varietal differences ofrutin in common buckwheat(fagopyrum

7/27/2019 Varietal Differences OfRutin in Common Buckwheat(Fagopyrum

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Proceedings a/the 9th International Symposium onBuckwheat, Prague2004

Varietal Differences of Rutin in Common Buckwheat (Fagopyrumesculentum Moench) Determined by MiceJlar Electrokinetic CapiHaryChromatographyJ. Kalinova, E. Dadakova

University ofSouth Bohemia, Faculty 0/Agriculture, Studentska 13, Ceske Budejovice, 37005ABSTRACT

In year 2000we evaluateddifferences in the rutin content amongvarietiesof commonbuckwheatandthe influence of nitrogen fertilization on rutin content in buckwheat and with help micellarelectrokinetic capillarychromatography. Themost of rutin was cumulated in flowers and slightly lessin leavesat stage of full flowering. The rutin contentwas influencedby variety in all plantparts. Somevarieties cumulated more rutin in flowers and the others in leaves. The nitrogen fertilization hadantagonistic influenceon rutin content in achenes . Therutin content in leaveshad positiverelationshipwith rutincontent in stemsandachenes.Keywords: rutin, variety, nitrogenfertilization, buckwheatINTRODUCTIONThe genus Fagopyrum is an important native source offlavonoids and good source for

rutin extraction. The rutin (quercetin 3 - ~ - r u t i n o s i d ) decreases the fragility of blood vesselsthat is connected with hypertense have antioxidative, anti-inflammatory, antimutagenic andanticarcerogenic activities and cause relaxation of smooth muscles (KAVACEVIC, RODE 1998).The rutin can influence digestible starch together with other polyphelols (KREFT et al. 1994).

The rutin content in buckwheat has very broad range of variability. Plants contain thehighest of rutin concentration in the period of beginning flowering (CHOI et al.1996,MICHALOVA, CEJKA 1997).

The aim of this contribution was to evaluate differences in the rutin content amongvarieties of common buckwheat (Fagopyrum esculentum Moench) and the influence ofnitrogen fertilization on rutin content in buckwheat and with help micellar electrokineticcapillary chromatography.

MATERIAL ANDMETHODSIn the year 2000 seven varieties of buckwheat - Pyra (CR), Aelita (Russia), Emka(Poland), Hruszowska (Poland), Kora (Poland) , Krupinka (Russia), Sumcanka (Russia) were grown in four repeated cycles on plots in Ceske Budejovice (380 m a.s.L, sand-loambrown soil gleyficated, precipitation of 620 mm, average temperature 7.gC). Seeds weresown in lines 12.5 em wide, growth density being 200 plants.m", During vegetation periodthere was no mechanical or chemical treatment of the growths.

The material for analysis (the whole plants of buckwheat - Fagopyrum esculentum)was harvested in flowering stage and divided into the parts for analysis (flowers, stems,leaves). This material was immediately frozen and freeze-dried (24 hours, - 46C, 0.25 mbar).The seeds were harvested in optimum maturity and were also freeze-dried (the sameconditions). The powdered sample was extracted with 50% methanol under reflux in a waterbath at 90 0 C for 2 hours. Ascorbic acid was used as the antioxidant. The solid material wasremoved by centrifugation and the extract was filtered through glass fibre filters. The rutinwas isolated using SPE columns (RP-18) and eluted by methanol. This solution was used assample for MECC analysis.The rutin content was calculated from analytical response and the equation of thecalibration function. The linear working range was proposed for concentration range of 102000 ug/ml, limit of quantification was considered to be 10 ug/ml.

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Proceedings of the 9th International Symposium on Buckwheat. Prague 2004

Tab.lBasic meteorological data in year 2000 (normal)

IV. - IX.

RESULTSTab . 2Rutin content in separated parts of buckwheat plant (g .lOOg! dry matter)Variety Stems Leaves Flowers Achenes

x s dif x s dif. x s dif. x s dif.Aelita 0.66 0.07 ab 5.50 0.61 a 6.63 0.73 ab 0.016 0.002 aEmka 0.57 0.06 ab 6.69 0.74 ab 5.93 0.66 a 0.019 0.002 abHruszowska 0.89 0.10 c 6.81 0.75 b 6.18 0.68 ab 0.016 0.002 aKora 0.77 0.09 bc 6.88 0.76 b 5.86 0.65 a 0.024 0.003 bKrupinka 0.55 0.06 a 5.75 0.64 ab 7.65 0.84 b 0.014 0.002 aPyra 0.66 0.07 ab 5.82 0.64 ab 5.90 0.65 a 0.014 0.002 aSumcanka 0.97 o.n c 6.21 0.68 ab 5.92 0.65 a 0.015 0.002 aOkgN 0.71 0.08 a 6.29 0.69 a 6.20 0.68 a 0.019 0.002 b50kgN 0.74 0.08 a 6.18 0.68 a 6.38 0.70 a 0.014 0.002 aAverage 0.72 0.08 6.24 0.69 6.30 0.69 0.017 0.002

The flowers and leaves contained the highest rutin content from different plant parts ofbuckwheat (table 2).

