Saponin Bitterness Reduction of Carica papaya …downloads.hindawi.com/journals/jfq/2018/5602729.pdfCarica papaya that belongs to Caricaceae family has long been known as a traditional

Research ArticleSaponin Bitterness Reduction of Carica papaya LeafExtracts through Adsorption of Weakly Basic IonExchange Resins

Sharifah Nuruljannah Syed Amran Noraziani Zainal Abidin Haslaniza Hashim and Saiful Irwan Zubairi

Centre for Biotechnology and Functional Food Faculty of Science and Technology Universiti Kebangsaan Malaysia (UKM)43600 Bangi Selangor Malaysia

Correspondence should be addressed to Saiful Irwan Zubairi saiful-zukmedumy

Received 2 March 2018 Accepted 14 August 2018 Published 24 September 2018

Academic Editor Efstathios Giaouris

Copyright copy 2018 Sharifah Nuruljannah Syed Amran et al is is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

Carica papaya that belongs to Caricaceae family has long been known as a traditional medicine for dengue fever as well as foranticancer and antiinammatory studies following identication of benecial phytochemicals such as saponins in the leavesUnfortunately the compound has been known to induce a bitter taste in the leaf extract for human consumption making themunpopular and nonconsumer friendly us this study aims to observe the potential adsorption of saponin compound from Cpapaya leaves using ion exchange resins as an adsorbent for the reduction of bitter-taste-inducing saponins is study uses threetypes of weakly basic ion exchanging resins namely Amberlite IRA-67 Diaion WA30 and DiaionWA21J at dierent adsorbentdoses of 5 (wv) and 10 (wv) e Peleg model suggests that the extraction of saponins from C papaya leaves lasted for 1250hours yield a maximum amount of saponins 931mgg Further study shows that there is a signicant dierence (plt 005) ofsaponin adsorption percentage between these three types of resins e Diaion WA30 resin showed the highest percentage ofadsorption at 8783 (ww) as compared to the other two 5 (wv) loaded resin e 10 (wv) resin-loaded Diaion WA30demonstrated the highest overall adsorption capacity as much as 9759 (ww) with the shortest exhaustive time of 499 hourse overall acceptance of samples in sensory evaluation treated with ion exchange resins gave good response in which the sampletreated with 10 (wv) resin-loaded DiaionWA30 demonstrated the highest overall acceptance in parallel with having the lowestbitterness score as compared to other samples and fresh samples (untreated) e Langmuir constant (RL) was less than one(0167ndash0398) indicating the adsorption of saponins onto Diaion WA30 was favourable

1 Introduction

C papaya locally known as papaya pear or pawpaworiginates from the Caricaceae family that stems into sixother genera including those of Carica Jarilla HorovitziaJacaraita Vasconcellea and Cylicomorphia [1 2] e planthas been used as traditional medicine to treat various diseasesthat aect human health In Nigeria alone it has been re-ported that the plant is widely used as a cure by the generalpopulation of those residing in selected local governmentareas of Kwara State including Moro Ifelodun Ekiti Afonand Oa for ailments such as typhoid malaria jaundicediarrhea measles and yellow fever [3] Meanwhile its

widespread uses in the Asia region are more likely to beassociated with treatment for dengue fever An array ofstudies have been reported on the use of C papaya plantparticularly the leaves in the treatment of dengue fever thatcontribute to an increased thrombocytes count in rats from9480times103 to 11985times103 after 72 hours of observationfollowing the administration of papaya leaf extracts as well asa signicant dierence (plt 005) in hematocrit levels betweenthe control group and the experimental group who were givencapsules made from papaya leaf extracts over the course of6ndash9 days [4 5]ere has also been a report on increased levelof platelet count from 55times103 microlminus1 to 168times103microlminus1 increasedwhite blood cell (WBC) count from 37times103microlminus1 to

HindawiJournal of Food QualityVolume 2018 Article ID 5602729 11 pageshttpsdoiorg10115520185602729

77times103microlminus1 as well as increased count of neutrophils from460 to 783 over the course of 5 days in dengue patientsfollowing administration of papaya leaf extracts [6]

e ability of the leaves to be used as a cure for thesediseases is most likely due to the presence of bioactive com-pounds and multiple phytochemicals in the leaves In 2007 atleast seven different bioactive compounds in the leaves of theCpapaya plant were identified including flavanols t tanninsalkaloids and carpaine [7 8] In fact the highest phyto-chemicals in C papaya leaves were also saponins which makeup to 0022plusmn 010 of all the phytochemicals tested present inthe same part [5] Although it bears the potential as a cure ormedicine for many ailments commercially available medicinalproducts from C papaya leaf extracts have hindered its po-tential due to its bitterness greatly believed to be caused bysaponins greatly believed to be caused by saponins [9 10]Previous research has found that one of the challenges of usingtraditional herbs as medicine for human health is the bittertaste associated with it particularly those that contain saponins[11] Recently many researchers have remarked that saponinsfrom a variety of plants including soybeans are very muchresponsible for the bitter taste of the extract even in lowconcentrations [12] It was found that there was indeeda positive correlation between the soya saponin concentrationin pea plant extracts (01 - 03 ww) and its bitterness levelanalysed via sensory evaluation [13] is was also true forsaponins that generally cause bitter tastes in dry pea samples[14]

In conjunction to this problem numerous attemptshave been made to remove saponins to prevent resur-facing of bitter taste in traditional medicinal herbs Ashighlighted over the years many methods have beendeveloped for the extraction isolation and quantificationof saponin compound [15] ese methods are usuallyseparated into two classes of conventional extractiontechnique and green extraction techniques with the lateremphasizing more on being environment friendly [16]However conventional extraction techniques are stillbeing preferred by 70 to 30 due to foregoing the cost ofheavy-equipment required for the extraction or isolationprocess Conventional methods for the extraction of sa-ponins are usually maceration (soaking) and the Soxhletmethod in reflux meanwhile the extraction techniquesinvolving green technology are those of ultrasound-assisted extraction techniques and microwave-assistedextraction techniques [17] is method manages toblock activation of the saponin compound through theaddition of phytosterols but this method is said to haveharmful effects when performed on animal feed as it willcause stunted animal growth and eventually death [18]

As such the objective of this research was to assess theeffectiveness of the saponin adsorption process of C papayaleaf extracts using three different types of weakly basic ionexchange resins to propose a new possible way of removingsaponins other than to determine the content percentage ofadsorption and exhaustive time of the overall adsorptionprocess as well as measuring the level of sensory acceptanceof C papaya leaf extracts which has undergone the ad-sorption process

2 Methodology

21 Materials and Chemicals Mature C papaya leaves (ofunspecified variety) were obtained from Banting Selangorarea in Malaysia ree weakly basic ion exchanger resins(Amberlite IRA-67 DiaionWA30 and DiaionWA21J) werepurchased from Sigma-Aldrich (St Louis MO USA) andtheir specifications are described in Table 1 e organicsolvent methanol of 99 (vv) analytical purity grade waspurchased from Bendosen Other analytical reagents andchemicals including vanillin sulphuric acid hydrochloricacid acetic acid and ethanol were also purchased fromBendosen Escin of 95 (ww) purity used as a standard wasobtained from Sigma-Aldrich (St Louis MO USA)

