (gmL) 060 plusmn 0039Hausnerrsquos ratio 117Carr index 1429Angle of repose 29∘441015840 plusmn 0032lowastThe values represent mean plusmn SD (119899 = 3)

3 Result and Discussion

The gum obtained from S tora seeds was an amorphous freeflowing odourless powder with dull brown colour (yield =35ww) The photograph of the gum powder was takenunder high intensity electron light microscope and usingdigital camera as depicted in Figures 1(a) and 1(b)

The results of micromeritic study are summarised inTable 2 The bulk density tapped density and angle ofrepose data reveal that S tora gum possesses good flowproperty Carr index is frequently used in pharmaceuticsas an indication of the compressibility and flowability of apowder In a free flowing powder the bulk density and tappeddensity would be close in value therefore Carr index wouldbe small A Carr index greater than 25 is considered to be anindication of poor flowability and a Carr index below 15 isconsidered to be of good flowability

Viscosity is the main parameter to assess the qualityof natural gums The applications of any natural gum aredependent on its viscosity For any polymer to be used in slowrelease hydrophilic matrix systems it should possess certaincharacteristics like fast hydration of the polymer and highgel strength and should be stable during the shelf life of theproduct Viscosity of 1wv solution of polysaccharide inwater was found to be 4520 cP at 50 revolutions per minute(rpm) The results indicated that the polysaccharide isolatedfrom S tora seeds possesses high viscosity with good swellingcapacity S tora seed polysaccharide hydrates quickly andswells rapidly and forms a thick viscous gel around it Sucha high viscosity indicates its utility in the development ofvarious modified release pharmaceutical dosage forms Theeffect of temperature on viscosity of S tora gum is depictedin Figure 2

4 International Journal of Biomaterials

(a) (b)

Figure 1 Photograph of gum powder taken (a) under high magnification microscope and (b) using digital camera

0000

0200

0400

0600

0800

1000

1200

1400

1600

0 20 40 60 80 100 120

Visc

osity

(cP)

Temperature (∘C)

Figure 2 Effect of temperature on viscosity of S tora gum

It has been reported in previous literature that S toragum contains galactomannans [11] Galactomannans areformed by several fractions of polymers with differentmannosegalactose ratios At low temperature moleculeswith low molecular weight and low mannosegalactose ratioare mainly dissolved At high temperatures galactomannanfractions with high molecular weight greater mannose togalactose relation and therefore zones without galactoseradicals are solved [17] The results indicated that viscosityof gum solution increases with increase in temperature Thismay be due to increase in solubility of galactomannans athigher temperature

The intrinsic viscosity was determined graphically fromHuggins plot (reduced viscosity versus concentration graphextrapolated to zero) Huggins plot is represented in Figure 3

The intrinsic viscosity obtained was 1568 dLg The aver-age molecular weight of purified S tora gum was found to be198 kDa by intrinsic viscosity method

Figure 4 represents FTIR spectrum of S tora polysaccha-ride

The FTIR spectrum of the initial sample (Figure 4) of iso-lated polysaccharide confirmed the presence of galactoman-nansThe interpretation was done using previously publishedliteratures [18ndash23] The FTIR data of the polysaccharide ispresented in Table 3

000

500

1000

1500

2000

2500

0 002 004 006

y = 36282x + 15684

R2 = 09575

Redu

ced

visc

osity

Concentration (gdL)

Figure 3 Huggins plot

The total microbial load was found to be 34023 plusmn1079 cfug of bacteria and 4439 plusmn 931 cfug of fungi Thetotal microbial load of the polysaccharide was thus withinthe acceptable limits (the acceptable limit for total microbialcount is 1000 cfug of bacteria and 100 cfug of fungi fornatural products) In specific tests for microorganisms it wasfound that Escherichia coli Salmonella typhi Pseudomonasaeruginosa and Staphylococcus aureus were absent which isessential criteria for their use as excipients in dosage forms

The results of accelerated stability studies on S tora seedpolysaccharide powder showed that there is no significantdifference between the initial and final samples withdrawnat time interval 6 months at 40∘C75 RH Slight changes inpH and moisture content values were observed The studiesproved that the S tora seed polysaccharide is stable for along period of time FTIR study revealed the absence ofdegradation or decomposition of polysaccharide at acceler-ated stability conditions for six months as shown in Figure 5

International Journal of Biomaterials 5

Table 3 Interpretation of FTIR data of polysaccharide

Wave length Peak assignments813 and 875 cmminus1 Anomeric configurations (120572 and 120573 conformers) and glycosidic linkages1198 and 983 cmminus1 Stretching vibration of CndashO in CndashOndashH bonds1149 cmminus1 Bending vibrational modes of CndashO present in the pyranose ring1134 and 983 cmminus1 CndashOH bending2800ndash3000 cmminus1 CndashH stretching3100ndash3500 cmminus1 OndashH stretching vibration

(1cm)

T(

)

90

80

70

60

4000 3500 3000 2500 2000 1750 1500 1250 1000 750 500

374190

354323

346222

339279

292409

235316

231844

179380

173979

167807

164528

153920

151412

145433

142732

133860

131545

114957

108785

102613

87568

81396

Figure 4 FTIR spectrum of the S tora polysaccharide (initial sample)

(1cm)

T(

)

4000 3500 3000 2500 2000 1750 1500 1250 1000 750

45

375

30

225

15

358567

354323

334650

298002

294337

288358

283150

235894

234158

215256

174751

173208

165107155655

153920

151991

147169

141768

131738

124602

115150

105892

102613

87375

81396

Figure 5 FTIR spectrum of the S tora polysaccharide (after 6 month)

6 International Journal of Biomaterials

4 Conclusion

The polysaccharide (gum) extracted from S tora seedspossesses good flow property The gum possesses excellentviscosity The gum polysaccharide was found to be stable ataccelerated stability condition Microbial studies confirmedits suitability as an excipient Based on the physicochemicalmicrobiological characteristics and stability studies it may bepointed out that the polysaccharide isolated from S tora seedshas the required properties and could be used as an excipientfor pharmaceutical dosage forms

Thus there is need to investigate further S tora polysac-charide as an excipient in different pharmaceutical dosageforms It may provide an alternative to synthetic or semisyn-thetic excipientspolymers currently used in the pharmaceu-tical industry

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors are thankful to Ultra College of PharmacyMadurai for providing the laboratory facilities to carry outthe present investigation

References

[1] A Ingle P Ranaware A Ladke and M Damle ldquoCassia toraphytochemical and pharmacological activityrdquo InternationalImperial Journal of Pharmacognosy amp Natural Products vol 2no 1 pp 14ndash23 2012

[2] C S Narayan and S Rangaswami ldquoIsolation of three crystallinesubstances from the seeds of Cassia torardquo Current Science vol25 article 559 1956

[3] I H Suba Jois and B L Manjunath ldquoChemical examination ofseeds of Cassia toralinnrdquo Journal of the Indian Chemical Societyvol 7 p 521 1930

[4] R E Wilkinson and W S Hardcastle ldquoComparative fatty acidcontent of various organs of Cassia tora Lrdquo Botanical Gazettevol 130 no 4 pp 254ndash258 1969

[5] H Pawar and PM DrsquoMello ldquoIsolation of seed gum fromCassiatora and preliminary studies of its application as binder fortabletsrdquo Indian Drugs vol 41 no 8 article 465 2004

[6] K R Kirtikar andBD Basu IndianMedicinal Plants PeriodicalExperts D-42 Vivek Vihar New Delhi India 1975

[7] T Nikaido T Ohmoto U Sankawa S Kitanaka and MTakido ldquoInhibitors of adenosine 3101584051015840-cyclic monophosphatephosphodiesterase in cassia seedrdquoChemical andPharmaceuticalBulletin vol 32 no 8 pp 3075ndash3078 1984

[8] H S Yun and IMChang ldquoPlantswith liver protective activities(I)rdquo Korean Journal of Pharmacognosy vol 8 pp 125ndash129 1977

[9] J D Hooker The Flora of British India vol 2 Reeve amp CoSuffolk UK 1879

[10] R S Thakur H S Puri and A HusainMajor Medicinal Plantsof India 1st edition 1989

[11] H A Pawar and K G Lalitha ldquoIsolation purification andcharacterization of galactomannans as an excipient from Senna

tora seedsrdquo International Journal of Biological Macromoleculesvol 65 pp 167ndash175 2014

[12] C V S Subrahmanyam Physical Pharmacy Vallabh PrakashanNew Delhi India 2nd edition 2000

