CN100360931C - A kind of quality control method of salvia miltiorrhiza and notoginseng preparation - Google Patents

Abstract

The present invention relates to a method for identifying the compound HPLC-DAD fingerprint of red sage root and notoginseng, which belongs to the field of the quality control of natural medicines, particularly to the field of finger prints. The present invention can be used as one of the indexes for controlling the quality of compound preparations containing the medicinal materials of danshen root and notoginseng, such as danqi tablets, compound danshen tablets, compound danshen dripping pills, etc.

Description

A kind of quality control method of salvia miltiorrhiza and notoginseng preparation

Technical field:

The invention relates to the field of quality control of natural medicines, in particular to the field of fingerprints, in particular to an HPLC-DAD fingerprint identification method of Danshen and Panax notoginseng compound prescriptions. It can be used as one of the indicators for the quality control of compound preparations containing Danshen and Panax notoginseng.

Background technique:

Preparations containing Danshen and Panax notoginseng, commonly known as compound Danshen preparations, are widely used clinically to treat cardiovascular diseases. The common ones are Compound Danshen Tablets, Compound Danshen Dripping Pills, Danqi Tablets, Guanxin Danshen Tablets, etc.

Salvia miltiorrhiza and Panax notoginseng are the two main components of compound salvia miltiorrhiza preparations. Salvia miltiorrhiza is the dry root of Salvia miltiorrhiza Bge., a plant of the Labiatae family. Tune, amenorrhea, dysmenorrhea, accumulation of lumps in the abdomen, tingling pain in the chest and abdomen, heat arthralgia pain, sore swelling and pain, upset and insomnia, hepatosplenomegaly, angina pectoris; Panax notoginseng is the Araliaceae plant Panax notoginseng (Burk.) F.H. The dried root of Chen has the effects of dispelling blood stasis and hemostasis, reducing swelling and relieving pain. For hemoptysis, hematemesis, epistaxis, traumatic hemorrhage, tumbling, swelling and pain embolisms.

Modern research shows that the active ingredients of Salvia miltiorrhiza are mainly fat-soluble diterpene quinones and water-soluble active ingredients of salvianolic acids, which have the functions of inhibiting platelet aggregation, antithrombotic formation, anti-oxidation and protecting cardiac microvascular endothelial cells; Fat-soluble components such as tanshinone can expand coronary arteries, improve coronary blood flow, protect myocardium and have broad-spectrum antibacterial effects. Panax notoginseng saponins are the main active ingredients of Panax notoginseng, which have anti-thrombosis, dilating blood vessels and protecting myocardium.

At present, most of the research reports about the fingerprint identification method of Danshen and Sanqi compound preparations are the identification of Danshen or Sanqi single medicinal material, such as the research on high performance liquid chromatography fingerprint of Danshen medicinal material ethanol extract (Chinese Journal of Pharmaceutical Sciences, August 2004, No. Volume 39, No. 8), research on fingerprints of Panax notoginseng (Strait Pharmaceutical Journal Vol 14 No52002). The prior art fails to simultaneously identify multiple types of active ingredients in Danshen and Notoginseng two herbs with fingerprints.

Invention content:

The purpose of the present invention is to establish a fingerprint that can simultaneously identify multiple types of active ingredients in Danshen and Sanqi compound prescriptions.

Another object of the present invention is to apply the fingerprint of the present invention to the quality control of the compound preparation containing Danshen and Panax notoginseng.

The present invention is achieved like this:

We simulated the prescription of Danqi Tablets (hereinafter referred to as Danqifang), and determined the HPLC-DAD fingerprint of Danqifang by studying the preparation method of Danqifang's test solution, the selection of chromatographic conditions and the methodology of chromatographic conditions. The fingerprint can meet the simultaneous detection and analysis of multiple types of active ingredients in Danshen and Panax notoginseng in Danqi prescription. We further used this fingerprint for the quality control of compound preparations containing Danshen and Panax notoginseng.

