CN101912428B - Method for detecting quality of traditional Chinese medicine indigowoad root granules - Google Patents
Method for detecting quality of traditional Chinese medicine indigowoad root granules Download PDFInfo
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Abstract
The invention provides a method for detecting the quality of traditional Chinese medicine indigowoad root granules. The method comprises the following steps of: detecting the traditional Chinese medicine indigowoad root granules by using a soft-ionization mass spectrometry technique; performing primary evaluation on the number and strength of characteristic fingerprint peaks in an electro-spray fingerprint; measuring by high performance liquid chromatography to obtain the peak area of a chromatographic peak corresponding to each of cytidine, adenine, uridine, guanosine and adenosine serving as standard reference substances; and measuring according to a standard curve to obtain the cytidine content, adenine content, uridine content, guanosine content and adenosine content of one gram of the traditional Chinese medicine indigowoad root granules: 0.234 to 0.238 milligrams of cytidine, 0.116 to 0.118 milligrams of adenine, 0.472 to 0.474 milligrams of uridine, 0.331 to 0.334 milligrams of guanosine and 0.447 to 0.450 milligrams of adenosine. The method has the advantages of ensuring the scientificity and progressiveness of the method for detecting the quality of the traditional Chinese medicine indigowoad root granules, along with qualitative and quantitative performance.
Description
Technical field
The invention belongs to the Natural Medicine Chemistry field, be specifically related to a kind of quality determining method of Chinese medicine banlangen granules, banlangen keli.
Background technology
Banlangen granules, banlangen keli is that the Pharmacopoeia of the People's Republic of China is kept the Chinese traditional patent formulation preparation that carries, and has clearing heat and detoxicatingly, and the effect of cool blood relieve sore throat is used to treat abscess of throat due to the lung and stomach excessive heat, pharyngoxerosis, cheek swelling, acute tonsillitis, mumps etc.
[1], modern pharmacodynamic study shows that banlangen granules, banlangen keli has stronger antivirus action
[2], at present, the method for quality control of banlangen granules, banlangen keli mainly is based on the quantitative test of compositions such as indigo red, adenosine
[3-5], be not enough to control on the whole the quality of banlangen granules, banlangen keli, as clinical common dosage forms; Therefore the influence of many factors such as the good and bad crude drug source of its true and false, production technology is necessary that the quality determining method of setting up a kind of science estimates the quality of banlangen granules, banlangen keli, (list of references: [1] Pharmacopoeia of People's Republic of China council. Pharmacopoeia of People's Republic of China [M]. Beijing: Chemical Industry Press; 2005:487-488. [2] easy to be strong, Jia Xiaobin, Chen Yan; Measure the content [J] of epigoitrin in the different manufacturers banlangen granules, banlangen keli Deng .RP-HPLC. Chinese traditional Chinese medicine magazine, 2009,24 (4): 529-531. [3] Yan Yizhou; Zou Pin literary composition .HPLC method is measured the content [J] of indigo red in the banlangen granules, banlangen keli. China Dispensary, 2008,19 (3): 211-212. [4] Hou Yaming; Wang Yulin, Hu Yibing. the content of adenosine [J] in the high effective liquid chromatography for measuring banlangen granules, banlangen keli. Hunan Journal of Traditional Chinese Medicine, 2005; 21 (3): 112-114. [5] Chen Mao, Liang Huiyu, Yang Dongsheng; Deng. the HPLC of adenosine analyzes [J] in the banlangen granules, banlangen keli. Guangzhou pharmacy, 2003,13 (6): 6-8.)
Summary of the invention
The quality determining method that the purpose of this invention is to provide a kind of Chinese medicine banlangen granules, banlangen keli; This method utilizes the electrospray ionization mass spectrum of Chinese medicine banlangen granules, banlangen keli to make finger-print; The content of nucleosides and purine constituents has guaranteed the science of Chinese medicine banlangen granules, banlangen keli quality determining method and the advance of quality testing in the Chinese medicine banlangen granules, banlangen keli that utilized high effective liquid chromatography for measuring.
