Rapid qualitative and quantitative detection method for quantitative analysis of multi-components by single marker of rhodiola rosea, astragalus membranaceus and safflower capsules

Abstract

The invention discloses a rapid qualitative and quantitative detection method for quantitative analysis of multiple components by single marker of rhodiola rosea, astragalus membranaceus and safflower capsules, and belongs to a detection and analysis technology. According to the method, three traditional Chinese medicines, namely rhodiola rosea, astragalus membranaceus and safflower carthamus, are identified on a thin-layer plate by adopting a thin-layer chromatography method and a same test solution, the pretreatment of a sample and a reference medicinal material only needs 3 hours, 265 mL of a solvent is adopted, spots are clear, and the method is rapid; according to the established single-detection double-evaluation method for salidroside and calycosin-7-glucoside, only an ultraviolet detector is needed, detection is carried out under the wavelength of 270 nm, chromatographic peaks do not interfere with one another, single-detection double-evaluation is achieved, the efficiency is high, and the cost is low. The invention breaks through a traditional identification mode that N traditional Chinese medicine varieties are identified, N test solutions and N thin-layer plates are prepared, N times of development are needed, and N medicinal materials are identified, and establishes a rapid thin-layer identification method for three traditional Chinese medicines on one thin-layer plate by adopting the same test solution and each reference medicinal material supernatant subjected to methanol ultrasonic treatment.

Description

Technical Field

[0001] The invention relates to a one-test-multiple-evaluation rapid qualitative and quantitative detection method for rhodiola rosea, astragalus and safflower capsules. The rapid qualitative method refers to a rapid thin-layer identification of three medicinal materials, rhodiola rosea, astragalus and safflower, using a thin-layer identification method; the rapid quantitative method refers to a high-performance liquid chromatography method, which only needs to use an ultraviolet detector to quantify two components, and has a lower cost than a DAD detector, and realizes a one-test-double-evaluation quantitative determination of salidroside and calycosin isoflavone glucoside. It belongs to the detection and analysis technology. Background Art

[0002] As people continue to deepen their research on Chinese medicinal materials, more and more Chinese medicinal materials are used in health products, especially Rhodiola Rosea Astragalus Safflower Capsules, which contain three Chinese medicinal materials, namely Rhodiola Rosea, Astragalus and Safflower. Its product instructions stipulate that the characteristic ingredient is salidroside, and its content per 100 grams should not be less than 0.4 grams. There is no content under the [Identification] item in its technical requirements. As the regulatory authorities continue to improve their requirements for product quality control, the regulatory authorities have proposed to establish product identification items for health products that are being re-registered, that is, to establish exclusive identification of each Chinese medicinal piece in the product, which will then be used to determine whether each Chinese medicinal piece in the product has been put into use, thereby ensuring the quality of the product.

[0003] The identification of traditional Chinese medicine in health products can refer to the thin layer identification method used in the quality inspection of Chinese medicinal materials and Chinese patent medicines. In the quality inspection of traditional Chinese medicine, thin layer identification is mostly the identification of one Chinese medicinal material. If there are N identifications, it is necessary to prepare N kinds of test sample solutions, N thin layer plates, and unfold them N times to identify N medicinal materials in the traditional identification mode. In order to eliminate interference, the sample pretreatment procedures are complicated and cumbersome, and a large amount of organic reagents are required for repeated purification, which is laborious, time-consuming, and reagent-consuming, pollutes the environment, endangers health, and has a long detection cycle. Therefore, finding a simple and fast detection method, improving detection efficiency, and reducing detection costs are also difficulties that must be overcome in the quality control of traditional Chinese medicinal materials in future health products. Taking the quality standard of Yiqing Capsules included in the 2020 edition of the Chinese Pharmacopoeia as an example, the analysis is as follows:

[0004] [Identification] (1) Take 0.5g of the content of this product, add 20ml of methanol, soak for 1 hour, filter, take 10ml of the filtrate, evaporate to dryness, add 10ml of water to the residue to dissolve it, add 1ml of hydrochloric acid, heat and reflux on a water bath for 30 minutes, cool immediately, shake and extract with ether twice, 20ml each time, combine the ether extracts, evaporate to dryness, add 1ml of chloroform to the residue to dissolve it, and use it as the test solution. Take 0.1g of rhubarb reference medicinal material and prepare the reference medicinal material solution in the same way. Take the rhubarb reference substance and add chloroform to prepare a solution containing 1mg per 1ml, which is used as the reference substance solution. According to the thin layer chromatography method (General Rule 0502), 4 μl of each of the three solutions mentioned above were taken and spotted on the same silica gel G thin layer plate, and the upper layer solution of petroleum ether (30-60°C)-ethyl formate-formic acid (15:5:1) was used as the developing agent. The plate was developed, taken out, dried, and examined under ultraviolet light (365nm). In the chromatogram of the test sample, the same orange-yellow fluorescent spot appeared at the corresponding position of the chromatogram of the reference medicinal material; the same orange-yellow fluorescent spot appeared at the corresponding position of the chromatogram of the reference substance; after being fumigated with ammonia vapor and examined under sunlight, the same red spot appeared.

[0005] (2) Take 1g of the content of this product, add 15ml of methanol, ultrasonically treat for 15 minutes, filter, evaporate the filtrate, add 20ml of water to the residue, heat to dissolve, adjust the pH to 3-4 with hydrochloric acid, shake and extract twice with ethyl acetate, 20ml each time, combine the ethyl acetate solution, wash with 30ml of 0.05mol / L sulfuric acid solution, and then wash with water until neutral, take the ethyl acetate solution, evaporate to dryness, add 1ml of methanol to the residue to dissolve, and use it as the test solution. Take another baicalin reference substance, add methanol to make a solution containing 1mg per 1ml, and use it as the reference solution. According to the thin layer chromatography method (General Rule 0502), take 5μl of each of the above two solutions and spot them on the same silica gel G thin layer plate with 4% sodium acetate sodium carboxymethyl cellulose solution as the adhesive, and use ethyl acetate-butanone-formic acid-water (5:3:1:1) as the developing solvent, develop, take out, dry, and spray with 2% ferric chloride ethanol solution. In the test sample chromatogram, a spot of the same color appears at the corresponding position in the reference sample chromatogram.

[0006] (3) Take 0.5g of the content of this product, add 10ml of methanol, soak for 30 minutes, shake from time to time, filter, add methanol to the filtrate to 10ml, as the test solution. Take another 50mg of the reference medicinal material of Coptis chinensis, add 5ml of methanol, soak for 30 minutes, shake from time to time, filter, add methanol to the filtrate to 5ml, as the reference medicinal material solution. Take berberine hydrochloride reference substance, add methanol to make a solution containing 0.5mg per 1ml, as the reference substance solution. According to the thin layer chromatography method (General Rule 0502), take 1μl of each of the above three solutions and spot them on the same silica gel G thin layer plate, use toluene-ethyl acetate-isopropanol-methanol-water (4:2:1:1:0.2) as the developing solvent, develop in a developing cylinder saturated with ammonia vapor, take out, dry, and examine under ultraviolet light (365nm). In the test sample chromatogram, the same yellow fluorescent spot appears at the corresponding position in the control medicinal material chromatogram; the same yellow fluorescent spot appears at the corresponding position in the control sample chromatogram.

[0007] Determination of Scutellaria baicalensis content

[0008] The chromatographic conditions and system suitability test used octadecylsilane bonded silica gel as filler, methanol-water-phosphoric acid (47:53:0.2) as mobile phase, and the detection wavelength was 280nm. The theoretical plate number calculated based on the baicalin peak should be no less than 3000.

[0009] Preparation of reference solution: Take about 12.5 mg of baicalin reference substance, weigh accurately, place in a 250 ml volumetric flask, dissolve with appropriate amount of methanol, dilute to scale with mobile phase, shake well, and obtain (each 1 ml contains 50 μg of baicalin).