22 ExperimentalDesign A simple 3times 2 factorial design wasemployed to determine the saponin adsorption capacity ()of two adsorbent dosesload based on the previous studywith respect to three types of ion exchange resins as shownin Table 2

23 Sample Preparation C papaya leaves were obtainedfrom the consistent source in Banting Only the matureddark green leaves without any damage were chosen for thisstudy Prior to processing all mature C papaya leaves werestored at minus20degC to minimize the deterioration of any bio-active compounds present e leaves were then cut intosmall pieces and freeze-dried C papaya leaves were thengrounded by using Kenwood grinder before sieved to le1mmparticle size using a 1mm stainless steel mesh sieve Brieflyground leaves were extracted in water under optimal con-ditions identified using Response Surface Methodology(RSM) at 85degC and a water-to-leaf ratio of 20 1mlg eextracted solution was filtered and then stored at minus18degC forthe next adsorption studies [19] Figure 1 shows the processundertaken for the preparation of saponin-enriched waterextracts from C papaya leaves

24 PelegModel e Peleg mathematical model was used toestimate the optimum value for saponin adsorption as wellas the exhaustive time of the overall adsorption process egraph obtained from this model is able to describe thecharacteristics of the sorption process well [20] In additionthe Peleg mathematical model is very advantageous tosorption studies involving limited preliminary data [21 22]e Peleg mathematical equation is shown as

Mt M0 plusmnt

k1 + k2t( 1113857 (1)

where Mt is the concentration of saponins at time t () M0is the initial concentration of saponins () t is time in hourbasis (h) and k1 and k2 describe the Peleg rate (hminus1) andthe Peleg capacity constant (minus1) respectively It should alsobe noted that a ldquoplusmnrdquo value is possible wherein a positive ldquo+rdquovalue describes the process of adsorption or absorption and

2 Journal of Food Quality

a negative ldquominusrdquo value describes the desorption and dryingprocess [22 23]e values for k1 and k2 are calculated basedon the equation of the linear graph of timeconcentration ofsaponins (minmg) vs time (min) where k1 is equal to the y-intercept of the graph and k2 is equal to the slope [24 25]

Rate of adsorption 1k1

(mgmin)

Maximum concentration of extract 1k2

(mg)

(2)

Table 1 Specification of each ion weakly basic anion exchange resin chosen for the adsorption of saponin from C papaya leaves

Product specificationsIon exchange resins

Amberlite IRA-67 Diaion WA21J Diaion WA30Resin type Weakly basic anion Weakly basic anion Weakly basic anionMatrix Acrylic gel Divinyl benzene-styrene Divinyl benzene-styreneFunctional group Polyamine Polyamine Tertiary amineIonic form Free base Free base Free baseIon exchange capacity (meqml) 16 20 15Moisture content () 56ndash64 40ndash52 43ndash55Min particle size (microm) 610 650 680Density 180 107 105Max operational temperature (degC) 60 100 100Operational pH 7ndash9 0ndash9 0ndash9Regeneration chemical Sodium hydroxide Sodium hydroxide Sodium hydroxide

Table 2 Experimental design of saponin adsorption by three different types of ion exchange resins

Adsorbent doseload (wv) Types of weakly basic ion exchange resins5 Diaion WA21J Amberlite IRA-67 Diaion WA3010e influence of dose adsorbent and type of resin on adsorption capacity () was assessed

C papaya leaves

Freeze drying

Grounding(using 1 mm nets)

Water extraction of saponin fromC papaya leaves

(85degC water-to-leaf ratio201 mlmiddotgndash1)

Filtration

Saponin-rich water extract fromC papaya leaves

Figure 1 Preparation of saponin-enriched water extracts from C papaya leaves

Journal of Food Quality 3

25 Adsorption Studies of Saponins Using Weakly Basic IonExchangeResins e 5 (wv) and 10 (wv) of each weaklytype basic ion exchange resins were conditioned as proposedby the manufacturer and then placed into a conical askbefore beingmixed directly withC papaya leaf extracts rich insaponins [26] e mixture was placed in an environmentalshaker at a rotation of 120 per minute and the temperaturewas set to 25 degC [27] e mixture was left until optimumadsorption was achieved reading of saponin content wastaken at every two hour interval in between the process eadsorption capacity S () was later calculated as

S C0 minusCC0

times 100 (3)

where C0 and C are the initial concentration of saponin andthe nal concentration of saponin respectively

26 Determination of Saponin Content Using Spectrophoto-metric Methods (UV Spectrophotometer) Determination ofthe saponin content of C papaya leaf extracts throughoutthe process was done based on the method used by Vuonget al [28] Approximately 05ml extracts were mixed with05ml of 8 (wv) vanillin and 5ml of 72 (vv) of sulphuricacid cooled on ice for 5mins and then incubated at 60degC for15min e mixture was then cooled on ice in room tem-perature before being measured at 560 nm using a UVspectrophotometer Escin was used as the standard for thecalibration curve with results expressed as mg of escinequivalents per g of sample (mg ASE gminus1)

27 Quantication of Saponin Compound Using High-Performance Liquid Chromatography Determination andquantication of saponins using chromatographic techniquein this study was carried out to make a comparison of thepresence compounds in each treated and untreated sample

e condition and method proposed for the chromatog-raphy were based on few references with slight modicationas described in Table 3 [29 30] Table 4 describes the solventgradient conditions of the HPLC used

28 CIE Llowastalowastblowast Color Test Color measurement (Llowast alowast alowast)was carried on the surface of the samples using a MinoltaChromameter (CR 400 Japan) Reading the analysis is lateron described using the CIE Llowast (lightness) alowast (redness) andalowast (yellowness) system e device was calibrated usinga provided white tile before measuring a colored sample

29 Sensory Evaluation A descriptive test was carried out toassess acceptance of four attributes associated with theobjective of the research mainly the aroma color taste(bitterness) and overall acceptance of the samples beforeand after treatment A total of 10 semi-trained panels wereselected to undergo the sensory evaluation Each panelevaluates eight type of samples consisting of six treatedsamples that has undergone the adsorption process oneuntreated sample (fresh sample) and one reference sample(RUJ) on a numerical scale with a seven-point gradingMinitab software (17th Version) was used for data analysis todetermine the standard deviation and mean as well as toproduce ANOVA test results that determine signicantdierences between the samples tested e signicantdierence is based on a 95 condence level (plt 005)

3 Results and Discussions

31 Exhaustive Saponin Extraction e kinetic graph inFigures 2 and 3 was obtained after the adsorption processwas left to set for 1440 minutes (24-hours) Previous study

Table 3 Specications and conditions of saponin detection in Cpapaya leaves extract using high-performance liquidchromatography

Specications ConditionsColumn Chromolith performance RP-18eDetector UV-VISWavelength 210 nmTemperature 23plusmn 1degCMobile phase (pump A) Deionized water +01 (vv) acetic acidMobile phase (pump B) Methanol (HPLC grade)Flow rate 10mlminElution mode Binary gradientInjection volume 20 microl

Table 4 Solvent gradient conditions of the high-performanceliquid chromatography for saponin quantication