The present invention utilizes the soft ionization mass-spectrometric technique that the Chinese medicine banlangen granules, banlangen keli is detected, and according to the number and the intensity at characteristic fingerprint peak in the electron spray finger-print of the Chinese medicine banlangen granules, banlangen keli that obtains, the quality of Chinese medicine banlangen granules, banlangen keli is carried out preliminary assessment; Consider and contain multiple nucleosides and purine constituents in the Chinese medicine banlangen granules, banlangen keli; And with cytidine, adenine, uridine, guanosine and adenosine is main; Therefore the present invention as standard reference material, through high-performance liquid chromatogram determination, obtains the peak area of the corresponding chromatographic peak of each standard reference material with these five kinds of compositions; According to the content of cytidine, adenine, uridine, guanosine and adenosine in the standard curve determination confession test agent, accomplish a kind of quality determining method of Chinese medicine banlangen granules, banlangen keli.
The technical scheme of embodiment of the present invention is following:
(1) banlangen granules, banlangen keli electron spray determining fingerprint pattern
1. banlangen granules, banlangen keli supplies the preparation of test agent solution
With the banlangen granules, banlangen keli is raw material, takes by weighing banlangen granules, banlangen keli 2-10g, adds that 10-50mL is water-soluble to be separated, and in the dry rearmounted 10mL volumetric flask, adds 50% (V/V) methanol constant volume to 10mL, filters, and is need testing solution;
2. banlangen granules, banlangen keli supplies the electron spray finger-print of test agent
The electron spray finger-print of test sample obtains through following steps: get 5-10 μ L need testing solution, be diluted to 100-500 μ L with methyl alcohol, do the electrospray ionization mass spectrum analysis; The electrospray ionization mass spectrum experiment condition is following: the electron spray ionisation source; Positive ion electrospray is from mode, and spray voltage is 4.0-5.0kV, capillary temperature 180-250 ℃; Injection pump sample introduction flow velocity 3-8 μ L/min flows; Sweep limit mass-to-charge ratio (m/z) 50-1000 in the m/z100-400 scope, exists m/z104, m/z116, m/z137, m/z175, m/z185, m/z203, m/z259, m/z264, m/z365, m/z381 and m/z397 ion;
(2) assay of purine and ucleosides composition in the banlangen granules, banlangen keli
1. high performance liquid chromatography experiment condition: C
18Chromatographic column: 4.6mm * 250mm, 5 μ m; Column temperature: 20-35 ℃; Moving phase: water and methyl alcohol; Water and methyl alcohol volume ratio are 90: 10-95: 5; Elution time: 15-30min; UV-detector detects wavelength: 230-280nm; Sample feeding amount: 5-20 μ L;
2. the preparation of purine and ucleosides ingredient standard reference substance solution: take by weighing cytidine, adenine, uridine, guanosine and each 1mg of adenosine standard reference material respectively, put in the 10mL volumetric flask, add 50% methyl alcohol (V/V) and be settled to 10mL, be made into standard reference material solution;
3. the investigation of purine and the sexual intercourse of ucleosides component lines
Cytidine, adenine, uridine, guanosine and adenosine standard reference material solution respectively with 2,4,6,8,10 μ L sample introductions, are recorded the high performance liquid chromatography peak area of each standard reference material; With each standard reference material peak area of gained is ordinate, and the sample introduction quality of corresponding each standard reference material is a horizontal ordinate, calculates equation of linear regression respectively;
4. the high-performance liquid chromatogram determination of banlangen granules, banlangen keli need testing solution: according to 1. high performance liquid chromatography experiment condition of step; The banlangen granules, banlangen keli need testing solution is carried out high-performance liquid chromatogram determination; Obtain the peak area of cytidine in the banlangen granules, banlangen keli need testing solution, adenine, uridine, guanosine and adenosine chromatographic peak respectively; Equation of linear regression according to cytidine, adenine, uridine, guanosine and adenosine standard reference material; Calculate the quality of cytidine in the need testing solution, adenine, uridine, guanosine and adenosine; And the ratio of the quality of the quality of combination test sample and Chinese medicine banlangen granules, banlangen keli, the content that obtains cytidine, adenine, uridine, guanosine and adenosine in the Chinese medicine banlangen granules, banlangen keli is respectively cytidine 0.234-0.238mg/g, adenine 0.116-0.118mg/g, uridine 0.472-0.474mg/g, guanosine 0.331-0.334mg/g, adenosine 0.447-0.450mg/g.
