45

沼津工業高等専門学校研究報告
第 54 号，2020 年 1 月

DART-TOF/MS ࡟ࡼࡿࢧࣛࢩ࢔ᒓ᳜≀ࡢಖ೺ᡂศࡢ᳨ฟⰾ㔝ᜤኈ *1
Determination of health beneficial components in Salacia plants using DART-TOF/MSYOSHINO Kyoji *1
Abstract: Direct analysis in real time (DART) coupled with time-of-flight mass spectrometry(TOF/MS), DART-TOF/MS, is one of the convenient methods to determine some active
components in natural products, because almost no pretreatment is needed. In this study, we
tried to determine salacinol and mangiferin, which could inhibit the activities ofsugar-digestive enzymes, in the stem extract of Salacia plant (Salacia reticulata) with
DART-TOF/MS. The ions at [M] mass of 334 and [M + H]+ of 335 were determined as salacinol,
and the ions at [M] mass of 244 and [M + H]+ of 245 were determined as the aglycone ofmangiferin. Then, this method could be useful to confirm the presence of these available
compounds in Salacia products. In the case of (-)-epicatechin, an ion at [M] mass of 290 was notdetected, so further investigation is necessary to research the suitable conditions for theanalysis of catechins with DART-TOF/MS.
Key Words: Salacia reticulata, DART-TOF/MS, Salacinol, Mangiferin䠍䠊䛿䛨䜑䛻ࢧࣛࢩ࢔ᒓ᳜≀࡜ࡋ࡚ࠊ(ᰴ)┒ගࡼࡾ౪୚ࡉࢀࡓࢫࣜࣛ
ࣥ࢝⏘ࡢ Salacia reticulata ࡢᖿࢆ⏝࠸ࡓࠋࡇࡢᖿࡢᚤ⢊ࢧࣛࢩ࢔ᒓ᳜≀ࡣࠊ࢖ࣥࢻࡸࢫࣜࣛࣥ࢝࡞࡝ࡢ⇕ᖏᆅᇦ○≀࡟ 9 ಸ㔞ࡢỈࢆຍ࠼ࠊ50Υࠊ1 ᫬㛫᣺┞ᢳฟࡋࡓࠋ࡟⏕ᜥࡋ࡚࠸ࡿࢹࢳ࣒ࣥࣝ⛉ࡢࢶࣝᛶᶞᮌ࡛࠶ࡿࠋ࢖ࣥࢻ㸪ᢳฟ≀ࢆ 10Υࠊ4000 rpmࠊ20 ศ㛫㐲ᚰศ㞳ࡋࡓᚋࠊࡑࢫࣜࣛࣥ࢝ࡢఏ⤫་Ꮫ࡛࠶ࡿ࢔࣮ࣦ࢙࣮ࣘࣝࢲ࡟グ㍕ࡢࡢୖΎࢆ෾⤖஝⇱ࡉࡏ࡚࢚࢟ࢫࢆᚓࡓࠋ཰㔞ࡣ⣙ 6㸣࡛࠶࠶ࡿࣁ࣮ࣈࡢ୍ࡘ࡛ࠊࡑࡢ᰿ࡸᖿࡀ⢾ᒀ⑓ࡢึᮇ἞⒪➼࡟ࡗࡓࠋࡇࡢ࢚࢟ࢫ࡟ࡘ࠸࡚ࠊ0.001%(w/v)ࡢỈ⁐ᾮࢆㄪ〇౑⏝ࡉࢀ࡚ࡁࡓ[1,2]ࠋస⏝ᶵᗎࡢ୍ࡘ࡜ࡋ࡚ࠊ஧⢾㢮ࡢᾘࡋ࡚ DART-TOF/MS ࡟ࡼࡿศᯒࢆ⾜ࡗࡓࠋ໬㓝⣲࡛࠶ࡿα-ࢢࣝࢥࢩࢲ࣮ࢮ㸦AGc㸧άᛶࢆ㜼ᐖࡍࡿస⏝ࡀ࠶ࡾ[3]ࠊࡑࡢ᭷ຠᡂศ࡜ࡋ࡚ࡣࢳ࢜⢾◲㓟ሷࡢࢧࣛࢩ㸰㸬㸰 DART-TOF/MS ἲ࡟ࡼࡿࢧࣛࢩࣀ࣮ࣝ࡜࣐ࣥࢠ