[13] M Emeje P Nwabunike C Isimi et al ldquoIsolation charac-terization and formulation properties of a new plant gumobtained from Cissus refescencerdquo International Journal of GreenPharmacy vol 3 no 1 pp 16ndash23 2009

[14] S E Gaisford S E Harding J R Mitchell and T D Bradley ldquoAcomparison between the hot and cold water soluble fractions oftwo locust bean gum samplesrdquo Carbohydrate Polymers vol 6no 6 pp 423ndash442 1986

[15] Ministry of Health and Family Welfare and Government ofIndia Indian Pharmacopoeia vol 556 Controller of Publica-tion New Delhi India 1996

[16] International Conference on Harmonization ICH GuidelinesQ1A (R2) Stability Testing of New Drug Substances and Products(Revision 2) InternationalConference onHarmonization 2003

[17] J A Casas A F Mohedano and F Garcıa-Ochoa ldquoViscosity ofguar gum and xanthanguar gum mixture solutionsrdquo Journal ofthe Science of Food and Agriculture vol 80 no 12 pp 1722ndash17272000

[18] S D Figueiro J C Goes R A Moreira and A S BSombra ldquoOn the physico-chemical and dielectric propertiesof glutaraldehyde crosslinked galactomannan-collagen filmsrdquoCarbohydrate Polymers vol 56 no 3 pp 313ndash320 2004

[19] BM Prado S Kim B F Ozen and L JMauer ldquoDifferentiationof carbohydrate gums and mixtures using fourier transforminfrared spectroscopy and chemometricsrdquo Journal of Agricul-tural and Food Chemistry vol 53 no 8 pp 2823ndash2829 2005

[20] M R S Prashanth K S Parvathy N S Susheelamma et alldquoGalactomannan estersmdashA simple cost-effective method ofpreparation and characterizationrdquo Food Hydrocolloids vol 20no 8 pp 1198ndash1205 2006

[21] S-N Yuen S-M Choi D L Phillips and C-Y Ma ldquoRamanand FTIR spectroscopic study of carboxymethylated non-starchpolysaccharidesrdquo Food Chemistry vol 114 no 3 pp 1091ndash10982009

[22] Y Peng L Zhang F Zeng and Y Xu ldquoStructure and antitumoractivity of extracellular polysaccharides from myceliumrdquo Car-bohydrate Polymers vol 54 no 3 pp 297ndash303 2003

[23] L Zhang L Yang Q Ding and X Chen ldquoStudies on molec-ular weights of polysaccharides of Auricularia auricula-judaerdquoCarbohydrate Research vol 270 no 1 pp 1ndash10 1995

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Page 2: Research Article Extraction, Characterization, and ...

2 International Journal of Biomaterials

antiasthenic and diuretic agent It is also given to improvevisual activity (eye diseases) and to treat liver disordersIn Korea the hot extract of seeds is taken orally for liverprotection Leaves and seeds are used in the treatment of skindisorders (ringworm and itch) Stem bark extract is used forvarious skin ailments and rheumatic diseases and as laxativeIn Ayurveda the plant is used in ldquoDadrughani Vatirdquo andldquoPamari Tailardquo [6ndash10]

The objective of the present research work was extractionof polysaccharide from Senna tora L seeds and furthercharacterization of it as pharmaceutical excipient Studyincludes evaluation of micromeritic properties viscosity andeffect of temperature on viscosity microbial bioburden andstability and determination of molecular weight of extractedpolysaccharide

2 Materials and Methods

21 Collection of Plant Material The pods of S tora werecollected fromThane district ofMaharashtra India Plantwasauthenticated by Dr Rajendra D Shinde Associate ProfessorBlatter Herbarium St Xavierrsquos College Mumbai and wasidentified as Senna tora (L) Roxb (herbarium specimennumber 8361) The herbarium specimen of Senna tora wasstored in Ultra College of Pharmacy Madurai for futurereference The seeds were separated from pods and stored incool place till further use

22 Isolation andPurification ofGum Thegumwas extractedfrom endosperm using solvent precipitation method asreported in literature [11] The dried seeds were dehuskedand degermed by mechanical treatment followed by millingand screening of the endosperm The powder was soakedin benzene-ethanol solution (1 1) overnight to remove lipidsand then it was dried in vacuum oven The endosperm pow-der of S tora seeds (10 g) was soaked in 200mL of distilledwater and stirred under overhead stirrer for 3-4 hours Theviscous solution obtainedwas passed through themuslinThemarc obtainedwas pressed to remove themucilage and boiledwith 200mL of water for one hour Viscous solution obtainedwas filtered through muslin cloth The marc obtained wasnot discarded but it was sent for multiple extractions withdecreasing quantity of extracting solvent that is water withthe increase of number of extractions The isolation wascontinued until the material becomes free frommucilage Allthe viscous solutions obtained weremixed together An equalquantity of 10 trichloroacetic acid was added to the mixtureto precipitate protein The solution was centrifuged and thesupernatant was precipitated out by addition of acetone inthe ratio 1 05 with continuous stirring The coagulatedmucilage which formed as a white mass was transferredto an evaporating dish and dried in vacuum oven at 40∘Cpowdered and stored in airtight containers

23 Characterization of Gum The yield of isolated polysac-charide was calculated and organoleptic features were notedThe polysaccharide was evaluated further for the followingproperties

24Micromeritic Properties Powder fluidity or flowpropertyis one of the important requirements of pharmaceuticalexcipient which decides its utility and application in thedevelopment of different dosage formsThe flowability of theextracted gumpowderwas evaluated by determining its angleof repose bulk and tapped density using methods reportedin the literature [12 13]

241 Angle of Repose The flow characteristics were mea-sured by angle of repose Improper flow of powder is due tofrictional forces between the particles These frictional forcesare quantified by angle of repose It can be calculated by thefollowing formula

tan 120579 = ℎ119903

or 120579 = tanminus1 ℎ119903

(1)

where ℎ is the height of pile 119903 is the radius of the base of thepile and 120579 is the angle of repose

A dry and clean funnel was fixed on to a burette standat particular height (2-3 cm) A graph paper was placed onthe flat surface and a sufficient quantity of the powder (10 g)was allowed to flow slowly through the funnel until theheap touched the tip of the funnel The circumference ofthe heap was drawn and the midpoint was located and itsradius was measured The experiment was repeated thriceand the average height and radius were calculated Usingthese readings and the above formula the angle of repose wascalculated

242 Bulk Density and Tapped Density A 10 g quantity ofgum powder was placed in a 50mL measuring cylinder andthe volume occupied by gum powder without tapping wasnoted After 100 taps the occupied volumewas readThe bulkand tap densities were calculated as the ratio of weight tovolume

243 Hausnerrsquos Ratio and Carr Index Hausnerrsquos ratio wascalculated as the ratio of tapped density to bulk density of thesamples

Carr index (compressibility index) was calculated usingthe following formula

Compressibility

=Tapped density minus bulk density

Tappeddensity times 100

(2)

25 Viscosity Measurement The viscosity of 1wv solutionof S tora gum was measured by Brookfield viscometer at50 RPM using spindle number 5

26 Molecular Weight Determination by Intrinsic ViscosityMethod Viscosity measurements were carried out with Ost-wald viscometer tubes after 24 hours of hydration of the gumFifty milligrams of the gum was dissolved in 100mL of waterto get concentration of 005wv The prepared solutionwas agitated vigorously for approximately 15min until thesolution become viscous and homogeneous

International Journal of Biomaterials 3

Table 1 Accelerated stability protocol of S tora gum

Product S tora seed polysaccharideQuantity loaded 20 g approx in each Petri dish

Quantity Sample 2 g approxSampling interval Storage conditions TestInitialFirst monthSecond monthThird monthSixth month

Temperature = 40∘CHumidity = 75 RH

(1) Organoleptic evaluation(2) pH of 1 wv solution(3) Moisture content(4) Microbial count

(5) FTIR

The relative viscosity of S tora gum dissolved in dis-tilled water (001ndash005 gdL) was measured using Ostwaldviscometer at room temperature Average molecular weight119872V was calculated using the Mark-Houwink relationship[Equation (3)] given by Doublier and Launay (1981) for guargum asmodified by Gaisford Harding Mitchell and Bradley(1986) to take into account the different values of mannose(M)galactose (G) of the galactomannans [14] Consider

120578 = 1155 times 10minus6

[(1 minus 120572) times119872V]098

(3)where 120572 = 1[(MG) + 1] and [120578] is expressed in dLg

27 Effect of Temperature on Viscosity of S tora Gum Effectof temperature on the viscosity of dilute solution (01wvin water) of S tora gum was studied experimentally withincrease in temperature from 36∘C to 100∘C The viscositymeasurements were performed using Ostwald viscometer