Beneficial effect: the present invention provides a kind of quality determining method of Chinese medicine banlangen granules, banlangen keli; Utilize the soft ionization mass-spectrometric technique that the Chinese medicine banlangen granules, banlangen keli is detected; According to the number and the intensity at characteristic fingerprint peak in the electron spray finger-print of the Chinese medicine banlangen granules, banlangen keli that obtains, the quality of Chinese medicine banlangen granules, banlangen keli is carried out preliminary assessment; Consider and contain multiple nucleosides and purine constituents in the Chinese medicine banlangen granules, banlangen keli; And with cytidine, adenine, uridine, guanosine and adenosine is main; Therefore the present invention with these five kinds of compositions as standard reference material; Pass through high-performance liquid chromatogram determination; Obtain the peak area of the corresponding chromatographic peak of each standard reference material, according to standard curve determination, the content that obtains cytidine, adenine, uridine, guanosine and adenosine in the Chinese medicine banlangen granules, banlangen keli is respectively cytidine 0.234-0.238mg/g, adenine 0.116-0.118mg/g, uridine 0.472-0.474mg/g, guanosine 0.331-0.334mg/g, adenosine 0.447-0.450mg/g.The present invention provides a kind of quality determining method of Chinese medicine banlangen granules, banlangen keli, and is both qualitative and quantitative to the quality testing of Chinese medicine banlangen granules, banlangen keli, guaranteed the scientific and advanced of Chinese medicine banlangen granules, banlangen keli quality determining method.
Embodiment
Embodiment 1
(1) banlangen granules, banlangen keli electron spray determining fingerprint pattern
1. banlangen granules, banlangen keli supplies the preparation of test agent solution
With the banlangen granules, banlangen keli is raw material, takes by weighing banlangen granules, banlangen keli 2g, adds that 10mL is water-soluble to be separated, and in the dry rearmounted 10mL volumetric flask, adds 50% (V/V) methanol constant volume to 10mL, filters, and is need testing solution;
2. banlangen granules, banlangen keli supplies the electron spray finger-print of test agent
The electron spray finger-print of test sample obtains through following steps: get 5 μ L need testing solutions, be diluted to 100 μ L with methyl alcohol, do the electrospray ionization mass spectrum analysis; The electrospray ionization mass spectrum experiment condition is following: the electron spray ionisation source; Positive ion electrospray is from mode, and spray voltage is 4.0kV, 180 ℃ of capillary temperatures; Injection pump sample introduction flow velocity 3 μ L/min flow; Sweep limit mass-to-charge ratio (m/z) 50-1000 in the m/z100-400 scope, exists m/z104, m/z116, m/z137, m/z175, m/z185, m/z203, m/z259, m/z264, m/z365, m/z381 and m/z397 ion.