Time (min) Pump A Pump B Flow rate (mlmin)0 296 704 1015 40 60 1030 40 60 1031 0 (stop) 0 (stop) mdash

y = ndash7 times 10ndash6 x2 + 00182x ndash 14622R2 = 09229

ndash4ndash2

02468

101214

0 200 400 600 800 1000 1200 1400 1600

Conc

entr

atio

n of

sapo

nins

(mg

g)

Time (min)

Figure 2 Kinetic equilibrium prole of saponin concentration(mgg) with respect to extraction time (min)

ed

cb b a

020406080

100120140160

840 960 1080 1200 1320 1440

Tim

eco

ncen

trat

ion

ofsa

poni

ns (m

inm

g)

Time (min)

Figure 3 Linear relationship between timeconcentration of sa-ponins (minmg) with respect to extraction time (min) andashe dif-ferent alphabets indicate a signicant dierence (plt 005)


has shown that the complete saponin extraction should notbe more than 41 hours as the extract was more likely todegrade rapidly which is indicated by the change of colorand smell [31]

For determining the exhaustive time and maximumamount of extract the Peleg model was carried out based onthe timeyield (minmg) versus the time interval (min)graph Figure 3 shows the linear relationship of thetimetotal saponin content (minmg) against time (min) inorder to determine the K1 and K2 coeplusmncients for the value ofextraction rate and the maximum amount of extract ob-tained e coeplusmncient of K1 value is obtained from the y-intercept value whereas the K2 coeplusmncient value is thegradient value through the time linear (t)concentrationversus time (t) graph Based on the equation above theaverage rate of saponin extraction were calculated to be005plusmn 0001mgmin for approximately 1250plusmn 108 hourswith a total yield of 931plusmn 126mgg e extraction wasrepeated (n 3) and the exhaustive extract concentrationwas obtained approximately with the predicted value

32 Saponin Adsorption Capacity Using Weakly Basic IonExchange Resin Adsorption studies of saponins using weaklybasic ion exchanger has previously been carried out elsewhereusing Amberlite IRA-67 at two known adsorbent doses of 01 gand 05 g However it was discovered that the particular resinshowed potential to be able to eectively adsorb saponins up to80 (ww) of the compound from the total concentration [32]Generally it is noted that resins with a microporous structureare able to adsorb organic compounds rather well [33] Figure 4shows the amount of saponins adsorbed () by three dierentweakly basic ion exchange resins at the adsorbent precondi-tioned dose of 5 (wv)

Figure 4 suggests that there was a signicant dierence(plt 005) between all three-ion exchange resins used at 5(wv) adsorbent dose in which the highest adsorption wasfrom samples treated with the resin Diaion WA30 is wasmarked by a few factors including the mean size of resin

particles (microm) and the resin structure Based on Table 1 itshould be noted that Diaion WA30 recorded the largestmean for its particle size at 680 microm in comparison withDiaion WA31J (650 microm) and Amberlite IRA-67 (610 microm)Previous study has shown that adsorption () of lead ions(Pb2+) and cadmium ions (Cd2+) increased proportionatelyto the mean size of adsorbent particles from 106 microm to500 microm [34] e results proved that an increase in theparticle size would mean an increase in micropores betweenthe particles as well thus eectively increasing the surfacearea for the overall adsorption process On another note thetype of matrix of the resin would also come to aect theadsorption process Based on Table 1 the Amberlite IRA-67was the only resin with an acrylic gel-type of matrix unlikeboth Diaion resins which possess a DVB-styrene matrixstructure is could very well contribute to the eectivenessof the adsorption process that nongel DVB-styrene typematrixes has larger porosity that serves for the adsorptionprocess of chromium ions (Cr2+) [35]

Between two adsorbent doses of 5 (wv) and 10 (wv)used it was found that each type of resin showed

a

b

c

65

70

75

80

85

90

Ads

orpt

ion

of sa

poni

ns (

)

WA30 5AB 5WA21 5

Types of ion exchange resins

Figure 4 Percentage of saponins adsorbed () by three dierentweakly basic ion exchange resins at 5 (wv) adsorbent dose andashcdierent alphabets indicate a signicant dierence (plt 005)

aa a

bb

b

0

20

40

60

80

100

120

WA30 AB WA21

Sapo

nins

adso

rptio

n (

)

Type of weakly basic ion exchange resins

5 (wv)10 (wv)

Figure 5 Eect of adsorbent dose at 5 (wv) and 10 (wv) onthe saponin adsorption () of three dierent types of weakly basicion exchange resins a b dierent alphabets indicate a signicantdierence (plt 005)

a

a

a

a

a a

02468

10121416

WA30 AB WA21

Exha

ustiv

e tim

e of t

head

sorp

tion

proc

ess (

hour

)

Types of weakly basic ion exchange resins

5 (wv)10 (wv)

Figure 6 Eect of adsorbent dose at 5 (wv) and 10 (wv) onthe saponin adsorption exhaustive time (hour) a the same al-phabet indicate no signicant dierence (pgt 005)


a significant difference (plt 005) on the adsorbed saponinsas shown in Figure 5 e adsorbent dose however did notproduce any significant difference (pgt 005) on the overalltime of the adsorption process as shown in Figure 6

Previous study has shown that an increase in adsorbentdose would unconditionally increase the amount of ad-sorption by ion exchange resins [36ndash38] is trend wasindeed in line with the textile dyes (Pb2+ and Cd2+ ionscaptures) bioadsorption waste treatment process and as itincreased proportionally with respect to the adsorbent dose of25 gL to 200 gL [39 40] is trend was subtly attributed tothe increase in the surface area for the adsorption process asthe dose increases e exhaustive time of the overall ad-sorption process however was not affected by the adsorbentdose as the adsorption process generally would not be sig-nificantly different from each other no matter how much thedose were given after 2-3 hours of the process is would bepossibly due to achieving a state of quasi-equilibrium at theearliest recorded maximum adsorption time

33 Quantification of Saponins Figure 7 shows the standardcurve produced by escin was used as an external standard forsaponin quantification Figure 8 shows the chromatogramprofiles of saponin availability of each untreated and treatedsamples after the overall adsorption process

As shown in Figure 8 the only visible peak obtained fromthe chromatogram was the fresh extract (untreated) at re-tention time of 2678plusmn 104mins Samples that were treatedusing ion exchange resin showed absolutely no apparentpeaking around the expected retention time of 20 to 27minmost likely due to saponins being adsorbed at almost 75ndash95(ww) from its total amount in detected prior to adsorptione absence of a peak during the expected retention timetherefore solidifies the fact that the adsorption process hadoccurred between the samples and the resin to remove theparticular saponin compound

34 CIE Llowastalowastblowast Color Test Figure 9 shows the result of theCIE Llowastalowastblowast color test for both fresh (untreated) and treatedsamples after the adsorption process was concluded Resintreatment was observed to have significant (plt 005) effect onthe color intensity of the samples compared to the fresh(untreated) sample It is noted that compared to the untreatedsample with an Llowast value of 5693plusmn 101 samples that weretreated particularly with Diaion WA30 10 (wv) showed thelightest color intensity of 8094plusmn 072 Llowast reading is is inline with the findings of the panel sensory evaluation thatconcluded samples treated with this particular resin gave outan impression of being weak in color compared to the freshsample e same is also true for the blowast reading that describesthe intensity of yellowish color in the sample Based on thesefindings it is probable that sensory acceptance of the sampleswas affected by the color representation of the sample whenserved to the sensory panels [34 35]

