Vitamin C, total polyphenols and antioxidant activity in raw, · PDF filetically processed and industrially processed Indian Chenopodium quinoa seeds Intelli1*, Dr.Beenu Tanwar2, Dr.Manju

Vitamin C total polyphenols and antioxidant activity in raw domes-tically processed and industrially processed Indian Chenopodium

quinoa seeds

Intelli1 DrBeenu Tanwar2 DrManju Reddy3 Ambika Chauhan1

1Research Scholar Lovely Professional University Jalandhar Punjab (INDIA)2Assistant Professor Food Science and Nutrition School of Agriculture and Food science Lovely Professional

University (INDIA)3Professor Food science and Human Nutrition Iowa State University (USA)

E-mail intellilpugmailcom

BACKGROUND Chenopodium quinoa an ancient crop sprung initially in Andean region of South America iswell recognized for its outstanding nutritional composition and versatility Quinoa from various geographicalregions like Bolivia Argentina Kenya Japan and Morocco has been studied however there is no study on IndianChenopodium quinoa Thus the present study aims to determine vitamin C antioxidant activity and total polyphe-nols in Indian quinoa Also it compares effect of domestic and industrial processing on the grainRESULTS The results show that domestically processed seeds have higher vitamin C total phenolic content(TPC) total flavonoid content (TFC) and also antioxidant activity as compared to the raw and industrially pro-cessed seeds Antioxidant activity was found significantly correlated to the total phenolic content in raw domes-tically processed and industrially processed seedsCONCLUSION Domestically processed quinoa mainly by germination is reported to be rich in antioxidantsvitamin C and higher phenolic content The results suggest use of domestic processing of quinoa seeds to retainnutrient value and also infer dietary importance of Indian Chenopodium quinoa Further studies are required onantioxidant and polyphenol profiling of Indian quinoa 2016 Trade Science Inc - INDIA

INTRODUCTION

Chenopodium quinoa an underutilized crop be-longing to family chenopodiaceae is an ancient cropwith modern perspectives Originated and sprung ini-tially in Andean region of South America with Peruand Bolivia being the main producers the crop isgaining popularity worldwide due to its extraordi-nary nutritional composition and versatility to adaptextreme environmental conditions Quinoa is an an-nual plant 1 to 4 m tall having erect cylindrical

stem green to pale yellow leaves flowers with nopetals and achene fruit Quinoa grain is dicotyle-donous and thus regarded as a pseudocereal Thegrain is found with wide range of colour variety likered black pink purple and white depending on thepresence of pigments and geographical area of cul-tivation Talking about the nutritional aspects ofquinoa it is found to have protein content (16)higher than maize and wheat Protein quality ofquinoa is found comparable to that of milk proteincasein It contains all essential amino acids with

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ABSTRACT

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presence of both lysine (54) and methionine(21) which gives it a unique feature and makesit a complete food[51] Quinoa is a gluten free grainappropriate for consumption by celiac patients Ithas ash content (3 - 4) higher than common cere-als like wheat and rice and fibre content (14-16total dietary fibre 2-3 crude fibre) which is morethan that present in corn and wheat In additionquinoa seeds and sprouts are found rich inisoflavones polyphenols and known to exhibit goodanti oxidant properties[17] Isoflavones like genisteinand daidzein have been found in quinoa It is foundto contain 11-17mg100g bound phenolics likecoumeric benzoic vanillic and ferulic acid and96-164mg100g free phenolics with quercetinacacetin and kaempferol being majorly presentGood antioxidant activity in quinoa may also beattributed to the presence of vitamin C and toco-pherols[50] which also protects against oxidation ofits fatty acids Ascorbic acid also known as Vita-min C has metabolic and therapeutic benefits whenincluded in human diet High nutritional and phytochemical content make quinoa a wonder grain withmany health benefits Successful propagation ofquinoa in field trials at worlds hottest and driest

place Arabian peninsula prove this crops versa-

tility to adapt in adverse climatic and ecologicalconditions[42] Thus astounding features of this cropsuch as tolerance to stressful environmental con-ditions like soil salinity drought frost high tem-perature etc ability to grow at up to 4500m ofaltitude from sea level and impeccable nutritionalbenefits permit advancement of quinoa cultivationout from the boundaries of its Andean motherlandto different parts of world In addition to the ex-traordinary features the increasing demand ofquinoa across the world has prompted many agri-culturists and researchers worldwide for quinoacultivation[49] After being successfully promotedin England (1970) Denmark Europe (1993) andKenya[13] quinoa has found its way to Asia withkeen interest for the crop mainly in the Indian sub-continent[8] Crop is more widespread in PakistanNepal and India India being a land of diverse cli-matic regions (tropical wet tropical dry subtropi-cal humid and mountains) and quinoa being a crop

profoundly known as well adapted to unusual en-vironmental conditions is found apt to grow in In-dian boundaries[8] Initially being found grown infoots of Himlayan hills the crop has been grownsuccessfully for the first time in year 2013 underthe project named Anantha in plains of drought

prone area of Anantpur region in Andhra Pradesh[18]The project has highly promoted cultivation ofquinoa among various private companies in SouthIndia The craze and demand for quinoa among In-dians has grown far more over than their demandfor the staple traditional crops and millets like sor-ghum pearl millet finger millet etc[49]