Concrete scheme of the present invention is as follows:

Using a high-performance liquid chromatograph, a DAD detector, and a chromatographic workstation, the compound solution of Salvia Miltiorrhiza and Radix Notoginseng was adopted with the following chromatographic conditions:

Chromatographic column: carbon octadecyl bonded silica gel packing column and pre-column;

Column temperature: 25-35°C;

Mobile phase: A is 0.1-0.5% phosphoric acid water; B is acetonitrile; A+B=100%, using gradient elution: 0-10min, 7-17%B, 10-12min, 17-20%B, 12 -16min, 20-21%B, 16-32min, 21%B, 32-40min, 21-29%B, 40-55min, 29-35%B, 55-65min, 35-70%B, 65-75min , 70-80% B, 75-80min, 80-97% B, 80-82min, 97-100% B, 82-85min, 100% B; or after 55 minutes the gradient is: 55-65min, 35-65 %B, 65-80min, 65-80%B, 80-85min, 80-100%B;

Flow rate: 1ml/min, at 22-28 minutes, the flow rate is 0.8ml/min;

The detection wavelength is 203, 281nm.

The preferred column temperature is 30°C.

The preferred concentration of phosphoric acid water is 0.1%, which is volume percentage.

The preparation method of the test solution is: take the pulverized Salvia miltiorrhiza and Notoginseng, add 85-95% ethanol to soak, extract in a water bath under reflux, let cool, and filter; Concentrate to near dryness, add 50-90% methanol to dissolve, and constant volume.

The preferred preparation method is as follows: take crushed salvia miltiorrhiza and Panax notoginseng in the same number, add 6-12 times the weight parts of 90% ethanol to soak, water bath reflux extraction, let cool, filter; 10-30 times the weight of the medicinal materials, let cool, filter, combine the filtrates, concentrate under reduced pressure at 50°C until nearly dry, add 70% methanol to dissolve, and constant volume.

The injection volume of the above-mentioned liquid chromatograph is preferably 10 μl.

The fingerprint identification method of the present invention needs a recording time of 85 minutes, which is 55-65 minutes for identifying Danqi Tablets and Compound Danshen Dripping Pills.

Below is the part test of the present invention:

1. Preparation of the test solution: Take an appropriate amount of crushed Salvia miltiorrhiza and Panax notoginseng, add 90% ethanol, soak, reflux in a water bath to extract for 2 hours, let cool, and filter; add appropriate amount of water to the dregs and continue to decoct for 2 hours, filter, and combine Filtrate, reclaim ethanol under reduced pressure to dryness, add 70% methanol to dissolve and transfer to volumetric flask and constant volume to mark, filter with microporous membrane, obtain need testing solution (0.024g crude drug/ml). Inject 10 μl for chromatographic analysis.

2. HPLC chromatographic analysis conditions:

Chromatographic column: carbon octadecyl bonded silica gel column (4.6×250mm ID, 5μm) and pre-column (4.6×12.5mmID, 5μm); column temperature: 30°C; mobile phase, A is 0.1% phosphoric acid water ; B is acetonitrile; A+B=100%, gradient elution: 0-10min, 7-17%B, 10-12min, 17-20%B, 12-16min, 20-21%B, 16-32min , 21%B, 32-40min, 21-29%B, 40-55min, 29-35%B, 55-65min, 35-70%B, 65-75min, 70-80%B, 75-80min, 80 -97%B, 80-82min, 97-100%B, 82-85min, 100%B; or after 55min the gradient is: 55-65min, 35-65%B, 65-80min, 65-80%B , 80-85min, 80-100% B. Flow rate: 1ml/min (22-28min, 0.8ml/min); detection wavelength 203, 281nm. The recording time is 85min.

3 Selection of chromatographic conditions and results

3.1 Elution system selection and results

Three mobile phase systems were selected in this experiment: (1) acetonitrile-water gradient elution; (2) acetonitrile-0.1%-0.5% phosphoric acid gradient elution; (3) acetonitrile-0.1% formic acid gradient elution.