35 Sensory Evaluation Figure 10 shows the result of thesensory evaluation responses consisting of four tested

attributes including color aroma taste (bitterness) andoverall acceptance Resin treatment was observed to havesignificant (plt 005) effect on all of the attributes testedcompared to the fresh (untreated) sample It was noted thatfor almost all of the attributes samples treated with DiaionWA30 resin at an adsorbent dose of 10 (wv) managed thelowest scores (for aroma intensity color intensity and bittertaste) while managing to also top overall acceptance amongstthe sensory panel compared to all other samples Samplestreated with the same resin at an adsorbent dose of 5 (wv)also threaded behind carefully however as Figure 5 ulti-mately described above the difference of total saponinadsorbed might attribute to the slight difference between thetwo It is noteworthy that in relation to the amount of saponinadsorbed throughout the process the overall acceptance in-creased in order as well as the bitterness of the sample fromAmberlite IRA-67ltDiaion WA21JltDiaion WA30

36 Adsorption Isotherm Modelling Data have been plottedbased on Langmuir adsorption isotherm in Figure 11 CeQegradient against Ce is aimed to obtain the Langmuir constantvalue (RL) e Langmuir constant value (RL) is used for thepurpose of connecting isotherm of adsorption of purecomponents [36] Another feature of the Langmuir equationcan be translated into a nondimensional constant (RL)equatione calculated RL value shown in Table 5 lies in therange 0 and 1 Based on previous study [37] the RL valuedetermines the isotherm whether it is unfavourable (RLgt 1)linear (RL 1) favourable (0ltRLlt 1) and nonreversible(RL 0) Based on the results in this study the dimensionlessseparation factor (RL) shows that the Resin Diaion WA30with absorbent dose 10 (wv) can be used as an alternativeto commercial adsorbent in bioactive removal in plants

4 Conclusion

Based on the research conducted it was found that theselected resins showed a great potential adsorbent for sa-ponin adsorption extracted from C papaya leavese studyfound that there were significant differences (plt 005) be-tween three types of resins used against saponin adsorption() in which Diaion WA30 showed the highest adsorptionas compared to other resins e differences were believed tobe due to the mean particle size of the resins (microm) and theresin matrix structure itself In addition the dry weight ofthe resin used showed significant differences (plt 005) onthe adsorption process in which the resin load of 10 (wv)showed higher adsorption as compared to that of 5 (wv)is is most likely due to the increase in dry weight resinwhich subsequently increases the amount of resin beads orarea for the efficient adsorption However the adsorbentdoseload did not literally affect the overall exhaustiveexposureincubation time (pgt 005) Moreover sensoryevaluation suggests that samples treated with weakly basicion exchange resins were in fact affected the aroma colortaste (bitterness) and overall acceptability compared to thefresh (untreated) samples (plt 005) e overall acceptanceof samples treated with ion exchange resins gave a consid-erable positive responses in reducing saponin bitterness in


e

d

c

b

ay = 94558x ndash 36564

R2 = 09798

0100000200000300000400000500000600000700000800000900000

1000000

0 20 40 60 80 100 120Pe

ak ar

eaConcentration (mgl)

Figure 7 Escin standard curve at concentration 20mgl to 100mgl (n 3) andashe dierent alphabets indicate a signicant dierence(plt 005)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(a)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(b)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(c)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(d)

Figure 8 Continued


which the sample treated with 10 (wv) resin load DiaionWA30 recorded the highest overall acceptance in parallelwith having the lowest bitterness score compared to other

samples including samples of fresh (without treatment)is result was also consistent with the earlier findings of theadsorption study that proved Diaion WA30 at an adsorbent

30(m

V)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(e)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(f )

(mV

)

250

200

150

100

50

0

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(g)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(h)

Figure 8 HPLC chromatogram profiles of C papaya leaves extract (a) standard (escin) (b) fresh (untreated) (cndashh) samples treated with ionexchange resins Red circle in (bndashh) shows the retention time of the saponin compound (a) Peak of standard (control escin) at con-centration of 100mgl using 210 nm detection wavelength (b) Peak of fresh sample (untreated) using 210 nm detection wavelength (c) Nopeak detected at the expected retention time of samples treated with DiaionWA21 resin at 5 (wv) adsorbent dose using 210 nm detectionwavelength (d) No peak detected at the expected retention time of samples treated with Diaion WA21 resin at 10 (wv) adsorbent doseusing 210 nm detection wavelength (e) No peak detected at the expected retention time of samples treated with Amberlite IRA-67 resin at5 (wv) adsorbent dose using 210 nm detection wavelength (f ) No peak detected at the expected retention time of samples treated withAmberlite IRA-67 resin at 10 (wv) adsorbent dose using 210 nm detection wavelength (g) No peak detected at the expected retention timeof samples treated with Diaion WA-30 resin at 5 (wv) adsorbent dose using 210 nm detection wavelength (h) No peak detected at theexpected retention time of samples treated with Diaion WA-30 resin at 10 (wv) adsorbent dose using 210 nm detection wavelength


dose of 10 (wv) had the highest percentage of saponinremoval after the adsorption process rough adsorptionstudies using the Langmuir isotherm model the results

showed adsorption using Resin Diaon WA30 was encour-aging with RL values ranging from 0 to 1 (0ltRLlt 1) (0167to 0398) is shows that Resin Diaon WA30 is eective inadsorption of the saponin compound

Data Availability

e data used to support the ndings of this study areavailable from the corresponding author upon request

Conflicts of Interest

e authors declare that they have no conicts of interest

e

d

a

b

d

c

d

b

d

c

c

c

a

b

e

a

b

f

f

a

b

ndash1000000

100020003000400050006000700080009000

Min

olta

chro

mam

eter

read

ing

WA21 5WA21 10AB 5

AB 10WA30 5

WA30 10FRESH

Llowast

Figure 9 CIE Llowastalowastblowast reading by the Minolta Chromameter on fresh (untreated) and treated C papaya leaf extracts andashf dierent alphabetsindicate a signicant dierence (plt 005)

000

100

200

300

400

500

600

700

800

Aroma Color Taste (bitterness) Overall acceptance

Mea

n sc

ores

Sensory attributes

RUJFRESHWA21 5

WA21 10AB 5AB 10

WA30 5WA30 10

abaab

ab

b

abab

bcb

a

c

a

bc bc

d

a a

b ba

a c

ca a

abab c

cbc

Figure 10 Mean scores of four tested sensory attributes (odor color taste and overall acceptance) of C papaya leaf extracts andashd dierentalphabets indicate a signicant dierence (plt 005)

R2 = 09686

0005

01015

02025

03035

0 02 04 06 08 1 12 14 16

C eQ

e

Ce

Figure 11 Isothermal adsorption of the saponin compound byResion Diaon WA30 with 10 (wv)