Post harvesting and pior to marketing grainsundergo industrial processing mainly the processof dehulling or decortications to remove the outerlayers of the grain Dehulling is known to improvegrain quality by lowering the content of anti nutri-ents[36] and enhancing the sensory parameters hencethe acceptance and pallatabilty of the grain Despitethese benefits of dehulling it reported to cause lossof nutrients from grains Thus to minimize loss nu-trients and increase bioavailabilty of nutrients re-searchers recommend use of common traditionaldomestic processing methods for grains[39 32] Soak-ing and germination are the commonly used methodsfor domestic processing of seed Antioxidant activ-ity and phenolic content have been reported to begreatly affected by domestic processing[23 20]

After many studies revealing nutritional and phy-tochemical composition of american quinoa[17 10 26

50] recently quinoa grown in Kenya Japan and Mo-rocco was studied for its seed quality yield levelantioxidative properties proximate flavonoid fattyacid and mineral compostion The studies demon-strate significant influence of grain variety and cli-matic conditions of area of cultivation on nutritionaland phytochemical composition of the seed Till nowno study has been done on Indian quinoa seeds Theaim of our study is to investigate antioxidant activ-ity content of vitamin c and total polyphenols in In-dian Chenopodium quinoa grains We also comparedthe effect of domestic and industrial processing ofIndian quinoa grain on antioxidant activity contentof vitamin c and total polyphenols which has neverbeen discussed before

Vitamin C total polyphenols and antioxidant activity in raw domestically processed32


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MATERIAL AND METHOD

Seed procurement

White Chenopodium quinoa seeds (Chenopo-dium quinoa Willd) raw and industrially processedwere procured from Andhra Pradesh Institute of Ru-ral Development (APARD) Hyderabad India Rawseeds were procured directly from the site of culti-vation while industrially processed seeds were pro-cured after the seeds had been dehulled by indus-trial method prior to packaging and marketing Bothraw and industrially processed seeds were initiallyrinsed with ethanol and soaked in 2 sodium hy-pochlorite solution[45] for 10 minutes for seed steril-ization They were then washed with deionized wa-ter until pH 7 was obtained The seeds were driedin vaccum drying oven at 40plusmn5degC and stored at 4degC

until further chemical analysis

Preparation and processing of seeds for analysis

Raw and industrially processed seeds

Dried raw and industrially processed seeds (500g) obtained after the process of surface sterilizationwere grinded to finely powdered flour with a labo-ratory grinder (Philips HL160603 500 W MixerGrinder) and stored at 4degC for further analysis

Domestic processing of raw seeds

Raw seeds were subjected to domestic process-ing by soaking and germination method

a Soaking

Raw Chenopodium quinoa (500g) seeds weresoaked for 24 hours in deionized water (15 wv)obtained through Millipore (Merck-Milli-Qreg Direct8 Water Purification System USA) Water used forsoaking was changed thrice at regular interval of 8hours After 24 hours the water was discarded andseeds were washed once with de ionized waterSoaked seeds were further dried in vacuum dryingoven at 40plusmn5degC Dried seeds were grinded to finely

powdered flour with laboratory grinder (PhilipsHL160603 500 W Mixer Grinder) Soaked quinoaseed flour was stored at 4degC for further analysis

b Germination

Raw Chenopdium quinoa (500g) seeds were

thoroughly washed with and soaked in de ionizedwater (15 wv) obtained through Millipore (Merck-Milli-Qreg Direct 8 Water Purification System USA)for 12 hours Seeds were then spread on to petridishes covered with autoclaved filter paper and in-cubated at 20degC in an incubator (Biotechnics India)

for 72 hours[10] Water was changed every 8 hoursGerminated seeds were then dried in vacuum dryingoven at 40plusmn5degC Dried germinated seeds were

grinded to flour with laboratory grinder (PhilipsHL160603 500 W Mixer Grinder) Finely pow-dered germinated quinoa seed flour was stored at4degC for further analysis