The results show that the acetonitrile-0.1%-0.5% phosphoric acid gradient elution system has more absorption peaks, better peak shape and stable baseline, which is better than acetonitrile-0.1% formic acid and acetonitrile-water gradient elution system. The gradient elution effect of acetonitrile-0.1%-0.5% phosphoric acid is equivalent, and the concentration of 0.1% phosphoric acid is lower, and the loss to the chromatographic column is small, so the gradient elution of acetonitrile-0.1% phosphoric acid is more optimal.

3.2 Selection and results of elution gradient, column temperature, and flow rate

Five elution gradients, column temperature (25-35°C) and flow rate were investigated. The result is OK:

Elution gradient: 0-10min, 7-17% B, 10-12min, 17-20% B, 12-16min, 20-21% B, 16-32min, 21% B, 32-40min, 21-29% B, 40-55min, 29-35% B, 55-65min, 35-70% B, 65-75min, 70-80% B, 75-80min, 80-97% B, 80-82min, 97-100% B, 82-85min, 100% B (mobile phase A is 0.1% phosphoric acid water; B is acetonitrile; A+B=100%). Or after 55 minutes the gradient is: 55-65min, 35-65%B, 65-80min, 65-80%B, 80-85min, 80-100%B. Preferred column temperature: 30°C

3.3 Determination of detection wavelength

The main components of Panax notoginseng in Danqifang extract are notoginseng saponins; the main components of Danshen are water-soluble salvianolic acids and lipid-soluble salvianolic quinones. According to the ultraviolet absorption characteristics of these components, five wavelengths of 203, 254, 270, 281 and 286nm were set to investigate the chromatographic conditions. There are different absorptions, 281nm is better than 254, 270 and 286nm to take into account the water-soluble and fat-soluble components of Danshen. Comprehensive consideration, choose two wavelengths of 203nm and 281nm.

3.4 The method and result of Danqifang test product solution preparation

In this experiment, the process of Danqifang was optimized, so that the test solution of Danqifang could fully reflect its active ingredients, and at the same time reduce the interference of impurities such as sugar and tannin.

Test solution 1: Take 0.6g of crushed salvia miltiorrhiza and 0.6g of notoginseng, add 30ml of water to soak for 1h, decoct for 2h, let cool, filter, and concentrate the filtrate at 50°C to nearly dryness under reduced pressure, add 70% methanol to dissolve and transfer Dilute to 50ml volumetric flask.

Test solution 2: Take 0.6g of crushed salvia miltiorrhiza and 0.6g of notoginseng, add 30ml of water to soak for 1h, decoct for 2h, let cool, filter, concentrate the filtrate to 2ml at 50°C under reduced pressure, add 3 times the amount of 95% ethanol Precipitate with alcohol, let stand in the refrigerator to pass through the liquid, filter, concentrate the filtrate at 50°C under reduced pressure to near dryness, add 70% methanol to dissolve and transfer to a 50ml volumetric flask to constant volume.

Test solution 3: Take 0.6g of crushed salvia miltiorrhiza and 0.6g of notoginseng, add 10ml of 90% ethanol to soak for 1h, reflux in a water bath to extract for 2h, let cool, filter, and concentrate the filtrate at 50°C to nearly dryness under reduced pressure, add 70% methanol Dissolve and transfer to a 50ml volumetric flask to volume.

Test solution 4: Take 0.6g of crushed salvia miltiorrhiza and 0.6g of notoginseng, add 10ml of 70% ethanol to soak for 1h, extract in a water bath for 2h under reflux, let cool, filter, continue to extract the dregs with 70% ethanol once, combine the filtrate, The filtrate was concentrated to near dryness under reduced pressure at 50°C, dissolved in 70% methanol and transferred to a 50ml volumetric flask to constant volume.

Test solution 5: take 0.6g of crushed salvia miltiorrhiza and 0.6g of notoginseng, add 10ml of 90% ethanol to soak for 1h, extract in a water bath for 2h under reflux, let cool, and filter; add 10ml of 50% ethanol to the dregs to continue extracting once, and combine the filtrate , The filtrate was concentrated to near dryness under reduced pressure at 50°C, dissolved in 70% methanol and transferred to a 50ml volumetric flask to constant volume.