28 Total Microbial Load The total microbial load is animportant parameter which decides the suitability of asubstance for use as excipient in pharmaceutical dosageforms According to many Pharmacopoeias for syntheticand semisynthetic substances the total aerobic count shouldnot be more than 100 colony forming units (cfu) per gramand the total fungal count (including yeasts and molds)should not exceed 50 cfug In the case of excipients fromnatural origin the total aerobic count should not be morethan 1000 cfug and total fungal count should not exceed100 cfug The total microbial load of the isolated gumsample was determined by plate count method as per IndianPharmacopoeia [15]

29 Tests for Presence of Specific Microorganisms Materialsto be used as excipients in dosage forms need to be free fromEscherichia coli Salmonella typhi Pseudomonas aeruginosaand Staphylococcus aureus according to PharmacopoeiasInternational Conference for Harmonization (ICH) has alsosupported this criterion The tests for these microorganismswere carried out as described in Indian Pharmacopoeia

210 Accelerated Stability Study S tora seed polysaccharidewas subjected to accelerated stability studies for six monthsaccording to ICH guidelines to predict the stability of Stora seed polysaccharide [16] The samples were analyzed atregular intervals as per the stability protocol (Table 1)

Table 2 Results of micromeritic properties of S tora seed gum

Parameters Resultslowast

Bulk density (gmL) 051 plusmn 0047Tapped density (gmL) 060 plusmn 0039Hausnerrsquos ratio 117Carr index 1429Angle of repose 29∘441015840 plusmn 0032lowastThe values represent mean plusmn SD (119899 = 3)

3 Result and Discussion

The gum obtained from S tora seeds was an amorphous freeflowing odourless powder with dull brown colour (yield =35ww) The photograph of the gum powder was takenunder high intensity electron light microscope and usingdigital camera as depicted in Figures 1(a) and 1(b)

The results of micromeritic study are summarised inTable 2 The bulk density tapped density and angle ofrepose data reveal that S tora gum possesses good flowproperty Carr index is frequently used in pharmaceuticsas an indication of the compressibility and flowability of apowder In a free flowing powder the bulk density and tappeddensity would be close in value therefore Carr index wouldbe small A Carr index greater than 25 is considered to be anindication of poor flowability and a Carr index below 15 isconsidered to be of good flowability

Viscosity is the main parameter to assess the qualityof natural gums The applications of any natural gum aredependent on its viscosity For any polymer to be used in slowrelease hydrophilic matrix systems it should possess certaincharacteristics like fast hydration of the polymer and highgel strength and should be stable during the shelf life of theproduct Viscosity of 1wv solution of polysaccharide inwater was found to be 4520 cP at 50 revolutions per minute(rpm) The results indicated that the polysaccharide isolatedfrom S tora seeds possesses high viscosity with good swellingcapacity S tora seed polysaccharide hydrates quickly andswells rapidly and forms a thick viscous gel around it Sucha high viscosity indicates its utility in the development ofvarious modified release pharmaceutical dosage forms Theeffect of temperature on viscosity of S tora gum is depictedin Figure 2

4 International Journal of Biomaterials

(a) (b)

Figure 1 Photograph of gum powder taken (a) under high magnification microscope and (b) using digital camera

0000

0200

0400

0600

0800

1000

1200

1400

1600

0 20 40 60 80 100 120

Visc

osity

(cP)

Temperature (∘C)

Figure 2 Effect of temperature on viscosity of S tora gum

It has been reported in previous literature that S toragum contains galactomannans [11] Galactomannans areformed by several fractions of polymers with differentmannosegalactose ratios At low temperature moleculeswith low molecular weight and low mannosegalactose ratioare mainly dissolved At high temperatures galactomannanfractions with high molecular weight greater mannose togalactose relation and therefore zones without galactoseradicals are solved [17] The results indicated that viscosityof gum solution increases with increase in temperature Thismay be due to increase in solubility of galactomannans athigher temperature

The intrinsic viscosity was determined graphically fromHuggins plot (reduced viscosity versus concentration graphextrapolated to zero) Huggins plot is represented in Figure 3

The intrinsic viscosity obtained was 1568 dLg The aver-age molecular weight of purified S tora gum was found to be198 kDa by intrinsic viscosity method

Figure 4 represents FTIR spectrum of S tora polysaccha-ride

The FTIR spectrum of the initial sample (Figure 4) of iso-lated polysaccharide confirmed the presence of galactoman-nansThe interpretation was done using previously publishedliteratures [18ndash23] The FTIR data of the polysaccharide ispresented in Table 3

000

500

1000

1500

2000

2500

0 002 004 006

y = 36282x + 15684

R2 = 09575

Redu

ced

visc

osity

Concentration (gdL)

Figure 3 Huggins plot

The total microbial load was found to be 34023 plusmn1079 cfug of bacteria and 4439 plusmn 931 cfug of fungi Thetotal microbial load of the polysaccharide was thus withinthe acceptable limits (the acceptable limit for total microbialcount is 1000 cfug of bacteria and 100 cfug of fungi fornatural products) In specific tests for microorganisms it wasfound that Escherichia coli Salmonella typhi Pseudomonasaeruginosa and Staphylococcus aureus were absent which isessential criteria for their use as excipients in dosage forms

The results of accelerated stability studies on S tora seedpolysaccharide powder showed that there is no significantdifference between the initial and final samples withdrawnat time interval 6 months at 40∘C75 RH Slight changes inpH and moisture content values were observed The studiesproved that the S tora seed polysaccharide is stable for along period of time FTIR study revealed the absence ofdegradation or decomposition of polysaccharide at acceler-ated stability conditions for six months as shown in Figure 5

International Journal of Biomaterials 5

Table 3 Interpretation of FTIR data of polysaccharide

Wave length Peak assignments813 and 875 cmminus1 Anomeric configurations (120572 and 120573 conformers) and glycosidic linkages1198 and 983 cmminus1 Stretching vibration of CndashO in CndashOndashH bonds1149 cmminus1 Bending vibrational modes of CndashO present in the pyranose ring1134 and 983 cmminus1 CndashOH bending2800ndash3000 cmminus1 CndashH stretching3100ndash3500 cmminus1 OndashH stretching vibration

(1cm)

T(

)

90

80

70

60

4000 3500 3000 2500 2000 1750 1500 1250 1000 750 500

374190

354323

346222

339279

292409

235316

231844

179380

173979

167807

164528

153920

151412

145433

142732

133860

131545

114957

108785

102613

87568

81396

Figure 4 FTIR spectrum of the S tora polysaccharide (initial sample)

(1cm)

T(

)

4000 3500 3000 2500 2000 1750 1500 1250 1000 750

45

375

30

225

15

358567

354323

334650

298002

294337

288358

283150

235894

234158

215256

174751

173208

165107155655

153920

151991

147169

141768

131738

124602

115150

105892

102613

87375

81396

Figure 5 FTIR spectrum of the S tora polysaccharide (after 6 month)

6 International Journal of Biomaterials

4 Conclusion

The polysaccharide (gum) extracted from S tora seedspossesses good flow property The gum possesses excellentviscosity The gum polysaccharide was found to be stable ataccelerated stability condition Microbial studies confirmedits suitability as an excipient Based on the physicochemicalmicrobiological characteristics and stability studies it may bepointed out that the polysaccharide isolated from S tora seedshas the required properties and could be used as an excipientfor pharmaceutical dosage forms

Thus there is need to investigate further S tora polysac-charide as an excipient in different pharmaceutical dosageforms It may provide an alternative to synthetic or semisyn-thetic excipientspolymers currently used in the pharmaceu-tical industry

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors are thankful to Ultra College of PharmacyMadurai for providing the laboratory facilities to carry outthe present investigation

References

[1] A Ingle P Ranaware A Ladke and M Damle ldquoCassia toraphytochemical and pharmacological activityrdquo InternationalImperial Journal of Pharmacognosy amp Natural Products vol 2no 1 pp 14ndash23 2012

[2] C S Narayan and S Rangaswami ldquoIsolation of three crystallinesubstances from the seeds of Cassia torardquo Current Science vol25 article 559 1956

[3] I H Suba Jois and B L Manjunath ldquoChemical examination ofseeds of Cassia toralinnrdquo Journal of the Indian Chemical Societyvol 7 p 521 1930

