A quality detection method for gynecological Baifeng oral liquid

Abstract

The invention discloses a quality detection method for gynecological Baifeng oral liquid, which specifically comprises the following steps: (a) preparing a reference solution; (b) preparing a test solution; (c) chromatographic conditions: the chromatographic column is C 18 Reverse phase chromatography column; mobile phase is a gradient eluent composed of acetonitrile and 0.09-0.11% phosphoric acid solution; detection wavelength is 260nm and 300nm, column temperature is 29-31°C; flow rate is 0.98ml / min-1.02ml / min; measurement time is 70 minutes. (d) Determination: The characteristic spectrum is obtained by high performance liquid chromatography. The detection method of the present invention is simple to operate, low in cost, retains the characteristic components intact, has good repeatability and stability, high precision, and strong specificity and durability. The separation effect of each characteristic peak in the obtained characteristic spectrum is good, and the various main active ingredients of the gynecological Baifeng oral liquid can be detected more comprehensively, the quality of the gynecological Baifeng oral liquid can be controlled, and the effectiveness, safety, stability and quality uniformity of the gynecological Baifeng oral liquid can be evaluated more completely and accurately.

Description

Technical Field

[0001] The invention belongs to the technical field of drug quality detection, and particularly relates to a quality detection method for gynecological Baifeng oral liquid. Background Art

[0002] Gynecological Baifeng Oral Liquid is composed of 20 kinds of medicinal materials, including black chicken, wormwood leaf, Achyranthes bidentata (salt-processed), Bupleurum root, dried ginger, white peony root (fried with wine), peony bark, Cyperus rotundus, Corydalis yanhusuo (vinegar-processed), Anemarrhena asphodeloides, Poria cocos, Coptis chinensis (wine-processed), Gentiana macrophylla, Angelica sinensis, Astragalus membranaceus (roasted), Psoralea corylifolia, Rehmannia root, Rehmannia glutinosa, Fritillaria cirrhosa, and Lycium bark. This prescription has the effect of replenishing qi and nourishing blood, and is mainly used for women with weak body and blood deficiency, irregular menstruation, and abdominal pain during menstruation.

[0003] As a Chinese patent medicine, the efficacy of Fuke Baifeng Oral Liquid is the result of the combined action of multiple active ingredients. In the formula, black chicken nourishes qi and blood, regulates menstruation and relieves pain. When blood is sufficient, qi is harmonious, and when qi and blood are regulated, menstruation is smooth. It is the main medicine; Rehmannia root, Angelica sinensis, White Peony Root, Cyperus rotundus, and Corydalis yanhusuo can greatly nourish the blood, nourish the liver and regulate menstruation, regulate qi and blood, and can treat abdominal pain during menstruation, and achieve the balance of qi and blood. They are all ministerial medicines; Cortex Moutan, Rhizoma Anemarrhenae, Radix Gentianae, Artemisiae Annua, Fritillaria, Coptis chinensis, Cortex Lycii, Dry Ginger, Artemisia argyi, Radix Astragali, and Poria clear and discharge deficiency heat, so that deficiency heat retreats and menstrual blood is calm, and can treat dry throat and dry mouth caused by deficiency heat burning body fluid. They are all adjuvant medicines; Radix Bupleuri and Radix Achyranthis Bidentatae regulate the liver and kidneys, and are all guiding medicines.

[0004] The current quality standard of Fuke Baifeng Oral Liquid included in the surgical gynecology volume of the National Compilation of Standards for Traditional Chinese Medicines has established five qualitative identification methods, including the TLC identification method for the reference medicinal materials of Achyranthes bidentata, dried ginger, Corydalis yanhusuo, paeonol reference substance, berberine hydrochloride reference substance and bamipine hydrochloride reference substance, as well as one component content determination method, paeoniflorin; Xia Jing, Sheng Liling, and Ji Shen studied the quality standard of Fuke Baifeng Oral Liquid and established five qualitative identification methods, including the TLC identification method for the reference medicinal materials of Achyranthes bidentata, dried ginger, Corydalis yanhusuo, paeonol reference substance, berberine hydrochloride reference substance and bamipine hydrochloride reference substance, as well as two component content determination methods, paeoniflorin and gentiopicroside; Song Guiping, Xiao Min, Chen Jun et al. used HPLC to determine the content of one component, paeoniflorin, in Fuke Baifeng Oral Liquid.