Extract preparation

Finely grounded seeds (in flour form) of IndianChenopodium quinoa (5g) ie raw industrially pro-cessed and domestically processed (soaked and ger-minated) were extracted at room temperature with50 ml of 80 ethanol and 02M hydrochloric acidin ratio 91 The mixture was sonicated in an ultrasonic bath sonicator (Fisher scientific UK) for about10 minutes It was then agitated for 1 hour at 120rpm in an orbital shaker Then it was transferred tocentrifugation tubes and centrifuged for 15 minutesat 9000rpm Subsequently the supernatant was sepa-rated and the residue was re-extracted in the sameway Both the supernatants were then combined andfiltered through syringe filters (Agilent premium sy-ringe filter 045microm) to obtain clear extracts Ex-

tracts prepared were stored at -80degC in a freezer for

further analysis

Vitamin C analysis

N- bromosuccinimide (NBS) method for deter-mination of vitamin C as given by Barakat et al1955and Miranda et al (2010) was used for determina-tion of vitamin C in quinoa samples Slight modifi-cations were made in analysis accordingly Themethod includes preparation of standard ascorbicacid solution standardization of NBS with ascorbicacid and estimation of ascorbic acid in sample ex-tract a) Preparation of standard ascorbic acid Stan-dard ascorbic acid of concentration04mg ml-1 wasprepared by dissolving 200mg ascorbic acid in 500ml distilled water b) Standardization of NBS Solu-

Intelli et al 33



tion Standard ascorbic acid solution (20ml) wasadded to a flask containing 4 ml of 4 potassiumiodide solution (KI) 16 ml of 10 acetic acid(CH

3COONa) 4 drops of 1 starch (used as an in-

dicator) and 25 ml distilled water It was then ti-trated with NBS (02mg ml-1) Appearance of per-manent blue colour was considered as end point oftitration c) Estimation of ascorbic acid in sampleQuinoa extracts acidified with 04 g oxalic acid wasadded to a flask containing 10 ml of 4 potassiumiodide solution (KI) 4 ml of 10 acetic acid(CH


dicator) and 40 ml distilled water Final vitamin Ccontent was expressed as mg 100-1 using followingequation

Determination of total phenolic content

Total phenolic content (TPC) was assessed ac-cording to method described by Ainsworth andGillespie (2007) using Folin-ciocalteau reagentSample extract (05ml) was diluted and volume wasmade upto 1ml After 2 minutes 2ml 0f 10 folin-ciocalteau reagent and vortex thoroughly At sixthminute add 8 ml of 700mM Na

2CO

3 and incubate

the mixture for 2 hours Transfer 2 ml of mixture toquartz cuvette and read the absorbance at 765nmusing spectrophotometer (Shimadzu Japan) Thereadings were compared to gallic acid standardcurve (linearity range 50-250 mgml and R2=0991)Final total phenolic content expressed as gallic acidequivalent (GAE) per 100 g in dry weight basis(dwb)

Determination of total flavonoid

Total flavonoid was determined according toprocedure followed by Carciochi et al (2014a)Quinoa extract (05ml) was taken in a test tube Tothe test tube 4 ml of distilled water and 05 ml of20 NaNO2 (Sodium nitrite) was added Mixturewas allowed to stand for about 5 minutes and then03ml of 10 AlCl

3 (Aluminium Chloride) was

added After 1 minute 05 ml 0f 2M NaOH (Sodiumhydroxide) was added to the reaction mixture Ab-

sorbance was read at 510nm using spectrophotom-eter (Shimadzu Japan) Quercitin was used as stan-dard Final results were expressed as mg of quer-citin equivalent100g ie mgQE100g

Determination of DPPH

DPPH ie 22-diphenyl-2-picryl-hydrazyl assaywas proceeded according to method followed byJubete et al (2010b) and Sun and Ho (2005) withsome modification Aliquots of quinoa extract inincreasing trend (ie 100 200 400 800 1000 microl)

were taken for serial dilution Diluted quinoa ex-tracts (1 ml) from each serial dilution was added tocuvettes respectively DPPH solution of concentra-tion 200microM (absorbance 14) was freshly prepared

and 1 ml of this solution was added to each cuvettecontaining quinoa extract The mixture was vortexedand incubated in dark for 30 minutes Absorbancewas measured at 517 nm using spectrophotometer(Shimadzu Japan) DPPH was expressed as mgTE100g Inhibitory concentration at 50 was also cal-culated (IC

50 values)

and denoted as TEAC (Trolox

equivalent antioxidant capacity) TEAC was calcu-lated according to method used by Jubete et al(2010b) using formula TEAC

50= IC

50troloxIC

50sample

times 105 Trolox (002M) was used as standard Theresult was expressed as trolox equivalents (mgTEg) Inhibitory concentration (IC