Test solution 6: Take 0.6g of crushed salvia miltiorrhiza and 0.6g of notoginseng, add 10ml of 90% ethanol to soak for 1h, reflux in a water bath to extract for 2h, let cool, and filter; add 30ml of water to decoct the dregs of the medicine for 2h, let cool, filter, Concentrate the filtrate to 2ml at 50°C under reduced pressure, add 3 times the amount of 95% ethanol for ethanol precipitation, let the liquid stand in the refrigerator, filter, combine the filtrate with 90% ethanol extraction filtrate, concentrate under reduced pressure at 50°C to nearly dryness, add 70% Dissolve in methanol and transfer to a 50ml volumetric flask to volume.

Test solution 7: Take 0.6g of crushed salvia miltiorrhiza and 0.6g of notoginseng, add 10ml of 90% ethanol to soak for 1h, reflux in a water bath to extract for 2h, let cool, and filter; add 30ml of water to decoct the dregs of the medicine for 2h, let cool, filter, The filtrates were combined, concentrated under reduced pressure at 50°C to near dryness, dissolved in 50% methanol and transferred to a 50ml volumetric flask to constant volume.

Test solution 8: Take 0.6g of crushed salvia miltiorrhiza and 0.6g of notoginseng, add 10ml of 90% ethanol to soak for 1h, reflux in a water bath to extract for 2h, let cool, and filter; add 30ml of water to decoct the dregs of the medicine for 2h, let cool, filter, The filtrates were combined, concentrated under reduced pressure at 50°C to near dryness, dissolved in 60% methanol and transferred to a 50ml volumetric flask to constant volume.

Test solution 9: take 0.6g of crushed salvia miltiorrhiza and 0.6g of notoginseng, add 10ml of 90% ethanol to soak for 1h, extract in a water bath for 2h under reflux, let cool, and filter; add 30ml of water to decoct the dregs of the medicine for 2h, let cool, filter, The filtrates were combined, concentrated under reduced pressure at 50°C to near dryness, dissolved in 70% methanol and transferred to a 50ml volumetric flask to constant volume.

Prepare each test solution of Danqifang, and automatically inject 10 μl into the liquid chromatograph for chromatographic analysis. The results show that: simple water extraction, 70% ethanol and 90% ethanol extraction can not take into account the water-soluble and fat-soluble components of Danqi Fang; The effect is good, but the alcohol precipitation of the water extraction part has a slight loss of protocatechualdehyde, so alcohol precipitation is not necessary: the sample is dissolved in 70% methanol than 50% and 60% methanol, which can better balance water solubility and fat solubility Element. In summary, the optimal preparation process for test solution 9 was selected.

3.5 Methodological investigation of detection methods

3.5.1 The test solution stability and instrument precision experiment

Prepare the Danqifang test product solution according to the method under "1", and detect the fingerprints at 0, 2, 4, 12, 18, and 24 hours respectively (see Figure 1-1, 1-2), and investigate the Danqifang for The stability of the test solution and the precision of the high performance liquid chromatography. The results showed that the test solution of Danqifang was stable within 24 hours, and the precision of the high performance liquid chromatography also met the requirements.

3.5.2 Repeatability experiment for the determination of the test solution

Take the same batch of Danshen and Panax notoginseng crude powder (40 mesh), prepare 3 parts of Danqifang test solutions in parallel according to the method under "1", detect the HPLC fingerprints (see Figure 2-1, 2-2), and investigate The extraction method of Danqifang and the stability and repeatability of the fingerprint detection method. The results showed that the preparation method of Danqifang test solution was stable and reliable, with good reproducibility.

3.5.3 Determination of recording time

The 2-hour record chart and the blank control chart (see Figure 3-1, 3-2) of the Danqifang test solution under the chromatographic condition of item "2" are used to investigate the best record time. The results show that after 80 minutes, there is basically no component in the sample, so the best recording time is determined to be 85 minutes.