[0005] The above methods only monitor one or two quality indicators. Due to the complex ingredients of Chinese patent medicines, the quality control of a single indicator is relatively limited, and it is difficult to fully control the quality of the product. The operation is also relatively complicated. In addition, no monitoring indicators and methods have been established for Junyao Black Chicken, and the quality level of Fuke Baifeng Oral Liquid cannot be fully reflected. Summary of the invention

[0006] In view of this, the main purpose of the present invention is to provide a quality detection method for gynecological Baifeng oral liquid, which solves the problems of complicated steps, high cost and inability to monitor multiple quality indicators at the same time in the prior art detection method.

[0007] To achieve the above object, the technical solution of the present invention is implemented as follows: a quality detection method for gynecological Baifeng oral liquid, the method specifically comprising the following steps:

[0008] The method specifically comprises the following steps:

[0009] Preparation of reference solution: Take appropriate amount of nicotinamide reference substance, 5-hydroxymethylfurfural reference substance, loganinic acid reference substance, chlorogenic acid reference substance, gentiopicroside reference substance, ferulic acid reference substance and potassium sorbate reference substance, add water to make a mixed solution, and obtain;

[0010] Preparation of test solution: Take Fuke Baifeng oral solution, filter it, and obtain;

[0011] Chromatographic conditions: Chromatographic column is C 18 Reverse phase chromatography column; gradient elution, mobile phase is a gradient eluent composed of mobile phase A and mobile phase B, mobile phase A is acetonitrile, mobile phase B is 0.09-0.11% phosphoric acid solution; detection wavelength is 260nm and 300nm, column temperature is 29-31°C; flow rate is 0.98ml / min-1.02ml / min; measurement time is 70 minutes;

[0012] Determination: Accurately aspirate the reference solution and the test solution and inject them into the high performance liquid chromatograph, determine them according to the high performance liquid chromatography method, obtain the characteristic spectrum, analyze the obtained chromatogram, and evaluate the quality of Fuke Baifeng Oral Liquid at the same time.

[0013] Preferably, the elution gradient table is as follows:

[0014]

[0015] Preferably, the injection volume of the reference solution and the test solution accurately injected into the high performance liquid chromatograph is 10 to 50 μl.

[0016] Preferably, the gynecological Baifeng oral liquid is made into an oral liquid from the following raw medicinal materials in parts by weight according to the existing conventional method: 108.8 parts of black chicken, 18.8 parts of mugwort, 9.8 parts of Achyranthes bidentata (salt-processed), 9.8 parts of Bupleurum chinense, 4.5 parts of dried ginger, 9.8 parts of white peony root (fried with wine), 9.8 parts of peony bark, 75 parts of Cyperus rotundus, 4.5 parts of Corydalis yanhusuo (vinegar-processed), 9.8 parts of Anemarrhena asphodeloides, 12 parts of Poria, 4.5 parts of Coptis chinensis (wine-processed), 6.8 parts of Gentiana macrophylla, 9.8 parts of Angelica sinensis, 9.8 parts of Astragalus membranaceus (roasted), 18.8 parts of Psoralea corylifolia, 9.8 parts of Rehmannia glutinosa, 18.8 parts of Rehmannia glutinosa, 9.8 parts of Fritillaria cirrhosa, and 4.5 parts of Lycium bark.

[0017] Compared with the prior art, by adopting the detection method of the present invention, not only the method is simple, low in cost, and has strong specificity and durability, but also one detection method can monitor seven components, namely nicotinamide, 5-hydroxymethylfurfural, loganinic acid, chlorogenic acid, gentiopicroside, ferulic acid and potassium sorbate, and monitor ten medicinal flavors, namely black chicken, wormwood leaf, white peony root, peony bark, cyperus rotundus, coptis root, gentiana macrophylla, angelica root, artemisia annua and rehmannia root, and two auxiliary materials, namely honey and potassium sorbate, thereby realizing the monitoring of multiple quality indicators at the same time, and comprehensively controlling and evaluating the intrinsic quality of the gynecological Baifeng oral liquid, so as to ensure the stability, effectiveness and safety of the drug quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The present invention will be further described in detail below in conjunction with the accompanying drawings.

