張婷婷等
1 引 言
中藥注射劑是在中醫藥制劑基礎上發展起來的一種劑型,具有注射劑所共同的優點,并在一定程度保留了中醫藥的特色,在醫療實踐中發揮了巨大作用。中藥注射劑中可能存在的大分子化合物(如蛋白質、淀粉、鞣質、樹脂等),這被認為是中藥注射劑不良反應的主要因素之一[ 1]。檢測和控制大分子物質已成為中藥注射劑質量標準的重要指標,因此非常有必要建立靈敏、準確、簡便的檢測大分子物質的檢測方法。
采用體積排阻除色譜法和示差折光檢測器對中藥注射劑大分子化合物進行分析,大大提高了測定結果的穩定性和靈敏度,但是與其它方法相比,示差檢測器的靈敏度仍然不高。電噴霧檢測器(Charged aerosol detector, CAD)作為一款通用型檢測器。具有靈敏度高、響應一致性、重現性出色和檢測范圍寬的優勢,因此具有廣泛的實際應用范圍[ 2,3]; 與紫外檢測器相比,CAD能夠很好地對無生色基團或弱紫外吸收的化合物進行測定,并且在同時檢測多種最大吸收波長差異較大的物質時,CAD具有響應一致性優勢[ 4]; 與示差折光檢測器相比,CAD兼容梯度洗脫,大大提高了分離效率,并且靈敏度也顯著提高[ 5]。本研究利用CAD的優勢,采用HPLCCAD方法測定中藥注射劑中的大分子化合物。
2 實驗部分
2.1 儀器與試劑 Dionex Ultimate 3000系統(美國賽默飛世爾科技公司); DGP3600SD雙三元液相色譜泵、WPS3000TSL自動進樣器、TCC3000SD柱溫箱、Corona Veo RS檢測器、Chromeleon Chromatography Data System7.2色譜軟件、MAbPac SEC1色譜柱(300 mm×4.0 mm,300 ,5 μm); Sartorius BS21S型電子天平(德國賽多利斯公司)。
乙腈(美國賽默飛世爾科技公司); 甲酸銨(江蘇永華精細化學品有限公司提供)。右旋糖酐對照品(右旋糖酐D1、右旋糖酐D2、右旋糖酐D3、右旋糖酐D6、右旋糖酐D7、右旋糖酐D8)、吐溫對照品(國藥集團化學試劑有限公司)。
苦黃注射液、血塞通注射液、參麥注射液、香丹注射液均由各藥物生產商提供。
References
1 CHEN YaFang. Antiinfection Pharmacy, 2008, 5(4) : 254-255
陳亞芳. 抗感染藥學, 2008, 5(4) : 254-255
2 Kou D, Maniusa G, Zhana S D, Chokshi H P. J. Chromatogr. A, 2009, 1216(28): 5424-5428
3 Sun P, Wang X D, Alquier L, Maryanoff C A. J. Chromatogr. A, 2008, 1177(1): 87-91
4 Huang Z, Richard M A, Zha Y, Francis R, Lozano R, Ruan J. J. Pharm. Biomed. Anal., 2009, 50(5): 809-814
5 Almeling S, Ilko D,Holzgrabe U. J. Pharm. Biomed. Anal., 2012, 69: 50-63
DualGradient Liquid ChromatographyTandem Charged Aerosol
Detector for Detection of Macromolecules in Traditional
Chinese Medicine Injections
ZHANG TingTing*1, WANG Yu2,3,, GU DanHua2, ZHANG Li3, JIN Yan1
1(Shanghai Application Lab, Thermofisher Scientific, Shanghai 200100, China)
2(China Pharmaceutical University, Nanjin 210009, China)
3(Jiangsu Institute for Food and Drug Control, Nanjin 210009, China)
Abstract A size exclusion chromatography method (SEC) was established for the screening of macromolecules in traditional Chinese medicine (TCM) injections. In the experiment, TCM was collected through 0.45 μm hybrid filter and separated by SEC with MAbPac SEC1 column. Acetonitrile was added by postcolumn mode with a flow rate of 0.2 mL/min to enhance the detection sensitivity, and 25 mmol/L ammonium formate was used as the mobile phase at flow rate of 0.2 mL/min. The charged aerosol detection was carried out with Dextran and Tween as the reference standards. The molecular weight distribution of Dextran ranged from 2500 to 133800 had a good linear relationship with the retention time. No macromolecules were detected out in the selected traditional Chinese medicine injections except Kuhuang injections.
