藿香正氣水中乙醇含量測定方法研究

2009年6月第1卷第6期中同現代中藥 Modem Chinese MedicineJun, 2009 Vol 11 No藿香正氣水中乙醇含量測定方法研究申放,黃婉峰,高衛東(佛山市藥品檢驗所,廣東佛山528000[摘要]目的:對霖香正氣水中乙醇的含量測定方法進(jìn)行改進(jìn)。方法:色譜柱為 phenomenex毛細管柱nZB- waxplus(30mx0.25mm,0.25μm);氮氣為載氣,流速為0.5 mL min,分流比為30:1;進(jìn)樣口溫度為90℃;柱溫為85℃;FID檢測器,溫度為200℃;進(jìn)樣量0.1μL;正丙醇為內標物。結果:在該色譜條件下,歡香正氣水中乙醇和內標物得到良好的分離,乙醇在0.010~0.1mL·mL線(xiàn)性良好(r=0.995,n=7);平均回收率為100.0%,RSD=1.18%(n=6)。結論:該法靈敏、準確,重復性好,可用于測定該制劑的乙醇含量關(guān)鍵香正氣水;乙醇;氣相色譜香正氣水具有解表化濕,理氣和中的功能,臨床常用于外感風(fēng)寒、內傷濕滯或夏傷暑濕所致的感冒等。對于制劑中乙醇含量的檢查,《中國藥典》中規定是以二乙烯苯乙基乙烯苯型高分子多孔小球作為載體的填充柱進(jìn)行測定,但是因為填充柱有不適宜做程序升溫以及相對理論塔板數比較低等不足,在藥品檢測工作中使用有減少的趨勢,而多用毛細管柱。氣相色譜毛細管柱因其高分離能力1.乙醉峰:2.丙酚峰高靈敏度、高分析速度等獨特優(yōu)點(diǎn)而得到迅速發(fā)展。A內標物正內醇:B.對照品:C香正氣水供試品查閱文獻未見(jiàn)有關(guān)于以聚乙二醇極性柱為色譜柱,圖1氣相色譜圖采用氫火焰離子化檢測器氣相色譜法測定制劑中乙醇含量的報道,故本實(shí)驗建立了相應方法,為乙醇2校正因子測定含量的測定提供參考。精密量取恒溫至20℃的無(wú)水乙醇4,5,6mL,1儀器與試藥分別置100m量瓶中,分別精密加入恒溫至20℃的正內醇(內標物)5mL,加水稀釋至刻度,取上述3Varian CP-3800氣相色譜儀,FD檢測器;phe種溶液適量,按上述色譜條件分別連續進(jìn)樣3次nonene聚乙二醇毛細管柱 zebron ZB-waxplus(30mx計算乙醇的校正因子為1.3904,RSD=1%6%。0.25mm,0.25μm)。無(wú)水乙醇,正丙醇均為分析2.3供試品溶液的制備純,水為超純水。藿香正氣水,四川蜀中制約有限23供試品溶液的制備精密吸取恒溫至20℃的本品溶液10mL,置公司(批號080302,080807,080923)。100mL量瓶中,精密加入恒溫至20℃的正丙醇5mL2方法和結果加水稀釋至刻度,搖勻,即得。2.1色譜條件與系統適應性試驗2.4線(xiàn)性關(guān)系氫氣流速為30 mL. min-',尾吹流速28mL·min精密量取恒溫至20℃的無(wú)水乙醇1,2,氮氣流速05mL·min1,空氣流速為300mLm8,10mL,分別置100mL量瓶中,分別精密加入柱溫為85℃;進(jìn)樣口溫度為90℃,檢測器溫度為恒溫至20℃的正丙醇(內標物)5mL,加水稀釋至刻200℃,分流比為30:1。在此色譜條件下,乙醇和正度,搖勻,按上述色譜條件進(jìn)樣,每次進(jìn)樣0.lμ,丙醇有較好的分離,色譜圖見(jiàn)圖1。以對照品含量為橫坐標,以對照品峰面積與內標峰[通訊作者]‘高衛東,E-mail:weidong@163.com009年6月第卷第6期中國現代中藥 Moclern Chinese medicineJun 2009 Vol 11 No 6面積之比為縱坐標,繪制標準曲線(xiàn)并得到回歸方程,表1加樣回收率試驗Y=14.352X-0.0472,r=0.9995。樣品含測得收率平均收RSD2.5精密度試驗99.5700.0L.18精密量取恒溫至20℃的無(wú)水乙醇5mL置100mL9.66100.87量瓶中,加入恒溫至20咒℃的正丙醇5mL,用水稀釋4.629624.629.5598.48刻度,搖勻。每次進(jìn)樣0.IμL,連續進(jìn)樣6次,計算校正因子,結果RSD=1.06%4.629.70101.73水乙醇加入量均為500mL2.6穩定性試驗取同一批號的供試品溶液,照2.3制備供試品表2樣品中乙醇含量測定溶液,精密吸取0.1μL,于0,1,2,3,5,6,12h乙醇平均含址08030245.5進(jìn)樣,測定,計算乙醇的含量,RSD=1.24%。結080807果表明,供試品溶液在12h內基本穩定。27重復性試驗精密量取同一批號的恒溫至20℃的樣品6份,每討論份10mL,加入恒溫至20℃的正丙醇5mL,用水稀釋至刻度,搖勻。按上述色譜條件測定,計算每份乙醇乙醇為極性物質(zhì),故選用極性柱。由于對照品溶液和樣品溶液的含水量較大,因此用較小的進(jìn)樣的含量,平均為4.2%,RSD=015%,重復性較好。