(2) assay of purine and ucleosides composition in the banlangen granules, banlangen keli
1. high performance liquid chromatography experiment condition: C
18Chromatographic column: 4.6mm * 250mm, 5 μ m; Column temperature: 20 ℃; Moving phase: water and methyl alcohol; Water and methyl alcohol volume ratio are 90: 10; Elution time: 15min; UV-detector detects wavelength: 230nm; Sample feeding amount: 20 μ L;
2. the preparation of purine and ucleosides ingredient standard reference substance solution: precision takes by weighing cytidine, adenine, uridine, guanosine and each 1mg of adenosine standard reference material respectively, puts in the 10mL volumetric flask, adds 50% methyl alcohol (V/V) and is settled to 10mL, is made into standard reference material solution;
3. the investigation of purine and the sexual intercourse of ucleosides component lines:
Cytidine, adenine, uridine, guanosine and adenosine standard reference material solution respectively with 2,4,6,8,10 μ L sample introductions, are recorded the high performance liquid chromatography peak area of each standard reference material; With each standard reference material peak area of gained is ordinate, and the sample introduction quality of corresponding each standard reference material is a horizontal ordinate, calculates equation of linear regression respectively;
4. the high-performance liquid chromatogram determination of banlangen granules, banlangen keli need testing solution: the banlangen granules, banlangen keli need testing solution is carried out high-performance liquid chromatogram determination; Obtain the peak area of cytidine in the banlangen granules, banlangen keli need testing solution, adenine, uridine, guanosine and adenosine chromatographic peak respectively; Equation of linear regression according to cytidine, adenine, uridine, guanosine and adenosine standard reference material; Calculate the quality of cytidine in the need testing solution, adenine, uridine, guanosine and adenosine; And the ratio of the quality of the quality of combination test sample and former banlangen granules, banlangen keli; The content that obtains cytidine in the banlangen granules, banlangen keli, adenine, uridine, guanosine and adenosine is respectively cytidine 0.235-0.237mg/g, adenine 0.117-0.118mg/g, uridine 0.473-0.474mg/g, guanosine 0.331-0.333mg/g, adenosine 0.447-0.448mg/g, accomplishes a kind of quality determining method of Chinese medicine banlangen granules, banlangen keli.
Embodiment 2
(1) banlangen granules, banlangen keli electron spray determining fingerprint pattern
1. banlangen granules, banlangen keli supplies the preparation of test agent solution
With the banlangen granules, banlangen keli is raw material, takes by weighing banlangen granules, banlangen keli 4g, adds that 20mL is water-soluble to be separated, and in the dry rearmounted 10mL volumetric flask, adds 50% (V/V) methanol constant volume to 10mL, filters, and is need testing solution;
2. banlangen granules, banlangen keli supplies the electron spray finger-print of test agent
The electron spray finger-print of test sample obtains through following steps: get 6 μ L need testing solutions, be diluted to 200 μ L with methyl alcohol, do the electrospray ionization mass spectrum analysis; The electrospray ionization mass spectrum experiment condition is following: the electron spray ionisation source; Positive ion electrospray is from mode, and spray voltage is 4.2kV, 250 ℃ of capillary temperatures; Injection pump sample introduction flow velocity 6 μ L/min flow; Sweep limit mass-to-charge ratio (m/z) 50-1000 in the m/z100-400 scope, exists m/z104, m/z116, m/z137, m/z175, m/z185, m/z203, m/z259, m/z264, m/z365, m/z381 and m/z397 ion.
(2) assay of purine and ucleosides composition in the banlangen granules, banlangen keli
1. high performance liquid chromatography experiment condition: C
18Chromatographic column: 4.6mm * 250mm, 5 μ m; Column temperature: 30 ℃; Moving phase: water and methyl alcohol; Water and methyl alcohol volume ratio are 92: 8; Elution time: 20min; UV-detector detects wavelength: 254nm; Sample feeding amount: 5 μ L;
2. the preparation of purine and ucleosides ingredient standard reference substance solution: precision takes by weighing cytidine, adenine, uridine, guanosine and each 1mg of adenosine standard reference material respectively, puts in the 10mL volumetric flask, adds 50% methyl alcohol (V/V) and is settled to 10mL, is made into standard reference material solution;
3. the investigation of purine and the sexual intercourse of ucleosides component lines:
Cytidine, adenine, uridine, guanosine and adenosine standard reference material solution respectively with 2,4,6,8,10 μ L sample introductions, are recorded the high performance liquid chromatography peak area of each standard reference material; With each standard reference material peak area of gained is ordinate, and the sample introduction quality of corresponding each standard reference material is a horizontal ordinate, calculates equation of linear regression respectively;
4. the high-performance liquid chromatogram determination of banlangen granules, banlangen keli need testing solution: the banlangen granules, banlangen keli need testing solution is carried out high-performance liquid chromatogram determination; Obtain the peak area of cytidine in the banlangen granules, banlangen keli need testing solution, adenine, uridine, guanosine and adenosine chromatographic peak respectively; Equation of linear regression according to cytidine, adenine, uridine, guanosine and adenosine standard reference material; Calculate the quality of cytidine in the need testing solution, adenine, uridine, guanosine and adenosine; And the ratio of the quality of the quality of combination test sample and former banlangen granules, banlangen keli; The content that obtains cytidine in the banlangen granules, banlangen keli, adenine, uridine, guanosine and adenosine is respectively cytidine 0.234-0.236mg/g, adenine 0.116-0.117mg/g, uridine 0.472-0.473mg/g, guanosine 0.332-0.334mg/g, adenosine 0.449-0.450mg/g, accomplishes a kind of quality determining method of Chinese medicine banlangen granules, banlangen keli.