Keywords Charged aerosol detection; Liquid chromatography; Size exclusion chromatography; Macromolecule screening
(Received 19 September 2014; accepted 11 October 2014)
1 引 言
中藥注射劑是在中醫藥制劑基礎上發展起來的一種劑型,具有注射劑所共同的優點,并在一定程度保留了中醫藥的特色,在醫療實踐中發揮了巨大作用。中藥注射劑中可能存在的大分子化合物(如蛋白質、淀粉、鞣質、樹脂等),這被認為是中藥注射劑不良反應的主要因素之一[ 1]。檢測和控制大分子物質已成為中藥注射劑質量標準的重要指標,因此非常有必要建立靈敏、準確、簡便的檢測大分子物質的檢測方法。
采用體積排阻除色譜法和示差折光檢測器對中藥注射劑大分子化合物進行分析,大大提高了測定結果的穩定性和靈敏度,但是與其它方法相比,示差檢測器的靈敏度仍然不高。電噴霧檢測器(Charged aerosol detector, CAD)作為一款通用型檢測器。具有靈敏度高、響應一致性、重現性出色和檢測范圍寬的優勢,因此具有廣泛的實際應用范圍[ 2,3]; 與紫外檢測器相比,CAD能夠很好地對無生色基團或弱紫外吸收的化合物進行測定,并且在同時檢測多種最大吸收波長差異較大的物質時,CAD具有響應一致性優勢[ 4]; 與示差折光檢測器相比,CAD兼容梯度洗脫,大大提高了分離效率,并且靈敏度也顯著提高[ 5]。本研究利用CAD的優勢,采用HPLCCAD方法測定中藥注射劑中的大分子化合物。
2 實驗部分
2.1 儀器與試劑 Dionex Ultimate 3000系統(美國賽默飛世爾科技公司); DGP3600SD雙三元液相色譜泵、WPS3000TSL自動進樣器、TCC3000SD柱溫箱、Corona Veo RS檢測器、Chromeleon Chromatography Data System7.2色譜軟件、MAbPac SEC1色譜柱(300 mm×4.0 mm,300 ,5 μm); Sartorius BS21S型電子天平(德國賽多利斯公司)。
乙腈(美國賽默飛世爾科技公司); 甲酸銨(江蘇永華精細化學品有限公司提供)。右旋糖酐對照品(右旋糖酐D1、右旋糖酐D2、右旋糖酐D3、右旋糖酐D6、右旋糖酐D7、右旋糖酐D8)、吐溫對照品(國藥集團化學試劑有限公司)。
苦黃注射液、血塞通注射液、參麥注射液、香丹注射液均由各藥物生產商提供。
References
1 CHEN YaFang. Antiinfection Pharmacy, 2008, 5(4) : 254-255
陳亞芳. 抗感染藥學, 2008, 5(4) : 254-255
2 Kou D, Maniusa G, Zhana S D, Chokshi H P. J. Chromatogr. A, 2009, 1216(28): 5424-5428
3 Sun P, Wang X D, Alquier L, Maryanoff C A. J. Chromatogr. A, 2008, 1177(1): 87-91
4 Huang Z, Richard M A, Zha Y, Francis R, Lozano R, Ruan J. J. Pharm. Biomed. Anal., 2009, 50(5): 809-814
5 Almeling S, Ilko D,Holzgrabe U. J. Pharm. Biomed. Anal., 2012, 69: 50-63
DualGradient Liquid ChromatographyTandem Charged Aerosol
Detector for Detection of Macromolecules in Traditional
Chinese Medicine Injections
ZHANG TingTing*1, WANG Yu2,3,, GU DanHua2, ZHANG Li3, JIN Yan1
1(Shanghai Application Lab, Thermofisher Scientific, Shanghai 200100, China)
2(China Pharmaceutical University, Nanjin 210009, China)
3(Jiangsu Institute for Food and Drug Control, Nanjin 210009, China)
Abstract A size exclusion chromatography method (SEC) was established for the screening of macromolecules in traditional Chinese medicine (TCM) injections. In the experiment, TCM was collected through 0.45 μm hybrid filter and separated by SEC with MAbPac SEC1 column. Acetonitrile was added by postcolumn mode with a flow rate of 0.2 mL/min to enhance the detection sensitivity, and 25 mmol/L ammonium formate was used as the mobile phase at flow rate of 0.2 mL/min. The charged aerosol detection was carried out with Dextran and Tween as the reference standards. The molecular weight distribution of Dextran ranged from 2500 to 133800 had a good linear relationship with the retention time. No macromolecules were detected out in the selected traditional Chinese medicine injections except Kuhuang injections.
