Page 22 - วารสารปีที่15ฉบับที่1

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“√¿“¬„μâ· ßÕ—≈μ√“‰«‚Õ‡≈μ∑’˧«“¡¬“«§≈◊Ëπ 254 2.5 °“√·¬°Õߧåª√–°Õ∫‡§¡’¥â«¬‡∑§π‘§ Col-

·≈– 366 π“‚π‡¡μ√ ·≈–μ√«® Õ∫°≈ÿà¡ “√¥â«¬ umn Chromatography
°“√æàπ¥â«¬πÈ”¬“æàπ (reagent) 3 ™π‘¥ ‰¥â·°à °“√·¬° “√ °—¥À¬“∫·Àâß∑’Ë °—¥¥â«¬μ—«∑”
anisaldehyde reagent, natural products re- ≈–≈“¬ acetone ®”π«π 13.5 °√—¡ (º≈º≈‘μ√âÕ¬≈–

agent ·≈– Dragendorffûs reagent 0.9) ¡“ °—¥¥â«¬μ—«∑”≈–≈“¬ (partition) ™π‘¥μà“ß Ê
2.4 °“√«‘‡§√“–ÀåÕߧåª√–°Õ∫∑“߇§¡’¢Õß “√ ‰¥â·°à hexane, ethyl acetate ·≈–πÈ” ·≈–π” “√

°—¥¥â«¬ GC-MS °—¥ à«ππÈ”¡“·¬° “√¥â«¬°√–∫«π°“√∑“ß‚§√¡“-
1) ‡μ√’¬¡μ—«Õ¬à“ß«‘‡§√“–Àå‚¥¬∫¥·≈– ‚∑°√“øï (Column Chromatography) ‚¥¬„™â
°—¥‚¥¬«‘∏’°“√À¡—°„π acetone ‡ªìπ‡«≈“ 48 ™—Ë«‚¡ß Sephadex LH-20 ª√‘¡“≥ 10 °√—¡ ‡ªìπ«—Ø¿“§ §ß∑’Ë

‡¡◊ËÕ§√∫‡«≈“ °√Õß ·≈–π” “√≈–≈“¬ acetone ¡“ (stationary phase) ·≈–™–¥â«¬«—Ø¿“§‡§≈◊ËÕπ∑’Ë
∑”„Àâ·Àâߥ⫬‡§√◊ËÕß√–‡À¬·Àâß·∫∫À¡ÿπ (rotary (mobile phase) ‰¥â·°à “√≈–≈“¬∑’ˇªìπ à«πº ¡
evaporator) ®“°π—Èπ≈–≈“¬ “√ °—¥À¬“∫ª√‘¡“≥ 10 √–À«à“ß methanol ·≈–πÈ”®“° 10% methanol ∂÷ß

¡‘≈≈‘°√—¡„π methanol 1 ¡‘≈≈‘≈‘μ√ °√Õߥ⫬ 100% methanol „πª√‘¡“μ√·μà≈– —¥ à«π 200 ml
Acrodisc 0.45 ‰¡§√Õπ §Õ≈—¡πå√ÿàπ, Thermo TG- ¥”‡π‘π°“√‡°Á∫ “√≈–≈“¬®“°§Õ≈—¡πå§√—Èß≈– 20 ml

5s-LMS (¬“« 30 ‡¡μ√ ‡ âπºà“π»Ÿπ¬å°≈“ß ¿“¬„π 𔉪√–‡À¬·Àâß ·≈–√«¡ fraction ‚¥¬μ√«® Õ∫
0.25 ¡‘≈≈‘‡¡μ√, §«“¡Àπ“¢Õßøî≈å¡ 0.25 ‰¡§√Õπ), “√∑’ˉ¥â®“° TLC „™â«—Ø¿“§‡§≈◊ËÕπ∑’Ë ethyl
„™â·°ä Œ’‡≈’¬¡‡ªìπ·°ä æ“Õ—μ√“°“√‰À≈ 1 ¡‘≈≈‘≈‘μ√ acetate:methanol:formic acid:water (50:3:3:6)