Table 5 Kinetic study on adsorption activity of the saponincompound by using resin Diaon WA30 10 (wv)C0 (mgL) 1750 1385 898 798RL 0167 0243 0312 0398


Acknowledgments

e authors would like to thank the Ministry of ScienceTechnology and Innovation (MOSTI) and Ministry of HigherEducation (MOE)Malaysia for providing financial support tothis project (06-01-02-SF1271 FRGS22013TK04UKM031 and GGPM-2013-078)

References

[1] D J Du Puy and I R H Telford ldquoChapter 30 CaricaceaeDlm George A S (pnyt)rdquo Flora of Australia Volume 50Oceanic Islands 2 pp 163-164 Australian GovernmentPublishing Service Canberra Australia 1993

[2] V M Badillo ldquoCarica L vs Vasconcella St Hil (Caricaceae)rdquoCon la Rehabilitacion de este Ultimo vol 10 pp 74ndash79 2000

[3] F A Bamisaye E O Ajani and J B Minari ldquoProspects ofethnobotanical uses of pawpaw (Carica papaya)rdquo Journal ofMedicinal Plant Studies vol 1 no 4 pp 171ndash177 2013

[4] K Sathasivam S Ramanathan S MMansorM R Haris andW H Wernsdorfer ldquorombocyte counts in mice after theadministration of papaya leaf suspensionrdquo Wiener KlinischeWochenschrift vol 121 no S3 pp 19ndash22 2009

[5] F Yunita E Hanani and J Kristiano ldquoe effect of Caricapapaya L Leaves extract capsules on platelets count andhaematocrit level in dengue fever patientrdquo InternationalJournal of Medicinal Aromatic Plants vol 2 pp 573ndash5782012

[6] N Ahmad H Fazal M Ayaz B H Abbasi I Mohammadand L Fazal ldquoDengue fever treatment with Carica papayaleaves extractrdquo Asian Pacific Journal of Tropical Biomedicinevol 1 no 4 pp 330ndash333 2011

[7] J Duke ldquoCarica papaya L (Caricaceae)rdquo May 2016 httpsunars-gringov8080npgspubxsqldukeplantdispxsqltaxon209

[8] P B Ayoola and A Adeyeye ldquoPhytochemical and nutrientevaluation of Carica papaya (pawpaw) leavesrdquo InternationalJournal of Recent Research and Applied Studies vol 5 no 3pp 325ndash328 2010

[9] T H Grenby ldquoIntense sweeteners for the food industry anoverviewrdquo Trends in Food Science and Technology vol 2pp 2ndash6 1991

[10] I Kitagawa ldquoLicorie root a natural sweetener and an im-portant ingredient in Chinese medicinerdquo Pure and AppliedChemistry vol 74 no 7 pp 1189ndash1198 2002

[11] A D Muir D Paton K Ballantyne and A A Aubin Processfor Recovery and Purification of Saponins and Sapogenins fromQuinoa (Chenopodium quinoa) United States Patent andTrademark Alexandria VA United States 2002

[12] E Aldin H A Reitmeier and P Murphy ldquoBitterness of soyextracts containing isoflavones and saponinsrdquo Journal of FoodScience vol 71 no 3 pp S211ndashS215 2006

[13] K R Price I T Johnson and G R Fenwick ldquoe chemistryand biological significance of saponins in foods and feedingstuffsrdquo CRC Critical Reviews in Food Science and Nutritionvol 26 pp 27ndash135 1987

[14] L Heng J Vincken G A van Koningsveld et al ldquoBitterness ofsaponins and their content in dry peasrdquo Journal of the Science ofFood and Agriculture vol 86 no 8 pp 1225ndash1231 2006

[15] S G Sparg M E Light and J van Staden ldquoBiological ac-tivities and distribution of plant saponinsrdquo Journal of Eth-nopharmacology vol 94 no 2-3 pp 219ndash243 2004

[16] C Y Cheok H A K Salman and R Sulaiman ldquoExtractionand quantification of saponins a reviewrdquo Food ResearchInternational vol 59 pp 16ndash40 2014

[17] J Azmir I S M Zaidul M M Rahman et al ldquoTechniques forextraction of bioactive compounds from plant materialsa reviewrdquo Journal of Food Engineering vol 117 no 4pp 426ndash436 2013

[18] E C M Coxworth and R E Salmon ldquoKochia seed asa component of the diet of turkey poults effects of differentmethods of saponin removal or activationrdquo Canadian Journalof Animal Science vol 52 no 4 pp 721ndash729 1972

[19] Q V Vuong S Hirun T L K Chuen et al ldquoAntioxidant andanticancer capacity of saponin-enriched Carica papaya leafextractsrdquo International Journal of Food Science and Tech-nology vol 50 no 1 pp 169ndash177 2015

[20] M D Vetal V G Lade and V K Rathod ldquoExtraction ofursolic acid from Ocimum sannctum by ultrasound processintensification and kinetic studiesrdquo Chemical Engineering andProcessing Process Intensification vol 69 pp 24ndash30 2013

[21] S M Shafaei A A Masoumi and H Roshan ldquoAnalysis ofwater absorption of bean and chickpea during soaking usingPeleg modelrdquo Journal of the Saudi Society of AgriculturalSciences vol 15 no 2 pp 135ndash144 2014

[22] F N Mohd Fazil N S Mohd Azzimia B Hisham YahayaN Atikah Kamalaldin and S I Zubairi ldquoKinetics extractionmodelling and antiproliferative activity of Clinacanthusnutans water extractrdquo Ce Scientific World Journal vol 2016Article ID 7370536 7 pages 2016

[23] O Corzo N Brancho and J Rodriguez ldquoComparison ofPeleg and Azura et al models in the modelling mass transferduring pile salting of goat sheetsrdquo LWT-Food Science andTechnology vol 4 no 2 pp 448ndash452 2012

[24] Y S Yien O Hassan and S I Zubairi ldquoDeodorizingmechanism of β-cyclodextrin-organic acids inclusion againststrong odor of Morinda citrifolia (Mengkudu) Juicerdquo JurnalTeknologi vol 79 no 10 pp 67ndash75 2016

[25] H Hashim S I Zubairi W A M Mustapha andM Y Maskat ldquoCharacterizing the deacidification adsorptionmodel of organic acids and phenolic compounds of noniextract using weak base ion exchangerrdquo Journal of Chemistryvol 2018 Article ID 6376929 10 pages 2018

[26] H Haslaniza W A Wan Yaacob Z Saiful Irwan andM Y Maskat ldquoPotential of Amberlite IRA-67 resin for de-acidification of organic acids in noni juicerdquo Der PharmaChemica vol 7 no 12 pp 62ndash69 2015

[27] J Kluczka T Korowais M Zolotajkin and J AdamekldquoBoron removal from water and wastewater using newpolystyrene-based resin grafted with glycidolrdquo Water Re-sources and Industry vol 11 pp 46ndash57 2015

[28] Q V Vuong S Hirun P D Roach M C BowyerP A Phillips and C J Scarlett ldquoEffect of extraction conditionon total phenolic compounds and antioxidant activities ofCarica papaya leaf aqueous extractrdquo Journal of HerbalMedicine vol 3 no 3 pp 104ndash111 2013