Data about rutin content in buckwheat (MICHALOVA, CEJKA 1997, KAvACEVIC, RODE1998, KREFT et al. 1999) are very different. Our values are similar as data of SHEVCHUK(1983) in case rutin content in flowers and stems. Differences are probably caused differentsites of growing, climatic conditions and the method of rutin determination too. MORISHITA,TETSUKA (2002) reported that very dry weather and high temperatures have positive influenceon rut in cumulat ion. Temperatures were strong above normal in Ceske Budejovice in Mayand June 2000 (table 1). Differences among sites can be caused day length too . HRADECKA(1997) confmned its influence on rutin formation. She established that plants reached rut inmaximum in conditions 16 short days and subsequently 16 long days after their emergence.The rutin content decreased influence of longer long day activity, it converted to pigments.

The rutin content was significant modified by variety in all plant parts and was thehighest in stems of variety Hruszowska and Sumcanka, in leaves of variety Hruszowska andKora, in flowers of variety Krupinka and achenes of variety Kora. Variety Krupinka andHruszowska had the highest total rutin content (13.95 % and 13.88 %) in abovegroundbiomass. MORISHITA, TETSUKA (2002) reported varietal differences in rutin content too .

The influence of nitrogen fertilization on rutin content in achenes was statisticalsignificant (table 3). Average rutin content was higher in control (without fertilization) (table2) as well as after subtraction increase of protein.

The nitrogen fertlilizer rate (50 kg per ha) did not influence the rutin content in leaves,stems and flowers. MAARCISZEWSKl et al . (1984) observed that nitrogen fertilization can

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Proceedings ofthe 9th International Symposium on Buckwheat, Prague 2004

increased rutin content in stems but its influence is considerably depend on ecologicalconditions, especially precipitation.Tab . 3Analysis of variance (MS)Source of variability N Stems Leaves Flowers AchenesVariety 6 0.156** 1.902** 2.577** 8.1E-05**Fertilization 1 0.008 0.117 0.324 2.5E-04**Variety x fertilization 6 0.058* 0.465 2.054 3.6E-05***p :50.05, *.p:5 0.Dl

We established positive correlation between rutin content in leaves and stems andbetween rutin content in leaves and achenes (table 4). SHEVCHUK (1983) reported that rutin iscreated in leaves and is transferred from here in other plant parts. It confirms us establishedpositive correlation among rutin content in leaves and stems and rutin content in leaves andachenes. We can explain non-significant relationship between flowers and leaves with theirsynchronous formation.Tab ACorrelation among rutin content in different parts of plant (r)

LeavesFlowersAchenes*p:5 0.05, **p:5 0.01

Stems0.599**0.1620.069

Leaves0.3240.565**

Flowers

-0.013

ACKNOWLEGDMENTSThis work was supported by grant of the Ministry ofEducation of the Czech Republic 12220002/2.

REFERENCESCHOI B.H. KIM S.L., KIM S.K. (1996): Rutin and functional ingredients of buckwheat and their

variations. Korean Journal ofCrop Science 41 : 69-93HRADECKA D. (1997): Pfispevek k ekologii pohanky. Pohanka seta-vyznam ve zdrave vYzive,vyzkurn, pestovani a trzni realizace. Sbornik referani z odborne konference, Praha. 36-47.KAVACEVIC M., RODE J. (1998) : Determination of rutin in buckwheat leaves. Chromatography and

Hyphenated techniques. Bled. 153.KREFT I., BONAFACCIA G., .lIGO A , (1994): Secondary metabolites of buckwheat and their

importance in human nutrition. Biotechnol. Rev. 32(4): 195-197.KREFT S., KNAPP M. KREFT 1. (1999): Extraction of rutin from buckwheat (Fagopyrum esculentum

Moench) seeds and determination by capillary electrophoresis. 1. Agric.Food Chem.47: 4649-4652.

MAARCISZEWSKl H., SUSZKO-PURZYCKA A., KALEMBASA S. (1984): The influence of nitrogenfertilization on flavonoid contents in buckwheat straw (Fagopyrum esculentum). Fagopyrum4: 14-16.MICHALovA A., CEJKA L. (1997): Variabili ta agronomickych a nutricnich znakU v genofondechpohanky, prosa a laskavce - moznosti jejiho vyuziti. Altemativni a maloobjemoveplodinypro lidskou vYzivu. VURV, Praha: 37-50.

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MORISHlTAT., TETSUKA T. (2002): Varietal differences of rutin, protein and oil content ofcommon buckwheat (Fagopyrum esculentum) grains in Kyushu area. Japanense Journal ofCrop Science 71(2): 192-197.

SHEVCHUK, T.E., 1983: The content of rutin in the different sorts of the buckwheat. Fagopyrum 3: 12.

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varietal differences ofrutin in common buckwheat(fagopyrum

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