[4] R E Wilkinson and W S Hardcastle ldquoComparative fatty acidcontent of various organs of Cassia tora Lrdquo Botanical Gazettevol 130 no 4 pp 254ndash258 1969

[5] H Pawar and PM DrsquoMello ldquoIsolation of seed gum fromCassiatora and preliminary studies of its application as binder fortabletsrdquo Indian Drugs vol 41 no 8 article 465 2004

[6] K R Kirtikar andBD Basu IndianMedicinal Plants PeriodicalExperts D-42 Vivek Vihar New Delhi India 1975

[7] T Nikaido T Ohmoto U Sankawa S Kitanaka and MTakido ldquoInhibitors of adenosine 3101584051015840-cyclic monophosphatephosphodiesterase in cassia seedrdquoChemical andPharmaceuticalBulletin vol 32 no 8 pp 3075ndash3078 1984

[8] H S Yun and IMChang ldquoPlantswith liver protective activities(I)rdquo Korean Journal of Pharmacognosy vol 8 pp 125ndash129 1977

[9] J D Hooker The Flora of British India vol 2 Reeve amp CoSuffolk UK 1879

[10] R S Thakur H S Puri and A HusainMajor Medicinal Plantsof India 1st edition 1989

[11] H A Pawar and K G Lalitha ldquoIsolation purification andcharacterization of galactomannans as an excipient from Senna

tora seedsrdquo International Journal of Biological Macromoleculesvol 65 pp 167ndash175 2014

[12] C V S Subrahmanyam Physical Pharmacy Vallabh PrakashanNew Delhi India 2nd edition 2000

[13] M Emeje P Nwabunike C Isimi et al ldquoIsolation charac-terization and formulation properties of a new plant gumobtained from Cissus refescencerdquo International Journal of GreenPharmacy vol 3 no 1 pp 16ndash23 2009

[14] S E Gaisford S E Harding J R Mitchell and T D Bradley ldquoAcomparison between the hot and cold water soluble fractions oftwo locust bean gum samplesrdquo Carbohydrate Polymers vol 6no 6 pp 423ndash442 1986

[15] Ministry of Health and Family Welfare and Government ofIndia Indian Pharmacopoeia vol 556 Controller of Publica-tion New Delhi India 1996

[16] International Conference on Harmonization ICH GuidelinesQ1A (R2) Stability Testing of New Drug Substances and Products(Revision 2) InternationalConference onHarmonization 2003

[17] J A Casas A F Mohedano and F Garcıa-Ochoa ldquoViscosity ofguar gum and xanthanguar gum mixture solutionsrdquo Journal ofthe Science of Food and Agriculture vol 80 no 12 pp 1722ndash17272000

[18] S D Figueiro J C Goes R A Moreira and A S BSombra ldquoOn the physico-chemical and dielectric propertiesof glutaraldehyde crosslinked galactomannan-collagen filmsrdquoCarbohydrate Polymers vol 56 no 3 pp 313ndash320 2004

[19] BM Prado S Kim B F Ozen and L JMauer ldquoDifferentiationof carbohydrate gums and mixtures using fourier transforminfrared spectroscopy and chemometricsrdquo Journal of Agricul-tural and Food Chemistry vol 53 no 8 pp 2823ndash2829 2005

[20] M R S Prashanth K S Parvathy N S Susheelamma et alldquoGalactomannan estersmdashA simple cost-effective method ofpreparation and characterizationrdquo Food Hydrocolloids vol 20no 8 pp 1198ndash1205 2006

[21] S-N Yuen S-M Choi D L Phillips and C-Y Ma ldquoRamanand FTIR spectroscopic study of carboxymethylated non-starchpolysaccharidesrdquo Food Chemistry vol 114 no 3 pp 1091ndash10982009

[22] Y Peng L Zhang F Zeng and Y Xu ldquoStructure and antitumoractivity of extracellular polysaccharides from myceliumrdquo Car-bohydrate Polymers vol 54 no 3 pp 297ndash303 2003

[23] L Zhang L Yang Q Ding and X Chen ldquoStudies on molec-ular weights of polysaccharides of Auricularia auricula-judaerdquoCarbohydrate Research vol 270 no 1 pp 1ndash10 1995

Submit your manuscripts athttpwwwhindawicom

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CorrosionInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Polymer ScienceInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CeramicsJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CompositesJournal of

NanoparticlesJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Biomaterials

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

NanoscienceJournal of

TextilesHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Journal of

NanotechnologyHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

CrystallographyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CoatingsJournal of

Advances in

Materials Science and EngineeringHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Smart Materials Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MetallurgyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

MaterialsJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Nano

materials

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal ofNanomaterials

Page 3: Research Article Extraction, Characterization, and ...

International Journal of Biomaterials 3

Table 1 Accelerated stability protocol of S tora gum

Product S tora seed polysaccharideQuantity loaded 20 g approx in each Petri dish

Quantity Sample 2 g approxSampling interval Storage conditions TestInitialFirst monthSecond monthThird monthSixth month

Temperature = 40∘CHumidity = 75 RH

(1) Organoleptic evaluation(2) pH of 1 wv solution(3) Moisture content(4) Microbial count

(5) FTIR

The relative viscosity of S tora gum dissolved in dis-tilled water (001ndash005 gdL) was measured using Ostwaldviscometer at room temperature Average molecular weight119872V was calculated using the Mark-Houwink relationship[Equation (3)] given by Doublier and Launay (1981) for guargum asmodified by Gaisford Harding Mitchell and Bradley(1986) to take into account the different values of mannose(M)galactose (G) of the galactomannans [14] Consider

120578 = 1155 times 10minus6

[(1 minus 120572) times119872V]098

(3)where 120572 = 1[(MG) + 1] and [120578] is expressed in dLg

27 Effect of Temperature on Viscosity of S tora Gum Effectof temperature on the viscosity of dilute solution (01wvin water) of S tora gum was studied experimentally withincrease in temperature from 36∘C to 100∘C The viscositymeasurements were performed using Ostwald viscometer

28 Total Microbial Load The total microbial load is animportant parameter which decides the suitability of asubstance for use as excipient in pharmaceutical dosageforms According to many Pharmacopoeias for syntheticand semisynthetic substances the total aerobic count shouldnot be more than 100 colony forming units (cfu) per gramand the total fungal count (including yeasts and molds)should not exceed 50 cfug In the case of excipients fromnatural origin the total aerobic count should not be morethan 1000 cfug and total fungal count should not exceed100 cfug The total microbial load of the isolated gumsample was determined by plate count method as per IndianPharmacopoeia [15]

29 Tests for Presence of Specific Microorganisms Materialsto be used as excipients in dosage forms need to be free fromEscherichia coli Salmonella typhi Pseudomonas aeruginosaand Staphylococcus aureus according to PharmacopoeiasInternational Conference for Harmonization (ICH) has alsosupported this criterion The tests for these microorganismswere carried out as described in Indian Pharmacopoeia

210 Accelerated Stability Study S tora seed polysaccharidewas subjected to accelerated stability studies for six monthsaccording to ICH guidelines to predict the stability of Stora seed polysaccharide [16] The samples were analyzed atregular intervals as per the stability protocol (Table 1)

Table 2 Results of micromeritic properties of S tora seed gum

Parameters Resultslowast

Bulk density (gmL) 051 plusmn 0047Tapped density (gmL) 060 plusmn 0039Hausnerrsquos ratio 117Carr index 1429Angle of repose 29∘441015840 plusmn 0032lowastThe values represent mean plusmn SD (119899 = 3)

3 Result and Discussion

The gum obtained from S tora seeds was an amorphous freeflowing odourless powder with dull brown colour (yield =35ww) The photograph of the gum powder was takenunder high intensity electron light microscope and usingdigital camera as depicted in Figures 1(a) and 1(b)

The results of micromeritic study are summarised inTable 2 The bulk density tapped density and angle ofrepose data reveal that S tora gum possesses good flowproperty Carr index is frequently used in pharmaceuticsas an indication of the compressibility and flowability of apowder In a free flowing powder the bulk density and tappeddensity would be close in value therefore Carr index wouldbe small A Carr index greater than 25 is considered to be anindication of poor flowability and a Carr index below 15 isconsidered to be of good flowability

