OBJECTIVE: To study the determination of chlorogenic acid and baicalin in Yinhuang oral liquid by dual-wavelength ratio spectroscopy. Methods: According to the spectral characteristics of chlorogenic acid and baicalin, 266 and 342mn were selected as the measurement wavelength. Results: The linear range of chlorogenic acid was 2～20μg/m1. The recovery rate was 98.9, the RSD was 1.58%, the baicalin linear range was 2-16 μg/ml, the recovery was 100.4%, and the RSD was 1.32%. Conclusion: This method is a sensitive, accurate and simple analytical method. It can be widely used in other traditional Chinese medicine preparations.
Key words: dual-wavelength ratio spectroscopy; Yinhuang oral liquid; chlorogenic acid; baicalin silver yellow oral liquid has heat-clearing and detoxifying, antibacterial and anti-inflammatory effects, attending upper respiratory tract infection, acute tonsillitis, pharyngitis and other symptoms. It is made from the processing of honeysuckle and Astragalus extract. Main chlorogenic acid (Chlorogenie acid) and baicalin (Baicalin). The Chinese Pharmacopoeia uses UV spectrophotometry to determine its content. There are reports on the determination of its content by high performance liquid chromatography . 2. The dual - wavelength ratio spectroscopy method was used to determine the content of chlorogenic acid and baicalin in Yinhuang oral liquid. The result is accurate.
1 Instruments and reagents Shimadzu UV-260 violet complement visible spectrophotometer; 752-C UV-visible spectrophotometer. Chlorogenic acid, baicalin reference substance (China National Institute for the Control of Pharmaceutical and Biological Products). Yinhuang Oral Liquid (Shandong Lunan Pharmaceutical Factory) Other reagents are of analytical grade.
2 Methods and results
2.1 Preparation of reference stock solution Precisely weigh 5mg of chlorogenic acid reference substance, put it in a 25ml volumetric flask, dilute the appropriate amount of water to dissolve the well and shake it up. Accurately weigh 5 mg of baicalin, place it in a 25 ml volumetric flask, add an appropriate amount of ethanol to the bath and warm it to dissolve. After adding to the room temperature, add ethanol to the mark and shake well.
2.2 Ratio spectrum drawing and measurement wavelength Select the appropriate amount of chlorogenic acid, baicalin reference substance stock solution, separate into 25ml volumetric flask, dilute to the degree with 0.2mol / L hydrochloric acid and shake; 0.2mol / L Hydrochloric acid was used as a blank and scanned in the wavelength range of 200 to 400 nm to obtain an absorption spectrum of chlorogenic acid and baicalin. Calculate the ratio of the chlorogenic acid absorbance to the baicalin absorbance at the wavelength λ, R green, and plot the R green versus λ to obtain the ratio spectrum of chlorogenic acid. Similarly, the ratio spectrum of baicalin can also be obtained. The acid has a large R value of zui at 342 mn and a small R value of zui at 266 nm; and baicalin is opposite to chlorogenic acid. There is a large R value of zui at 266 nm and a small R value of zui at 342 nm; therefore, 266 and 342 nm were selected as the measurement wavelengths of chlorogenic acid and baicalin.
2.3 The drawing of the standard curve accurately draws the appropriate amount of chlorogenic acid and baicalin reference substance stock solution. The mixture was placed in a 25 ml volumetric flask, diluted to a degree with O.2 mol/L hydrochloric acid, and shaken to obtain a standard reference solution (8.29 μg/ml for chlorogenic acid and 8.12 μg/ml for baicalin). The absorbance was measured at 266 and 342 mn, respectively.
Precisely absorb the chlorogenic acid and baicalin reference substance reserves, and dilute to a certain concentration with 0.2mol/L hydrochloric acid to prepare a series of mixed standard solutions containing chlorogenic acid and baicalin. The absorbance was measured at 266 and 342 tun, respectively. Calculate ΔR and perform linear regression analysis on ΔR with concentration C to obtain a regression equation.