50) was estimated by

interpolation to 50 inhibition and expressed asTEAC

50

Determination of FRAP

FRAP ie Ferric reducing ability of plasma as-say was proceeded according to procedure followedby Jubete et al (2010b) and Benzie and Strain(1996) with some modification FRAP reagent wasprepared by mixing 25 ml of 001M TPTZ in 004Mhydrochloric acid 25ml of 002M ferric chlorideand 25 ml of 03M sodium acetate buffer (pH 36)Quinoa extracts (100microl-300microl) and 2ml of FRAP

reagent was taken in a 5ml volumetric flask Dis-tilled water was used to make up the volume Solu-tions were kept in dark at 37degC for 60 minutes Ab-

sorbance was read at 595nm using spectrophotom-eter (Shimadzu Japan) Trolox stock solution of002M was used as standard for the assay FRAP




reagent (2ml) made up to 5 ml in a volumetric flaskwas used as blank

Statistical analysis

The experiments were performed in triplicatesand the data was expressed as meanplusmnstandard de-

viation The data was analyzed on Microsoft officeexcel (2007) and Graphpad prism 5 software (Lajolla CA USA) Means were compared using oneway analysis of variance (ANOVA) followed byTukeys multiple comparison test for comparison be-tween means The with values considered signifi-cant at p005

RESULT AND DISCUSSION

Vitamin C content of seeds

Vitamin C content in Indian Chenopodiumquinoa is shown in TABLE 1 Vitamin C content ofraw quinoa seeds in our study was found to be 13mg100g-1 which is within the range of vitamin c contentas reported by Miranda et al (2010) (12-23mg 100-

1) and corresponds closely to Cahuil variety (138mg 100g-1) among six chilean quinoa ecotypes stud-ied in his study The value reported in our study isgreater than the values reported by Koziol (1992)(4mg 100g-1) and less than as reported by Rualesand Nair (1993) (164 mg 100g-1) Miranda et al(2013) (22-31 mg 100g-1) in two quinoa genotypesfrom Temuco and Vacuna localities in Chile Thisdifference in vitamin C content may be due to dif-ferent environmental and storage conditions as fac-tors like light intensity amount of nitrogen fertiliz-ers frequency of irrigation and temperature of theregion strongly affect the vitamin C content in

crops[35]Also there is significant difference in vitamin C

content of raw and domestically processed Indianquinoa seeds (plt005) As depicted in our study vi-tamin C content increased by 15 in soaked quinoaseeds and by 46 in germinated quinoa seedsHigher increase in germinated seeds was observedmight be due to synthesis of vitamin C in process ofgermination (Sattar et al (1995) and Fernandez-Orozco et al (2006) Yang et al (2001) reportedvitamin C content in sprouted mungbean where asnil vitamin C content was reported in raw seedswhich confirms vitamin C synthesis during germi-nation process Increased vitamin C content in ger-minated indian quinoa seeds is also supported byfindings of Khattak et al (2007) where linear rela-tionship was observed between germination time andcontent of vitamin C in chickpea seeds Also sig-nificant difference in Vitamin C content of raw andindustrially processed seeds was observed Indus-trial processing decreased the vitamin C content by30 De hulling pearling shelling etc are post har-vest industrial treatments applied to cereal grainswhich lead to loss of their nutritional content [47]

Total phenolic content

Total phenolic content (TPC) of Indian Chenopo-dium quinoa is shown in TABLE 2 TPC of rawIndian quinoa seeds in our study was reported tobe 43 mgGAE 100g-1 which corresponds well toTPC content reported by Repo-Carrasco-Valenciaet al (2010) (42 mg GAE 100-1) and Vollamannovaet al (2013) (45mgGAE 100g-1) in Carmen varietyof quinoa

Also the value of TPC content found in our study

Indian Chenopodium quinoa Vitamin C (mg 100g)

Raw 1343plusmn04a

Domestically Processed

Soaked 1509plusmn017b

Germinated 1938plusmn028c

Industrially processed 945plusmn035d

TABLE 1 Vitamin C content of raw domestically andindustrially processed Indian Chenopodium quinoa seeds(mg 100g-1) A

A values are meanplusmn SD n=3 Vales followed by same letter in

same coloumn are not significantly different (plt 005)

Indian Chenopodium quinoa Total Phenolic Content (mg GAE100g)

Raw 432plusmn028a




Industrially processed 346plusmn033a

TABLE 2 Total phenolic content (TPC) of raw domes-tically and industrially processed Indian Chenopodiumquinoa seeds A

A values are meanplusmn SD n=3 Values denoted by same letter within

a column are not significantly different (plt 005)

Intelli et al 35



lies close to reported the values of TPC in raw quinoaseeds by Carciochi et al (2014a) (39 mgGAE 100g-