Embodiment 3
(1) banlangen granules, banlangen keli electron spray determining fingerprint pattern
1. banlangen granules, banlangen keli supplies the preparation of test agent solution
With the banlangen granules, banlangen keli is raw material, takes by weighing banlangen granules, banlangen keli 6g, adds that 30mL is water-soluble to be separated, and in the dry rearmounted 10mL volumetric flask, adds 50% (V/V) methanol constant volume to 10mL, filters, and is need testing solution;
2. banlangen granules, banlangen keli supplies the electron spray finger-print of test agent
The electron spray finger-print of test sample obtains through following steps: get 7 μ L need testing solutions, be diluted to 300 μ L with methyl alcohol, do the electrospray ionization mass spectrum analysis; The electrospray ionization mass spectrum experiment condition is following: the electron spray ionisation source; Positive ion electrospray is from mode, and spray voltage is 4.5kV, 250 ℃ of capillary temperatures; Injection pump sample introduction flow velocity 6 μ L/min flow; Sweep limit mass-to-charge ratio (m/z) 50-1000 in the m/z100-400 scope, exists m/z104, m/z116, m/z137, m/z175, m/z185, m/z203, m/z259, m/z264, m/z365, m/z381 and m/z397 ion.
(2) assay of purine and ucleosides composition in the banlangen granules, banlangen keli
1. high performance liquid chromatography experiment condition: C
18Chromatographic column: 4.6mm * 250mm, 5 μ m; Column temperature: 35 ℃; Moving phase: water and methyl alcohol; Water and methyl alcohol volume ratio are 93: 7; Elution time: 22min; UV-detector detects wavelength: 265nm; Sample feeding amount: 10 μ L;
2. the preparation of purine and ucleosides ingredient standard reference substance solution: precision takes by weighing cytidine, adenine, uridine, guanosine and each 1mg of adenosine standard reference material respectively, puts in the 10mL volumetric flask, adds 50% methyl alcohol (V/V) and is settled to 10mL, is made into standard reference material solution;
3. the investigation of purine and the sexual intercourse of ucleosides component lines:
Cytidine, adenine, uridine, guanosine and adenosine standard reference material solution respectively with 2,4,6,8,10 μ L sample introductions, are recorded the high performance liquid chromatography peak area of each standard reference material; With each standard reference material peak area of gained is ordinate, and the sample introduction quality of corresponding each standard reference material is a horizontal ordinate, calculates equation of linear regression respectively;
4. the high-performance liquid chromatogram determination of banlangen granules, banlangen keli need testing solution: the banlangen granules, banlangen keli need testing solution is carried out high-performance liquid chromatogram determination; Obtain the peak area of cytidine in the banlangen granules, banlangen keli need testing solution, adenine, uridine, guanosine and adenosine chromatographic peak respectively; Equation of linear regression according to cytidine, adenine, uridine, guanosine and adenosine standard reference material; Calculate the quality of cytidine in the need testing solution, adenine, uridine, guanosine and adenosine; And the ratio of the quality of the quality of combination test sample and former banlangen granules, banlangen keli; The content that obtains cytidine in the banlangen granules, banlangen keli, adenine, uridine, guanosine and adenosine is respectively cytidine 0.237-0.238mg/g, adenine 0.116-0.117mg/g, uridine 0.472-0.473mg/g, guanosine 0.333-0.334mg/g, adenosine 0.448-0.449mg/g, accomplishes a kind of quality determining method of Chinese medicine banlangen granules, banlangen keli.