3.5.4 Standard control experiment:

Protocatechualdehyde, salvianolic acid B, cryptotanshinone, tanshinone IIA, notoginsenoside R1, ginsenoside Rg1, and ginsenoside Rb1 were used as reference substances, and an appropriate amount was accurately weighed, dissolved in 70% methanol to prepare a standard solution. Record the chromatogram (see Figure 4). in conclusion:

Through the research on the preparation method of Danqifang's test solution, the selection of chromatographic conditions and the methodology of chromatographic conditions, the determined chromatographic analysis conditions of Danqifang can meet the simultaneous detection of multiple types of active ingredients in Danshen and Panax notoginseng and analyse.

We further used this fingerprint condition for the quality control of compound preparations containing Danshen and Panax notoginseng. details as follows:

1. Preparation of the test solution of the compound salvia miltiorrhiza preparation:

Take an appropriate amount of Danqi Tablets (remove sugar coating), Compound Danshen Tablets and Compound Danshen Dropping Pills (remove film coating), grind into powder, dissolve with 70% methanol and transfer to a volumetric flask to constant volume. Ultrasonic extraction, let cool, add 70% methanol to make up the weight. The solution is obtained by passing through a 0.45 μm filter membrane.

2. Use the fingerprint conditions of the present invention to carry out HPLC-DAD chromatographic analysis on the test solution of the compound Danshen preparation to obtain the fingerprint of the compound Danshen preparation.

Description of drawings

Figure 1-1 Experimental chromatograms (203nm) of test solution stability and instrument precision, in which chromatograms WDQ00135～WDQ00140 are 0～24 hours chromatograms respectively

Figure 1-2 Experimental chromatograms (281nm) for the stability of the test product solution and instrument precision, in which the chromatograms WDQ00135～WDQ00140 are the chromatograms of 0～24 hours respectively

The repeatability experiment chromatogram (203nm) that Fig. 2-1 need testing solution measures

The repeatability experiment chromatogram (281nm) that Fig. 2-2 need testing solution measures

Figure 3-1 Chromatogram of recording time experiment (203nm: where WDQ00120 is blank; WDQ00121 is the test product)

Figure 3-2 Chromatogram of recording time experiment (281nm: where WDQ00120 is blank; WDQ00121 is the test product)

The chromatogram of Fig. 4 standard control test

Fig. 5 is Dan Qifang fingerprint collection in embodiment 1

Fig. 6 is the fingerprint collection of commercially available Danqi tablets in embodiment 2

Fig. 7 is the fingerprint of the Compound Danshen Tablets whose commercially available batch number is 040602 in Example 3

Fig. 8 is the fingerprint of the Compound Danshen Tablets whose commercially available batch number is 050308 in Example 3

Fig. 9 is the fingerprint of the Compound Danshen Tablets whose commercially available batch number is 03121024 in Example 3

Fig. 10 is the fingerprint of the Compound Danshen Dripping Pills whose batch number is 20020921 in Example 4

Fig. 11 is the fingerprint of the Compound Danshen Dripping Pills whose batch number is 20030604 in Example 4

Fig. 12 is the fingerprint of the Compound Danshen Dripping Pills whose batch number is 20040303 in Example 4

Fig. 13 is the fingerprint of Dan Qifang in Example 5.

Fig. 14 is the fingerprint spectrum of Danqi tablets in Example 5.

Fig. 15 is the fingerprint of Compound Danshen Tablets in Example 5.

Fig. 16 is the fingerprint of Compound Danshen Dripping Pills in Example 5.

Fig. 17 is the fingerprint spectrum of Danqi tablets in Example 6.

Fig. 18 is the fingerprint of Compound Danshen Dripping Pills in Example 6.

The numbers in the above drawings represent: 1. Protocatechualdehyde 2. Notoginsenoside R1 3. Ginsenoside Rg1 4. Salvianolic acid B 5. Ginsenoside Rb1 6. Cryptotanshinone 7. Tanshinone IIA

Detailed ways:

Example 1

1 Experimental materials

1.1 Instruments and equipment:

Agilent 1100 series high performance liquid chromatography, including G1312A quaternary gradient pump, G1313A autosampler, G1316A column oven, G1315A DAD detector, HP Chemstation chromatography workstation (U.S. Hewlett-Packard Company); rotary thin film evaporator (Switzerland) Büchi company).