1) Pasko et al (2009) (38mgGAE 100g-1) higherthan as reported by Gorenstein et al (2007) (30mgGAE 100g-1) and Miranda et al (2010) (28mgGAE 100g -1) Higher reported values in our studycan be explained as raw seeds (direct from the field)used were with the seed coat while the quinoa seedsprocured by these authors were as available in thelocal market which might be industrially processedfor removal of seed coat which leads to decrease inphenolic content[40] The values obtained are lowerthan values reported by Repo-Carrasco-Valencia etal (2011) (142-197mg GAE 100-1) Palombini etal (2013) (63 mgGAE 100g-1) Carciochi et al(2014b) (67 to 103 mgGAE 100g-1) Chlopika(2012) (280mg GAE 100g-1) and Tang et al (2015)(518 mgGAE 100g-1) Significant difference in phe-nolic contents reported by various other authors maybe due to different environment conditions forgrowth[43 34] extraction solvents[11] quinoa varietieswith coloured testa[9 50] Results of our study are alsosignificantly different than as reported by Nsimba etal (2008) due to different origin of quinoa varietiesand different standard solutions used for analysis oftotal phenols Total phenolic content of quinoa ishigher than other wheat[2] and some Peruvian Andeanseeds like kwicha haba and tuber oca[16] Total phe-nolic content in soaked quinoa seeds was reportedas 311 mgGAE 100g-1 which is significantly less(28 Plt 005) as compared to raw seeds The re-sult corresponds to 26-56 loss in total phenoliccontent of black beans (Phaseolus vulgaris L) re-ported by Xu and Chang (2008) Lower phenoliccontent was also reported in soaked faba beans bySiah et al (2015) They explain hydrolysis and leach-ing of some condensed polyphenols during prolongedsoaking period into water used for soaking as rea-son for reduction in TPC in soaked seeds Germi-nated seeds were found to exhibit 134 increase intotal phenolic content as compared to the raw quinoaseeds Germination leads to increase in phenoliccontent of seeds[22] as synthesis of phenolic acid isenhanced by seed growth during germination[14] In-crease in total phenolic content in germinated quinoahas also been reported by Carciochi et al 2014a

(56 after 48 hours and 1012 after 72 hours ofgermination) and Jubete et al 2010b (107 after82 hours of germination) Difference in total phe-nolic increase can be explained on the basis of vary-ing germination time and techniques Industrial pro-cessed quinoa seeds exhibited 20 decrease in to-tal phenolic content (34 mgGAE 100g-1) Commonprocess involved in industrial seed processing isdecortication also known as pearling which re-moves its saponins present mainly in outer layer ofquinoa[15] Similar decrease in phenolic compoundsof pearled quinoa (abrasion degree of 30) wasreported by Gomez-Caravaca et al (2014) with215 and 352 decrease in free and bound phe-nolic compounds respectively Decrease in TPC af-ter decortication (industrial abrasive dehulling) hasalso been reported in soyabean (11) chickpeas(37) and yellow peas (34) By Xu et al (2007)Han and Baik (2008) (in wheat) and Madhujith etal (2006) (in barley) Cardador-Martyacuteacutenez et al

(2002) (in common beans) and Price et al (1998)(in green beans) have also reported decrease in TPCafter decortication of seeds

Total flavonoids

Total flavonoid content (TFC) of quinoa seedsis shown in TABLE 3 In our study TFC of rawIndian quinoawas reported as 114 mg QE 100g-1Results agree with the findings of Carciochi et al2014a (1106 mg QE 100g-1) and Chirinos et al 2013(11 mg QE 100g-1) Similar value (116 mg QE 100g-

1) was reported by Repo-carrasco-Valencia (2010)in an ecotype of quinoa (Salcedo INIA)

The result reported in our study also lie withinthe range of values of total flavonoid reported byMiranda et al 2014 (77- 1437 mg QE 100g-1) stud-

Indian Chenopodium quinoa Total Flavonoids (mg QE100g)

Raw 114plusmn008a


Soaked 72plusmn008b



TABLE 3 Total flavonoid content (TFC) of raw domes-tically and industrially processed Indian Chenopodiumquinoa seeds






ied in six different ecotypes of quinoa The total fla-vonoid content reported in our study is significantlydifferent to the values reported by Marmouzi et al(2015) Carciochi et al (2014a) Pasko et al (2008)and Chlopika et al (2012) This might be due todifferent solvents used for extraction difference intemperature during extraction process and differentmethods of flavonoid analysis used (HPLC orspectrophotometry) Intensity of seed coat colourmajorly affects total flavonoid content in quinoa Redand black quinoa have 50-100 more flavonoidcontent as compared to white quinoa seeds[50] Highertotal flavonoid content in quinoa than other Peru-vian Andean grains (Haba Kwicha) fruits(TunaGuinda Granadilla Tumbo) and tubers (OcaMashua) has been reported[16] Total flavonoid con-tent of soaked quinoa seeds decreased by 36 (72mg QE 100g-1) Similarly decrease in flavonoid con-tent after soaking has been found in pinto bean chickpea[46] and white sorgum Although quinoa is reportedto be rich in quercitin derivatives but Hirose et al2010 could not detect any flavanols like quercetinand kaempferol upon quantitative analysis in hy-drolysed quinoa seeds which may be due to leach-ing of these flavanols on hydrolysis Although theflavonoid content decreased after soaking of quinoaseeds but the difference is not significant (plt005)as significant decrease are found at higher soakingtemperature and not at room temperature[46] Germi-nation of quinoa seeds lead to significant increaseby 56 in flavoniod content (18 mg QE 100g-1)Similarly increase in flavonoid content by 59 hasbeen reported by Carciochi et al 2014a in germi-nated quinoa seeds The result agrees with the find-ings of Guajardo-Flores et al (2014) where two fold