[29] H O A Ahmed and C Wang ldquoDetermination of tea saponinin Camellia seed oil with UV and HPLC analysisrdquo WorldJournal of Engineering and Technology vol 3 no 4 pp 30ndash37

[30] M J Ahn and J Kim ldquoIdentification and quantification ofsteroidal saponins in Polyganatum species by HPLCESIMSrdquoArchives of Pharmacal Research vol 28 no 5 pp 592ndash5972005

[31] A J Empungan ldquoPenjerapan sebatian saponin dalam ekstrakdaun Carica papaya menggunakan resin penukar ion berbeslemahrdquo Tesis Ijazah Sarjana Muda Program Sains Makanan


Pusat Pengajian Sains Kimia dan Teknologi Makanan FakultiSains dan Teknologi Universiti Kebangsaan Malaysia BangiMalaysia 2015

[32] J Jiang Z Wu W Liu Y Gao S Guo and S KangldquoSeparation of soybean saponins from soybean meal bya technology of foam fractionation and resin adsorptionrdquoPreparative Biochemistry and Biotechnology vol 46 no 4pp 346ndash353 2015

[33] O L Charles and S A Odomelam ldquoStudies on adsorbentdosage particle sizes and Ph constraints on biosorption of Pb(II) and Cd(II) Ions from aqueous solution using modifiedCrasstrotrea gasar (Bivalve) biomassrdquo International Archive ofApplied Sciences and Technology vol 1 pp 62ndash68 2010

[34] P Koujalagi R Kulkarni S Divekar and R Nugaraie ldquoKi-netics thermodynamic and adsorption studies on removal ofchromium (VI) using Tulsion A-27(MP) resinrdquo DesalinationandWater Treatment vol 51 no 16ndash18 pp 3273ndash3283 2013

[35] A M Aljeboree A N Alshirifi and A F Alkaim ldquoKineticsand equilibrium study for the adsorption of textile dyes oncoconut shell activated carbonrdquo Arabian Journal of Chem-istry vol 10 no S2 pp S3381ndashS3393 2014

[36] C S Gulipalli B Prasad and K L Wasewar ldquoBatch studyequilibrium and kinetics of adsorption of selenium using ricehusk ash (RHA)rdquo Journal of Engineering Science and Tech-nology vol 6 no 5 pp 586ndash605 2011

[37] F M Clydesdale ldquoColor as a factor in food choicerdquo CriticalReviews in Food Science and Nutrition vol 33 no 1pp 83ndash101 1993

[38] C Spence ldquoOn the psychological impact of food colourrdquoFlavour vol 4 no 1 pp 1ndash9 2015

[39] J N Putro A Kurniawan S Ismadji and Y H JuldquoNanocelluluse based biosorbents for wastewater treatmentStudy of isotherm kinetic thermodynamic and reusabilityrdquoEnvironmental Nanotechnology Monitoring and Manage-ment vol 8 pp 134ndash149 2017

[40] A Babarinde and G O Onyiaocha ldquoEquilibrium sorption ofdivalent metal ions onto groundnut (Arachis hypogaea) shellkinetics isotherm and thermodynamicsrdquo Chemistry In-ternational vol 2 no 3 2016


Hindawiwwwhindawicom

International Journal of

Volume 2018

Zoology

Hindawiwwwhindawicom Volume 2018

Anatomy Research International

PeptidesInternational Journal of



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GenomicsInternational Journal of


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The Scientific World Journal

Volume 2018


BioinformaticsAdvances in

Marine BiologyJournal of



Neuroscience Journal


BioMed Research International

Cell BiologyInternational Journal of



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ArchaeaHindawiwwwhindawicom Volume 2018


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MicrobiologyHindawiwwwhindawicom

Nucleic AcidsJournal of

Volume 2018

Submit your manuscripts atwwwhindawicom

77times103microlminus1 as well as increased count of neutrophils from460 to 783 over the course of 5 days in dengue patientsfollowing administration of papaya leaf extracts [6]

e ability of the leaves to be used as a cure for thesediseases is most likely due to the presence of bioactive com-pounds and multiple phytochemicals in the leaves In 2007 atleast seven different bioactive compounds in the leaves of theCpapaya plant were identified including flavanols t tanninsalkaloids and carpaine [7 8] In fact the highest phyto-chemicals in C papaya leaves were also saponins which makeup to 0022plusmn 010 of all the phytochemicals tested present inthe same part [5] Although it bears the potential as a cure ormedicine for many ailments commercially available medicinalproducts from C papaya leaf extracts have hindered its po-tential due to its bitterness greatly believed to be caused bysaponins greatly believed to be caused by saponins [9 10]Previous research has found that one of the challenges of usingtraditional herbs as medicine for human health is the bittertaste associated with it particularly those that contain saponins[11] Recently many researchers have remarked that saponinsfrom a variety of plants including soybeans are very muchresponsible for the bitter taste of the extract even in lowconcentrations [12] It was found that there was indeeda positive correlation between the soya saponin concentrationin pea plant extracts (01 - 03 ww) and its bitterness levelanalysed via sensory evaluation [13] is was also true forsaponins that generally cause bitter tastes in dry pea samples[14]

In conjunction to this problem numerous attemptshave been made to remove saponins to prevent resur-facing of bitter taste in traditional medicinal herbs Ashighlighted over the years many methods have beendeveloped for the extraction isolation and quantificationof saponin compound [15] ese methods are usuallyseparated into two classes of conventional extractiontechnique and green extraction techniques with the lateremphasizing more on being environment friendly [16]However conventional extraction techniques are stillbeing preferred by 70 to 30 due to foregoing the cost ofheavy-equipment required for the extraction or isolationprocess Conventional methods for the extraction of sa-ponins are usually maceration (soaking) and the Soxhletmethod in reflux meanwhile the extraction techniquesinvolving green technology are those of ultrasound-assisted extraction techniques and microwave-assistedextraction techniques [17] is method manages toblock activation of the saponin compound through theaddition of phytosterols but this method is said to haveharmful effects when performed on animal feed as it willcause stunted animal growth and eventually death [18]

As such the objective of this research was to assess theeffectiveness of the saponin adsorption process of C papayaleaf extracts using three different types of weakly basic ionexchange resins to propose a new possible way of removingsaponins other than to determine the content percentage ofadsorption and exhaustive time of the overall adsorptionprocess as well as measuring the level of sensory acceptanceof C papaya leaf extracts which has undergone the ad-sorption process

2 Methodology

21 Materials and Chemicals Mature C papaya leaves (ofunspecified variety) were obtained from Banting Selangorarea in Malaysia ree weakly basic ion exchanger resins(Amberlite IRA-67 DiaionWA30 and DiaionWA21J) werepurchased from Sigma-Aldrich (St Louis MO USA) andtheir specifications are described in Table 1 e organicsolvent methanol of 99 (vv) analytical purity grade waspurchased from Bendosen Other analytical reagents andchemicals including vanillin sulphuric acid hydrochloricacid acetic acid and ethanol were also purchased fromBendosen Escin of 95 (ww) purity used as a standard wasobtained from Sigma-Aldrich (St Louis MO USA)