Viscosity is the main parameter to assess the qualityof natural gums The applications of any natural gum aredependent on its viscosity For any polymer to be used in slowrelease hydrophilic matrix systems it should possess certaincharacteristics like fast hydration of the polymer and highgel strength and should be stable during the shelf life of theproduct Viscosity of 1wv solution of polysaccharide inwater was found to be 4520 cP at 50 revolutions per minute(rpm) The results indicated that the polysaccharide isolatedfrom S tora seeds possesses high viscosity with good swellingcapacity S tora seed polysaccharide hydrates quickly andswells rapidly and forms a thick viscous gel around it Sucha high viscosity indicates its utility in the development ofvarious modified release pharmaceutical dosage forms Theeffect of temperature on viscosity of S tora gum is depictedin Figure 2

4 International Journal of Biomaterials

(a) (b)

Figure 1 Photograph of gum powder taken (a) under high magnification microscope and (b) using digital camera

0000

0200

0400

0600

0800

1000

1200

1400

1600

0 20 40 60 80 100 120

Visc

osity

(cP)

Temperature (∘C)

Figure 2 Effect of temperature on viscosity of S tora gum

It has been reported in previous literature that S toragum contains galactomannans [11] Galactomannans areformed by several fractions of polymers with differentmannosegalactose ratios At low temperature moleculeswith low molecular weight and low mannosegalactose ratioare mainly dissolved At high temperatures galactomannanfractions with high molecular weight greater mannose togalactose relation and therefore zones without galactoseradicals are solved [17] The results indicated that viscosityof gum solution increases with increase in temperature Thismay be due to increase in solubility of galactomannans athigher temperature

The intrinsic viscosity was determined graphically fromHuggins plot (reduced viscosity versus concentration graphextrapolated to zero) Huggins plot is represented in Figure 3

The intrinsic viscosity obtained was 1568 dLg The aver-age molecular weight of purified S tora gum was found to be198 kDa by intrinsic viscosity method

Figure 4 represents FTIR spectrum of S tora polysaccha-ride

The FTIR spectrum of the initial sample (Figure 4) of iso-lated polysaccharide confirmed the presence of galactoman-nansThe interpretation was done using previously publishedliteratures [18ndash23] The FTIR data of the polysaccharide ispresented in Table 3

000

500

1000

1500

2000

2500

0 002 004 006

y = 36282x + 15684

R2 = 09575

Redu

ced

visc

osity

Concentration (gdL)

Figure 3 Huggins plot

The total microbial load was found to be 34023 plusmn1079 cfug of bacteria and 4439 plusmn 931 cfug of fungi Thetotal microbial load of the polysaccharide was thus withinthe acceptable limits (the acceptable limit for total microbialcount is 1000 cfug of bacteria and 100 cfug of fungi fornatural products) In specific tests for microorganisms it wasfound that Escherichia coli Salmonella typhi Pseudomonasaeruginosa and Staphylococcus aureus were absent which isessential criteria for their use as excipients in dosage forms

The results of accelerated stability studies on S tora seedpolysaccharide powder showed that there is no significantdifference between the initial and final samples withdrawnat time interval 6 months at 40∘C75 RH Slight changes inpH and moisture content values were observed The studiesproved that the S tora seed polysaccharide is stable for along period of time FTIR study revealed the absence ofdegradation or decomposition of polysaccharide at acceler-ated stability conditions for six months as shown in Figure 5

International Journal of Biomaterials 5

Table 3 Interpretation of FTIR data of polysaccharide

Wave length Peak assignments813 and 875 cmminus1 Anomeric configurations (120572 and 120573 conformers) and glycosidic linkages1198 and 983 cmminus1 Stretching vibration of CndashO in CndashOndashH bonds1149 cmminus1 Bending vibrational modes of CndashO present in the pyranose ring1134 and 983 cmminus1 CndashOH bending2800ndash3000 cmminus1 CndashH stretching3100ndash3500 cmminus1 OndashH stretching vibration

(1cm)

T(

)

90

80

70

60

4000 3500 3000 2500 2000 1750 1500 1250 1000 750 500

374190

354323

346222

339279

292409

235316

231844

179380

173979

167807

164528

153920

151412

145433

142732

133860

131545

114957

108785

102613

87568

81396

Figure 4 FTIR spectrum of the S tora polysaccharide (initial sample)

(1cm)

T(

)

4000 3500 3000 2500 2000 1750 1500 1250 1000 750

45

375

30

225

15

358567

354323

334650

298002

294337

288358

283150

235894

234158

215256

174751

173208

165107155655

153920

151991

147169

141768

131738

124602

115150

105892

102613

87375

81396

Figure 5 FTIR spectrum of the S tora polysaccharide (after 6 month)

6 International Journal of Biomaterials

4 Conclusion

The polysaccharide (gum) extracted from S tora seedspossesses good flow property The gum possesses excellentviscosity The gum polysaccharide was found to be stable ataccelerated stability condition Microbial studies confirmedits suitability as an excipient Based on the physicochemicalmicrobiological characteristics and stability studies it may bepointed out that the polysaccharide isolated from S tora seedshas the required properties and could be used as an excipientfor pharmaceutical dosage forms

Thus there is need to investigate further S tora polysac-charide as an excipient in different pharmaceutical dosageforms It may provide an alternative to synthetic or semisyn-thetic excipientspolymers currently used in the pharmaceu-tical industry

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors are thankful to Ultra College of PharmacyMadurai for providing the laboratory facilities to carry outthe present investigation

References

[1] A Ingle P Ranaware A Ladke and M Damle ldquoCassia toraphytochemical and pharmacological activityrdquo InternationalImperial Journal of Pharmacognosy amp Natural Products vol 2no 1 pp 14ndash23 2012

[2] C S Narayan and S Rangaswami ldquoIsolation of three crystallinesubstances from the seeds of Cassia torardquo Current Science vol25 article 559 1956

[3] I H Suba Jois and B L Manjunath ldquoChemical examination ofseeds of Cassia toralinnrdquo Journal of the Indian Chemical Societyvol 7 p 521 1930

[4] R E Wilkinson and W S Hardcastle ldquoComparative fatty acidcontent of various organs of Cassia tora Lrdquo Botanical Gazettevol 130 no 4 pp 254ndash258 1969

[5] H Pawar and PM DrsquoMello ldquoIsolation of seed gum fromCassiatora and preliminary studies of its application as binder fortabletsrdquo Indian Drugs vol 41 no 8 article 465 2004

[6] K R Kirtikar andBD Basu IndianMedicinal Plants PeriodicalExperts D-42 Vivek Vihar New Delhi India 1975

[7] T Nikaido T Ohmoto U Sankawa S Kitanaka and MTakido ldquoInhibitors of adenosine 3101584051015840-cyclic monophosphatephosphodiesterase in cassia seedrdquoChemical andPharmaceuticalBulletin vol 32 no 8 pp 3075ndash3078 1984

[8] H S Yun and IMChang ldquoPlantswith liver protective activities(I)rdquo Korean Journal of Pharmacognosy vol 8 pp 125ndash129 1977

[9] J D Hooker The Flora of British India vol 2 Reeve amp CoSuffolk UK 1879

[10] R S Thakur H S Puri and A HusainMajor Medicinal Plantsof India 1st edition 1989

[11] H A Pawar and K G Lalitha ldquoIsolation purification andcharacterization of galactomannans as an excipient from Senna

tora seedsrdquo International Journal of Biological Macromoleculesvol 65 pp 167ndash175 2014

[12] C V S Subrahmanyam Physical Pharmacy Vallabh PrakashanNew Delhi India 2nd edition 2000

[13] M Emeje P Nwabunike C Isimi et al ldquoIsolation charac-terization and formulation properties of a new plant gumobtained from Cissus refescencerdquo International Journal of GreenPharmacy vol 3 no 1 pp 16ndash23 2009

[14] S E Gaisford S E Harding J R Mitchell and T D Bradley ldquoAcomparison between the hot and cold water soluble fractions oftwo locust bean gum samplesrdquo Carbohydrate Polymers vol 6no 6 pp 423ndash442 1986

[15] Ministry of Health and Family Welfare and Government ofIndia Indian Pharmacopoeia vol 556 Controller of Publica-tion New Delhi India 1996

[16] International Conference on Harmonization ICH GuidelinesQ1A (R2) Stability Testing of New Drug Substances and Products(Revision 2) InternationalConference onHarmonization 2003

[17] J A Casas A F Mohedano and F Garcıa-Ochoa ldquoViscosity ofguar gum and xanthanguar gum mixture solutionsrdquo Journal ofthe Science of Food and Agriculture vol 80 no 12 pp 1722ndash17272000

[18] S D Figueiro J C Goes R A Moreira and A S BSombra ldquoOn the physico-chemical and dielectric propertiesof glutaraldehyde crosslinked galactomannan-collagen filmsrdquoCarbohydrate Polymers vol 56 no 3 pp 313ndash320 2004