increase in flavonoid content of germinated blackbeans was reported Uchegbu (2015) also reportedincrease in flavonoid content of germinated Africanyam bean HPLC analysis of flavonoids by Carciochiet al (2014a) reported 44 times increase in totalflavonoid content of germinated quinoa seeds withquercitin (increased by 56) in abundant amountand kaempferol glycosides in moderate amounts[2]The increase in flavonoid content on germination ofseeds is due to synthesis of metabolites like fla-vonoids[41] by phenylproponoid pathway commonto all plants during process of seed germination[52

21] Industrial processing of quinoa seeds lead to re-duction in flavonoid content by 47 Result is sup-ported by reduction in flavonoid content of buck-wheat on processing reported by Dietrych-Szostak(1999) The findings may be attributed to the factthat most of the flavonoids are contained in the seedcoat and industrial processing involves removal ofouter layer of seed thus causing decrease in the fla-vonoid content[53]

Antioxidant acivity (FRAP and DPPH)

Antioxidant activity of Indian Chenopodiumquinoa seeds is shown in TABLE 4 Raw quinoaseeds in our study were reported to have 596 mgTE100g-1 and 373 TEAC as calculated by DPPHmethod and 844 mg TE 100g-1 as calculated by FRAPmethod The results were close to antioxidant activ-ity according to Jubete et al (2010b) (577 mg TE100g and 348 TEAC by DPPH method and 841 mgTE 100g-1 by FRAP method) Antioxidant activity ofquinoa reported by Ranilla et al (2009) (by DPPH)was much higher (86 mg TE 100g-1) than as reportedin our study (596 mgTE 100g-1) This is because

TABLE 4 Antioxidant activity (FRAP and DPPH) of Raw domestically and Industrially processed Indian Che-nopodium quinoa seeds A

A values are meanplusmn SD n=3 Values denoted by same letter within a column are not significantly different (plt 005)

Indian Chenopodium quinoa

Antioxidant activity

FRAP mg TEg

DPPH

TEAC (IC50TroloxIC50) x 105 mg TEg Raw 8446plusmn59 a 373plusmn045a 5961plusmn039a


Soaked 9646plusmn15a 3499plusmn142

b 5351plusmn056b

Germinated 15923plusmn0b 7048plusmn03c 6141plusmn189c

Industrially processed 7235plusmn182a 3073plusmn19d 4969plusmn15a

Intelli et al 37



red quinoa were used by Ranilla et al (2009) anddifference in color of seeds strongly effects antioxi-dant activity with dark colored seed coats exhibit-ing highest antioxidants activities Quinoa seeds ex-hibit higher antioxidant activity (evaluated by FRAPand DPPH) as compared to grain Amaranth[5517] andsome Peruvian Andean fruit like Tuna and grainKwicha[16] It is found to exhibit lower antioxidantactivity as compared to buckwheat[2] oat and rice[29]higher than amaranth[2] and almost similar to wheat[2]

Antioxidant activity of quinoa also depends onintensity of colour of seed coat Soaked quinoa seedsexhibited 7 decrease in antioxidant activity ascompared to raw seeds The results is supported byfindings of Afiffy et al (2012) who reported de-crease in antioxidant activity in soaked white sor-ghum Xu et al (2008) also reported decrease inantioxidant activity of soaked green pea (9) yel-low pea (8) and lentil (7) As phenols and fla-vonoids contribute significantly to antioxidant ac-tivity[48] the decrease may be due to leaching ofphenols and flavonoids in water used for soakingthe seeds Quinoa seeds germinated in day light for7 days exhibit significantly high antioxidant activitythan raw seeds[17] In our study antioxidant activityof germinated seeds (after 4 days or 48 hours) wasfound to increase by 90 (calculated by DPPHmethod) The result is supported by findings ofCarciochi et al (2014a) which showed 100 in-crease in antioxidant activity of germinated quinoaseeds as evaluated by DPPH method Similarly in-crease in antioxidant activity of germinated quinoa

