Determination of chlorogenic acid and Baicalin in Yinhuang Oral Liquid by dual wavelength ratio spectrometry

Abstract: Objective: To study the determination of chlorogenic acid and Baicalin in Yinhuang Oral Liquid by dual wavelength ratio spectroscopy Methods: 266 and 342mn were selected as the wavelength according to the ratio spectrum characteristics of chlorogenic acid and baicalin Results: the linear range of chlorogenic acid was 2-20 μ g / ml, the recovery rate was 98.9, RSD was 1.58%; the linear range of baicalin was 2-16 μ g / ml, the recovery rate was 100.4%, RSD was 1.32% Conclusion: this method is sensitive, accurate and simple It can be widely used in other traditional Chinese medicine preparations Key words: dual wavelength ratio spectrometry; Yinhuang Oral Liquid; chlorogenic acid; baicalin Yinhuang Oral Liquid has the functions of clearing away heat and detoxification, bacteriostasis and antiphlogistic, mainly treating upper respiratory tract infection, acute tonsillitis, pharyngitis and other diseases It is processed and refined from honeysuckle and scutellaria extract Chlorogenic acid and baicalin were determined by UV Spectrophotometry in Chinese Pharmacopoeia It has been reported that liquid chromatography is used to determine its content In this paper, the content of chlorogenic acid and Baicalin in Yinhuang Oral Liquid was determined by dual wavelength ratio spectrometry 1 Instrument and reagent: Shimadzu UV-260 ultraviolet supplement visible spectrophotometer; 752-c ultraviolet visible spectrophotometer Chlorogenic acid and baicalin reference substance (China Institute for the control of pharmaceutical and biological products) Other reagents of Yinhuang Oral Liquid (Shandong Lunan pharmaceutical factory) are analytical pure 2 methods and results 2.1 preparation of control stock solution accurately weigh 5mg of chlorogenic acid control solution, put it into a 25ml measuring bottle, add water to dissolve the solution well and dilute it to the scale, shake well Weigh 5mg of baicalin precisely, put it into a 25ml measuring flask, add some ethanol, put it on a water bath and warm it to dissolve it When it reaches room temperature, add ethanol to the scale, and shake well 2.2 ratio spectrum drawing and determination wavelength selection: accurately absorb a proper amount of chlorogenic acid and baicalin reference stock solution, put them into a 25ml measuring flask, dilute them to the degree with 0.2mol/l hydrochloric acid and shake them well; take 0.2mol/l hydrochloric acid as a blank, scan them in the wavelength range of 200-400nm to obtain the absorption spectra of chlorogenic acid and baicalin Calculate the ratio R green of chlorogenic acid absorbance to baicalin absorbance at wavelength λ, and make a graph of R green to λ to get the ratio spectrum of chlorogenic acid; similarly, we can also get the ratio spectrum of baicalin It can be seen that there is a big r value of Zui at 342mn and a small R value of Zui at 266mn; while baicalin is the opposite of chlorogenic acid There is a big r value of Zui at 266nm and a small R value of Zui at 342nm; therefore, 266 and 3 are selected 42mn was used as the wavelength of chlorogenic acid and baicalin 2.3 draw the standard curve accurately and draw a proper amount of chlorogenic acid and baicalin reference stock solution Put them into a 25ml volumetric flask, dilute them to the degree with o.2mol/l hydrochloric acid and shake them well, as the standard reference solution (chlorogenic acid is 8.29 μ g / ml, baicalin is 8.12 μ g / M1) The absorbance was measured at 266 and 342 Mn, respectively A series of mixed standard solutions containing chlorogenic acid and Huangling glycoside were prepared by accurately absorbing chlorogenic acid and baicalin with proper amount and diluting them to a certain concentration with 0.2mol/l hydrochloric acid The absorbance was measured at 266 and 342tun respectively Δ r was calculated, and Δ r was analyzed by linear regression with concentration C to obtain the regression equation 2 4 recovery test Accurately suck 4m1 of the diluent of Yinhuang Oral Liquid with known content, put a proper amount of chlorogenic acid and baicalin reference stock solution into a 10ml measuring flask, dilute to the scale with 0.2mol/l hydrochloric acid, and shake it well The concentration of chlorogenic acid added into Qi preparation is 1.5, 2.5, 6.0, 11.0 and 16.0 μ g / ml, and the concentration of baicalin added is 2.5, 5.0, 7.0, 9.0 and 11.0 μ g / ml, and measure according to the method under the standard curve The recovery of chlorogenic acid and baicalin was calculated 25 Repeat the tripe test, accurately suck 2ml of Yinhuang Oral Liquid (batch No 970307), place it in a 100ml measuring bottle, add water to dilute it to the scale and shake it well; then precisely suck 2M1 of upper liquid, place it in a 100ml measuring bottle, dilute it to the scale with 0.2mol/l hydrochloric acid and shake it well The content of chlorogenic acid and baicalin was calculated at 266 and 342 nm The RSD was 0.51% and 0.44% respectively 2.6 sample determination: take 2ml of Yinhuang Oral liquid precisely and put it into 100ml measuring bottle Measure the content of chlorogenic acid and baicalin according to the operation under 2.5, and compare with Pharmacopoeia 3 The analysis of Yinhuang Oral Liquid by liquid chromatography shows that there are 5-6 unknown components in the ultraviolet region in Yinhuang Oral liquid These UN removed components may have some influence on spectrometric determination, but the influence can be accepted The results of this method are consistent with those of Pharmacopoeia method (P > 0.05) It shows that the dual wavelength ratio spectrum method has the ability of anti-interference to coexisting components, which can be further applied to the quality control of other Chinese patent medicines Reference 1 Chinese Pharmacopoeia, Vol 1, 1995:594 2 Yang Yuechun, Dai Qichang, determination of chlorogenic acid and Baicalin in Yinhuang Oral Liquid by liquid chromatography, China Modern Applied Pharmacy, 1998, 15 (4) 54