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´¢Á¢°¦rš¸É ¡¸Á° 4 7 ¨³ 9 ‹µ„œ´Ê œ˜´ÊŠš·ÊŠÅªo¦³¥³Áª¨µœµœ 1 ´ÉªÃ¤Š ‹µ„œ´ÊœšÎµ„µ¦„µ¦ª·Á‡¦µ³®r ‡ªµ¤µ¤µ¦™Äœ„µ¦
˜oµœ°œ» ¤¼¨°· ¦³ DPPH —oª¥ª·›¸„µ¦ °Š Hatano et al. (1988) ¨³‡ªµ¤µ¤µ¦™Äœ„µ¦Áž} œ˜´ª¦¸—·ªŽr —oª¥ª·›¸„µ¦ °Š
Oyaizu (1996)
„µ¦ª·Á‡¦µ³®rž¦·¤µ–µ¦ž¦³„°¢eœ°¨· „š´ÊŠ®¤— (Total phenolic content)
„µ¦ª· Á‡¦µ³®rž¦· ¤µ–µ¦ž¦³„°¢eœ°¨·„š´ÊŠ®¤—čoª·›¸„µ¦ °Š Singleton ¨³ Rossi (1965) ×¥œÎµµ¦
„´—Á°šµœ°¨‹µ„¦Îµ oµª´Š r®¥—‡ªµ¤Á o ¤ oœ 0.5 ¤·¨¨·„¦´¤/¤·¨¨· ¨·˜¦ ž¦·¤µ˜¦ 200 ŤÇ¦¨·˜¦ Än¨ŠÄœ®¨°—
š—¨°Š œµ— 10 ¤· ¨¨·¨·˜¦ Á˜·¤ Folin-Ciocalteu ž¦·¤µ˜¦ 1.5 ¤·¨¨· ¨·˜¦ š·Ê ŠÅªo š¸É°»–®£¼¤·®o °Š œµœ 5 œµš¸ Á˜· ¤
µ¦¨³¨µ¥ÃŽÁ—¸¥¤Å‡µ¦r°Áœ˜ ‡ªµ¤Á o ¤ oœ 0.566 䨵¦r ž¦·¤µ˜¦ 1.5 ¤·¨¨·¨·˜¦ Á ¥nµµ¦¨³¨µ¥Ä®o Á oµ„´œ š·ÊŠÅªo
š¸É °»–®£¼¤· ®o°Š œµœ 90 œµš¸ ª´ —‡n µ„µ¦—¼—„¨ºœÂŠš¸É 725 œµÃœÁ¤˜¦ —o ª¥Á‡¦ºÉ°Š UV/Vis spectrophotometer
‡Îµœª–ž¦·¤µ–µ¦ž¦³„°¢eœ°¨¨·„š´ÊŠ®¤— ×¥„µ¦Áž¦¸¥Áš¸ ¥„´„¦µ¢¤µ˜¦“µœ °Šµ¦¨³¨µ¥„¦—„¨¨·„
(gallic acid)
„µ¦ª· Á‡¦µ³®r‡ªµ¤µ¤µ¦™Äœ„µ¦Áž}œ˜´ª¦¸—·ªŽr (Reducing power)
„µ¦ª· Á‡¦µ³®r‡ªµ¤µ¤µ¦™Äœ„µ¦Áž}œ˜´ª¦¸—·ªŽr (reducing power) čoª·›¸„µ¦ °Š Oyaizu (1996) ×¥œÎµµ¦
„´—Á°šµœ°¨‹µ„¦Îµ oµª´ Š r®¥—‡ªµ¤Á o¤ oœ 2 ¤·¨¨· „¦´¤/¤·¨¨·¨·˜¦ ž¦·¤µ˜¦ 2.5 ¤· ¨¨·¨·˜¦ Än¨ŠÄœ®¨°—š—¨°Š
œµ— 25 ¤·¨¨· ¨·˜¦ Á˜·¤ 0.2 䨵¦r¢°Á¢˜´Á¢°¦r ¡¸ Á° 6.6 ž¦·¤µ˜¦ 2.5 ¤·¨¨·¨·˜¦ Á˜· ¤µ¦¨³¨µ¥ÃžÂšÁŽ¸¥¤
