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Method of quercetin extraction from dry scales of onion.

Article  in  Vegetable Crops Research Bulletin · October 2002

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HORBOWICZ M. 2002. Method of quercetin extraction from dry scales of onion. Veget.
Crops Res. Bull. 57: 119-124.

METHOD OF QUERCETIN EXTRACTION

FROM DRY SCALES OF ONION

Marcin HORBOWICZ
Research Institute of Vegetable Crops
95-100 Skierniewice, Konstytucji 3 Maja 1/3, Poland

Summary
Dry outer scales of brown onion is one of the reachest known sources of free
quercetin. In other plant tissues quercetin is present as glycosides only. Dry scales of onion is
a waste during production and processing of onion. A method of extraction of quercetin from
powdered onion scales was elaborated. During research effects of time, temperature and
solvents used for extraction process were studied. It was found that 4-hours extraction by
shaking with cold ethyl acetate is effective, fast and simple method of isolation crude
quercetin from dry scales of onion. Obtained product is a powder of yellow colour, and its
purity is equal 70%. Hot extraction improved slightly yield of the process, but product purity
was lower. Extraction with ethanol containing various concentration of water caused that
crude quercetin is oily and contain up to 78% of contaminants. Such product needs further
purification.

Key words: onion, dry scales, quercetin, extraction method

INTRODUCTION
Flavonoids are part of a large group naturally occurring plant phenolics. They occur in
plants mainly as a glycosides - conjugates of several types of aglycones with carbohydrates
(Herrmann 1988). Flavonols are flavonoids of particular importance as they have been found
to posses antioxidant and free radical scavenging activity in foods (Hertog et al. 1993;
Hollman 1997). Among them quercetin is most widely distributed within the plant kingdom
(Horbowicz 2000). Quercetin is derivative of flavan-3,4-diols, containing 5 hydroxyl groups
(Fig. 1). Quercetin is a dietary antioxidant that prevents oxidation of low-density lipoproteins
in vitro, and increases absorption of vitamin C (Rimm et al. 1996). The quercetin works in a
synergistic way with vitamin C to rebuild small blood vessels and many other structural parts
of the body (Das & Rey 1988; Formica & Regelson 1995).
Vegetables, fruits and beverages are the main dietary sources of flavonols, primarily as
quercetin and kaempferol (Herrmann 1988; Hertog et al. 1992). The epidemiological studies
have indicated that high consumption of flavonols is associated with reduced risk of cancer
and coronary diseases (Horbowicz 2000). One of the major sources of flavonols in the
European diet is the onion and some other Allium crops (Hertog et al. 1992; Horbowicz 2000).
Edible part of onion contain several type of quercetin glucosides, and onion leaves glucosides
of quercetin and kaempferol (Horbowicz and Kotlińska 1998). Many fruits, vegetables, nuts,
and seeds contain bioflavonoids (mainly quercetin), and a diet rich in these ingredients
provides adequate their amounts; however, if the diet does not allow for proper intake of these
substances or if bodily injury or malfunction is experienced, a supplement which provides 500
mg or more per day is recommended.
There are hundreds of commercially available preparations containing quercetin and
other bioflavonoids. One tablet usually contains 100 to 500 mg of quercetin, rutin, hesperedin and
other flavonoids and vitamin C. The quercetin and other flavonoids are isolated from different
plant sources: seeds, nuts, leaves and fruits. One of the best source of natural free quercetin
can be dry scales of onion. The main aim of present research was to elaborate the simple and
sufficient method for isolation of crude quercetin, which can be used for further purification
and/or production of capsules, tablets and powdered form.