1.2 Reagents and reagents:

Medicinal materials: Salvia miltiorrhiza (Shaanxi Tasly Plant Pharmaceutical Co., Ltd., Shangluo, Shaanxi), identified by Professor Li Ping as the dry root of Salvia miltiorrhiza Bge. The dried root of Panax notoginseng (Burk.) F.H.Chen.

2. Experimental method

2.1 Chromatographic conditions

Chromatographic column: C18 chromatographic column (ZORBAX ODS 4.6×250mm ID, 5μm) and C18 pre-column (4.6×12.5mmID, 5μm); column temperature: 30°C; mobile phase, A is 0.1% phosphoric acid water; B is acetonitrile; A+B=100%, gradient elution: 0-10min, 7-17%B, 10-12min, 17-20%B, 12-16min, 20-21%B, 16-32min, 21%B, 32-40min, 21-29%B, 40-55min, 29-35%B, 55-65min, 35-70%B, 65-75min, 70-80%B, 75-80min, 80-97%B, 80-82min, 97-100%B, 82-85min, 100%B; flow rate: 1ml/min (22-28min, 0.8ml/min); detection wavelength 203, 281nm. The recording time is 85min.

2.2 Preparation of the test solution

Take crushed salvia miltiorrhiza 1.2g, panax notoginseng 1.2g, add 25ml of 90% ethanol, soak for 1h, reflux in a water bath for 2h, let cool, and filter; add 40ml of water to the dregs and continue to decoct for 2h, filter, combine the filtrate, and put in 50 Recover ethanol under reduced pressure at ℃ to dryness, add 70% methanol to dissolve and transfer to a 100ml volumetric flask to set the volume to the mark, and filter through a microporous membrane to obtain the test solution.

3. Experimental results: 10 μl of the test solution was injected for chromatographic analysis, and the fingerprint of Dan Qifang was obtained, and the recording time was 85 minutes. See Figure 5.

Example 2

Take 20 commercially available Danqi Tablets (Hunan Anbang Pharmaceutical Co., Ltd., batch number: 040803), remove the sugar coating and grind them into powder, accurately weigh 0.5g, put in a 25ml brown volumetric flask, and dilute to volume with 70% methanol. Ultrasonic extraction was performed for 30 min, allowed to cool, and 70% methanol was added to make up the weight. The solution was passed through a 0.45 μm membrane filter. Other conditions are with embodiment 1. Inject 10 μl of sample for chromatographic analysis, and get the HPLC-DAD fingerprint of seven tablets of Dandan, as shown in Figure 6.

Example 3

Get commercially available Compound Danshen Tablets (Guangdong Baiyunshan Pharmaceutical Factory, batch number: 03121024; Nanjing Tongrentang Pharmaceutical Co., Ltd., batch number: 040602; Zhejiang Huqing Yutang Pharmaceutical Co., Ltd., batch number: 050308) 20 tablets, ground into powder, and accurately weighed Take 0.5g, put it in a 25ml brown volumetric flask, and dilute to volume with 70% methanol. Ultrasonic extraction was performed for 30 min, allowed to cool, and 70% methanol was added to make up the weight. The solution was passed through a 0.45 μm membrane filter. Other conditions are with embodiment 1. Inject 10 μl of sample for chromatographic analysis, and obtain the HPLC-DAD fingerprints of Compound Danshen Tablets from three manufacturers, as shown in Figure 7-9.

Example 4

Get 75 commercially available compound danshen dripping pills (Tianjin Tasly Pharmaceutical Co., Ltd., batch number: 20020921, 20030604, 20040303), accurately weighed, grind into powder, remove film coat, dissolve with 70% methanol and transfer to 25ml brown capacity Make up to volume in the bottle. Ultrasonic extraction was performed for 30 min, allowed to cool, and 70% methanol was added to make up the weight. The solution was passed through a 0.45 μm membrane filter. Other conditions are with embodiment 1. Inject 10 μl for chromatographic analysis, and obtain the HPLC-DAD fingerprints of 3 batches of Compound Danshen Dripping Pills. See Figure 10-12.