μàÕπ“∑’, splitless mode, Õÿ≥À¿Ÿ¡‘¢Õß à«π©’¥ ‡¡◊ËÕ√«¡ fraction ∑’Ë¡’Õߧåª√–°Õ∫‡§¡’§≈⓬°—π
“√ 200 Õß»“‡´≈‡´’¬ , ª√‘¡“μ√∑’Ë©’¥ 4 ‰¡‚§√≈‘μ√, ‡¡◊ËÕ„™â à«πº ¡¢Õß methanol ·≈–πÈ”‡ªìπ«—Ø¿“§
Õÿ≥À¿Ÿ¡‘ oven ‡√‘Ë¡μâπ∑’Ë 60 Õß»“‡´≈‡´’¬ §ß∑’Ë ‡§≈◊ËÕπ∑’Ë„π°“√™– “√ ∑’Ë fraction 15-20 æ∫ “√ 4-

‡ªìπ‡«≈“ 5 π“∑’ ®“°π—Èπ‡æ‘Ë¡Õÿ≥À¿Ÿ¡‘®π∂÷ß coumaric acid (8 ¡‘≈≈‘°√—¡ §‘¥‡ªìπ√âÕ¬≈– 0.059
300 Õß»“‡´≈‡´’¬ ¥â«¬Õ—μ√“ 8 Õß»“‡´≈‡´’¬ μàÕ μàÕπ”Àπ—° “√ °—¥) ·≈– fraction ∑’Ë 23-25 æ∫ “√
π“∑’ 5,7,4'-trimethoxyflavone (20 ¡‘≈≈‘°√—¡ §‘¥‡ªìπ

2) ‡μ√’¬¡μ—«Õ¬à“ߧ«—π∑’ˉ¥â®“°°“√‡º“ √âÕ¬≈– 0.148 μàÕπÈ”Àπ—° “√ °—¥)
À≠⓬ÿà¡ ‚¥¬„™âºßÀ≠⓬ÿà¡·Àâß 6 °√—¡ ¡“‡º“„π 2.6 °“√À“ª√‘¡“≥øïπÕ≈‘°√«¡ (total phe-

¿“™π–∑’Ë¡’Ω“ªî¥´÷Ëß∫√√®ÿπÈ”°≈—Ëπ 1 ≈‘μ√ μ—Èß∑‘È߉«â 30 nolic content) ·≈– ‡ø≈‚«πÕ¬¥å√«¡ (total fla-
π“∑’ ®“°π—Èπ‡°Á∫§«—π à«π∑’Ë≈–≈“¬„ππÈ” (grass vonoid content)
smoke) ‚¥¬π”πÈ”¡“∑”„Àâ·Àâߥ⫬‡§√◊ËÕß rotary °“√«‘‡§√“–Àåª√‘¡“≥øïπÕ≈‘°√«¡ ‚¥¬«‘∏’∑“ß

evaporator ·≈–™–§√“∫‡À≈◊Õß∑’ˇ°‘¥¢÷Èπ®“°°“√‡º“ ‡§¡’´÷Ëߪ√—∫«‘∏’«‘‡§√“–Àåμ“¡√“¬ß“π¢Õß Kaisoon et
[20]
(smoke residue) ®“°Ω“ªî¥¥â«¬ methanol al. „™âμ—«Õ¬à“ß “√ °—¥ 4 μ—«Õ¬à“ß®“°μ—«∑”
·≈–π”¡“√–‡À¬·Àâß ‡æ◊ËÕ𔉪«‘‡§√“–Àå‚¥¬‡∑§π‘§ ≈–≈“¬ acetone, ethanol, πÈ” ·≈–πÈ”μâ¡ ‚¥¬™—Ëß
GC-MS μ“¡«‘∏’«‘‡§√“–Àå¢â“ßμâπ ºßÀ≠⓬ÿà¡·Àâß 5 °√—¡ ·≈–‡μ‘¡μ—«∑”≈–≈“¬·μà≈–