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°“√‡μ√’¬¡¬“ª√– –‰æ≈™π‘¥≈Ÿ°°≈Õπ ·≈–¬“ ¿“«–¢Õß√–∫∫ HPLC ∑’Ë„™â«‘‡§√“–Àå “√∫àß

ª√– –‰æ≈™π‘¥·§ª´Ÿ≈ ™’È∑—Èß 4 ™π‘¥
¬“ª√– –‰æ≈™π‘¥≈Ÿ°°≈Õπ®–∂Ÿ°‡μ√’¬¡¢÷Èπ 2 „π°“√»÷°…“§√—Èßπ’ȉ¥â¡’°“√„™â HPLC condi-
·∫∫ §◊Õ·∫∫∑’Ë„™âπÈ”º÷Èß∏√√¡¥“‡ªìππÈ”°√– “¬¬“ tions ”À√—∫∑”°“√«‘‡§√“–Àåª√‘¡“≥ “√ D °—∫ “√

·≈–·∫∫∑’Ë„™âπÈ”º÷Èß∑’ˇ§’ˬ«®π‡À𒬫‡ªìππÈ”°√– “¬¬“ 1, 2 ·≈– 3 ¥—ßμ“√“ß∑’Ë 1
ªíôπ‡ªìπ≈Ÿ°°≈Õπ·∫∫≈– 100 ‡¡Á¥ ¢π“¥‡ âπºà“π Pump : Water 600 Controllor
»Ÿπ¬å°≈“ß 1 ‡´π쑇¡μ√ à«π¬“ª√– –‰æ≈™π‘¥ Column : Hypersil gold 5 ‰¡§√Õπ C-

·§ª´Ÿ≈‰¥â∂Ÿ°‡μ√’¬¡¢÷Èπ®“°‡§√◊ËÕß∫√√®ÿ·§ª´Ÿ≈ 18, (250 x 4.6 ¡‘≈≈‘‡¡μ√)
‚¥¬„™â·§ª´Ÿ≈‡∫Õ√å 0 ®”π«π 100 ·§ª´Ÿ≈ Flow rate : 1.0 ¡‘≈≈‘≈‘μ√/π“∑’

°“√À“ª√‘¡“≥πÈ”„ππÈ”º÷Èß∑’Ëπ”¡“∑”¬“≈Ÿ° Detector : Water 2996 PDA ∑’Ë 254 π“-
°≈Õπ ‚π‡¡μ√
°“√À“ª√‘¡“≥πÈ”„ππÈ”º÷Èß®–„™â«‘∏’°“√«—¥§à“¥—™π’ Autosampler : Water 717 plus (10
7
À—°‡À (refractive index) ∑’ËÕÿ≥À¿Ÿ¡‘ÀâÕß ‚¥¬„™â ‰¡‚§√≈‘μ√)
‡§√◊ËÕß refractometer ATAGO PAL-3 ‚¥¬°“√ 1. Linearity ·≈– Range: ®—¥∑”°√“ø

ÕÿàππÈ”º÷Èß„À⇪ìπ¢Õ߇À≈«„πμŸâÕ∫∑’Ë 45 Õß»“‡´≈‡´’¬ ¡“μ√∞“π·≈–°”Àπ¥™à«ß∑’Ë„™â„π°“√«‘‡§√“–Àå
·≈⫪≈àÕ¬„À⇬Áπ≈ß∑’ËÕÿ≥À¿Ÿ¡‘ÀâÕß ∑”°“√À¬¥πÈ” ¡°“√‡ âπμ√ߢÕß°√“ø¡“μ√∞“π·≈–§à“ cor-
2
º÷Èß≈߉ª„Àâ∑—Ë«º‘«¢Õ߇´≈≈å‚¥¬‡¡◊Ëժ´≈≈å·≈â«μâÕß relation coefficient (r ) ‡ªìπ ¡°“√§≥‘μ»“ μ√å∑’Ë
‰¡à¡’øÕßÕ“°“» ª√‘¡“≥πÈ”®–«—¥·≈–§”π«≥Õ¬Ÿà„π ®–„™â„π°“√·ª√º≈°“√«‘‡§√“–À宓°§à“¢âÕ¡Ÿ≈æ◊Èπ∑’Ë
Àπ૬¢Õß√âÕ¬≈–¢Õß¡«≈ (mass percentage) „μâ°√“ø¢Õß “√·μà≈–™π‘¥ „À⇪ìπ§à“§«“¡‡¢â¡¢âπ
°“√«—¥®–∑”´È” 3 §√—Èß π”§à“∑—ÈßÀ¡¥¡“À“§à“‡©≈’ˬ ¢Õß “√π—Èπ Ê ‚¥¬‡μ√’¬¡™à«ß§«“¡‡¢â¡¢âπ∑’Ë„™â„π

·≈–§à“‡∫’ˬ߇∫π¡“μ√∞“π °“√®—¥∑”°√“ø¡“μ√∞“π¢Õß “√ D “√ 1, 2 ·≈– 3

μ“√“ß∑’Ë 1 · ¥ß gradient elution condition

‡«≈“ (π“∑’) Flow rate (¡≈./π“∑’) 1% Acetic acid Acetonitrile Curve
0.01 0.80 85.00 15.00 6
8.00 1.00 70.00 30.00 6
25.00 1.00 55.00 45.00 6
30.00 1.00 50.00 50.00 6
55.00 1.00 30.00 70.00 6
80.00 1.00 15.00 85.00 6
100.00 1.00 0.00 100.00 6
105.00 1.00 85.00 15.00 6