ࣀ࣮ࣝ[4]࠾ࡼࡧࢥࢱࣛࣀ࣮ࣝ[5]࡜ࡑࡢ㢮⦕໬ྜ≀ࠊ࢟ࢧࣇ࢙ࣜࣥࡢᐃᛶศᯒࣥࢺࣥ㓄⢾యࡢ࣐ࣥࢠࣇ࢙ࣜࣥ[6]➼ࡀ▱ࡽࢀ࡚࠸ࡿࠋࡲࡓࠊࢧࣛࢩ࢔ᒓ᳜≀ࡢᖿ࢚࢟ࢫ࡟ࡘ࠸࡚ࠊࡑࢀ࡟ྵ᭷ࡉࢀࡿ࣏ࣜࣇ࢙ࣀ࣮ࣝᡂศ࡜ࡋ࡚ࠊ(-)-࢚ࣆ࢝ࢸ࢟ࣥ㸦EC㸧㸪 (-)-

ࢧࣛࢩࣀ࣮ࣝ࡜࣐ࣥࢠࣇ࢙ࣜࣥࡢᐃᛶศᯒࢆ࢚ࣆ࢞ࣟ࢝ࢸ࢟ࣥ࡜ࡑࡢ࣓ࢳࣝ໬≀ࡸ 2 㔞య࡜࠸ࡗࡓ࢝DART-TOF/MS ἲࢆ⏝࠸࡚⾜ࡗࡓࠋ⿦⨨ࡣ JEOL ♫〇ࢸ࢟ࣥ㢮࡜ࡑࡢㄏᑟయࡶ᳨ฟࡉࢀ࡚࠸ࡿ[7]ࠋ

DART ࢖ ࢜ ࣥ ※ ⿦ ╔ JMS-T100LC AccuTOF τᮏ◊✲࡛ࡣࠊࣜ࢔ࣝࢱ࢖࣒┤᥋ศᯒ㸦DART㸧-㣕⾜᫬㛫

LC-TOFMSࠊ࢖࢜ࣥ໬࢞ࢫࡣ࣒࣊ࣜ࢘ࠊ࢞ࢫ ᗘࡣ 350Υࠊ

ᆺ㉁㔞ศᯒ㸦TOF/MS㸧ἲࢆ⏝࠸࡚ࠊࢧࣛࢩ࢔ᒓ᳜≀୰ࡢṇ࢖࣮࢜ࣥࣔࢻ࡛ ᐃࡋࡓࠋ࢜ࣜࣇ࢕ࢫ 1 㟁ᅽࡣ 85 Vࠊࡇࢀࡽࡢᡂศࢆ᳨ฟࡍࡿࡓࡵࡢ⡆౽࡞᪉ἲ࡟ࡘ࠸᳨࡚ウ࢜ࣜࣇ࢕ࢫ 2 㟁ᅽࡣ 5 Vࠊࣜࣥࢢࣞࣥࢬ㟁ᅽࡣ 15 Vࠊࣆ࣮ࢆ⾜ࡗࡓࠋࢡ㟁ᅽࡣ 500 V ࡜ࡋࡓࠋ ᐃ⠊ᅖࡣ m/z 50-1000 Da ࢆ⏝࠸ࡓࠋ䠎䠊ᮦᩱ䛚䜘䜃᪉ἲ㸰㸬㸯 ࢧࣛࢩ࢔ᒓ᳜≀ࡢᖿ࢚࢟ࢫࡢㄪ〇䠏䠊⤖ᯝ䛚䜘䜃⪃ᐹࢧࣛࢩ࢔ᒓ᳜≀ᬑཬ༠఍࡛ࡣࠊࢧࣛࢩ࢔ᒓ᳜≀ࢆྵ᭷ࡍ*1 ≀㉁ᕤᏛ⛉Department of Chemistry & Biochemistryࡿ〇ရࡢရ㉁ࢆ☜ㄆࡍࡿࡓࡵࡢᇶ‽ࡢ୍ࡘ࡜ࡋ࡚ࠊࢧࣛࢩ࢔ᒓ᳜≀ࡢ࢚࢟ࢫ࡟ࡘ࠸࡚ࠕࢧࣛࢩ࢔ᒓ᳜≀≉᭷ࡢᡂศ࡛