Keywords Charged aerosol detection; Liquid chromatography; Size exclusion chromatography; Macromolecule screening
(Received 19 September 2014; accepted 11 October 2014)
1 引 言
中藥注射劑是在中醫藥制劑基礎上發展起來的一種劑型,具有注射劑所共同的優點,并在一定程度保留了中醫藥的特色,在醫療實踐中發揮了巨大作用。中藥注射劑中可能存在的大分子化合物(如蛋白質、淀粉、鞣質、樹脂等),這被認為是中藥注射劑不良反應的主要因素之一[ 1]。檢測和控制大分子物質已成為中藥注射劑質量標準的重要指標,因此非常有必要建立靈敏、準確、簡便的檢測大分子物質的檢測方法。
采用體積排阻除色譜法和示差折光檢測器對中藥注射劑大分子化合物進行分析,大大提高了測定結果的穩定性和靈敏度,但是與其它方法相比,示差檢測器的靈敏度仍然不高。電噴霧檢測器(Charged aerosol detector, CAD)作為一款通用型檢測器。具有靈敏度高、響應一致性、重現性出色和檢測范圍寬的優勢,因此具有廣泛的實際應用范圍[ 2,3]; 與紫外檢測器相比,CAD能夠很好地對無生色基團或弱紫外吸收的化合物進行測定,并且在同時檢測多種最大吸收波長差異較大的物質時,CAD具有響應一致性優勢[ 4]; 與示差折光檢測器相比,CAD兼容梯度洗脫,大大提高了分離效率,并且靈敏度也顯著提高[ 5]。本研究利用CAD的優勢,采用HPLCCAD方法測定中藥注射劑中的大分子化合物。
2 實驗部分
2.1 儀器與試劑 Dionex Ultimate 3000系統(美國賽默飛世爾科技公司); DGP3600SD雙三元液相色譜泵、WPS3000TSL自動進樣器、TCC3000SD柱溫箱、Corona Veo RS檢測器、Chromeleon Chromatography Data System7.2色譜軟件、MAbPac SEC1色譜柱(300 mm×4.0 mm,300 ,5 μm); Sartorius BS21S型電子天平(德國賽多利斯公司)。
乙腈(美國賽默飛世爾科技公司); 甲酸銨(江蘇永華精細化學品有限公司提供)。右旋糖酐對照品(右旋糖酐D1、右旋糖酐D2、右旋糖酐D3、右旋糖酐D6、右旋糖酐D7、右旋糖酐D8)、吐溫對照品(國藥集團化學試劑有限公司)。
苦黃注射液、血塞通注射液、參麥注射液、香丹注射液均由各藥物生產商提供。
References
1 CHEN YaFang. Antiinfection Pharmacy, 2008, 5(4) : 254-255
陳亞芳. 抗感染藥學, 2008, 5(4) : 254-255
2 Kou D, Maniusa G, Zhana S D, Chokshi H P. J. Chromatogr. A, 2009, 1216(28): 5424-5428
3 Sun P, Wang X D, Alquier L, Maryanoff C A. J. Chromatogr. A, 2008, 1177(1): 87-91
4 Huang Z, Richard M A, Zha Y, Francis R, Lozano R, Ruan J. J. Pharm. Biomed. Anal., 2009, 50(5): 809-814
5 Almeling S, Ilko D,Holzgrabe U. J. Pharm. Biomed. Anal., 2012, 69: 50-63
DualGradient Liquid ChromatographyTandem Charged Aerosol
Detector for Detection of Macromolecules in Traditional
Chinese Medicine Injections
ZHANG TingTing*1, WANG Yu2,3,, GU DanHua2, ZHANG Li3, JIN Yan1
1(Shanghai Application Lab, Thermofisher Scientific, Shanghai 200100, China)
2(China Pharmaceutical University, Nanjin 210009, China)
3(Jiangsu Institute for Food and Drug Control, Nanjin 210009, China)
Abstract A size exclusion chromatography method (SEC) was established for the screening of macromolecules in traditional Chinese medicine (TCM) injections. In the experiment, TCM was collected through 0.45 μm hybrid filter and separated by SEC with MAbPac SEC1 column. Acetonitrile was added by postcolumn mode with a flow rate of 0.2 mL/min to enhance the detection sensitivity, and 25 mmol/L ammonium formate was used as the mobile phase at flow rate of 0.2 mL/min. The charged aerosol detection was carried out with Dextran and Tween as the reference standards. The molecular weight distribution of Dextran ranged from 2500 to 133800 had a good linear relationship with the retention time. No macromolecules were detected out in the selected traditional Chinese medicine injections except Kuhuang injections.
Keywords Charged aerosol detection; Liquid chromatography; Size exclusion chromatography; Macromolecule screening
(Received 19 September 2014; accepted 11 October 2014)