量,以免將FD滅火和對色譜柱產(chǎn)生損壞。因未見(jiàn)2.8回收率試驗有文獻報道采用FID檢測器,以聚乙二醇極性毛細精密量取恒溫至20℃的已知含量的相同樣品6管色譜柱測定藿香正氣水中乙醇的含量,故本文報份,每份10mL,置100mL量瓶中,精密加入恒溫至道之。方法學(xué)考察證實(shí)此方法可行。內標法加校正20℃的無(wú)水乙醇5mL,再加入恒溫至20的正內醇因子測定樣品中乙醇的含量,操作較簡(jiǎn)便,分離效5mL,用水稀釋至10mL,搖勻。按上述色譜條件測果好,分析時(shí)間短,結果準確可靠。cM定,記錄色譜圖,計算回收率,結果見(jiàn)表I。2.9樣品測定參考文獻精密吸取恒溫至20℃的樣品l0mL,加入恒溫至[]國家藥典委員會(huì ),中國藥典(一部)[S].北京:化學(xué)工業(yè)20℃的正丙醇5mL,用水稀釋至刻度,搖勻。按上出版社,2005:662述色譜條件測定并計算,結果見(jiàn)表2。(收稿日期2008-1203Improved Method for Determining Ethanol in Huo Xiang ZhengQiShuiFang, Huang Wanfeng, GaFoshan Institute for Drug control, Foshan Guangdong 528000, China)[Abstract]Objective: To improve the method for determining ethanol in Huo Xiang Zheng QiShui by GC.Methods: Chromatographic column was Phenomenex Zebron ZB-Waxplus(30m x0. 25mm, 0. 25 um), the carrier gaswas nitrogen, Alow rate was 0 5mL.min", split ratio was 30: 1. The column temperature was at 85C, inlet tempera-ture was 90C, FID detection was 200C, injection volume was 0. IuL The n-propanol was used as intemal substance. Results: The linearity of ethanol were good in the range of 0. 01-0. 1 mL.- with r=0. 999 5. The aver-age recovery of ethanol was 100. 0%, RSD was 1. 18%. Conclusion: This method is sensitive, rapid, accurate andreproducible, and can be used for the test of ethanol in Huo Xiang ZhengQiShui.I Key words] Huo Xiang ZhengQiShui; Ethanol; GC(E4%24 A)in characteristic fingerprints, and Similarity Evaluation system was applied to evaluate fingerprint of theten batches of Herba Andrographis, the total content of andrographolide and dehydroandrographolide was between1. 09% and 4. 47%. Conclusion: The method is sensitive, repeatable and accurate, which can be used as qualitycontrol for Herba Andrographis and has been used in Hong Kong Chinese Materia Medica Standards. We suggest thatthe HPLC fingerprinting should be involved in the quality control of Herba Andrographis, and the total content of andrographolide and dehydroandrographolide in Herba Andrographis is not less than 1. 0%Key words] Quality assessment; Herba Andrographis; HPLC fingerprint; Content determination