46

沼津工業高等専門学校研究報告࠶ࡿࢧࣛࢩࣀ࣮ࣝࡢᐃᛶศᯒࠖ࡟ࡼࡿࢧࣛࢩ࢔ᒓ᳜≀࡛࠶ࡿࡇ࡜ࡢ☜ㄆヨ㦂ࢆ⾜࠺ࡇ࡜ࢆ᥎ዡࡋ࡚࠸ࡿ[8]ࠋࢧࣛࢩࣀ

第 54 号[M + H]+࣮ࣝࡀ㑅ᢥࡉࢀࡓ⌮⏤࡜ࡋ࡚ࡣࠊAGc άᛶ㜼ᐖస⏝ࢆ♧ࡍ୺せᡂศ࡛࠶ࡿࡇ࡜࡜ࠊࡑࡢᶆ‽≀㉁ࡀᕷ㈍ࡉࢀ࡚࠸ࡿࡇ࡜ࡀ⪃࠼ࡽࢀࡿࠋ⌧ᅾࠊࢧࣛࢩࣀ࣮ࣝࡢᐃ㔞ศᯒἲ࡟ࡣࠊ

[M]ᾮయࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮/㉁㔞ศᯒ㸦LC/MS㸧ἲ[9]ࡸ࢟ࣕ
ࣆ࣮ࣛࣜࢰ࣮ࣥ㟁ẼὋືἲ[10]ࡀ▱ࡽࢀ࡚࠸ࡿࠋ୍᪉ࠊࡑࡢᐃᛶศᯒἲ࡜ࡋ࡚ࡣࠊ⡆౽࡞᪉ἲࡀぢฟࡉࢀ࡚࠸࡞࠸ࠋDART-TOF/MS ἲࡣࠊศᯒࡢࡓࡵࡢヨᩱࡢ๓ฎ⌮ࡀ࡯ࡰᚲせ࡞ࡃࠊ⢒〇ࡢኳ↛≀࢚࢟ࢫ➼ࢆࡑࡢࡲࡲ౑⏝࡛ࡁࡿ⡆౽࡞ᐃᛶศᯒࡢᡭἲࡢ୍ࡘ࡛࠶ࡿࠋࡋ࠿ࡋࠊࡇࢀࡲ࡛࡟ࡇࡢ᪂ࡓ࡞ศᯒ᪉ἲࡢࡓࡵࡢศᯒ᮲௳ࡘ࠸࡚ࡣࠊ༑ศ࡟᳨ᅗ 㻞 㻌 䝃 䝷 䝅 䜰 ᒓ᳜ ≀䛾 ᖿ 䜶 䜻 䝇 ୰䛾 䝃 䝷 䝅 䝜 䞊 䝹 䛾㻌ウࡉࢀ࡚ࡣ࠸࡞࠸ࡶࡢ࡜ᛮࢃࢀࡿࠋᡃࠎࡣࠊ࢔࢝ࢨ࢝ࢬࣛ