MATERIAL AND METHODS

Material
Dry outer scales of onion were collected from different brown cultivars of onion,
mainly Sochaczewska and Blonska. The scales were pulverized in laboratory mill and sieved.
Particles smaller then 0.2 mm was used to experiments with extraction of quercetin.
Extraction methods:
1. Extraction by shaking for 4 h of material in laboratory shaker using:
a. 60% ethanol; b. 80% of ethanol; c. 96% of ethanol; d. ethyl acetate
e. extraction by 2 hours heating under reflux condenser with ethyl acetate
f. extraction by 2 hours heating under reflux condenser with 60% ethanol
Proportion of solvent to material was: 50 ml of solvent per 1 g of powder.
2. Extraction by shaking of material in laboratory shaker with ethyl cold acetate during:
a. 0.5 h; b. 1.0 h; c. 4.0 h; d. 24 hours
3. Extraction by shaking of material in laboratory shaker with cold ethyl actete during 4 hours
in proportion volumes of solvent (ml) to weight of powdered onion scales (g):
a. 10 : 1; b. 20 : 1; c. 30 : 1; d. 50 : 1; e. 100 : 1.
Extracts were filtered under vacuum, and 20 microliters of clear supernatants were
taken to analysis. Solvent was evaporated in rotary evaporator under vacuum in 40°C. Weight
of residue after evaporation of solvent was used for calculation of extraction efficiency (in %).
Analysis
To analyse the quercetin content in obtained extracts a LKB (Sweden) HPLC apparatus
equipped with Rheodyne 7125 injection system (loop 20 µl), UV detector (2151 Variable
Wavelenght Monitor) set at 370 nm, and Shimadzu C-R6A Chromatopac integrator was used
(Patil et al., 1995; Horbowicz 1999). The quercetin was isocratically separated on Lichrosorb
RP18 (4 x 250 mm, 10 µm) column, and a mobile phase was methanol:water mixture (55:45,
v/v) contained 0.2% ortho-phosphoric acid. The flow rate was 0.8 ml/min. In such conditions
retention time of quercetin was 13.1 min. The standard curve was prepared for concentration
range 0.2 to 10.0 µg/ml. Analyses were done in three replicates, and results were statistically
calculated.

RESULTS AND DISCUSSION

Highest yield of extraction of quercetin from powdered dry scales of onion was
obtained by hot extraction with 60% of ethanol, although content of quercetin in crude extract
was only 21% (Table 1). Obtained through such method product was liquid and oily. In all
cases when ethanol solutions were used crude quercetin was always oily. Such crude quercetin
was contaminated up to 78% with soluble carbohydrates, phenolic acids and non-volatile
organic acids. Addition of water to ethanol increased efficiency of extraction process, but
decreased clearly its purity (Table 1).

When quercetin was extracted from powdered onion scales by shaking with cold ethyl
acetate the yield of crude product was ranged from 3.57% to 4.56%, and purity reached level
70% (Tables 2 and 3). According to results from Table 2 half hour, and 1 hour of extraction
process is too short time to get high yield of crude quercetin. The process should be
continuing for 4 hours, and longer time did not increase the extraction efficiency. When hot
ethyl acetate was used the yield of crude quercetin was higher - 4.66%, but its purity declined
to value 60% (Table 1).

Table 3 contains results of investigation on effect of volume of used ethyl acetate on

efficiency of extraction process. The results indicated that for proper extraction process 50
volumes of solvent on one weight of powdered onion scales have to be used. There is no
information in available literature concerning extraction and purification methods of quercetin
from natural sources.

CONCLUSIONS

a. Cold extraction with ethyl acetate is effective, fast and simple method of isolation of crude
quercetin from powdered dry scales of onion. Using such procedure yellow, powdered
quercetin can be obtained with 70% purity.
b. Hot extraction with ethyl acetate improved non-significantly yield of crude quercetin, but
decreased its purity.
c. Yield of crude quercetin obtained by its extraction with ethanol-water solutions was much
higher, but product was oily, and contained up to 78% of contaminants. It needs further
purification.

Acknowledgement
The financial support obtained from The State Committee for Scientific Research (KBN),
Poland, grant no. 5 P06C 021 12, is greatfully acknowledged.

REFERENCES

Das M., Rey P.K. 1988. Lipid antioxidant properties of quercetin in vitro. Biochem. Int. 17:203-208.
Formica J.V., Regelson W. 1995. Review of the biology of quercetin and related bioflavonoids. Fd.
Chem. Toxic. 33:1061-1080.
Herrmann K. 1988. On the occurrance of flavonol and flavone glycosides in vegetables. Z. Lebensm.
Unters. Forsch. 186:1-5.
Hertog M.G.L., Feskens E.J.M., Hollmann P.C.H., Katan M.B., Kromhout D. 1993. Dietary
antioxidants flavonoids and risk of coronary heart disease: The Zutphen Elderly Study. Lancet
342:1007-1011.

Hertog M.G.L., Hollman P.C.H., Katan M.B. 1992. Content of potentially anticarcinogenic flavonoids
of 28 vegetables and 9 fruits commonly consumed in the Netherlands. J. Agr. Food Chem.
40:2379-2383.