[0010] Preparation of test solution Take the contents of this product under the item of filling volume difference, mix well, take out about 0.1g, accurately weigh, place in a 100ml volumetric flask, add 50ml of mobile phase, ultrasonically treat (power 250W, frequency 50kHz) for 30 minutes, cool, add mobile phase to the scale, shake well, centrifuge, accurately measure 25ml of supernatant, place in a 50ml volumetric flask, add mobile phase to the scale, shake well, and obtain.

[0011] Determination method: Accurately pipette 10μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0012] Each capsule of this product contains baicalin C 21 H 18 O 11 It shall not be less than 30.0mg.

[0013] Determination of rhubarb content

[0014] The chromatographic conditions and system suitability test used octadecylsilane bonded silica gel as filler, methanol-0.1% phosphoric acid (85:15) as mobile phase, and the detection wavelength was 254 nm. The theoretical plate number calculated based on the rhein peak should not be less than 2000.

[0015] Preparation of reference solution: Take appropriate amount of rhubarb reference substance and chrysophanol reference substance, weigh accurately, add methanol to make a solution containing 10 μg of rhubarb reference substance and chrysophanol per 1 ml.

[0016] Preparation of test solution Take the contents of this product under the item of filling amount difference, mix well, take about 0.1g, weigh accurately, place in a 100ml conical flask, add 10ml of 2.5mol / L sulfuric acid solution, ultrasonically treat (power 250W, frequency 50kHz) for 5 minutes, then add 15ml of chloroform, heat reflux in a 70℃ water bath for 30 minutes, cool, transfer to a separating funnel, take chloroform solution, add chloroform to the acid solution and heat reflux twice (10ml, 10ml), each time for 20 minutes, take chloroform solution, and extract the acid solution with 10ml of chloroform by shaking, combine the chloroform solutions, evaporate to dryness, dissolve the residue with methanol, transfer to a 10ml volumetric flask, add methanol to the scale, shake well, filter, and take the filtrate to obtain.

[0017] Determination method: Accurately pipette 10μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0018] Each capsule of this product contains rhubarb and rhubarb C 15 H 10 O5 and Chrysophanol C 15 H 10 The total amount of O4 shall not be less than 0.70 mg.

[0019] For the three thin-layer identifications in the above example, a rough calculation shows that the sample pretreatment time alone for the three thin-layer identifications will take 8 hours, with about 3g of sample, 360ml of organic solvent (including developing agent), and 3 hours of developing time. The thin-layer identification of a batch of samples will take about 2 days and nearly 400ml of organic solvent. Adding the 3-day quantitative determination time of Scutellaria baicalensis and Rhubarb, 465ml of sample treatment solvent, 1600ml of organic phase in the mobile phase, it will take 5 days to complete the thin-layer identification and quantitative determination of a batch of samples, and 2500ml of organic reagent. In case of retesting, it will take half a month to get the test results, and the detection speed cannot match the mechanized large-scale production. There are many quality control methods for Chinese herbal compound preparations like the analysis examples, so improving detection efficiency, reducing detection costs, and reducing environmental pollution have become urgent goals for testers. Under this background, we established thin layer identification items for Rhodiola rosea, Astragalus and Safflower Capsules, and at the same time added the content determination of characteristic ingredients in Rhodiola rosea and Astragalus. We comprehensively carried out research on improving product standards, improving and simplifying standards, and invented this one-test-two-evaluation rapid qualitative and quantitative detection method.

[0020] The prescription composition and preparation method of Rhodiola rosea, Astragalus and Safflower Capsules are product confidential of the company and are not convenient to describe. Summary of the invention

[0021] Using thin layer chromatography method and the same test solution, three Chinese medicinal herbs, Rhodiola rosea, Astragalus membranaceus and Carthamus tinctorius, were identified on a thin layer plate. The pretreatment of samples and control herbs only required 3 hours, 265 mL of solvent, clear spots and a quick method. The established one-measurement, two-evaluation quantitative method for salidroside and calycosin isoflavone glucoside only required an ultraviolet detector and detection at a wavelength of 270 nm. The chromatographic peaks did not interfere with each other, and one-measurement, two-evaluation was highly efficient and low cost.