[19] BM Prado S Kim B F Ozen and L JMauer ldquoDifferentiationof carbohydrate gums and mixtures using fourier transforminfrared spectroscopy and chemometricsrdquo Journal of Agricul-tural and Food Chemistry vol 53 no 8 pp 2823ndash2829 2005

[20] M R S Prashanth K S Parvathy N S Susheelamma et alldquoGalactomannan estersmdashA simple cost-effective method ofpreparation and characterizationrdquo Food Hydrocolloids vol 20no 8 pp 1198ndash1205 2006

[21] S-N Yuen S-M Choi D L Phillips and C-Y Ma ldquoRamanand FTIR spectroscopic study of carboxymethylated non-starchpolysaccharidesrdquo Food Chemistry vol 114 no 3 pp 1091ndash10982009

[22] Y Peng L Zhang F Zeng and Y Xu ldquoStructure and antitumoractivity of extracellular polysaccharides from myceliumrdquo Car-bohydrate Polymers vol 54 no 3 pp 297ndash303 2003

[23] L Zhang L Yang Q Ding and X Chen ldquoStudies on molec-ular weights of polysaccharides of Auricularia auricula-judaerdquoCarbohydrate Research vol 270 no 1 pp 1ndash10 1995

Submit your manuscripts athttpwwwhindawicom

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CorrosionInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Polymer ScienceInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CeramicsJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CompositesJournal of

NanoparticlesJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Biomaterials

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

NanoscienceJournal of

TextilesHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Journal of

NanotechnologyHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

CrystallographyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CoatingsJournal of

Advances in

Materials Science and EngineeringHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Smart Materials Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MetallurgyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

MaterialsJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Nano

materials

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal ofNanomaterials

Page 4: Research Article Extraction, Characterization, and ...

4 International Journal of Biomaterials

(a) (b)

Figure 1 Photograph of gum powder taken (a) under high magnification microscope and (b) using digital camera

0000

0200

0400

0600

0800

1000

1200

1400

1600

0 20 40 60 80 100 120

Visc

osity

(cP)

Temperature (∘C)

Figure 2 Effect of temperature on viscosity of S tora gum

It has been reported in previous literature that S toragum contains galactomannans [11] Galactomannans areformed by several fractions of polymers with differentmannosegalactose ratios At low temperature moleculeswith low molecular weight and low mannosegalactose ratioare mainly dissolved At high temperatures galactomannanfractions with high molecular weight greater mannose togalactose relation and therefore zones without galactoseradicals are solved [17] The results indicated that viscosityof gum solution increases with increase in temperature Thismay be due to increase in solubility of galactomannans athigher temperature

The intrinsic viscosity was determined graphically fromHuggins plot (reduced viscosity versus concentration graphextrapolated to zero) Huggins plot is represented in Figure 3

The intrinsic viscosity obtained was 1568 dLg The aver-age molecular weight of purified S tora gum was found to be198 kDa by intrinsic viscosity method

Figure 4 represents FTIR spectrum of S tora polysaccha-ride

The FTIR spectrum of the initial sample (Figure 4) of iso-lated polysaccharide confirmed the presence of galactoman-nansThe interpretation was done using previously publishedliteratures [18ndash23] The FTIR data of the polysaccharide ispresented in Table 3

000

500

1000

1500

2000

2500

0 002 004 006

y = 36282x + 15684

R2 = 09575

Redu

ced

visc

osity

Concentration (gdL)

Figure 3 Huggins plot

The total microbial load was found to be 34023 plusmn1079 cfug of bacteria and 4439 plusmn 931 cfug of fungi Thetotal microbial load of the polysaccharide was thus withinthe acceptable limits (the acceptable limit for total microbialcount is 1000 cfug of bacteria and 100 cfug of fungi fornatural products) In specific tests for microorganisms it wasfound that Escherichia coli Salmonella typhi Pseudomonasaeruginosa and Staphylococcus aureus were absent which isessential criteria for their use as excipients in dosage forms

The results of accelerated stability studies on S tora seedpolysaccharide powder showed that there is no significantdifference between the initial and final samples withdrawnat time interval 6 months at 40∘C75 RH Slight changes inpH and moisture content values were observed The studiesproved that the S tora seed polysaccharide is stable for along period of time FTIR study revealed the absence ofdegradation or decomposition of polysaccharide at acceler-ated stability conditions for six months as shown in Figure 5

International Journal of Biomaterials 5

Table 3 Interpretation of FTIR data of polysaccharide

Wave length Peak assignments813 and 875 cmminus1 Anomeric configurations (120572 and 120573 conformers) and glycosidic linkages1198 and 983 cmminus1 Stretching vibration of CndashO in CndashOndashH bonds1149 cmminus1 Bending vibrational modes of CndashO present in the pyranose ring1134 and 983 cmminus1 CndashOH bending2800ndash3000 cmminus1 CndashH stretching3100ndash3500 cmminus1 OndashH stretching vibration

(1cm)

T(

)

90

80

70

60

4000 3500 3000 2500 2000 1750 1500 1250 1000 750 500

374190

354323

346222

339279

292409

235316

231844

179380

173979

167807

164528

153920

151412

145433

142732

133860

131545

114957

108785

102613

87568

81396

Figure 4 FTIR spectrum of the S tora polysaccharide (initial sample)

(1cm)

T(

)

4000 3500 3000 2500 2000 1750 1500 1250 1000 750

45

375

30

225

15

358567

354323

334650

298002

294337

288358

283150

235894

234158

215256

174751

173208

165107155655

153920

151991

147169

141768

131738

124602

115150

105892

102613

87375

81396

Figure 5 FTIR spectrum of the S tora polysaccharide (after 6 month)

6 International Journal of Biomaterials

4 Conclusion

The polysaccharide (gum) extracted from S tora seedspossesses good flow property The gum possesses excellentviscosity The gum polysaccharide was found to be stable ataccelerated stability condition Microbial studies confirmedits suitability as an excipient Based on the physicochemicalmicrobiological characteristics and stability studies it may bepointed out that the polysaccharide isolated from S tora seedshas the required properties and could be used as an excipientfor pharmaceutical dosage forms

Thus there is need to investigate further S tora polysac-charide as an excipient in different pharmaceutical dosageforms It may provide an alternative to synthetic or semisyn-thetic excipientspolymers currently used in the pharmaceu-tical industry

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors are thankful to Ultra College of PharmacyMadurai for providing the laboratory facilities to carry outthe present investigation

References

[1] A Ingle P Ranaware A Ladke and M Damle ldquoCassia toraphytochemical and pharmacological activityrdquo InternationalImperial Journal of Pharmacognosy amp Natural Products vol 2no 1 pp 14ndash23 2012

[2] C S Narayan and S Rangaswami ldquoIsolation of three crystallinesubstances from the seeds of Cassia torardquo Current Science vol25 article 559 1956

[3] I H Suba Jois and B L Manjunath ldquoChemical examination ofseeds of Cassia toralinnrdquo Journal of the Indian Chemical Societyvol 7 p 521 1930

[4] R E Wilkinson and W S Hardcastle ldquoComparative fatty acidcontent of various organs of Cassia tora Lrdquo Botanical Gazettevol 130 no 4 pp 254ndash258 1969

[5] H Pawar and PM DrsquoMello ldquoIsolation of seed gum fromCassiatora and preliminary studies of its application as binder fortabletsrdquo Indian Drugs vol 41 no 8 article 465 2004

[6] K R Kirtikar andBD Basu IndianMedicinal Plants PeriodicalExperts D-42 Vivek Vihar New Delhi India 1975

[7] T Nikaido T Ohmoto U Sankawa S Kitanaka and MTakido ldquoInhibitors of adenosine 3101584051015840-cyclic monophosphatephosphodiesterase in cassia seedrdquoChemical andPharmaceuticalBulletin vol 32 no 8 pp 3075ndash3078 1984

[8] H S Yun and IMChang ldquoPlantswith liver protective activities(I)rdquo Korean Journal of Pharmacognosy vol 8 pp 125ndash129 1977

[9] J D Hooker The Flora of British India vol 2 Reeve amp CoSuffolk UK 1879

[10] R S Thakur H S Puri and A HusainMajor Medicinal Plantsof India 1st edition 1989

[11] H A Pawar and K G Lalitha ldquoIsolation purification andcharacterization of galactomannans as an excipient from Senna

tora seedsrdquo International Journal of Biological Macromoleculesvol 65 pp 167ndash175 2014

