Page 26 - วารสารปีที่16ฉบับที่2
P. 26
198
many), chloroform AR (Merck, Germany), DI
Water, anisaldehyde (Fluka, Switzerland), sul-
furic acid (Labscan, Thailand)
4. ‡§√◊ËÕß¡◊Õ
TM
Acquity Ultra Performance Liquid √Ÿª∑’Ë 1 ‚§√ß √â“ß∑“߇§¡’¢Õß ‚°‚æ‡≈∑‘π
Chromatograph-PDA detector (Waters, USA)
·≈– Empower 2 software, ‡§√◊ËÕß™—Ë߉øøÑ“ §«“¡ 2. «‘∏’«‘‡§√“–Àåª√‘¡“≥ ‚°‚æ‡≈∑‘π„πº≈¬Õ
≈–‡Õ’¬¥ 0.01 ¡‘≈≈‘°√—¡ (Mettler-Toledo, Swit- 2.1 °“√æ—≤π“«‘∏’«‘‡§√“–Àåª√‘¡“≥ “√
[10]
zerland) ‚°‚æ‡≈∑‘π„πº≈¬Õ
5. «— ¥ÿ«‘∑¬“»“ μ√å·≈–‡§√◊ËÕß·°â« °. °“√‡≈◊Õ°™π‘¥¢Õßμ—«∑”≈–≈“¬„π°“√
TM
Acquity UPLC BEH C18, 2.1 × 100 °—¥
mm, 1.7 μm (Waters, USA), PTFE syringe fil- ∑”°“√»÷°…“º≈¢Õß°“√ °—¥μ—«Õ¬à“ߺ≈¬Õ
ter ”À√—∫°√Õß “√≈–≈“¬μ—«Õ¬à“ß, TLC plate ™π‘¥ 500 ¡‘≈≈‘°√—¡ ¥â«¬μ—«∑”≈–≈“¬Õ‘π∑√’¬å™π‘¥μà“ß Ê
Silica gel GF254 precoated (E. Merck, Ger- ‰¥â·°à ‡¡∑“πÕ≈ ‡Õ∑‘≈Õ–´’‡μ∑ ·≈–πÈ” ª√‘¡“μ√ 30
many), °√–¥“…°√Õß ‡∫Õ√å 4 (Whatman, UK), ¡‘≈≈‘≈‘μ√ ∑’ˇ«≈“ 60 π“∑’ °√Õß ®“°π—Èπ “√ °—¥
PVDF membrane filter 0.2 μm ”À√—∫°√Õßμ—« ‡¡∑“πÕ≈π”¡“ª√—∫ª√‘¡“μ√¥â«¬‡¡∑“πÕ≈®π§√∫ 50
∑”≈–≈“¬, Nylon membrane filter 0.45 μm (Pall ¡‘≈≈‘≈‘μ√ “√ °—¥‡Õ∑‘≈Õ–´’‡μ∑π”¡“√–‡À¬·Àâß
Corporation, USA) ”À√—∫°√Õß DI water, Tu- ·≈â«≈–≈“¬°≈—∫·≈–ª√—∫ª√‘¡“μ√¥â«¬‡¡∑“πÕ≈®π
berculin syringe ”À√—∫¥Ÿ¥ “√≈–≈“¬μ—«Õ¬à“ß §√∫ 50 ¡‘≈≈‘≈‘μ√ à«π “√ °—¥πÈ”π”¡“ª√‘¡“μ√
„π¢—ÈπμÕπ°“√‡μ√’¬¡μ—«Õ¬à“ß, volumetric flask, ¥â«¬πÈ”®π§√∫ 50 ¡‘≈≈‘≈‘μ√ ·≈⫇ª√’¬∫‡∑’¬∫º≈
pipette ·≈–‡§√◊ËÕß·°â« ¢Õßμ—«∑”≈–≈“¬μàÕ§«“¡ “¡“√∂„π°“√ °—¥ “√
‚°‚æ‡≈∑‘π®“°º≈¬Õ ‚¥¬°“√©’¥ “√≈–≈“¬
«‘∏’°“√»÷°…“
μ—«Õ¬à“߇¢â“ Ÿà√–∫∫ UPLC ·≈⫇ª√’¬∫ peak area
1. «‘∏’«‘‡§√“–À凙‘ߧÿ≥¿“æ¢Õߺ≈¬Õ¥â«¬«‘∏’ ¢Õß “√ ‚°‚æ‡≈∑‘π®“° “√ °—¥μà“ß Ê ª√–°Õ∫°—∫
√ߧ‡≈¢º‘«∫“ß (TLC) peak shape ‘Ë߇®◊ÕªπÕ◊Ëπ Ê (impurity) ·≈–
«‘∏’«‘‡§√“–À凙‘ߧÿ≥¿“æ¥â«¬«‘∏’ TLC ‰¥â∂Ÿ° §«“¡§ßμ—«¢Õß “√≈–≈“¬μ—«Õ¬à“ß
æ—≤π“¢÷Èπ‡æ◊ËÕ„™â ”À√—∫°“√μ√«®‡Õ°≈—°…≥å∑“߇§¡’ ¢. °“√»÷°…“√–¬–‡«≈“∑’ˇÀ¡“– ¡„π°“√
¢Õߺ≈¬Õ ‚¥¬„™â “√ ‚°‚æ‡≈∑‘π ´÷Ë߇ªìπ “√ ”§—≠ °—¥
∑’ËÕÕ°ƒ∑∏‘Ï„πº≈¬Õ‡ªìπ chemical marker ∑”°“√»÷°…“º≈¢Õß°“√ °—¥μ—«Õ¬à“ߺ≈¬Õ
‚§√ß √â“ß∑“߇§¡’¢Õß “√ ‚°‚æ‡≈∑‘π ¥—ß· ¥ß„π 1 °√—¡ ¥â«¬‡¡∑“πÕ≈ª√‘¡“μ√ 30 ¡‘≈≈‘≈‘μ√ ‚¥¬«‘∏’
√Ÿª∑’Ë 1 μâ¡ °—¥‡ªìπ‡«≈“ 30, 60 ·≈– 120 π“∑’ °√Õß