㻰㻭㻾㼀㻙㼀㻻㻲㻛㻹㻿 䝇䝨䜽䝖䝹㻌

㸦Anredera cordifolia㸧ࡢⴥ࡟ྵࡲࢀࡿ⢾྾཰ᢚไస⏝ࢆᣢࡘࢺࣜࢸࣝ࣌ࣀ࢖ࢻࢧ࣏ࢽࣥࡢ boussingoside A1 ࡟ࡘ࠸࡚ࠊDART-TOF/MS ࡟ࡼࡿᐃᛶศᯒࢆሗ࿌ࡋࡓ[11]ࠋ
ᮏᐇ㦂࡛ࡣࠊࢧࣛࢩ࢔ᒓ᳜≀ࡢᖿ࢚࢟ࢫ࡟ࡘ࠸࡚ࠊࡲࡎ

[M][M + H]+ࢧࣛࢩࣀ࣮ࣝࡢᏑᅾࢆ DART-TOF/MS ᳨࡛ฟࡍࡿࡇ࡜ࢆヨࡳࡓࠋᮏ◊✲࡛ศᯒࢆ⾜ࡗࡓ໬ྜ≀ࡢ໬Ꮫᵓ㐀ࢆᅗ 1 ࡟♧ࡍࠋࢧࣛࢩࣀ࣮ࣝࡢศᏊ㔞ࡣ 334 ࡛࠶ࡿࡓࡵࠊm/z 330
̺355 ࡢ⠊ᅖ࡛ࡢ DART-TOF/MS ࡢࢫ࣌ࢡࢺࣝࢆᅗ 2 ࡟♧ࡍࠋm/z 334 ࠾ࡼࡧ m/z 335 ࡟ࠊࢧࣛࢩࣀ࣮ࣝࡢศᏊ࢖
࢜ࣥ[M]࡜ࡑࡢࣉࣟࢺࣥ௜ຍయศᏊ࢖࢜ࣥ[M + H]+࡟┦ᙜࡍࡿࣆ࣮ࢡࡀぢࡽࢀࠊࢧࣛࢩࣀ࣮ࣝࡢᏑᅾࡀ♧၀ࡉࢀࡓࠋḟ࡟ࠊ࣐ࣥࢠࣇ࢙ࣜࣥ࡟ࡘ࠸᳨࡚ウࡋࡓࡀࠊ࣐ࣥࢠࣇ࢙ࣜ
ࣥࡢ[M]࠾ࡼࡧ[M + H]+࡟┦ᙜࡍࡿ఩⨨࡟ࡣࠊᑠࡉ࡞ࣆ࣮ᅗ 㻟㻌 䝃䝷䝅䜰ᒓ᳜≀䛾ᖿ䜶䜻䝇୰䛾䝬䞁䜼䝣䜵䝸䞁䜰䜾䝸㻌ࢡࡋ࠿ㄆࡵࡽࢀ࡞࠿ࡗࡓࠋࡑࡇ࡛ࠊࡑࡢ࢔ࢢࣜࢥࣥࢆ☜ㄆ䝁䞁䛾 㻰㻭㻾㼀㻙㼀㻻㻲㻛㻹㻿 䝇䝨䜽䝖䝹㻌ࡍࡿࡓࡵࠊm/z 235̺255 ࡢ⠊ᅖ࡛ࡢ DART-TOF/MS ࡢࢫ࣌ࢡࢺࣝࢆᅗ 3 ࡟♧ࡍࠋm/z 244 ࠾ࡼࡧ m/z 245 ࡟ࠊ࣐ࣥOH