22 ExperimentalDesign A simple 3times 2 factorial design wasemployed to determine the saponin adsorption capacity ()of two adsorbent dosesload based on the previous studywith respect to three types of ion exchange resins as shownin Table 2

23 Sample Preparation C papaya leaves were obtainedfrom the consistent source in Banting Only the matureddark green leaves without any damage were chosen for thisstudy Prior to processing all mature C papaya leaves werestored at minus20degC to minimize the deterioration of any bio-active compounds present e leaves were then cut intosmall pieces and freeze-dried C papaya leaves were thengrounded by using Kenwood grinder before sieved to le1mmparticle size using a 1mm stainless steel mesh sieve Brieflyground leaves were extracted in water under optimal con-ditions identified using Response Surface Methodology(RSM) at 85degC and a water-to-leaf ratio of 20 1mlg eextracted solution was filtered and then stored at minus18degC forthe next adsorption studies [19] Figure 1 shows the processundertaken for the preparation of saponin-enriched waterextracts from C papaya leaves

24 PelegModel e Peleg mathematical model was used toestimate the optimum value for saponin adsorption as wellas the exhaustive time of the overall adsorption process egraph obtained from this model is able to describe thecharacteristics of the sorption process well [20] In additionthe Peleg mathematical model is very advantageous tosorption studies involving limited preliminary data [21 22]e Peleg mathematical equation is shown as

Mt M0 plusmnt

k1 + k2t( 1113857 (1)

where Mt is the concentration of saponins at time t () M0is the initial concentration of saponins () t is time in hourbasis (h) and k1 and k2 describe the Peleg rate (hminus1) andthe Peleg capacity constant (minus1) respectively It should alsobe noted that a ldquoplusmnrdquo value is possible wherein a positive ldquo+rdquovalue describes the process of adsorption or absorption and




(mgmin)


(mg)

(2)






C papaya leaves

Freeze drying




Filtration





S C0 minusCC0

times 100 (3)














ndash4ndash2

02468

101214

0 200 400 600 800 1000 1200 1400 1600

Conc

entr

atio

n of

sapo

nins

(mg

g)

Time (min)


ed

cb b a

020406080

100120140160

840 960 1080 1200 1320 1440

Tim

eco

ncen

trat

ion

ofsa

poni

ns (m

inm

g)

Time (min)









a

b

c

65

70

75

80

85

90

Ads

orpt

ion

of sa

poni

ns (

)

WA30 5AB 5WA21 5



aa a

bb

b

0

20

40

60

80

100

120

WA30 AB WA21

Sapo

nins

adso

rptio

n (

)


5 (wv)10 (wv)


a

a

a

a

a a

02468

10121416

WA30 AB WA21

Exha

ustiv

e tim

e of t

head

sorp

tion

proc

ess (

hour

)


5 (wv)10 (wv)











4 Conclusion



e

d

c

b

ay = 94558x ndash 36564

R2 = 09798

0100000200000300000400000500000600000700000800000900000

1000000

0 20 40 60 80 100 120Pe

ak ar



30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(a)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(b)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(c)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(d)

Figure 8 Continued




30(m

V)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(e)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(f )

(mV

)

250

200

150

100

50

0

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(g)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(h)





Data Availability




e

d

a

b

d

c

d

b

d

c

c

c

a

b

e

a

b

f

f

a

b

ndash1000000

100020003000400050006000700080009000

Min

olta

chro

mam

eter

read

ing

WA21 5WA21 10AB 5

AB 10WA30 5

WA30 10FRESH

Llowast


000

100

200

300

400

500

600

700

800


Mea

n sc

ores

Sensory attributes

RUJFRESHWA21 5

WA21 10AB 5AB 10

WA30 5WA30 10

abaab

ab

b

abab

bcb

a

c

a

bc bc

d

a a

b ba

a c

ca a

abab c

cbc


R2 = 09686

0005

01015

02025

03035

0 02 04 06 08 1 12 14 16

C eQ

e

Ce




Acknowledgments


References














































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018




(mgmin)


(mg)

(2)






C papaya leaves

Freeze drying




Filtration





S C0 minusCC0

times 100 (3)














ndash4ndash2

02468

101214

0 200 400 600 800 1000 1200 1400 1600

Conc

entr

atio

n of

sapo

nins

(mg

g)

Time (min)


ed

cb b a

020406080

100120140160

840 960 1080 1200 1320 1440

Tim

eco

ncen

trat

ion

ofsa

poni

ns (m

inm

g)

Time (min)









a

b

c

65

70

75

80

85

90

Ads

orpt

ion

of sa

poni

ns (

)

WA30 5AB 5WA21 5



aa a

bb

b

0

20

40

60

80

100

120

WA30 AB WA21

Sapo

nins

adso

rptio

n (

)


5 (wv)10 (wv)


a

a

a

a

a a

02468

10121416

WA30 AB WA21

Exha

ustiv

e tim

e of t

head

sorp

tion

proc

ess (

hour

)


5 (wv)10 (wv)











4 Conclusion



e

d

c

b

ay = 94558x ndash 36564

R2 = 09798

0100000200000300000400000500000600000700000800000900000

1000000

0 20 40 60 80 100 120Pe

ak ar



30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(a)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(b)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(c)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(d)

Figure 8 Continued




30(m

V)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(e)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(f )

(mV

)

250

200

150

100

50

0

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(g)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(h)





Data Availability




e

d

a

b

d

c

d

b

d

c

c

c

a

b

e

a

b

f

f

a

b

ndash1000000

100020003000400050006000700080009000

Min

olta

chro

mam

eter

read

ing

WA21 5WA21 10AB 5

AB 10WA30 5

WA30 10FRESH

Llowast


000

100

200

300

400

500

600

700

800


Mea

n sc

ores

Sensory attributes

RUJFRESHWA21 5

WA21 10AB 5AB 10

WA30 5WA30 10

abaab

ab

b

abab

bcb

a

c

a

bc bc

d

a a

b ba

a c

ca a

abab c

cbc


R2 = 09686

0005

01015

02025

03035

0 02 04 06 08 1 12 14 16

C eQ

e

Ce




Acknowledgments


References














































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018



S C0 minusCC0

times 100 (3)














ndash4ndash2

02468

101214

0 200 400 600 800 1000 1200 1400 1600

Conc

entr

atio

n of

sapo

nins

(mg

g)

Time (min)


ed

cb b a

020406080

100120140160

840 960 1080 1200 1320 1440

Tim

eco

ncen

trat

ion

ofsa

poni

ns (m

inm

g)

Time (min)









a

b

c

65

70

75

80

85

90

Ads

orpt

ion

of sa

poni

ns (

)

WA30 5AB 5WA21 5



aa a

bb

b

0

20

40

60

80

100

120

WA30 AB WA21

Sapo

nins

adso

rptio

n (

)


5 (wv)10 (wv)


a

a

a

a

a a

02468

10121416

WA30 AB WA21

Exha

ustiv

e tim

e of t

head

sorp

tion

proc

ess (

hour

)


5 (wv)10 (wv)











4 Conclusion



e

d

c

b

ay = 94558x ndash 36564

R2 = 09798

0100000200000300000400000500000600000700000800000900000

1000000

0 20 40 60 80 100 120Pe

ak ar



30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(a)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(b)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(c)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(d)