[0019] Figure 1 This is a full wavelength scan of the characteristic spectrum detection method of Fuke Baifeng Oral Liquid;

[0020] Figure 2 It is the characteristic spectrum matching diagram of 15 batches of Fuke Baifeng oral liquid (300nm);

[0021] Figure 3 It is the standard characteristic spectrum of Fuke Baifeng oral liquid (300nm);

[0022] Figure 4 It is the characteristic spectrum matching diagram of 15 batches of Fuke Baifeng oral liquid (260nm);

[0023] Figure 5 It is the standard characteristic spectrum of Fuke Baifeng oral liquid (260nm). DETAILED DESCRIPTION

[0024] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0025] The embodiment of the present invention provides a quality detection method for Fuke Baifeng oral liquid, which specifically comprises the following steps:

[0026] Preparation of reference solution: Take appropriate amount of nicotinamide reference substance, 5-hydroxymethylfurfural reference substance, loganinic acid reference substance, chlorogenic acid reference substance, gentiopicroside reference substance, ferulic acid reference substance and potassium sorbate reference substance, add water to make a mixed solution containing 0.05 mg of nicotinamide, 0.02 mg of 5-hydroxymethylfurfural, 0.1 mg of loganinic acid, 0.03 mg of chlorogenic acid, 0.2 mg of gentiopicroside, 0.02 mg of ferulic acid and 0.5 mg of potassium sorbate per 1 ml;

[0027] Preparation of test solution: Take Fuke Baifeng oral solution, filter it, and obtain;

[0028] Chromatographic conditions: Chromatographic column is C 18 Reverse phase chromatography column; gradient elution, mobile phase is a gradient eluent composed of mobile phase A and mobile phase B, mobile phase A is acetonitrile, mobile phase B is 0.09-0.11% phosphoric acid solution; detection wavelength is 260nm and 300nm, column temperature is 29-31°C; flow rate is 0.98ml / min-1.02ml / min; measurement time is 70 minutes;

[0029] Determination: Accurately aspirate the reference solution and the test solution and inject them into the high performance liquid chromatograph, determine them according to the high performance liquid chromatography method, obtain the characteristic spectrum, analyze the obtained chromatogram, and evaluate the quality of Fuke Baifeng Oral Liquid at the same time.

[0030] Further, the elution gradient table is as follows:

[0031]

[0032] Furthermore, the gynecological Baifeng oral liquid is made into an oral liquid from the following raw medicinal materials in parts by weight according to the existing conventional method: 108.8 parts of black chicken, 18.8 parts of mugwort, 9.8 parts of Achyranthes bidentata (salt-processed), 9.8 parts of Bupleurum chinense, 4.5 parts of dried ginger, 9.8 parts of white peony root (fried with wine), 9.8 parts of peony bark, 75 parts of Cyperus rotundus, 4.5 parts of Corydalis yanhusuo (vinegar-processed), 9.8 parts of Anemarrhena asphodeloides, 12 parts of Poria, 4.5 parts of Coptis chinensis (wine-processed), 6.8 parts of Gentiana macrophylla, 9.8 parts of Angelica sinensis, 9.8 parts of Astragalus membranaceus (roasted), 18.8 parts of Psoralea corylifolia, 9.8 parts of Rehmannia root, 18.8 parts of Rehmannia root prepared, 9.8 parts of Fritillaria cirrhosa, and 4.5 parts of Lycium bark.