ࢠࣇ࢙ࣜࣥࡢ࢔ࢢࣜࢥࣥ㒊ศࡢ [M]࡜ [M + H]+࡟┦ᙜࡍ-HOOH

OSO3HOHOS +OOH

HOO

OH

OHOH

OH

ࡋ࠿ࡋࠊIonSense ♫ࡢ࢔ࣉࣜࢣ࣮ࢩࣙࣥࣀ࣮ࢺ[14]࡛ࡣࠊ
ࢳࣕ㸦Camellia sinensis㸧ࡢ࢝ࢸ࢟ࣥࢆ DART-TOF/MSO

OHEC ➼ࡢ࢝ࢸ࢟ࣥ㢮࠶ࡿ࠸ࡣࡑࡢ㢮⦕໬ྜ≀࡟ࡘ࠸࡚ࡶࠊ

ࢀࡿ࢚ࣆ࢔ࣇࢮࣞ࢟ࣥࢆ DART-TOF/MS ࡛ ᐃࡋ࡚࠸ࡿࠋ䝃䝷䝅䝜䞊䝹㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻌 㻔㻙㻕㻙䜶䝢䜹䝔䜻䞁㻌HOࡿศᯒ࡟ࡘ࠸࡚ࡣࠊ
ࡇࢀࡲ࡛࡟ Wang ࡽ[12]ࡢሗ࿌ࡀ࠶ࡿࠋ

[13]ࡣࠊ࢝ࢩ࢔ᒓࡢ᳜≀㸦Cassia sieberiana㸧ࡢ᰿࡟ྵࡲOHOH

ࣇࣛ࣎ࣀ࢖ࢻ㢮࡜ࡑࡢ࢔ࢢࣜࢥࣥࡢ DART-TOF/MS ࡟ࡼ

DART-TOF/MS ࡟ࡼࡿศᯒࡢሗ࿌ࡣᑡ࡞࠸ࠋKpegba ࡽOHHOࡿࣆ࣮ࢡࡀぢࡽࢀࠊ࣐ࣥࢠࣇ࢙ࣜࣥࡢᏑᅾࡀ♧၀ࡉࢀࡓࠋ࡛ ᐃࡍࡿሙྜࠊࡑࡢỈ㓟ᇶࢆࢺ࣓ࣜࢳࣝࢩࣜࣝ໬࡟ࡼࡾOಖㆤࡋ࡞࠸࡜⇕ศゎ࡟ࡼࡾ[M]ࡢࣆ࣮ࢡࡀ᳨ฟࡉࢀ࡞࠸ྍ䝬䞁䜼䝣䜵䝸䞁㻌⬟ᛶࡀ࠶ࡿࡇ࡜ࡀ♧ࡉࢀ࡚࠸ࡿࠋᐇ㝿ࠊᮏ◊✲࡛⏝࠸ࡓᐇ㦂᮲௳ୗ࡛ EC ࡢᶆ‽ရࡢ 0.1%(w/v)Ỉ⁐ᾮࢆศᯒࡋࡓ࡜ᅗ 㻝㻌 䝃䝷䝅䝜䞊䝹䚸䝬䞁䜼䝣䜵䝸䞁䚸㻔㻙㻕㻙䜶䝢䜹䝔䜻䞁䛾໬㻌ࡇࢁࠊࡑࡢศᏊ㔞࡛࠶ࡿ 290 ࡟┦ᙜࡍࡿࣆ࣮ࢡࡣぢࡽࢀ࡞Ꮫᵓ㐀㻌࠿ࡗࡓ㸦ᅗ 4㸧ࠋ࣐ࣥࢠࣇ࢙ࣜࣥࡸ࢝ࢸ࢟ࣥ㢮ࡣࠊ㧗㏿ᾮయ

DART-TOF/MS によるサラシア属植物の保健成分の検出

47jp/approach.html, 2019.9.29 ྲྀᚓ.
[9] O. Muraoka, T. Morikawa, et al. (2010): J. Pharm.

Biomed. Anal., 52, 770-773.
[10] W. F. Zandberg, S. Mohan, et al. (2010): Anal.