[0022] The invention uses 20g of test sample for thin layer identification of three medicinal materials and quantitative determination of two effective ingredients, and the pretreatment time is 4h plus the time and solvent for thin layer development and quantitative determination, and the total cost of solvent is 2000ml, and the time is 2 days. Compared with the above-mentioned Yiqing capsule, the thin layer identification and quantitative determination of a batch of samples can be completed, which can save 3 days and 1000ml of organic reagent. Therefore, the invention greatly improves the social benefits of the controllability of preparation quality, and further improves the detection efficiency, saves 60% of the detection time, reduces the detection cost, reduces environmental pollution, and forms a set of simple, fast, efficient, low-consumption, low-pollution, multi-index modern standards for controlling drug quality.

[0023] The solution adopted by the present invention to solve the technical problem is:

[0024] Rapid Thin Layer Identification Method

[0025] Take 10g of the contents of Rhodiola rosea, Astragalus and Safflower capsules, grind them into powder, add 50mL of 50% ethanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, and add 30mL of water to the residue to dissolve it. Shake and extract with ethyl acetate twice, 30mL each time, combine the ethyl acetate extracts, evaporate to dryness, and add 2mL of methanol to the residue to dissolve it as the test solution. Take 1g of Rhodiola rosea control medicinal material, 1g of Astragalus control medicinal material, and 0.1g of Safflower control medicinal material, add 10mL of 50% ethanol respectively, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, and add 20mL of water to the residue to suspend it. Extract twice with ethyl acetate by shaking, 20 mL each time, combine the ethyl acetate extracts, evaporate to dryness, add 1 mL of methanol to the residue to dissolve, take 2-4 μL of the above test solution and reference medicinal material solution, respectively, and spot them on the same silica gel G thin layer plate containing 1% NaOH, use chloroform-methanol (15:1) as the developing agent, develop, take out, dry, and examine under ultraviolet light (365 nm). The test sample shows the same color fluorescent spot at the chromatographic position as the reference medicinal material of Rhodiola rosea, reference medicinal material of Astragalus, and reference medicinal material of Carthamus tinctorius, and there is no interference with the negative.

[0026] (2) Rapid determination method of salidroside and calycosin with one test and two evaluations

[0027] The chromatographic conditions and system suitability test used octadecylsilane bonded silica gel as the filler; acetonitrile (A)-0.2% phosphoric acid solution (B) as the mobile phase for gradient elution: 0-15 min, 13% A, 15-30 min, 13%-17% A, 30-35 min, 17% A; detection wavelength: 270 nm; column temperature of 35°C; the number of theoretical plates calculated based on the salidroside peak should be no less than 3000.

[0028] Preparation of reference solution: Take appropriate amount of salidroside and calycosin glucoside reference substances respectively, weigh accurately, add methanol to prepare a mixed solution containing 289.48 μg salidroside and 254.98 μg calycosin glucoside per 1 mL;

[0029] Preparation of test solution: Take an appropriate amount of the contents of Rhodiola rosea, Astragalus and Safflower Capsules, grind them into powder, take 10 g, weigh it accurately, put it in a stoppered conical flask, accurately add 50 mL of 50% ethanol, stopper it tightly, weigh it, and treat it ultrasonically at a power of 100 W and a frequency of 40 kHz for 30 minutes. Let it cool, weigh it again, make up the lost weight with 50% methanol, shake it well, filter it, evaporate the filtrate to dryness, dissolve the residue in 30 mL of water, extract it twice with n-butanol by shaking, 30 mL each time, combine the n-butanol extracts, evaporate them to dryness, dissolve the residue in 2 mL of methanol as the test solution, and prepare the negative sample solution in the same way.

[0030] The determination method is to accurately aspirate 10 μL of the reference solution, the test solution, and the negative sample solution, inject them into the liquid chromatograph, and determine the result.