[12] C V S Subrahmanyam Physical Pharmacy Vallabh PrakashanNew Delhi India 2nd edition 2000

[13] M Emeje P Nwabunike C Isimi et al ldquoIsolation charac-terization and formulation properties of a new plant gumobtained from Cissus refescencerdquo International Journal of GreenPharmacy vol 3 no 1 pp 16ndash23 2009

[14] S E Gaisford S E Harding J R Mitchell and T D Bradley ldquoAcomparison between the hot and cold water soluble fractions oftwo locust bean gum samplesrdquo Carbohydrate Polymers vol 6no 6 pp 423ndash442 1986

[15] Ministry of Health and Family Welfare and Government ofIndia Indian Pharmacopoeia vol 556 Controller of Publica-tion New Delhi India 1996

[16] International Conference on Harmonization ICH GuidelinesQ1A (R2) Stability Testing of New Drug Substances and Products(Revision 2) InternationalConference onHarmonization 2003

[17] J A Casas A F Mohedano and F Garcıa-Ochoa ldquoViscosity ofguar gum and xanthanguar gum mixture solutionsrdquo Journal ofthe Science of Food and Agriculture vol 80 no 12 pp 1722ndash17272000

[18] S D Figueiro J C Goes R A Moreira and A S BSombra ldquoOn the physico-chemical and dielectric propertiesof glutaraldehyde crosslinked galactomannan-collagen filmsrdquoCarbohydrate Polymers vol 56 no 3 pp 313ndash320 2004

[19] BM Prado S Kim B F Ozen and L JMauer ldquoDifferentiationof carbohydrate gums and mixtures using fourier transforminfrared spectroscopy and chemometricsrdquo Journal of Agricul-tural and Food Chemistry vol 53 no 8 pp 2823ndash2829 2005

[20] M R S Prashanth K S Parvathy N S Susheelamma et alldquoGalactomannan estersmdashA simple cost-effective method ofpreparation and characterizationrdquo Food Hydrocolloids vol 20no 8 pp 1198ndash1205 2006

[21] S-N Yuen S-M Choi D L Phillips and C-Y Ma ldquoRamanand FTIR spectroscopic study of carboxymethylated non-starchpolysaccharidesrdquo Food Chemistry vol 114 no 3 pp 1091ndash10982009

[22] Y Peng L Zhang F Zeng and Y Xu ldquoStructure and antitumoractivity of extracellular polysaccharides from myceliumrdquo Car-bohydrate Polymers vol 54 no 3 pp 297ndash303 2003

[23] L Zhang L Yang Q Ding and X Chen ldquoStudies on molec-ular weights of polysaccharides of Auricularia auricula-judaerdquoCarbohydrate Research vol 270 no 1 pp 1ndash10 1995

Submit your manuscripts athttpwwwhindawicom

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CorrosionInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Polymer ScienceInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CeramicsJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CompositesJournal of

NanoparticlesJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Biomaterials

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

NanoscienceJournal of

TextilesHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Journal of

NanotechnologyHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

CrystallographyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CoatingsJournal of

Advances in

Materials Science and EngineeringHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Smart Materials Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MetallurgyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

MaterialsJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Nano

materials

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal ofNanomaterials

Page 5: Research Article Extraction, Characterization, and ...

International Journal of Biomaterials 5

Table 3 Interpretation of FTIR data of polysaccharide

Wave length Peak assignments813 and 875 cmminus1 Anomeric configurations (120572 and 120573 conformers) and glycosidic linkages1198 and 983 cmminus1 Stretching vibration of CndashO in CndashOndashH bonds1149 cmminus1 Bending vibrational modes of CndashO present in the pyranose ring1134 and 983 cmminus1 CndashOH bending2800ndash3000 cmminus1 CndashH stretching3100ndash3500 cmminus1 OndashH stretching vibration

(1cm)

T(

)

90

80

70

60

4000 3500 3000 2500 2000 1750 1500 1250 1000 750 500

374190

354323

346222

339279

292409

235316

231844

179380

173979

167807

164528

153920

151412

145433

142732

133860

131545

114957

108785

102613

87568

81396

Figure 4 FTIR spectrum of the S tora polysaccharide (initial sample)

(1cm)

T(

)

4000 3500 3000 2500 2000 1750 1500 1250 1000 750

45

375

30

225

15

358567

354323

334650

298002

294337

288358

283150

235894

234158

215256

174751

173208

165107155655

153920

151991

147169

141768

131738

124602

115150

105892

102613

87375

81396

Figure 5 FTIR spectrum of the S tora polysaccharide (after 6 month)

6 International Journal of Biomaterials

4 Conclusion

The polysaccharide (gum) extracted from S tora seedspossesses good flow property The gum possesses excellentviscosity The gum polysaccharide was found to be stable ataccelerated stability condition Microbial studies confirmedits suitability as an excipient Based on the physicochemicalmicrobiological characteristics and stability studies it may bepointed out that the polysaccharide isolated from S tora seedshas the required properties and could be used as an excipientfor pharmaceutical dosage forms

Thus there is need to investigate further S tora polysac-charide as an excipient in different pharmaceutical dosageforms It may provide an alternative to synthetic or semisyn-thetic excipientspolymers currently used in the pharmaceu-tical industry

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors are thankful to Ultra College of PharmacyMadurai for providing the laboratory facilities to carry outthe present investigation

References

[1] A Ingle P Ranaware A Ladke and M Damle ldquoCassia toraphytochemical and pharmacological activityrdquo InternationalImperial Journal of Pharmacognosy amp Natural Products vol 2no 1 pp 14ndash23 2012

[2] C S Narayan and S Rangaswami ldquoIsolation of three crystallinesubstances from the seeds of Cassia torardquo Current Science vol25 article 559 1956

[3] I H Suba Jois and B L Manjunath ldquoChemical examination ofseeds of Cassia toralinnrdquo Journal of the Indian Chemical Societyvol 7 p 521 1930

[4] R E Wilkinson and W S Hardcastle ldquoComparative fatty acidcontent of various organs of Cassia tora Lrdquo Botanical Gazettevol 130 no 4 pp 254ndash258 1969

[5] H Pawar and PM DrsquoMello ldquoIsolation of seed gum fromCassiatora and preliminary studies of its application as binder fortabletsrdquo Indian Drugs vol 41 no 8 article 465 2004

[6] K R Kirtikar andBD Basu IndianMedicinal Plants PeriodicalExperts D-42 Vivek Vihar New Delhi India 1975

[7] T Nikaido T Ohmoto U Sankawa S Kitanaka and MTakido ldquoInhibitors of adenosine 3101584051015840-cyclic monophosphatephosphodiesterase in cassia seedrdquoChemical andPharmaceuticalBulletin vol 32 no 8 pp 3075ndash3078 1984

[8] H S Yun and IMChang ldquoPlantswith liver protective activities(I)rdquo Korean Journal of Pharmacognosy vol 8 pp 125ndash129 1977

[9] J D Hooker The Flora of British India vol 2 Reeve amp CoSuffolk UK 1879

[10] R S Thakur H S Puri and A HusainMajor Medicinal Plantsof India 1st edition 1989

[11] H A Pawar and K G Lalitha ldquoIsolation purification andcharacterization of galactomannans as an excipient from Senna

tora seedsrdquo International Journal of Biological Macromoleculesvol 65 pp 167ndash175 2014

[12] C V S Subrahmanyam Physical Pharmacy Vallabh PrakashanNew Delhi India 2nd edition 2000

[13] M Emeje P Nwabunike C Isimi et al ldquoIsolation charac-terization and formulation properties of a new plant gumobtained from Cissus refescencerdquo International Journal of GreenPharmacy vol 3 no 1 pp 16ndash23 2009

[14] S E Gaisford S E Harding J R Mitchell and T D Bradley ldquoAcomparison between the hot and cold water soluble fractions oftwo locust bean gum samplesrdquo Carbohydrate Polymers vol 6no 6 pp 423ndash442 1986

[15] Ministry of Health and Family Welfare and Government ofIndia Indian Pharmacopoeia vol 556 Controller of Publica-tion New Delhi India 1996

[16] International Conference on Harmonization ICH GuidelinesQ1A (R2) Stability Testing of New Drug Substances and Products(Revision 2) InternationalConference onHarmonization 2003

[17] J A Casas A F Mohedano and F Garcıa-Ochoa ldquoViscosity ofguar gum and xanthanguar gum mixture solutionsrdquo Journal ofthe Science of Food and Agriculture vol 80 no 12 pp 1722ndash17272000

