Method for simultaneously determining nine index components in Yanbeng initial antibacterial liquid and constructing characteristic chromatogram and application of method for simultaneously determining nine index components and characteristic chromatogram in Yanbeng initial antibacterial liquid

Abstract

The invention discloses a method for simultaneously determining nine index components in a Zhibeng bacteriostatic liquid and constructing a characteristic chromatogram and application, and belongs to the technical field of detection and analysis. Octadecylsilane chemically bonded silica is used as a filling agent; 0.1% phosphoric acid water is used as a mobile phase A, and acetonitrile is used as a mobile phase B; the detection wavelength 316 nm is used for establishing a characteristic spectrum and determining the content of cimicifugin and liquiritin, the detection wavelength 402 nm is used for determining the content of hydroxysafflor pigment A and dehydrated safflower yellow B, the detection wavelength 254 nm is used for determining the content of ganoderic acid A and schizandrin, and the detection wavelength 275 nm is used for determining the content of lozastilbin, salidroside and 5-hydroxymethylfurfural. According to the method, separation and accurate detection are realized by adopting a gradient elution procedure. So far, no report about simultaneous determination of the nine components is found, and the method makes up the blank of quality control and fingerprint spectrum research in prescriptions containing the medicinal materials.

Description

Technical Field

[0001] This invention relates to a method and application for simultaneously determining nine indicator components and constructing characteristic chromatograms in Yanbenchu ​​antibacterial solution. Specifically, using high-performance liquid chromatography (HPLC) with gradient elution, different wavelengths can be switched under the same elution conditions to simultaneously determine the contents of cimicifugoside, glycyrrhizin, hydroxysafflower pigment A, dehydrated safflower yellow pigment B, ganoderic acid A, schisandrol A, astilbene, rhodioloside, and 5-hydroxymethylfurfural in Yanbenchu ​​antibacterial solution and construct characteristic chromatograms. In addition to the components measured above, ferulic acid, gallic acid, caffeic acid, and rhodioloside were identified as characteristic components of the chromatogram. This invention belongs to the field of detection and analysis technology. Background Technology

[0002] A literature review revealed the following chemical structural formulas for cimicifugoside, glycyrrhizin, hydroxysafflower pigment A, dehydrated safflower yellow pigment B, ganoderic acid A, schisandrol A, astilbene, rhodioloside, and 5-hydroxymethylfurfural:

[0003]

[0004]

[0005]

[0006]

[0007] As can be seen from the above structural formulas, a gradient elution procedure can be used to separate and accurately detect the above components based on the differences in polarity of each substance. To date, no reports have been found regarding the simultaneous determination of these nine medicinal material components (cicadaein glycoside, glycyrrhizin, hydroxysafflower pigment A, dehydrated safflower yellow pigment B, ganoderic acid A, schisandrol A, astilbene glycoside, rhodioloside, and 5-hydroxymethylfurfural). Summary of the Invention

[0008] This invention, based on the aforementioned background, aims to improve detection efficiency, reduce detection costs, and minimize environmental pollution. For the first time, it employs a simple and rapid pretreatment method to obtain the test solution. High-performance liquid chromatography (HPLC) with a diode array detector is used, employing a gradient elution with A (0.1% phosphoric acid)-B (acetonitrile) as the mobile phase. The elution program is as follows: 0–45 min 2–7% B, 45–60 min 7–9% B, 60–105 min 9–15% B, 105–150 min 15–17% B, 150–155 min 17–20% B, 155–195 min 20–55% B, 195–205 min 55–2% B, 205–210 min 2% B. Under the same elution conditions, by switching wavelengths, nine indicator components can be simultaneously determined and characteristic chromatograms established. The mobile phase was 0.1% phosphoric acid aqueous solution (A) and acetonitrile (B). The detection wavelength was 316 nm for establishing characteristic chromatograms and determining the content of cimicifugoside and glycyrrhizin; 402 nm for determining the content of hydroxysafflower pigment A and dehydrated safflower yellow pigment B; 254 nm for determining the content of ganoderic acid A and schisandrol A; and 275 nm for determining the content of astilbin, rhodioloside, and 5-hydroxymethylfurfural. The column temperature was 30℃, and the flow rate was 0.7 mL / min. -1 Injection volume 10µL; The calculated peak values ​​of the theoretical plate for cimicifugin, glycyrrhizin, hydroxysafflower pigment A, dehydrated safflower yellow pigment B, ganoderic acid A, schisandrol A, astilbene, rhodioloside, and 5-hydroxymethylfurfural are all not less than 5000. The peak separation was excellent, achieving the goals of multi-evaluation with a single test, high efficiency, low cost, and easy popularization. This is the first report of simultaneous determination of nine indicator components in these formulated medicinal materials, demonstrating low cost and efficient multi-indicator detection.