seeds has also been reported by Pasko et al (2008)FRAP values of germinated quinoa seeds increasedby 89 The result is supported by increase in FRAPvalues of quinoa sprouts (79) as reported by Jubeteet al (2010b) Quinoa (Chenopodium quinoa)sprouts have lower antioxidant activity (evaluatedby FRAP) as compared to Amaranth (Chenopodiumalbum) sprouts[17] However industrial processingof the seeds lead to decline in antioxidant activityProcessed quinoa seeds showed decline of 14 and19 antioxidant activity as evaluated by FRAP andDPPH respectively The result is supported by de-crease in antioxidant activity of rye Zielinski ampKozlowska (2004) wheat[33] barley[31] after under-going industrial processing like decortications andpearling Decline in antioxidant activity after pro-cessing can be due to removal of hulls which aremajorly responsible for antioxidant activity[56] andCardador-Martyacuteacutenez et al (2002) As illustrated in

Figure 2 a strong linear correlation was observedbetween total phenolic content (TPC) and DPPH ofraw soaked germinated and industrially processedIndian quinoa Observed correlation is in agreementwith previously reported literature by Hirose et al(2010) The significant correlation observed in ourstudy is supported by observations of Jubete et al(2010b) where similar correlation (r=099) wasobserved between TPC and DPPH antioxidant ac-tivity of quinoa seeds and sprouts Palombini et al(2013) observed good correlation (r=087) betweentotal phenolic content and antioxidant activity inamaranth and quinoa seeds

Figure 1 Vitamin C TPC FRAP and DPPH for raw and processed Indian quinoa seeds Vitamin C in mg100gTPC in mg GAE100g FRAP and DPPH in mgTE100g




CONCLUSION

Indian Chenopodium quinoa seeds grown inAnantpur district of Hyderabad India were studiedfor the first time for its vitamin C content totalpolyphenols and antioxidant activity The study in-volved comparison of effect of domestic and indus-trial processing techniques on total polyphenols vi-tamin C content and antioxidant activity The resultsreveal domestic processing of quinoa seeds mainlyby the process of germination enriches its vitamin Cpolyphenol content and antioxidant activity Germi-nated quinoa being rich in antioxidants can be used indaily diets for cure of various degenerative diseasesDomestically processed quinoa by germination shouldbe preferred over industrially processed quinoa

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Figure 2 Correlations between TPC and DPPH parameters of Indian quinoa seeds (A) raw seeds (Rsup2 = 09951

y=05395 x - 15851) (B) soaked seeds (Rsup2 = 09854 y = 24204x - 53323) (C) germinated seeds (Rsup2 = 09924 y=15343x

- 98008 (D) industrially processed seeds (Rsup2 = 09982 y = 14373x - 4071)

Intelli et al 39



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Intelli et al 31



presence of both lysine (54) and methionine(21) which gives it a unique feature and makesit a complete food[51] Quinoa is a gluten free grainappropriate for consumption by celiac patients Ithas ash content (3 - 4) higher than common cere-als like wheat and rice and fibre content (14-16total dietary fibre 2-3 crude fibre) which is morethan that present in corn and wheat In additionquinoa seeds and sprouts are found rich inisoflavones polyphenols and known to exhibit goodanti oxidant properties[17] Isoflavones like genisteinand daidzein have been found in quinoa It is foundto contain 11-17mg100g bound phenolics likecoumeric benzoic vanillic and ferulic acid and96-164mg100g free phenolics with quercetinacacetin and kaempferol being majorly presentGood antioxidant activity in quinoa may also beattributed to the presence of vitamin C and toco-pherols[50] which also protects against oxidation ofits fatty acids Ascorbic acid also known as Vita-min C has metabolic and therapeutic benefits whenincluded in human diet High nutritional and phytochemical content make quinoa a wonder grain withmany health benefits Successful propagation ofquinoa in field trials at worlds hottest and driest

place Arabian peninsula prove this crops versa-

tility to adapt in adverse climatic and ecologicalconditions[42] Thus astounding features of this cropsuch as tolerance to stressful environmental con-ditions like soil salinity drought frost high tem-perature etc ability to grow at up to 4500m ofaltitude from sea level and impeccable nutritionalbenefits permit advancement of quinoa cultivationout from the boundaries of its Andean motherlandto different parts of world In addition to the ex-traordinary features the increasing demand ofquinoa across the world has prompted many agri-culturists and researchers worldwide for quinoacultivation[49] After being successfully promotedin England (1970) Denmark Europe (1993) andKenya[13] quinoa has found its way to Asia withkeen interest for the crop mainly in the Indian sub-continent[8] Crop is more widespread in PakistanNepal and India India being a land of diverse cli-matic regions (tropical wet tropical dry subtropi-cal humid and mountains) and quinoa being a crop

profoundly known as well adapted to unusual en-vironmental conditions is found apt to grow in In-dian boundaries[8] Initially being found grown infoots of Himlayan hills the crop has been grownsuccessfully for the first time in year 2013 underthe project named Anantha in plains of drought

prone area of Anantpur region in Andhra Pradesh[18]The project has highly promoted cultivation ofquinoa among various private companies in SouthIndia The craze and demand for quinoa among In-dians has grown far more over than their demandfor the staple traditional crops and millets like sor-ghum pearl millet finger millet etc[49]