[0031] The principle of the present invention is as follows:

[0032] 1. According to the different chemical structures and polarities of each active ingredient of traditional Chinese medicine, as the developing agent moves, the adsorption and desorption abilities on the thin layer plate are different, so that the spots of each active ingredient can be separated. In addition, by taking advantage of the polarity of each ingredient, active ingredients with similar polarity are selected, and under different inspection conditions, different spot colors are presented, overlapping each other, but not interfering with each other. Several medicinal materials and the same test solution can be tested on the same thin layer plate for multiple identification applications.

[0033] 2. By adjusting the components and proportions of the mobile phase, effective ingredients with different properties and contents can all show good chromatographic peak separation, no interference, appropriate peak area in the same mobile phase, and a good linear relationship between the chromatographic peak area and the content, so that quantitative determination can be achieved.

[0034] The innovative features and beneficial effects of the present invention are as follows:

[0035] (1) A breakthrough was made in the traditional identification method that one Chinese medicine variety has N items for identification, and N test sample solutions, N thin layer plates, and N times of development to identify N medicinal materials. A rapid thin layer identification method for three Chinese medicines was established using the same test sample solution and the supernatant of each control medicinal material by methanol ultrasound. This method not only has the innovation of breaking the routine, but also has the effects of improving detection efficiency, saving detection costs, and reducing environmental pollution. (2) The present invention also quantifies the specific components of each Chinese medicinal material in the Rhodiola rosea, Astragalus and Safflower Capsules. Two components of the three Chinese medicines were quantified, and the determination was carried out at the same detection wavelength in a small gradient elution method. The detection wavelength: Salidroside and calycosin glucoside are both 270nm, the size of each peak is appropriate, Salidroside and calycosin glucoside do not interfere, one test and two evaluations are simple, fast, accurate, low cost, high efficiency, and conducive to market supervision.

[0036] (3) Compared with the current quality inspection methods for Chinese medicine preparations with more than three ingredients, the present invention not only greatly improves the quality controllability of the preparations, but also improves the inspection efficiency by more than 5 times, saves 80% of the inspection time, reduces the inspection cost by 80%, and reduces environmental pollution by 90%, thus forming a set of simple, fast, efficient, low-cost, low-pollution, multi-index modern standards for controlling the quality of drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1The TLC diagram of Rhodiola rosea, Astragalus membranaceus and Carthamus tinctorius in ... capsules under 365 nm; wherein, 1-Rhodiola rosea control medicinal material; 2-sample; 3-Rhodiola rosea negative control; 4-Astragalus membranaceus control medicinal material; 5-sample; 6-Astragalus membranaceus negative control; 7-Carthamus tinctorius control medicinal material; 8-sample; 9-Carthamus tinctorius negative control;

[0038] Figure 2 is the injection amount and peak area of ​​salidroside, where the horizontal axis is the injection amount ug and the vertical axis is the peak area;

[0039] Figure 3 is the injection amount and peak area of ​​calycosin isoflavone glucoside, wherein the abscissa is the injection amount ug and the ordinate is the peak area;

[0040] Figure 4 This is the chromatogram of salidroside reference substance;

[0041] Figure 5 This is the chromatogram of Rhodiola Rosea Astragalus Safflower Capsule;

[0042] Figure 6 This is the chromatogram of the blank sample lacking Rhodiola rosea;

[0043] Figure 7 This is the chromatogram of the reference substance of calycosin isoflavone glucoside;

[0044] Figure 8 This is the chromatogram of Rhodiola Rosea Astragalus Safflower Capsule;

[0045] Fig. 9 This is the chromatogram of the blank sample lacking Astragalus.

[0046] Specific embodiment

[0047] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments. The following embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0048] A one-test-multiple-evaluation rapid qualitative and quantitative detection method for Rhodiola rosea, Astragalus membranaceus and Safflower Capsules comprises the following steps:

[0049] (1) One-test-multiple-evaluation rapid qualitative identification method

[0050] Take 10g of the contents of Rhodiola rosea, Astragalus membranaceus and Safflower capsules, grind them into powder, add 50mL of 50% ethanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, and dissolve the residue in 30mL of water; shake and extract with ethyl acetate twice, 30mL each time, combine the ethyl acetate extracts, evaporate to dryness, and dissolve the residue in 2mL of methanol as the test solution; take 1g of Rhodiola rosea control medicinal material, 1g of Astragalus control medicinal material, and 0.1g of Safflower control medicinal material, add 10mL of 50% ethanol respectively, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, and dissolve the residue in 20mL of water; shake and extract with ethyl acetate twice, 20mL each time, combine the ethyl acetate extracts, evaporate to dryness, and dissolve the residue in 1mL of methanol, and draw 2-4μL of the above test solution and control medicinal material solution, respectively, and apply them on the same surface containing 1% On the silica gel G thin layer plate with NaOH, chloroform-methanol (15:1) was used as the developing agent, developed, taken out, dried, and examined under ultraviolet light (365nm); the test sample showed the same color fluorescent spots at the chromatographic position as the control medicinal materials of Rhodiola rosea, Astragalus membranaceus, and Carthamus tinctorius;

[0051] (2) Rapid determination method of salidroside and calycosin with one test and two evaluations

[0052] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler; acetonitrile (A)-0.2% phosphoric acid solution (B) was used as mobile phase for gradient elution: 0-15min, 13% A, 15-30min, 13%-17% A, 30-35min, 17% A; detection wavelength: 270nm; column temperature: 35°C; theoretical plate number calculated based on salidroside peak should be not less than 3000;

[0053] Preparation of reference solution: Take appropriate amount of salidroside and calycosin glucoside reference substances, weigh accurately, add methanol to prepare a mixed solution containing 289.48 μg salidroside and 254.98 μg calycosin glucoside per 1 ml respectively;

[0054] Preparation of test solution Take an appropriate amount of the contents of Rhodiola rosea, Astragalus and Safflower Capsules, grind them into powder, take 10 g, weigh accurately, put it in a stoppered conical flask, accurately add 50 ml of 50% ethanol, stopper it tightly, weigh it, treat it with ultrasound at a power of 100 W and a frequency of 40 kHz for 30 minutes, let it cool, weigh it again, make up the lost weight with 50% methanol, shake it well, filter it, evaporate the filtrate to dryness, add 30 mL of water to dissolve the residue, shake and extract it twice with n-butanol, 30 mL each time, combine the n-butanol extracts, evaporate to dryness, add 2 mL of methanol to dissolve the residue, and use it as the test solution. Prepare the negative sample solution in the same way;

[0055] The determination method is to accurately aspirate 10 μL of the reference solution, the test solution, and the negative sample solution, inject them into the liquid chromatograph, and determine the result.

[0056] The rapid qualitative method refers to the thin layer identification method; the rapid quantitative method refers to the high performance liquid chromatography method, which only needs to use an ultraviolet detector to quantify the two components, and has lower cost than the DAD detector.

[0057] Through methodological investigation, the injection amount of salidroside was between 3.4737 and 34.7374 μg, which showed a good linear relationship with the peak area. The regression equation was: y = 136311x + 504598, r = 0.9998 (Table 1, Figure 2 ); the injection amount of calycosin isoflavone glucoside was 0.7649-7.6493 μg, which showed a good linear relationship with the peak area, and the regression equation was: Y=111476X+5875.9, r=0.9998 (Table 2, Figure 3 ); The results of the sample recovery experiment showed that the average recovery rate of salidroside was 100.30% for 6 determinations, and the RSD was 1.47% (Table 3); the average recovery rate of calycosin glucoside was 99.95% for 6 determinations, and the RSD was 1.10% (Table 4); the precision (Table 5), stability (Table 6), repeatability (Table 7), specificity ( Figure 4 , 5 ,6, Figure 7 , 8 , 9) The data all meet the methodological requirements. Table 8 shows the content determination results of two components in Hongjingtian Huangqi Honghua Capsule (mg / g). It is applicable to the one-test-two-evaluation quantitative determination requirements of salidroside and calycosin glucoside in Hongjingtian Huangqi Honghua Capsule.