[0033] The following is the process and principle of analyzing the chromatogram obtained by detection (specifically, Figure 2-Figure 5 shown):

[0034] According to the characteristic spectra of 15 batches of Fuke Baifeng oral liquid obtained, the characteristic spectrum at 300nm should have 12 characteristic peaks, among which peak No. 6 is gentiopicroside, which is S peak and belongs to Gentiana macrophylla; peak No. 1 is 5-hydroxymethylfurfural, which belongs to Rehmannia glutinosa and honey; peak No. 3 is chlorogenic acid, which belongs to Artemisia argyi and Artemisia annua; peak No. 8 is ferulic acid, which belongs to Cyperus rotundus and Angelica sinensis; peak No. 9 is potassium sorbate, which belongs to potassium sorbate; the components of other characteristic peaks are not clear, among which peaks 2, 5, 10, 11 and 12 all belong to Artemisia argyi and Artemisia annua, peak No. 4 belongs to Coptis chinensis, and peak No. 7 belongs to Cyperus rotundus.

[0035] The characteristic spectrum at 260nm should have 7 characteristic peaks, of which peak 7 is gentiopicroside as the reference peak, which is the S peak and belongs to Gentiana macrophylla; peak 1 is nicotinamide, which belongs to black chicken; peak 6 is loganin acid, which belongs to Gentiana macrophylla; the other characteristic peaks have no clear components, of which peak 2 belongs to black chicken, peak 3 belongs to white peony root and peony bark, peak 4 belongs to Rehmannia glutinosa, and peak 5 belongs to peony bark and Cyperus rotundus; the relative retention time and relative standard deviation of each characteristic peak are:

[0036] Peak 1: relative retention time 0.114 ± 0.001, relative standard deviation 0.18%;

[0037] Peak 2: relative retention time 0.197 ± 0.001, relative standard deviation 0.17%;

[0038] Peak 3: relative retention time 0.362 ± 0.001, relative standard deviation 0.13%;

[0039] Peak 4: relative retention time 0.477 ± 0.001, relative standard deviation 0.09%;

[0040] Peak 5: relative retention time 0.762 ± 0.001, relative standard deviation 0.07%;

[0041] Peak 6: relative retention time 0.785 ± 0.009, relative standard deviation 0.32%;

[0042] Peak 7: relative retention time 1.000±0.000, relative standard deviation 0.00%.

[0043] The characteristic spectrum at a wavelength of 300 nm has 12 characteristic peaks, of which peak 1 is 5-hydroxymethylfurfural, peak 3 is chlorogenic acid, peak 6 is gentiopicroside as a reference peak, peak 8 is ferulic acid, and peak 9 is potassium sorbate. The relative retention time and relative standard deviation of each characteristic peak are:

[0044] Peak 1: relative retention time 0.433 ± 0.001, relative standard deviation 0.08%;

[0045] Peak 2: relative retention time 0.692 ± 0.001, relative standard deviation 0.06%;

[0046] Peak 3: relative retention time 0.897 ± 0.001, relative standard deviation 0.04%;

[0047] Peak 4: relative retention time 0.920 ± 0.001, relative standard deviation 0.03%;

[0048] Peak 5: relative retention time 0.945 ± 0.001, relative standard deviation 0.04%;

[0049] Peak 6: relative retention time 1.000±0.000, relative standard deviation 0.00%;

[0050] Peak 7: relative retention time 1.194 ± 0.001, relative standard deviation 0.05%;

[0051] Peak 8: relative retention time 1.309 ± 0.006, relative standard deviation 0.13%;

[0052] Peak 9: relative retention time 1.365 ± 0.001, relative standard deviation 0.04%;

[0053] Peak 10: relative retention time 1.546 ± 0.002, relative standard deviation 0.07%;

[0054] Peak 11: relative retention time 1.583 ± 0.001, relative standard deviation 0.06%;

[0055] Peak 12: relative retention time 1.682±0.002, relative standard deviation 0.07%.