Embodiment 4
(1) banlangen granules, banlangen keli electron spray determining fingerprint pattern
1. banlangen granules, banlangen keli supplies the preparation of test agent solution
With the banlangen granules, banlangen keli is raw material, takes by weighing banlangen granules, banlangen keli 8g, adds that 40mL is water-soluble to be separated, and in the dry rearmounted 10mL volumetric flask, adds 50% (V/V) methanol constant volume to 10mL, filters, and is need testing solution;
2. banlangen granules, banlangen keli supplies the electron spray finger-print of test agent
The electron spray finger-print of test sample obtains through following steps: get 8 μ L need testing solutions, be diluted to 400 μ L with methyl alcohol, do the electrospray ionization mass spectrum analysis; The electrospray ionization mass spectrum experiment condition is following: the electron spray ionisation source; Positive ion electrospray is from mode, and spray voltage is 4.8kV, 220 ℃ of capillary temperatures; Injection pump sample introduction flow velocity 7 μ L/min flow; Sweep limit mass-to-charge ratio (m/z) 50-1000 in the m/z100-400 scope, exists m/z104, m/z116, m/z137, m/z175, m/z185, m/z203, m/z259, m/z264, m/z365, m/z381 and m/z397 ion.
(2) assay of purine and ucleosides composition in the banlangen granules, banlangen keli
1. high performance liquid chromatography experiment condition: C
18Chromatographic column: 4.6mm * 250mm, 5 μ m; Column temperature: 20 ℃; Moving phase: water and methyl alcohol; Water and methyl alcohol volume ratio are 94: 6; Elution time: 25min; UV-detector detects wavelength: 270nm; Sample feeding amount: 8 μ L;
2. the preparation of purine and ucleosides ingredient standard reference substance solution: precision takes by weighing cytidine, adenine, uridine, guanosine and each 1mg of adenosine standard reference material respectively, puts in the 10mL volumetric flask, adds 50% methyl alcohol (V/V) and is settled to 10mL, is made into standard reference material solution;
3. the investigation of purine and the sexual intercourse of ucleosides component lines:
Cytidine, adenine, uridine, guanosine and adenosine standard reference material solution respectively with 2,4,6,8,10 μ L sample introductions, are recorded the high performance liquid chromatography peak area of each standard reference material; With each standard reference material peak area of gained is ordinate, and the sample introduction quality of corresponding each standard reference material is a horizontal ordinate, calculates equation of linear regression respectively;
4. the high-performance liquid chromatogram determination of banlangen granules, banlangen keli need testing solution: the banlangen granules, banlangen keli need testing solution is carried out high-performance liquid chromatogram determination; Obtain the peak area of cytidine in the banlangen granules, banlangen keli need testing solution, adenine, uridine, guanosine and adenosine chromatographic peak respectively; Equation of linear regression according to cytidine, adenine, uridine, guanosine and adenosine standard reference material; Calculate the quality of cytidine in the need testing solution, adenine, uridine, guanosine and adenosine; And the ratio of the quality of the quality of combination test sample and former banlangen granules, banlangen keli; The content that obtains cytidine in the banlangen granules, banlangen keli, adenine, uridine, guanosine and adenosine is respectively cytidine 0.234-0.235mg/g, adenine 0.117-0.118mg/g, uridine 0.473-0.474mg/g, guanosine 0.331-0.332mg/g, adenosine 0.447-0.448mg/g, accomplishes a kind of quality determining method of Chinese medicine banlangen granules, banlangen keli.