Ŏ¥µÅœ—r 1% ž¦·¤µ˜¦ 2.5 ¤·¨¨·¨·˜¦ n¤Äœ°nµŠœÊε¦o°œ (water bath) š¸É°»–®£¼¤· 50 °Š«µÁŽ¨ÁŽ¸¥œµœ 20 œµš¸ Á˜· ¤
µ¦¨³¨µ¥„¦—ئ‡¨°Ã¦°³Ž·˜·„‡ªµ¤Á o¤ oœ 10% ž¦·¤µ˜¦ 2.5 ¤·¨¨· ¨·˜¦ œÎµÅžž{~œÁ®ª¸É¥Š (centrifuge) š¸É 650g
œµœ 10 œµš¸ œÎµn ªœÄž¦·¤µ˜¦ 5 ¤· ¨¨·¨·˜¦ Än¨ŠÄœ®¨°—š—¨°Š œµ— 25 ¤·¨¨·¨·˜¦ Á˜·¤œÊÎ µ„¨´Éœ ž¦· ¤µ˜¦ 5
¤·¨¨·¨·˜¦Á˜·¤µ¦¨³¨µ¥Á¢°¦r ¦·‡‡¨°Å¦—r‡ªµ¤Á o ¤ oœ 0.1% ž¦·¤µ˜¦ 1 ¤·¨¨·¨·˜¦ Á ¥nµµ¦¨³¨µ¥Ä®oÁ oµ„´œ ª´—‡nµ„µ¦
—¼—„¨ºœÂŠš¸É 700 œµÃœÁ¤˜¦ —oª¥Á‡¦ºÉ°Š UV/Vis spectrophotometer
„µ¦ª· Á‡¦µ³®r‡ªµ¤µ¤µ¦™Äœ„µ¦˜oµœ°œ»¤¼¨°· ¦³ DPPH (DPPH radical scavenging activity)
„·‹„¦¦¤„µ¦˜oµœ°œ»¤¼¨°· ¦³Äœ¦¼ž °Š radical scavenging activity čo®¨´ „„µ¦ª´—‡nµ—¼—„¨ºœÂŠ °Š
DPPH š¸ÉÁž¨¸É¥œÂž¨ŠÅž (Hatano et al., 1988) ×¥œÎµµ¦„´—Á°šµœ°¨‹µ„¦Îµ oµª´Š r®¥—‡ªµ¤Á o¤ oœ 2 ¤·¨¨·„¦´ ¤
˜n°¤· ¨¨·¨·˜¦ ž¦·¤µ˜¦ 0.1 ¤·¨¨·¨·˜¦ Än¨ŠÄœ®¨°—š—¨°Š œµ— 10 ¤·¨¨·¨·˜¦ Á˜·¤µ¦¨³¨µ¥ DPPH (2,3-diphenyl-1-
picrylhydrazyl) ‡ªµ¤Á o¤ oœ 6 x 10-5 䨵¦rĜµ¦¨³¨µ¥Á¤šµœ°¨ ž¦·¤µ˜¦ 3.9 ¤·¨¨·¨·˜¦ Á ¥nµµ¦¨³¨µ¥Ä®oÁ o µ
„´œ ˜´ÊŠš·ÊŠÅªoĜš¸É¤º—œµœ 30 œµš¸ ª´—‡nµ„µ¦—¼—„¨ºœÂŠ °Š˜´ª°¥nµŠÂ¨³ blank š¸É‡ªµ¤¥µª‡¨ºÉœ 517 œµÃœÁ¤˜¦ —oª¥
Á‡¦ºÉ°Š UV/Vis spectrophotometer ‡Îµœª–®µ % °Š radical scavenging activity ˜µ¤¤„µ¦
DPPH-scavenging activity (%) = (OD 517 »—‡ª‡»¤ - OD517 ˜´ª°¥nµŠ) X 100
(OD 517 »—‡ª‡»¤)