[0056] In addition, the constructed HPLC standard characteristic spectrum of the gynecological Baifeng oral liquid at a wavelength of 260nm is composed of 7 characteristic peaks. Compared with the reference substance, peak 1 is nicotinamide, peak 6 is loganic acid, peak 7 is gentiopicroside, gentiopicroside is used as the reference substance, peak 7 is the reference peak, and the peak corresponding to the reference peak is the S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time is within ±5% of the specified value, and the specified value is peak 1: 0.11, peak 2: 0.20, peak 3: 0.36, peak 4: 0.48, peak 5: 0.76, peak 6: 0.79, peak 7 (S): 1.00; The constructed HPLC standard characteristic spectrum of the gynecological Baifeng oral liquid at a wavelength of 300nm The spectrum is composed of 12 characteristic peaks. Compared with the reference substance, peak 1 is 5-hydroxymethylfurfural, peak 3 is chlorogenic acid, peak 6 is gentiopicroside, peak 8 is ferulic acid, and peak 9 is potassium sorbate. Gentiopicroside is used as the reference substance, peak 6 is the reference peak, and the peak corresponding to the reference peak is the S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time is within ±5% of the specified value. The specified value is peak 1: 0.43, peak 2: 0.69, peak 3: 0.90, peak 4: 0.92, peak 5: 0.95, peak 6 (S): 1.00, peak 7: 1.19, peak 8: 1.31, peak 9: 1.37, peak 10: 1.55, peak 11: 1.58, peak 12: 1.68.

[0057] In addition, the present invention adopts a dual-wavelength detection method to achieve the purpose of increasing the quality coverage, wherein 13 characteristic peaks at a wavelength of 300nm clarify five components, namely 5-hydroxymethylfurfural, chlorogenic acid, gentiopicroside, ferulic acid, and potassium sorbate, and monitor seven medicinal flavors, namely wormwood, cyperus rotundus, coptis chinensis, gentiana macrophylla, angelica sinensis, artemisia annua, and rehmannia root, and two auxiliary materials, namely honey and potassium sorbate; 7 characteristic peaks at a wavelength of 260nm clarify three components, namely nicotinamide, loganinic acid, and gentiopicroside, and monitor six medicinal flavors, namely black chicken, white peony root, moutan bark, cyperus rotundus, gentiana macrophylla, and rehmannia root. One detection method can comprehensively, observably, and scientifically evaluate the quality of the gynecological Baifeng oral liquid.

[0058] Instrument precision test: Take the same reference solution and inject it 6 times continuously according to the above chromatographic conditions. Record the retention time and peak area of ​​each chromatographic peak. The RSD value of the retention time of each chromatographic peak is less than 1.0%, and the RSD value of the peak area is less than 2.0%, indicating that the instrument is stable and has good precision.

[0059] Injection precision test: Take the test solution of the same batch of Fuke Baifeng oral liquid, and inject it continuously for 6 times according to the above chromatographic conditions. Record the retention time and peak area of ​​each chromatographic peak. The RSD value of the retention time of each chromatographic peak is less than 1.0%, and the RSD value of the peak area is less than 2.0%, indicating that the method has good injection precision.

[0060] Repeatability test: Take the same batch of Fuke Baifeng oral liquid and prepare 6 portions according to the test solution preparation method. According to the above chromatographic conditions, inject the samples and record the relative retention time and relative peak area of ​​each characteristic peak. The RSD value of the relative retention time of each characteristic peak is less than 2.0%, and the RSD value of the relative peak area is less than 3.0%, indicating that the method has good repeatability.

[0061] Solution stability test: Take the test solution of the same batch of Fuke Baifeng oral liquid, and inject samples at 0, 4, 8, 16, 32, 48, 64, 80, and 96 hours according to the above chromatographic conditions, and record the relative retention time and relative peak area of ​​each characteristic peak. The RSD value of the relative retention time of each characteristic peak is less than 2.0%, and the RSD value of the relative peak area is less than 3.0%, indicating that the test solution is stable within 96 hours.

[0062] Column temperature durability test: The same batch of gynecological Baifeng oral solution was used to test the effects of column temperatures of 29°C, 30°C, and 31°C on the separation of each characteristic peak, and the relative retention time and relative peak area of ​​each characteristic peak were recorded. The results showed that different temperatures had little effect on the separation of each characteristic peak, the RSD value of the relative retention time of each characteristic peak was less than 3.0%, and the RSD value of the relative peak area was less than 5.0%, indicating that the column temperature was 29-31°C, and the method was feasible.