Embodiment 5
(1) banlangen granules, banlangen keli electron spray determining fingerprint pattern
1. banlangen granules, banlangen keli supplies the preparation of test agent solution
With the banlangen granules, banlangen keli is raw material, takes by weighing banlangen granules, banlangen keli 10g, adds that 50mL is water-soluble to be separated, and in the dry rearmounted 10mL volumetric flask, adds 50% (V/V) methanol constant volume to 10mL, filters, and is need testing solution;
2. banlangen granules, banlangen keli supplies the electron spray finger-print of test agent
The electron spray finger-print of test sample obtains through following steps: get 10 μ L need testing solutions, be diluted to 500 μ L with methyl alcohol, do the electrospray ionization mass spectrum analysis; The electrospray ionization mass spectrum experiment condition is following: the electron spray ionisation source; Positive ion electrospray is from mode, and spray voltage is 5.0kV, 190 ℃ of capillary temperatures; Injection pump sample introduction flow velocity 8 μ L/min flow; Sweep limit mass-to-charge ratio (m/z) 50-1000 in the m/z100-400 scope, exists m/z104, m/z116, m/z137, m/z175, m/z185, m/z203, m/z259, m/z264, m/z365, m/z381 and m/z397 ion.
(2) assay of purine and ucleosides composition in the banlangen granules, banlangen keli
1. high performance liquid chromatography experiment condition: C
18Chromatographic column: 4.6mm * 250mm, 5 μ m; Column temperature: 35 ℃; Moving phase: water and methyl alcohol; Water and methyl alcohol volume ratio are 95: 5; Elution time: 30min; UV-detector detects wavelength: 280nm; Sample feeding amount: 12 μ L;
2. the preparation of purine and ucleosides ingredient standard reference substance solution: precision takes by weighing cytidine, adenine, uridine, guanosine and each 1mg of adenosine standard reference material respectively, puts in the 10mL volumetric flask, adds 50% methyl alcohol (V/V) and is settled to 10mL, is made into standard reference material solution;
3. the investigation of purine and the sexual intercourse of ucleosides component lines:
Cytidine, adenine, uridine, guanosine and adenosine standard reference material solution respectively with 2,4,6,8,10 μ L sample introductions, are recorded the high performance liquid chromatography peak area of each standard reference material; With each standard reference material peak area of gained is ordinate, and the sample introduction quality of corresponding each standard reference material is a horizontal ordinate, calculates equation of linear regression respectively;
4. the high-performance liquid chromatogram determination of banlangen granules, banlangen keli need testing solution: the banlangen granules, banlangen keli need testing solution is carried out high-performance liquid chromatogram determination; Obtain the peak area of cytidine in the banlangen granules, banlangen keli need testing solution, adenine, uridine, guanosine and adenosine chromatographic peak respectively; Equation of linear regression according to cytidine, adenine, uridine, guanosine and adenosine standard reference material; Calculate the quality of cytidine in the need testing solution, adenine, uridine, guanosine and adenosine; And the ratio of the quality of the quality of combination test sample and former banlangen granules, banlangen keli; The content that obtains cytidine in the banlangen granules, banlangen keli, adenine, uridine, guanosine and adenosine is respectively cytidine 0.236-0.238mg/g, adenine 0.116-0.117mg/g, uridine 0.472-0.473mg/g, guanosine 0.331-0.332mg/g, adenosine 0.448-0.449mg/g, accomplishes a kind of quality determining method of Chinese medicine banlangen granules, banlangen keli.