[0058] Table 1 Salidroside injection amount and peak area

[0059]

[0060] Table 2 Injection amount and peak area of ​​calycosin isoflavone glucoside

[0061]

[0062] Table 3 Results of the recovery test of salidroside in samples

[0063]

[0064] Table 4 Results of the test on the recovery of calycosin isoflavone glucoside in samples

[0065]

[0066] Table 5 Precision test results

[0067]

[0068]

[0069] Table 6 Stability test results

[0070]

[0071] Table 7 Repeatability test results

[0072]

[0073]

[0074] Table 8 Determination results of the contents of two components in Rhodiola rosea, Astragalus and Safflower Capsules (mg / g)

[0075]

[0076] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A one-test-multiple-evaluation rapid qualitative and quantitative detection method for Rhodiola rosea, Astragalus and Safflower Capsules, characterized in that: The following steps are involved: (1) One-test-multiple-evaluation rapid qualitative identification method Take 10g of the contents of Rhodiola rosea, Astragalus membranaceus and Safflower capsules, grind them into powder, add 50mL of 50% ethanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, and add 30mL of water to the residue to dissolve it; shake and extract with ethyl acetate twice, 30mL each time, combine the ethyl acetate extracts, evaporate to dryness, and add 2mL of methanol to the residue to dissolve it as the test solution; take 1g of Rhodiola rosea control medicinal material, 1g of Astragalus control medicinal material, and 0.1g of Safflower control medicinal material, add 10mL of 50% ethanol respectively, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, and add 20mL of water to the residue. L to dissolve; shake and extract with ethyl acetate twice, 20 mL each time, combine the ethyl acetate extracts, evaporate to dryness, add 1 mL of methanol to the residue to dissolve, take 2 to 4 μL of the above test solution and reference medicinal material solution, respectively, spot them on the same silica gel G thin layer plate containing 1% NaOH, use chloroform-methanol (15:1) as the developing agent, develop, take out, dry, and examine under ultraviolet light (365 nm); the test sample shows the same color fluorescent spots at the chromatographic position as the reference medicinal materials of Rhodiola rosea, Astragalus membranaceus, and Carthamus tinctorius; (2) Rapid determination method of salidroside and calycosin with one test and two evaluations Chromatographic conditions and system suitability tests were conducted using octadecylsilane bonded silica gel as filler; Gradient elution with acetonitrile (A)-0.2% phosphoric acid solution (B) as mobile phase: 0-15min, 13% A, 15-30min, 13%-17% A, 30-35min, 17% A; detection wavelength: 270nm; column temperature: 35°C; theoretical plate number calculated based on salidroside peak should be not less than 3000; Preparation of reference solution: Take appropriate amount of salidroside and calycosin glucoside reference substances, weigh accurately, add methanol to prepare a mixed solution containing 289.48 μg salidroside and 254.98 μg calycosin glucoside per 1 ml respectively; Preparation of test solution Take an appropriate amount of the contents of Rhodiola rosea, Astragalus and Safflower Capsules, grind them into powder, take 10 g, weigh accurately, put it in a stoppered conical flask, accurately add 50 ml of 50% ethanol, stopper it tightly, weigh it, treat it with ultrasound at a power of 100 W and a frequency of 40 kHz for 30 minutes, let it cool, weigh it again, make up the lost weight with 50% methanol, shake it well, filter it, evaporate the filtrate to dryness, add 30 mL of water to dissolve the residue, shake and extract it twice with n-butanol, 30 mL each time, combine the n-butanol extracts, evaporate to dryness, add 2 mL of methanol to dissolve the residue, and use it as the test solution. Prepare the negative sample solution in the same way; The determination method is to accurately aspirate 10 μL of the reference solution, the test solution, and the negative sample solution, inject them into the liquid chromatograph, and determine the result.

2. The one-test-multiple-evaluation rapid qualitative and quantitative detection method for Rhodiola rosea, Astragalus and Safflower Capsules according to claim 1 is characterized in that: The rapid qualitative method refers to the thin layer identification method; the rapid quantitative method refers to the high performance liquid chromatography method, which only needs to use an ultraviolet detector to quantify the two components, and has lower cost than the DAD detector.

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