Nutr Res Pract.  2011 Aug;5(4):281-287.

Antioxidant activity and analysis of proanthocyanidins from pine (Pinus densiflora) needles

• Park YS
• Jeon MH
• Hwang HJ
• Park MR
• Lee SH
• Kim SG
• Kim M

Abstract

In this study, we evaluated the antioxidant activity of pine needle extracts prepared with hot water, ethanol, hexane, hot water-hexane (HWH), and hot water-ethanol (HWE), using the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical method. The hot water extract possessed superior antioxidant activity than the other extracts. We also compared the antioxidant activity of pine needle extracts through ROS inhibition activity in a cellular system using MC3T3 E-1 cells. The hot water extract exhibited the lowest ROS production. The pattern of HPLC analysis of each extract indicated that the hot water extract contained the highest proanthocyanidin level. The pine needle hot-water extract was then isolated and fractionated with Sephadex LH-20 column chromatography to determine the major contributor to its antioxidant activity. The No.7 and 12 fractions had high antioxidant activities, that is, the highest contents of proanthocyanidins and catechins, respectively. These results indicate that the antioxidant activity of procyanidins from the hot water extract of pine needles is positively related to not only polymeric proanthocyanidins but also to monomeric catechins. Moreover, the antioxidant activity of the pine needle hot water extract was similar to well-known antioxidants, such as vitamin C. This suggests that pine needle proanthocyanidins and catechins might be of interest for use as alternative antioxidants.

Keyword

DPPH; ROS; HPLC analysis; proanthocyanidin; pine needle

MeSH Terms

Antioxidants
Ascorbic Acid
Catechin
Chromatography
Chromatography, High Pressure Liquid
Dextrans
Ethanol
Needles
Polymers
Proanthocyanidins
Water
Antioxidants
Ascorbic Acid
Catechin
Dextrans
Ethanol
Polymers
Proanthocyanidins
Water

Figure

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  Fig. 1 DPPH radical scavenging activities of pine needle extracts

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  Fig. 2 Effects of pine needle extracts on H2O2-induced reactive oxygen species (ROS) production in MC3T3-E1 osteoblastic cells. MC3T3-E1 cells loaded with DCF-DA were treated with or without pine needle extracts, followed by addition of 0.3 mM H2O2 in a cuvette for the quantitation of fluorescence intensity, which was detected by a multi-detection microplate.

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  Fig. 3 HPLC chromatograms for proanthocyanidin standards of grape seed analyzed by the (B) Federico Peterlongo 1999 method and a (A) newly developed method

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  Fig. 4 DPPH radical scavenging activity of hot water extract fractions

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  Fig. 5 Analysis of a HPLC for fractions No. 7 and 12 separated from hot water extract

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