Claims (1)
1. the detection method of a Chinese medicine banlangen granules, banlangen keli is characterized in that, step and condition are following:
(1) banlangen granules, banlangen keli electron spray determining fingerprint pattern
1. banlangen granules, banlangen keli supplies the preparation of test agent solution
With the banlangen granules, banlangen keli is raw material, takes by weighing banlangen granules, banlangen keli 2-10g, adds that 10-50mL is water-soluble to be separated, and in the dry rearmounted 10mL volumetric flask, adds 50% (V/V) methanol constant volume to 10mL, filters, and is need testing solution;
2. banlangen granules, banlangen keli supplies the electron spray finger-print of test agent
The electron spray finger-print of test sample obtains through following steps: get 5-10 μ L need testing solution, be diluted to 100-500 μ L with methyl alcohol, do the electrospray ionization mass spectrum analysis; The electrospray ionization mass spectrum experiment condition is following: the electron spray ionisation source; Positive ion electrospray is from mode, and spray voltage is 4.0-5.0kV, capillary temperature 180-250 ℃; Injection pump sample introduction flow velocity 3-8 μ L/min flows; Sweep limit m/z50-1000 in the m/z100-400 scope, exists m/z104, m/z116, m/z137, m/z175, m/z185, m/z203, m/z259, m/z264, m/z365, m/z381 and m/z397 ion;
(2) assay of purine and ucleosides composition in the banlangen granules, banlangen keli
1. high performance liquid chromatography experiment condition: C
18Chromatographic column: 4.6mm * 250mm, 5 μ m; Column temperature: 20-35 ℃; Moving phase: water and methyl alcohol; Water and methyl alcohol volume ratio are 90: 10-95: 5; Elution time: 15-30min; UV-detector detects wavelength: 230-280nm; Sample feeding amount: 5-20 μ L;
2. the preparation of purine and ucleosides ingredient standard reference substance solution: take by weighing cytidine, adenine, uridine, guanosine and each 1mg of adenosine standard reference material respectively, put in the 10mL volumetric flask, add 50% methyl alcohol (V/V) and be settled to 10mL, be made into standard reference material solution;
3. the investigation of purine and the sexual intercourse of ucleosides component lines
Cytidine, adenine, uridine, guanosine and adenosine standard reference material solution respectively with 2,4,6,8,10 μ L sample introductions, are recorded the high performance liquid chromatography peak area of each standard reference material; With each standard reference material peak area of gained is ordinate, and the sample introduction quality of corresponding each standard reference material is a horizontal ordinate, calculates equation of linear regression respectively;
4. the high-performance liquid chromatogram determination of banlangen granules, banlangen keli need testing solution: according to 1. high performance liquid chromatography experiment condition of step; The banlangen granules, banlangen keli need testing solution is carried out high-performance liquid chromatogram determination; Obtain the peak area of cytidine in the banlangen granules, banlangen keli need testing solution, adenine, uridine, guanosine and adenosine chromatographic peak respectively; Equation of linear regression according to cytidine, adenine, uridine, guanosine and adenosine standard reference material; Calculate the quality of cytidine in the need testing solution, adenine, uridine, guanosine and adenosine; And the ratio of the quality of the quality of combination test sample and Chinese medicine banlangen granules, banlangen keli, the content that obtains cytidine, adenine, uridine, guanosine and adenosine in the Chinese medicine banlangen granules, banlangen keli is respectively cytidine 0.234-0.238mg/g, adenine 0.116-0.118mg/g, uridine 0.472-0.474mg/g, guanosine 0.331-0.334mg/g, adenosine 0.447-0.450mg/g.
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| CN106442801B (en) * | 2016-11-10 | 2018-12-28 | 陕西师范大学 | Method that is a kind of while measuring 11 kinds of active constituent contents in folium isatidis |
| CN107192829A (en) * | 2017-05-18 | 2017-09-22 | 南京中医药大学 | A kind of method of Radix Isatidis identification of proteins |
| CN108333280A (en) * | 2018-01-26 | 2018-07-27 | 上海上药第生化药业有限公司 | The separation method of nucleosides material and its application in a kind of Pericarpium Trichosanthis injection |
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| CN101732406A (en) * | 2010-01-28 | 2010-06-16 | 昆明中药厂有限公司 | Quality detecting method for indigowoad root heat removing pellet |
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