2 4 Recovery test precision draws the known content of Yinhuang oral liquid dilution 4m1. Place the appropriate amount of chlorogenic acid and baicalin reference substance in a 10ml volumetric flask, dilute to the mark with 0.2mol/L hydrochloric acid and shake. A series of solutions containing chlorogenic acid concentrations of 1.5, 2.5, 6.0, 11.0 and 16.0 μg/ml and addition of baicalin at concentrations of 2.5, 5.0, 7.0, 9.0 and 11.0 μg/ml, according to the standard curve The method was determined to calculate the recovery rate of chlorogenic acid and baicalin.
2 5 Reproduce the belly test and accurately absorb 2ml of Yinhuang Oral Liquid (batch 970307), place it in a 100ml volumetric flask, dilute to the mark with water and shake it up: then accurately absorb the upper liquid 2m1, set it in a 100ml volumetric flask, use 0.2mol/L Dilute hydrochloric acid to the mark and shake well. The absorbance was measured at 266 and 342 nm. The contents of chlorogenic acid and baicalin were calculated and tested in parallel for 5 times. The RSD calculated from the results was 0.51% and 0.44%, respectively.
2.6 Sample determination Precision extraction of 2ml of Yinhuang oral solution, placed in a 100ml volumetric flask. The contents of chlorogenic acid and baicalin were determined according to the operation of 2.5 and compared with the pharmacopoeia method.
3 Discussion High performance liquid chromatography analysis of Yinhuang oral liquid shows that Yinhuang oral liquid still contains 5-6 unknown components in the ultraviolet region. These unremoved components may have a certain effect on spectrometry, but this effect can be accepted. Moreover, the method is consistent with the results of the pharmacopoeia method (P>0.05), indicating that the anti-interference ability of the dual-wavelength ratio spectroscopy on coexisting components can be further extended to the quality control of other proprietary Chinese medicine preparations.
references
1. Chinese Pharmacopoeia. One 1995:594
2. Yang Yuechun, Dai Qichang, Determination of Chlorogenic Acid and Baicalin in Yinhuang Oral Liquid by High Performance Liquid Chromatography, China Modern Applied Pharmacy, 1998, 15(4) 54
Key words: dual-wavelength ratio spectroscopy; Yinhuang oral liquid; chlorogenic acid; baicalin silver yellow oral liquid has heat-clearing and detoxifying, antibacterial and anti-inflammatory effects, attending upper respiratory tract infection, acute tonsillitis, pharyngitis and other symptoms. It is made from the processing of honeysuckle and Astragalus extract. Main chlorogenic acid (Chlorogenie acid) and baicalin (Baicalin). The Chinese Pharmacopoeia uses UV spectrophotometry to determine its content. There are reports on the determination of its content by high performance liquid chromatography . 2. The dual - wavelength ratio spectroscopy method was used to determine the content of chlorogenic acid and baicalin in Yinhuang oral liquid. The result is accurate.
1 Instruments and reagents Shimadzu UV-260 violet complement visible spectrophotometer; 752-C UV-visible spectrophotometer. Chlorogenic acid, baicalin reference substance (China National Institute for the Control of Pharmaceutical and Biological Products). Yinhuang Oral Liquid (Shandong Lunan Pharmaceutical Factory) Other reagents are of analytical grade.
2 Methods and results
2.1 Preparation of reference stock solution Precisely weigh 5mg of chlorogenic acid reference substance, put it in a 25ml volumetric flask, dilute the appropriate amount of water to dissolve the well and shake it up. Accurately weigh 5 mg of baicalin, place it in a 25 ml volumetric flask, add an appropriate amount of ethanol to the bath and warm it to dissolve. After adding to the room temperature, add ethanol to the mark and shake well.