Post harvesting and pior to marketing grainsundergo industrial processing mainly the processof dehulling or decortications to remove the outerlayers of the grain Dehulling is known to improvegrain quality by lowering the content of anti nutri-ents[36] and enhancing the sensory parameters hencethe acceptance and pallatabilty of the grain Despitethese benefits of dehulling it reported to cause lossof nutrients from grains Thus to minimize loss nu-trients and increase bioavailabilty of nutrients re-searchers recommend use of common traditionaldomestic processing methods for grains[39 32] Soak-ing and germination are the commonly used methodsfor domestic processing of seed Antioxidant activ-ity and phenolic content have been reported to begreatly affected by domestic processing[23 20]

After many studies revealing nutritional and phy-tochemical composition of american quinoa[17 10 26

50] recently quinoa grown in Kenya Japan and Mo-rocco was studied for its seed quality yield levelantioxidative properties proximate flavonoid fattyacid and mineral compostion The studies demon-strate significant influence of grain variety and cli-matic conditions of area of cultivation on nutritionaland phytochemical composition of the seed Till nowno study has been done on Indian quinoa seeds Theaim of our study is to investigate antioxidant activ-ity content of vitamin c and total polyphenols in In-dian Chenopodium quinoa grains We also comparedthe effect of domestic and industrial processing ofIndian quinoa grain on antioxidant activity contentof vitamin c and total polyphenols which has neverbeen discussed before




MATERIAL AND METHOD

Seed procurement








a Soaking



b Germination





Extract preparation



further analysis

Vitamin C analysis


Intelli et al 33










2CO

3 and incubate











50 values)



50= IC

50troloxIC

50sample




50






















Raw 1343plusmn04a









Raw 432plusmn028a








Intelli et al 35







Total flavonoids





Raw 114plusmn008a


Soaked 72plusmn008b


















FRAP mg TEg

DPPH




b 5351plusmn056b



Intelli et al 37











CONCLUSION


REFERENCES







Intelli et al 39

































































Intelli et al 41




















MATERIAL AND METHOD

Seed procurement








a Soaking



b Germination





Extract preparation



further analysis

Vitamin C analysis


Intelli et al 33










2CO

3 and incubate











50 values)



50= IC

50troloxIC

50sample




50






















Raw 1343plusmn04a









Raw 432plusmn028a








Intelli et al 35







Total flavonoids





Raw 114plusmn008a


Soaked 72plusmn008b


















FRAP mg TEg

DPPH




b 5351plusmn056b



Intelli et al 37











CONCLUSION


REFERENCES







Intelli et al 39

































































Intelli et al 41

















Intelli et al 33










2CO

3 and incubate











50 values)



50= IC

50troloxIC

50sample




50






















Raw 1343plusmn04a









Raw 432plusmn028a








Intelli et al 35







Total flavonoids





Raw 114plusmn008a


Soaked 72plusmn008b


















FRAP mg TEg

DPPH




b 5351plusmn056b



Intelli et al 37











CONCLUSION


REFERENCES







Intelli et al 39

































































Intelli et al 41


































Raw 1343plusmn04a









Raw 432plusmn028a








Intelli et al 35







Total flavonoids





Raw 114plusmn008a


Soaked 72plusmn008b


















FRAP mg TEg

DPPH




b 5351plusmn056b



Intelli et al 37











CONCLUSION


REFERENCES







Intelli et al 39

































































Intelli et al 41

















Intelli et al 35







Total flavonoids





Raw 114plusmn008a


Soaked 72plusmn008b


















FRAP mg TEg

DPPH




b 5351plusmn056b



Intelli et al 37











CONCLUSION


REFERENCES







Intelli et al 39

































































Intelli et al 41





























FRAP mg TEg

DPPH




b 5351plusmn056b



Intelli et al 37











CONCLUSION


REFERENCES







Intelli et al 39

































































Intelli et al 41

















Intelli et al 37











CONCLUSION


REFERENCES







Intelli et al 39

































































Intelli et al 41




















CONCLUSION


REFERENCES







Intelli et al 39

































































Intelli et al 41

















Intelli et al 39

































































Intelli et al 41



















































Intelli et al 41

















Intelli et al 41

















Documents

Vitamin C, total polyphenols and antioxidant activity in raw, · PDF filetically processed and industrially processed Indian Chenopodium quinoa seeds Intelli1*, Dr.Beenu Tanwar2, Dr.Manju