[18] S D Figueiro J C Goes R A Moreira and A S BSombra ldquoOn the physico-chemical and dielectric propertiesof glutaraldehyde crosslinked galactomannan-collagen filmsrdquoCarbohydrate Polymers vol 56 no 3 pp 313ndash320 2004

[19] BM Prado S Kim B F Ozen and L JMauer ldquoDifferentiationof carbohydrate gums and mixtures using fourier transforminfrared spectroscopy and chemometricsrdquo Journal of Agricul-tural and Food Chemistry vol 53 no 8 pp 2823ndash2829 2005

[20] M R S Prashanth K S Parvathy N S Susheelamma et alldquoGalactomannan estersmdashA simple cost-effective method ofpreparation and characterizationrdquo Food Hydrocolloids vol 20no 8 pp 1198ndash1205 2006

[21] S-N Yuen S-M Choi D L Phillips and C-Y Ma ldquoRamanand FTIR spectroscopic study of carboxymethylated non-starchpolysaccharidesrdquo Food Chemistry vol 114 no 3 pp 1091ndash10982009

[22] Y Peng L Zhang F Zeng and Y Xu ldquoStructure and antitumoractivity of extracellular polysaccharides from myceliumrdquo Car-bohydrate Polymers vol 54 no 3 pp 297ndash303 2003

[23] L Zhang L Yang Q Ding and X Chen ldquoStudies on molec-ular weights of polysaccharides of Auricularia auricula-judaerdquoCarbohydrate Research vol 270 no 1 pp 1ndash10 1995

Submit your manuscripts athttpwwwhindawicom

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CorrosionInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Polymer ScienceInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CeramicsJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CompositesJournal of

NanoparticlesJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Biomaterials

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

NanoscienceJournal of

TextilesHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Journal of

NanotechnologyHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

CrystallographyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CoatingsJournal of

Advances in

Materials Science and EngineeringHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Smart Materials Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MetallurgyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

MaterialsJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Nano

materials

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal ofNanomaterials

Page 6: Research Article Extraction, Characterization, and ...

6 International Journal of Biomaterials

4 Conclusion

The polysaccharide (gum) extracted from S tora seedspossesses good flow property The gum possesses excellentviscosity The gum polysaccharide was found to be stable ataccelerated stability condition Microbial studies confirmedits suitability as an excipient Based on the physicochemicalmicrobiological characteristics and stability studies it may bepointed out that the polysaccharide isolated from S tora seedshas the required properties and could be used as an excipientfor pharmaceutical dosage forms

Thus there is need to investigate further S tora polysac-charide as an excipient in different pharmaceutical dosageforms It may provide an alternative to synthetic or semisyn-thetic excipientspolymers currently used in the pharmaceu-tical industry

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors are thankful to Ultra College of PharmacyMadurai for providing the laboratory facilities to carry outthe present investigation

References

[1] A Ingle P Ranaware A Ladke and M Damle ldquoCassia toraphytochemical and pharmacological activityrdquo InternationalImperial Journal of Pharmacognosy amp Natural Products vol 2no 1 pp 14ndash23 2012

[2] C S Narayan and S Rangaswami ldquoIsolation of three crystallinesubstances from the seeds of Cassia torardquo Current Science vol25 article 559 1956

[3] I H Suba Jois and B L Manjunath ldquoChemical examination ofseeds of Cassia toralinnrdquo Journal of the Indian Chemical Societyvol 7 p 521 1930

[4] R E Wilkinson and W S Hardcastle ldquoComparative fatty acidcontent of various organs of Cassia tora Lrdquo Botanical Gazettevol 130 no 4 pp 254ndash258 1969

[5] H Pawar and PM DrsquoMello ldquoIsolation of seed gum fromCassiatora and preliminary studies of its application as binder fortabletsrdquo Indian Drugs vol 41 no 8 article 465 2004

[6] K R Kirtikar andBD Basu IndianMedicinal Plants PeriodicalExperts D-42 Vivek Vihar New Delhi India 1975

[7] T Nikaido T Ohmoto U Sankawa S Kitanaka and MTakido ldquoInhibitors of adenosine 3101584051015840-cyclic monophosphatephosphodiesterase in cassia seedrdquoChemical andPharmaceuticalBulletin vol 32 no 8 pp 3075ndash3078 1984

[8] H S Yun and IMChang ldquoPlantswith liver protective activities(I)rdquo Korean Journal of Pharmacognosy vol 8 pp 125ndash129 1977

[9] J D Hooker The Flora of British India vol 2 Reeve amp CoSuffolk UK 1879

[10] R S Thakur H S Puri and A HusainMajor Medicinal Plantsof India 1st edition 1989

[11] H A Pawar and K G Lalitha ldquoIsolation purification andcharacterization of galactomannans as an excipient from Senna

tora seedsrdquo International Journal of Biological Macromoleculesvol 65 pp 167ndash175 2014

[12] C V S Subrahmanyam Physical Pharmacy Vallabh PrakashanNew Delhi India 2nd edition 2000

[13] M Emeje P Nwabunike C Isimi et al ldquoIsolation charac-terization and formulation properties of a new plant gumobtained from Cissus refescencerdquo International Journal of GreenPharmacy vol 3 no 1 pp 16ndash23 2009

[14] S E Gaisford S E Harding J R Mitchell and T D Bradley ldquoAcomparison between the hot and cold water soluble fractions oftwo locust bean gum samplesrdquo Carbohydrate Polymers vol 6no 6 pp 423ndash442 1986

[15] Ministry of Health and Family Welfare and Government ofIndia Indian Pharmacopoeia vol 556 Controller of Publica-tion New Delhi India 1996

[16] International Conference on Harmonization ICH GuidelinesQ1A (R2) Stability Testing of New Drug Substances and Products(Revision 2) InternationalConference onHarmonization 2003

[17] J A Casas A F Mohedano and F Garcıa-Ochoa ldquoViscosity ofguar gum and xanthanguar gum mixture solutionsrdquo Journal ofthe Science of Food and Agriculture vol 80 no 12 pp 1722ndash17272000

[18] S D Figueiro J C Goes R A Moreira and A S BSombra ldquoOn the physico-chemical and dielectric propertiesof glutaraldehyde crosslinked galactomannan-collagen filmsrdquoCarbohydrate Polymers vol 56 no 3 pp 313ndash320 2004

[19] BM Prado S Kim B F Ozen and L JMauer ldquoDifferentiationof carbohydrate gums and mixtures using fourier transforminfrared spectroscopy and chemometricsrdquo Journal of Agricul-tural and Food Chemistry vol 53 no 8 pp 2823ndash2829 2005

[20] M R S Prashanth K S Parvathy N S Susheelamma et alldquoGalactomannan estersmdashA simple cost-effective method ofpreparation and characterizationrdquo Food Hydrocolloids vol 20no 8 pp 1198ndash1205 2006

[21] S-N Yuen S-M Choi D L Phillips and C-Y Ma ldquoRamanand FTIR spectroscopic study of carboxymethylated non-starchpolysaccharidesrdquo Food Chemistry vol 114 no 3 pp 1091ndash10982009

[22] Y Peng L Zhang F Zeng and Y Xu ldquoStructure and antitumoractivity of extracellular polysaccharides from myceliumrdquo Car-bohydrate Polymers vol 54 no 3 pp 297ndash303 2003

[23] L Zhang L Yang Q Ding and X Chen ldquoStudies on molec-ular weights of polysaccharides of Auricularia auricula-judaerdquoCarbohydrate Research vol 270 no 1 pp 1ndash10 1995

Submit your manuscripts athttpwwwhindawicom

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CorrosionInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Polymer ScienceInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CeramicsJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CompositesJournal of

NanoparticlesJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Biomaterials

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

NanoscienceJournal of

TextilesHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Journal of

NanotechnologyHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

CrystallographyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CoatingsJournal of

Advances in

Materials Science and EngineeringHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Smart Materials Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MetallurgyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

MaterialsJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Nano

materials

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal ofNanomaterials

Page 7: Research Article Extraction, Characterization, and ...

Submit your manuscripts athttpwwwhindawicom

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CorrosionInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Polymer ScienceInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CeramicsJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CompositesJournal of

NanoparticlesJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

International Journal of

Biomaterials

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

NanoscienceJournal of

TextilesHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Journal of

NanotechnologyHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

CrystallographyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CoatingsJournal of

Advances in

Materials Science and EngineeringHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Smart Materials Research

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MetallurgyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

MaterialsJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Nano

materials

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal ofNanomaterials