[0063] Flow rate durability test: Take the same batch of gynecological Baifeng oral solution test solution, and investigate the effect of flow rates of 0.98ml / min, 1.00ml / min, and 1.02ml / min on the separation of each characteristic peak, and record the relative retention time and relative peak area of ​​each characteristic peak. The results show that different flow rates have little effect on the separation of each characteristic peak, and the RSD value of the relative retention time of each characteristic peak is less than 3.0%, and the RSD value of the relative peak area is less than 5.0%, indicating that the flow rate is 0.98-1.02ml / min, and the method is feasible.

[0064] Durability test of mobile phase composition: Take the same batch of Fuke Baifeng oral liquid test solution, investigate the effect of mobile phase B of 0.09%, 0.10%, 0.11% phosphoric acid solution on the separation of each characteristic peak, and record the relative retention time and relative peak area of ​​each characteristic peak. The results show that different mobile phase compositions have little effect on the separation of each characteristic peak, the RSD value of the relative retention time of each characteristic peak is less than 3.0%, and the RSD value of the relative peak area is less than 5.0%, indicating that the mobile phase B is 0.09% to 0.11% phosphoric acid solution, and the method is feasible.

[0065] Chromatographic column durability test: The same batch of gynecological Baifeng oral liquid test solution was taken to investigate the effects of three brands of C18 reversed-phase chromatographic columns on the separation of each characteristic peak, and the relative retention time and relative peak area of ​​each characteristic peak were recorded. The results showed that different brands of chromatographic columns had little effect on the separation of each characteristic peak, the RSD value of the relative retention time of each characteristic peak was less than 3.0%, and the RSD value of the relative peak area was less than 5.0%, indicating that the method was highly durable.

[0066] In conclusion, by adopting the detection method of the present invention, not only the method is simple, low-cost and durable, but also multiple quality indicators can be detected at the same time, which can comprehensively reflect the quality level of Fuke Baifeng Oral Liquid.

[0067] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A method for detecting gynecological Baifeng oral liquid, characterized in that: The method specifically comprises the following steps: Preparation of reference solution: Take appropriate amount of nicotinamide reference substance, 5-hydroxymethylfurfural reference substance, loganinic acid reference substance, chlorogenic acid reference substance, gentiopicroside reference substance, ferulic acid reference substance and potassium sorbate reference substance, add water to make a mixed solution, and obtain; Preparation of test solution: Take Fuke Baifeng oral solution, filter it, and obtain; Chromatographic conditions: The chromatographic column is a C18 reverse phase column; gradient elution is adopted, the mobile phase is a gradient eluent composed of mobile phase A and mobile phase B, mobile phase A is acetonitrile, and mobile phase B is 0.09-0.11% phosphoric acid solution; the detection wavelength is 260nm and 300nm, the column temperature is 29-31°C; the flow rate is 0.98ml / min-1.02ml / min; the measurement time is 70 minutes; Wherein, the procedure of the gradient elution is: ； Determination: Accurately pipette the reference solution and the test solution into a high performance liquid chromatograph, determine according to the high performance liquid chromatography method, obtain a characteristic spectrum, and analyze the obtained chromatogram.

2. The detection method of a gynecological Baifeng oral liquid according to claim 1, characterized in that: The injection volume of the reference solution and the test solution accurately injected into the HPLC is 10 to 50 μl.

3. A detection method for gynecological Baifeng oral liquid according to claim 1 or 2, characterized in that, The gynecological Baifeng oral liquid is prepared into an oral liquid from the following raw medicinal materials in parts by weight according to an existing conventional method: 108.8 parts of black chicken, 18.8 parts of wormwood leaves, 9.8 parts of Achyranthes bidentata, 9.8 parts of Bupleurum chinense, 4.5 parts of dried ginger, 9.8 parts of white peony root, 9.8 parts of peony bark, 75 parts of Cyperus rotundus, 4.5 parts of Corydalis yanhusuo, 9.8 parts of Anemarrhena asphodeloides, 12 parts of Poria, 4.5 parts of Coptis chinensis, 6.8 parts of Gentiana macrophylla, 9.8 parts of Angelica sinensis, 9.8 parts of Astragalus membranaceus, 18.8 parts of Psoralea corylifolia, 9.8 parts of Rehmannia root, 18.8 parts of Rehmannia root prepared, 9.8 parts of Fritillaria cirrhosa, and 4.5 parts of Lycium bark.

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