【口服液中麻黄碱含量】 麻黄碱口服液

Two-dimensional high performance liquid chromatography

analysis of oral liquid content of ephedrine alkaloids

Abstract Objective : To investigate the oral solution in children Qingrezhike the content of ephedrine alkaloids analysis. Methods : based on Chinese herbal medicinal ingredients of the complexity and diversity, through the development of off-line two-dimensional high performance liquid chromatography system, to solve the complex components of Chinese medicine sample separation and quantification of trace components of the problem. One-dimensional column of Agilent"s Asahipak ODP50 (4.0 mm * 125 mm), 5 μ m particle size, two -dimensional column for the Octagon"s Aichrom BondAQ C18 (4.6 mm * 250 mm), 5 μ m particle size; buffer solution as the 20Mmol KH2PO4, adjusted with H3PO4 and triethylamine pH = 2.7; one-dimensional and two-dimensional mobile phase were buffer solution / acetonitrile (97 / 3, V / V), flow rate of 0.8 ml / min; column temperature is 30 ℃ ; DAD detector detection wavelength was 210 nm. Result: ephedrine, pseudoephedrine, ephedrine and methyl respectively in the injection volume 0.36 ~ 3.6 μ g, 0.34 ~ 3.5 μ g, 0.32 ~ 3.3 μ g sample volume within the framework of a good linear relationship with the peak area, the average recoveries were 97.4% , 98.6% and 100.2%. Conclusion significant improvement in the separation of samples of traditional Chinese medicine, and this system can be used for direct injection analysis of Chinese medicine.

Keywords ：Multi-dimensional high-performance liquid chromatography; off-line; ephedrine; pseudo-ephedrine; methyl ephedrine

Multidimensional HPLC (Multidimensional HPLC, MD - HPLC) using two or more components to be analysed in a chromatography column of complex samples, between the column using the switch valve will after all or part of th e separation of components for the first time selectively moved to another root or more different types of further in the chromatographic column separation

[1 ~ 6].Compared with conventional single column separation, the technology h as higher selectivity, the bigger the peak capacity and better resolution. The tec hnique, the operation is simple, make up for the deficiency of the conventional single column chromatography, is a kind of potential separation analysis meth od. Ephedrine a drug, ephedrine and pseudoephedrine and methyl ephedrine wit h similar structure, including ephedrine and pseudoephedrine belongs to the iso mers is the main active ingredients of TCM ephedra. An Ephedra drug in clini c is widely used in the treatment of colds, asthma and allergic disease. Many of the proprietary Chinese medicine, Chinese and western compound preparatio ns containing ephedra contain ephedrine drugs (7 ~ 10]. At present it has a lo t of reports of ephedrine class alkaloid, but due to the complexity of traditiona l Chinese medicine samples contained in the material composition, drug content is very low, direct sampling analysis difficult. Traditional Chinese medicine (TCM) is realized by using two-dimensional chromatography can sample directly into the analysis of the process is simple, the result is accurate.

1. Equipment

Agilent 1100 series high performance liquid chromatograph by to take off the angry machine, dual pumps, automatic sampler, column temperature and dio de array detector and the chemical workstation. The column temperature box w ith six switch valve, column switching can be performed. Reference substance: ephedrine and pseudoephedrine and methyl ephedrine offices shall purchase fro m Chinese pharmaceutical and biological products; Samples of Zhengzhou city Concorde pharmaceutical factory production of infantile heat cough oral liquid;

Experiments for Wahaha purified water, water with 0.45 mu m micro porous fi lter mempane filtration; KH2PO4, H3PO4, triethylamine as the analysis of pur e; Pure methanol and acetonitrile as chromatography. 2. Methods and results 2.1 Reference and sample preparation

Weigh accurately amount of ephedrine, pseudoephedrine and methylephedrine control products, buy 50 ml volumetric flask and add pure water to scale, shake, preparation of standard solution concentration 450,436,412 mg/l, respectively. Take 10 ml of qingre zhike oral solution in children, buy 50 ml volumetric flask and add pure water to scale, shake, and filtered through 0.45 μm pore filter mempane. All the samples stored in the 4 c refrigerator spare. 2.2 chromatographic conditions

Color spectrum column for a dimension column for Agilent company of Asahipak ODP50 (4.0 mmx125 mm), 5 μ m grain diameter, II dimension column for Octagon company of Aichrom BondAQ C18 (4.6 mmx250 mm), 5 μ m grain diameter; buffer for 20Mmol of KH2PO4, with H3PO4 and three ethylamine adjustable pH value for 2.7; a dimension and the II dimension of flow phase are for buffer/b nitrile (97/3,V/V), Flow rate of 0.8 mL/min column temperature 30 ℃; DAD detector, the detection wavelength of 210 nm. First put the samples in the first dimension column for separation, then column using six-way valve switch, after the separation of the 1th interested constituents to switch to the second column, 1th dimension column components are not interested in being cleared away, and then in the second column will be further separated the components of interest. 2.3 Sample determination and chromatogram Chromatographic conditions as described above for oral liquid 2DHPLC analysis sample and standard sample, get two dimensional chromatogram with the chromatogram of the sample mixture, peak and 1~3 correspond to ephedrine, pseudoephedrine and methylephedrine. While online with diode array detector scans

all peaks of UV-vis spectroscopy, found various peaks of UV absorption peaks meet the ultraviolet absorption characteristics of ephedrine alkaloids. 2.4 The working curve of ephedrine alkaloids 0.5,1,3,4,5 ml were obtained from the above standard solutions prepared into a series of mixed standard solution concentration, using peak areas (Y) on the mass concentration (X,mg/L) 3 species of ephedrine alkaloids derived from regression analysis regression equation, correlation coefficient, the linear range and lowest detection limit (S/N=3). Therefore, good linear correlation of the ephedrine alkaloids. 2.5 Determination of ephedrine alkaloids in oral liquid

By oral liquid of chromatographic conditions on preparation of samples for HPLC Analysis, measurements of ephedrine, pseudoephedrine and methylephedrine content are as follows: 470.01, 145.33, 41.46 mg/l. 2.6 Precision experiments Under the chromatographic conditions, use the same alkaloids ephedrine mixed standard solution injection total sample 5, sample 40 μ l/, determination of their peak area and the relative standard deviation of the peak areas (RSD). 2.7 recovery experiments

Recovery by adding sample experiments. In the known content of samples, precise respectively to join 3 different concentration of the standard solution of ephedrine, also according to the sample analysis methods for determination, the measured sample recovery. Thus, samples high. 3. Brief summary

This experiment adopts two DHPLC methods to determine the pediatric class ephedra alkaloid content in the heat cough oral liquid. Due to the complexity of traditional Chinese medicine samples contained in the material composition, drug content is very low, direct sampling analysis difficult. Using our own developed offline multidimensional HPLC technology, can realize direct injection analysis of TCM samples, simple process, results are accurate.

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