2.2 Ratio spectrum drawing and measurement wavelength Select the appropriate amount of chlorogenic acid, baicalin reference substance stock solution, separate into 25ml volumetric flask, dilute to the degree with 0.2mol / L hydrochloric acid and shake; 0.2mol / L Hydrochloric acid was used as a blank and scanned in the wavelength range of 200 to 400 nm to obtain an absorption spectrum of chlorogenic acid and baicalin. Calculate the ratio of the chlorogenic acid absorbance to the baicalin absorbance at the wavelength λ, R green, and plot the R green versus λ to obtain the ratio spectrum of chlorogenic acid. Similarly, the ratio spectrum of baicalin can also be obtained. The acid has a large R value of zui at 342 mn and a small R value of zui at 266 nm; and baicalin is opposite to chlorogenic acid. There is a large R value of zui at 266 nm and a small R value of zui at 342 nm; therefore, 266 and 342 nm were selected as the measurement wavelengths of chlorogenic acid and baicalin.
2.3 The drawing of the standard curve accurately draws the appropriate amount of chlorogenic acid and baicalin reference substance stock solution. The mixture was placed in a 25 ml volumetric flask, diluted to a degree with O.2 mol/L hydrochloric acid, and shaken to obtain a standard reference solution (8.29 μg/ml for chlorogenic acid and 8.12 μg/ml for baicalin). The absorbance was measured at 266 and 342 mn, respectively.
Precisely absorb the chlorogenic acid and baicalin reference substance reserves, and dilute to a certain concentration with 0.2mol/L hydrochloric acid to prepare a series of mixed standard solutions containing chlorogenic acid and baicalin. The absorbance was measured at 266 and 342 tun, respectively. Calculate ΔR and perform linear regression analysis on ΔR with concentration C to obtain a regression equation.
2 4 Recovery test precision draws the known content of Yinhuang oral liquid dilution 4m1. Place the appropriate amount of chlorogenic acid and baicalin reference substance in a 10ml volumetric flask, dilute to the mark with 0.2mol/L hydrochloric acid and shake. A series of solutions containing chlorogenic acid concentrations of 1.5, 2.5, 6.0, 11.0 and 16.0 μg/ml and addition of baicalin at concentrations of 2.5, 5.0, 7.0, 9.0 and 11.0 μg/ml, according to the standard curve The method was determined to calculate the recovery rate of chlorogenic acid and baicalin.
2 5 Reproduce the belly test and accurately absorb 2ml of Yinhuang Oral Liquid (batch 970307), place it in a 100ml volumetric flask, dilute to the mark with water and shake it up: then accurately absorb the upper liquid 2m1, set it in a 100ml volumetric flask, use 0.2mol/L Dilute hydrochloric acid to the mark and shake well. The absorbance was measured at 266 and 342 nm. The contents of chlorogenic acid and baicalin were calculated and tested in parallel for 5 times. The RSD calculated from the results was 0.51% and 0.44%, respectively.
2.6 Sample determination Precision extraction of 2ml of Yinhuang oral solution, placed in a 100ml volumetric flask. The contents of chlorogenic acid and baicalin were determined according to the operation of 2.5 and compared with the pharmacopoeia method.
3 Discussion High performance liquid chromatography analysis of Yinhuang oral liquid shows that Yinhuang oral liquid still contains 5-6 unknown components in the ultraviolet region. These unremoved components may have a certain effect on spectrometry, but this effect can be accepted. Moreover, the method is consistent with the results of the pharmacopoeia method (P>0.05), indicating that the anti-interference ability of the dual-wavelength ratio spectroscopy on coexisting components can be further extended to the quality control of other proprietary Chinese medicine preparations.
references
1. Chinese Pharmacopoeia. One 1995:594
2. Yang Yuechun, Dai Qichang, Determination of Chlorogenic Acid and Baicalin in Yinhuang Oral Liquid by High Performance Liquid Chromatography, China Modern Applied Pharmacy, 1998, 15(4) 54
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