Figure 8 Continued




30(m

V)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(e)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(f )

(mV

)

250

200

150

100

50

0

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(g)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(h)





Data Availability




e

d

a

b

d

c

d

b

d

c

c

c

a

b

e

a

b

f

f

a

b

ndash1000000

100020003000400050006000700080009000

Min

olta

chro

mam

eter

read

ing

WA21 5WA21 10AB 5

AB 10WA30 5

WA30 10FRESH

Llowast


000

100

200

300

400

500

600

700

800


Mea

n sc

ores

Sensory attributes

RUJFRESHWA21 5

WA21 10AB 5AB 10

WA30 5WA30 10

abaab

ab

b

abab

bcb

a

c

a

bc bc

d

a a

b ba

a c

ca a

abab c

cbc


R2 = 09686

0005

01015

02025

03035

0 02 04 06 08 1 12 14 16

C eQ

e

Ce




Acknowledgments


References














































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018








a

b

c

65

70

75

80

85

90

Ads

orpt

ion

of sa

poni

ns (

)

WA30 5AB 5WA21 5



aa a

bb

b

0

20

40

60

80

100

120

WA30 AB WA21

Sapo

nins

adso

rptio

n (

)


5 (wv)10 (wv)


a

a

a

a

a a

02468

10121416

WA30 AB WA21

Exha

ustiv

e tim

e of t

head

sorp

tion

proc

ess (

hour

)


5 (wv)10 (wv)











4 Conclusion



e

d

c

b

ay = 94558x ndash 36564

R2 = 09798

0100000200000300000400000500000600000700000800000900000

1000000

0 20 40 60 80 100 120Pe

ak ar



30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(a)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(b)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(c)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(d)

Figure 8 Continued




30(m

V)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(e)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(f )

(mV

)

250

200

150

100

50

0

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(g)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(h)





Data Availability




e

d

a

b

d

c

d

b

d

c

c

c

a

b

e

a

b

f

f

a

b

ndash1000000

100020003000400050006000700080009000

Min

olta

chro

mam

eter

read

ing

WA21 5WA21 10AB 5

AB 10WA30 5

WA30 10FRESH

Llowast


000

100

200

300

400

500

600

700

800


Mea

n sc

ores

Sensory attributes

RUJFRESHWA21 5

WA21 10AB 5AB 10

WA30 5WA30 10

abaab

ab

b

abab

bcb

a

c

a

bc bc

d

a a

b ba

a c

ca a

abab c

cbc


R2 = 09686

0005

01015

02025

03035

0 02 04 06 08 1 12 14 16

C eQ

e

Ce




Acknowledgments


References














































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018










4 Conclusion



e

d

c

b

ay = 94558x ndash 36564

R2 = 09798

0100000200000300000400000500000600000700000800000900000

1000000

0 20 40 60 80 100 120Pe

ak ar



30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(a)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(b)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(c)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(d)

Figure 8 Continued




30(m

V)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(e)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(f )

(mV

)

250

200

150

100

50

0

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(g)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(h)





Data Availability




e

d

a

b

d

c

d

b

d

c

c

c

a

b

e

a

b

f

f

a

b

ndash1000000

100020003000400050006000700080009000

Min

olta

chro

mam

eter

read

ing

WA21 5WA21 10AB 5

AB 10WA30 5

WA30 10FRESH

Llowast


000

100

200

300

400

500

600

700

800


Mea

n sc

ores

Sensory attributes

RUJFRESHWA21 5

WA21 10AB 5AB 10

WA30 5WA30 10

abaab

ab

b

abab

bcb

a

c

a

bc bc

d

a a

b ba

a c

ca a

abab c

cbc


R2 = 09686

0005

01015

02025

03035

0 02 04 06 08 1 12 14 16

C eQ

e

Ce




Acknowledgments


References














































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018


e

d

c

b

ay = 94558x ndash 36564

R2 = 09798

0100000200000300000400000500000600000700000800000900000

1000000

0 20 40 60 80 100 120Pe

ak ar



30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(a)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(b)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(c)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(d)

Figure 8 Continued




30(m

V)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(e)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(f )

(mV

)

250

200

150

100

50

0

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(g)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(h)





Data Availability




e

d

a

b

d

c

d

b

d

c

c

c

a

b

e

a

b

f

f

a

b

ndash1000000

100020003000400050006000700080009000

Min

olta

chro

mam

eter

read

ing

WA21 5WA21 10AB 5

AB 10WA30 5

WA30 10FRESH

Llowast


000

100

200

300

400

500

600

700

800


Mea

n sc

ores

Sensory attributes

RUJFRESHWA21 5

WA21 10AB 5AB 10

WA30 5WA30 10

abaab

ab

b

abab

bcb

a

c

a

bc bc

d

a a

b ba

a c

ca a

abab c

cbc


R2 = 09686

0005

01015

02025

03035

0 02 04 06 08 1 12 14 16

C eQ

e

Ce




Acknowledgments


References














































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018




30(m

V)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(e)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375(min)

(f )

(mV

)

250

200

150

100

50

0

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(g)

30

(mV

)

25201510

50

00 25 50 75 100 125 150 175 200 225 250 275 300(min)

(h)





Data Availability




e

d

a

b

d

c

d

b

d

c

c

c

a

b

e

a

b

f

f

a

b

ndash1000000

100020003000400050006000700080009000

Min

olta

chro

mam

eter

read

ing

WA21 5WA21 10AB 5

AB 10WA30 5

WA30 10FRESH

Llowast


000

100

200

300

400

500

600

700

800


Mea

n sc

ores

Sensory attributes

RUJFRESHWA21 5

WA21 10AB 5AB 10

WA30 5WA30 10

abaab

ab

b

abab

bcb

a

c

a

bc bc

d

a a

b ba

a c

ca a

abab c

cbc


R2 = 09686

0005

01015

02025

03035

0 02 04 06 08 1 12 14 16

C eQ

e

Ce




Acknowledgments


References














































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018




Data Availability




e

d

a

b

d

c

d

b

d

c

c

c

a

b

e

a

b

f

f

a

b

ndash1000000

100020003000400050006000700080009000

Min

olta

chro

mam

eter

read

ing

WA21 5WA21 10AB 5

AB 10WA30 5

WA30 10FRESH

Llowast


000

100

200

300

400

500

600

700

800


Mea

n sc

ores

Sensory attributes

RUJFRESHWA21 5

WA21 10AB 5AB 10

WA30 5WA30 10

abaab

ab

b

abab

bcb

a

c

a

bc bc

d

a a

b ba

a c

ca a

abab c

cbc


R2 = 09686

0005

01015

02025

03035

0 02 04 06 08 1 12 14 16

C eQ

e

Ce




Acknowledgments


References














































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018


Acknowledgments


References














































Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018















Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018




Volume 2018

Zoology











Volume 2018

















Advances in




Enzyme Research





Volume 2018


Documents

Saponin Bitterness Reduction of Carica papaya …downloads.hindawi.com/journals/jfq/2018/5602729.pdfCarica papaya that belongs to Caricaceae family has long been known as a traditional