Example 5

Use the same method as the above example, but change the elution gradient to 55 minutes later: 55-65min, 35-65%B, 65-80min, 65-80%B, 80-85min, 80-100%B. Chromatographic analysis was carried out on Danqifang, Danqi Tablets, Compound Danshen Tablets (Guangdong Baiyunshan Pharmaceutical Factory, batch number: 03121024) and Compound Danshen Dripping Pills (Tianjin Tasly Pharmaceutical Co., Ltd., batch number: 20020921), and the fingerprints are shown in the figure 13-16.

Example 6

With the method of Example 1, 2, 4, only the recording time was changed to 60 minutes, and the chromatographic analysis was carried out to Danqi Tablets and Compound Danshen Dripping Pills (Tianjin Tasly Pharmaceutical Co., Ltd., batch number: 20020921) respectively. The obtained fingerprints are shown in Figure 17-18.

In summary, the HPLC-DAD fingerprints of the Danshen and Panax notoginseng compound established in the present invention can be used to simultaneously detect the water-soluble components of phenolic acids such as salvianolic acid B and the lipid-soluble components of diterpene quinones such as tanshinone IIA and Panax notoginseng The composition of saponins is reliable and stable, and it can be used for quality control of compound preparations containing Danshen and Panax notoginseng.

Claims (7)

1, the finger print atlas identifying method of the red sage root and notoginseng preparation: comprise the standard of foundation finger-print, with same procedure measure the finger-print of testing sample, with the finger-print and the standard finger-print contrast of product to be tested, its feature adopts following method acquisition finger-print:

Use high performance liquid chromatograph, the DAD detecting device, chromatographic work station, with the red sage root and notoginseng solution, adopt following chromatographic condition:

Chromatographic column: carbon octadecyl silane filler chromatographic column and pre-column;

Column temperature: 25-35 ℃;

Moving phase: A is the phosphoric acid water of 0.1-0.5%; B is an acetonitrile; A+B=100% adopts gradient elution: 0-10min, 7-17%B, 10-12min, 17-20%B, 12-16min, 20-21%B, 16-32min, 21%B, 32-40min, 21-29%B, 40-55min, 29-35%B, 55-65min, 35-70%B, 65-75min, 70-80%B, 75-80min, 80-97%B, 80-82min, 97-100%B, 82-85min, 100%B; Perhaps gradient is after 55 minutes: 55-65min, 35-65%B, 65-80min, 65-80%B, 80-85min, 80-100%B;

Flow velocity: 1ml/min, in the time of 22～28 minutes, flow velocity is 0.8ml/min;

Detect wavelength: 203,281nm.

2, the finger print atlas identifying method of claim 1, wherein column temperature is 30 ℃.

3, the finger print atlas identifying method of claim 1, wherein the concentration of phosphoric acid water is 0.1%.

4, the finger print atlas identifying method of claim 1, wherein the preparation method of standard diagram need testing solution is: get the red sage root and the pseudo-ginseng of pulverizing, add the 85-95% alcohol immersion, the water-bath refluxing extraction is put coldly, filters; Dregs of a decoction boiling is put coldly, filters, and merging filtrate is concentrated near doing, and adds the 50-90% dissolve with methanol, constant volume.

5, the finger print atlas identifying method of claim 4, wherein the preparation method of standard diagram need testing solution is: get the red sage root and the pseudo-ginseng of the same umber of pulverizing, add 6-12 times of parts by weight 90% alcohol immersion, the water-bath refluxing extraction is put coldly, filters; The dregs of a decoction add poach, amount of water be dregs of a decoction weight 10-30 doubly, put coldly, filter, merging filtrate is evaporated near doing in 50 ℃, adds 70% dissolve with methanol, constant volume.

6, the finger print atlas identifying method of claim 1, wherein the preparation method of testing sample is: with testing sample preparation desaccharification clothing or removal film-coating, pulverize, with 70% dissolve with methanol and constant volume, ultrasonic Extraction, put cold, add 70% methyl alcohol and supply weight, filter, promptly.

7, the finger print atlas identifying method of claim 1, liquid chromatograph sample size are 10 μ l.

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