高效能液相層析法對液體製劑中水溶性維生素及咖啡因之同時分析

Simultaneous determination of some water-soluble vitamins and caffeine in liquid pharmaceuticals by high performance liquid chromatography

謝淑紋(Shu-Wen Hsieh)

指導教授 ： 柯黃盛

高雄醫學大學/藥學院/藥學研究所碩士在職專班/碩士(2008年)

摘要

本研究目的為建立一經分析確效的高效能液相層析法(HPLC)，以離子對-逆相層析配合紫外光偵測器分析水溶性維生素vitamin B1 (thiamine HCl), vitamin B2 (riboflavin), vitamin B3 (nicotinamide), vitamin B6 (pyridoxine HCl) 與咖啡因(caffeine)，應用於市售液體製劑之品質管制。本方法為以HPLC串接兩支RP-C18逆相層析管柱 (4.6 mm×100 mm, particle size 6μm) 為固定相，以 6%甲醇-1.5%四氫呋喃-2%醋酸(v/v,含2.23 mM sodium 1-hexanesulfonate; pH =2.6 )為移動相；流速為1.2 mL/min，配合紫外光偵測器(波長280 nm)，以雙層析管柱恆溫控制箱(28℃)中管柱切換閥(column switching)控制移動相於兩支層析管柱間的流路，可將實驗的分析週期時間(cycle time) 由65分鐘縮短為在約40分鐘內分析完成。經測定線性範圍分別為10-30 (B1)、2-6 (B2)、15-45 (B3)、1-3 (B6)及50-150 (caffeine) μg/mL，且線性相關係數R2值皆在0.999以上，各成份回收率介於97.8% - 100.8%，中間精密度(同日間與異日間分析)皆小於2 %。此方法以低組成的有機相(7.5%，含甲醇6%及四氫呋喃1.5%)為移動相，並藉助於管柱切換閥節省分析週期時間(cycle time)，並成功應用所建立的分析法於市售的能量飲料或營養補充製劑中水溶性維生素及咖啡因之含量分析。

關鍵字

高效液相層析法 ； 管柱切換閥 ； 水溶性維生素 ； 咖啡因

並列摘要

A reversed-phase ion-pair high performance liquid chromatographic method (HPLC) has been developed and validated for the routine analysis of vitamin B1 (thiamine hydrochloride), vitamin B2 (riboflavin), vitamin B3 (nicotinamide), vitamin B6 (pyridoxine hydrochloride) and caffeine in liquid pharmaceuticals. HPLC separation of the vitamins and caffeine was performed on a tandem combination of two same stationary phases: Inertsil C18 column from Vercopack ® (4.6 mm×100 mm, particle size 6μm) and detected by ultra-violet detector at 280 nm. The use of 6% methanol - 1.5 % tetrahydrofuran - 2% acetic acid solution (v/v; containing 2.23 mM sodium 1-hexanesulfonate, pH = 2.6) as the mobile phase at a flow-rate of 1.2 mL/min. enables separation of the five analytes free from interferences with isocratic elution at 28°C. The cycle time was saved from 65 to 40 min in a single run by using a column switching to control the mobile phase flow regime. The method was good linearity (R2＞0.999) in the ranges of 10-30 (B1), 2-6 (B2), 15-45 (B3), 1-3 (B6) and 50-150 (caffeine) μg/mL. The coefficients of variation of intermediate precision (intraday and interday assays) were less than 2%. The recoveries were between 97.8% and 100.8%. The method was successfully applied a column switching to save cycle time; used of mobile phases consisting of ion pair buffer and low concentration of organic solvent (7.5%; methano1 6.0% and tetrahydrofuran 1.5%) that are fully compatible with HPLC technique.

並列關鍵字

HPLC ； column switching ； water soluble vitamins ； caffeine

參考文獻

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