Chem., 82, 5323-5330.
[11] ⰾ㔝ᜤኈ, ჆ᓥᗣ஧, ௚ (2018): ἟ὠ㧗ᑓ◊✲ሗ࿌,52, 33-38.
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Anal., 121, 30-38.
[13] K. Kpgba, A. Agbonon, et al. (2011): J. Nat. Prod.,74, 455-459.
ᅗ 㻠㻌 㻔㻙㻕㻙䜶䝢䜹䝔䜻䞁䛾 㻰㻭㻾㼀㻙㼀㻻㻲㻛㻹㻿 䝇䝨䜽䝖䝹㻌[14] Green tea analysis with DART,IonSense

Application Notes, IonSense Inc., https://www.ionsense.
ࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮㸦HPLC㸧ࢆ⏝࠸࡚ᐜ᫆࡟ศᯒ࡛ࡁࡿ

com/internal/appNotesDetails.cfm/30/en/2, 2019.9.29 ྲྀࡀ[15]ࠊ࢝ࢸ࢟ࣥ㢮ࢆ DART-TOF/MS ࡛ ᐃࡍࡿሙྜ࡟ᚓ.

ࡣศᯒ᮲௳ࡸヨᩱࡢ๓ฎ⌮ࡢ᳨ウࢆ⾜࠺ᚲせࡀ࠶ࡿࡶࡢ

[15] ⰾ㔝ᜤኈ, ᓊ ⏤⣖஀, ௚ (2012): ᪥ᮏᰤ㣴࣭㣗⣊Ꮫ࡜⪃࠼ࡽࢀࡿࠋ

఍ㄅ, 65, 221-227.

䠐䠊䜎䛸䜑ᮏ◊✲ࡢ⤖ᯝ࠿ࡽࠊࢧࣛࢩ࢔ᒓ᳜≀࡟ྵࡲࢀࡿಖ೺ᡂศࡢ࠺ࡕࠊࢧࣛࢩࣀ࣮ࣝ࡜࣐ࣥࢠࣇ࢙ࣜࣥ࡟ࡘ࠸࡚ࡣDART-TOF/MS ࢆ⏝࠸࡚ࡑࡢᐃᛶศᯒࢆ⾜࠺ࡇ࡜ࡀྍ⬟࡛࠶ࡿࡇ࡜ࡀࢃ࠿ࡗࡓࠋDART-TOF/MS ࡣࠊ ᐃᑐ㇟ࡢヨᩱࢆ⡆༢࡞๓ฎ⌮ࡢࡳ࡛ศᯒ࡛ࡁࡿࡓࡵࠊ௒ᚋࠊ་⸆ရࡸಖ೺ᶵ⬟㣗ရࠊ࠶ࡿ࠸ࡣẘ≀➼࡟ྵࡲࢀࡿ≉ᐃᡂศࡢ᳨ฟࡸྠᐃ࡬ࡢ฼⏝ࡀᮇᚅࡉࢀ࡚࠸ࡿࠋ䠑䠊ཧ⪃ᩥ⊩[1] M. Modak, P. Dixit, et al. (2007): J. Clin. Biochem.

Nutr., 40, 163-173.
[2] ྜྷᕝ㞞அ (2002): ໬Ꮫ࡜⏕≀, 40, 172-178.
[3] ྜྷᕝ㞞அ㸪Y. Pongpiriyadacha㸪௚ (2003): YakugakuZasshi, 123, 871-880.
[4] M. Yoshikawa, T. Morikawa, et al. (2002): Bioorg.

Med. Chem., 10, 1547-1554.
[5] M. Yoshikawa, T. Murakami, et al. (1998): Chem.

Pharm. Bull., 46, 1339-1340.
[6] ྜྷᕝ㞞அ㸪す⏣඾Ọ㸪௚ (2001): Yakugaku Zasshi, 121,371-378.
[7] M. Yoshikawa, K. Ninomiya, et al. (2002): Biol.

Pharm. Bull., 25, 72-76.
[8] ࢧࣛࢩ࢔ᒓ᳜≀ つ᱁ᇶ‽⟇ᐃ࡟ྥࡅ࡚ࡢ༠఍ࡢྲྀࡾ⤌ࡳ, ࢧࣛࢩ࢔ᒓ᳜≀ᬑཬ༠఍, http://salacia-association.