Hollman P.C.H. 1997. Determinants of the absorption of the dietary antioxidant flavonoid quercetin in
man. PhD Thesis. Landbouwuniversiteit Wageningen: pp.1-187.
Horbowicz M. 1999. Changes of flavonol content during vegetation period and storage of
onion. Veget. Crops Res. Bull. 50: 81-91.

Horbowicz M. 2000. [Occurence, biosynthesis and biological characteristcs of flavonols]

Post.
Nauk. Rol., 2/2000:3-18. [in Polish with English summary].

Horbowicz M., Kotlińska T. 1998. [Flavonols content in wild and cultivated Allium species]. Zesz.
Prob. Post. Nauk Rol. 463:529-537. [in Polish with English summary].

Patil B.S., Pike L.M., Yoo K.S. 1995. Variation in the quercetin content in different coloured onions
(Allium cepa L.). J. Amer. Soc. Hort. Sci. 120:909-913.

Rimm E.B., Katan M.B., Asherio A., Stampfer M.J., Willet W.C. 1996. Relation between intake of
flavonoids and risk of coronary heart disease in male health professionals. Ann. Intern. Med.
125:384-389.

METODA EKSTRAKCJI KWERCETYNY Z SUCHYCH ŁUSEK CEBULI

Streszczenie
Sucha łuska okrywowa brązowych odmian cebuli jest jednym z najbogatszych,
znanych źródeł kwercetyny w stanie wolnym. W innych tkankach roślinnych kwercetyna
występuje jedynie w postaci glikozydów. Sucha łuska jest surowcem odpadowym podczas
produkcji i przetwórstwa cebuli. Podjęto badania dotyczące wyboru rodzaju rozpuszczalnika i
warunków prowadzenia procesu ekstrakcji na jej wydajność i czystość uzyskiwanej
kwercetyny. Stwierdzono, że 4-godzinna ekstrakcja na zimno poprzez wytrząsanie z octanem
etylu jest efektywną, szybką i prostą metodą wyodrębniania kwercetyny z suchych łusek
cebuli. Otrzymuje się produkt w postaci żółtego proszku o czystości 70%. Ekstrakcja na
gorąco nie poprawia wydajności procesu, natomiast otrzymany produkt jest niższej czystości.
Ekstrakcja z różnymi roztworami wodnymi etanolu pozwala uzyskać kwercetynę techniczną
w postaci oleistej zawierającej do 78% domieszek. Wymaga ona dalszego oczyszczania.
Table 1. Efficiency of quercetin extraction from powdered dry onion scales and purity of obtained
crude quercetin

Method of extraction and time of the process Efficiency of Quercetin content in

extraction residue after solvent
*
(% of dry powder ) evaporation (%*)
Shaking with cold ethyl actetate , 4 hours 4.40 e 70 a
Shaking with cold 60% of ethanol, 4 hours 10.82 b 45 d
Shaking with cold 80% of ethanol, 4 hours 8.82 c 55 c
Shaking with cold 96% of ethanol, 4 hours 6.80 d 62 b
Extraction with ethyl acetate by heating under reflux 4.66 e 61 b
condenser, 2 hours
Extraction with 60% of ethanol by heating under 22.05 a 21 e
reflux condenser, 2 hours

*
-results marked with the same letters do not differ significantly
Table 2. Effect of extraction time on efficiency and purity of obtained crude quercetin.

Extraction time Efficiency of extraction Quercetin content in residue

(hours) (% of dry powder*) after solvent evaporation
(% *)
0.5 3.57 c 69 a
1.0 3.86 b 70 a
4.0 4.40 a 70 a
24 4.30 a 69 a

*
- results marked with the same letters do not differ significantly
 Table 3. Effect of amount of used ethyl acetate on efficiency and purity of obtained crude quercetin.

Volume of ethyl acetate (ml) Efficiency of extraction Quercetin content in residue

used per weight (g) of powdered (% of dry powder *) after solvent evaporation
dry onion scales (% *)
10 : 1 3.30 c 70 a
20 : 1 4.20 b 69 a
30 : 1 4.32 b 71 a
50 : 1 4.56 a 70 a
100 : 1 4.66 a 68 a

*
- results marked with the same letters do not differ significantly

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