[0009] Methodological studies showed that the injection amounts of cimicifugoside (89.98–1799.60 ng), glycyrrhizin (214.95–2149.50 ng), hydroxysafflower pigment A (53.74–1074.80 ng), dehydrated safflower yellow pigment B (51.01–1020.10 ng), ganoderic acid A (19.25–385.00 ng), schisandrin A (14.00–280.08 ng), astilbin (29.44–294.40 ng), rhodioloside (47.92–958.40 ng), and 5-hydroxymethylfurfural (111.19–2223.80 ng) exhibited good linearity with peak area (see Tables 1–9). Figures 1-9The regression equations are as follows: for cimicifugain glycoside, Y = 338.45x + 2447.8, R² = 0.9998; for glycyrrhizin, Y = 24.289x + 463.41, R² = 0.9995; for hydroxysafflower pigment A, Y = 200.87x - 381.31, R² = 0.9995; for dehydrated safflower yellow pigment B, Y = 462.14x + 3924, R² = 0.9997; for ganoderic acid A, Y = 806.24x - 498.17, R² = 0.9995; for schisandrol A, Y = 1721.7x + 1461.9, R² = 0.9998; and for astilbin, Y = 288.55x - 348.83, R² = 0.9998. 0.9998; for rhodioloside, Y = 264.99x + 1255.2, R² = 0.9999; for 5-hydroxymethylfurfural, Y = 1562x + 8769.9, R² = 0.9998. Recovery experiments were conducted, and the results showed that: the average recovery rate of cimicifugoside in 6 determinations was 98.86%, with an RSD of 1.20% (see Table 10); the average recovery rate of glycyrrhizin in 6 determinations was 97.90%, with an RSD of 1.50% (see Table 11); the average recovery rate of hydroxysafflower pigment A in 6 determinations was 99.27%, with an RSD of 1.17% (see Table 12); the average recovery rate of dehydrated safflower yellow pigment B in 6 determinations was 97.88%, with an RSD of 1.72% (see Table 13); ganoderic acid A... The average recovery rate of the six determinations was 98.54%, with an RSD of 1.81% (see Table 14); the average recovery rate of schisandrol A was 99.10%, with an RSD of 0.83% (see Table 15); the average recovery rate of astilbin was 98.56%, with an RSD of 1.01% (see Table 16); the average recovery rate of rhodioloside was 98.85%, with an RSD of 1.14% (see Table 17); and the average recovery rate of 5-hydroxymethylfurfural was 98.38%, with an RSD of 1.43% (see Table 18). Precision (see Table 19), stability (see Table 20), repeatability (see Table 21), and UV spectra of the determined components (see Table 20) were also evaluated. Figures 10-18 ), reference standard chromatogram (see Figures 19-27 ), sample chromatogram (see Figures 28-31 ), Exclusivity (see Figures 32-38 The experimental methods and data all met the methodological requirements. This method is simple, accurate, and efficient, and can be used for the simultaneous quantitative determination of cimicifugain glycoside, glycyrrhizin, hydroxysaffron pigment A, dehydrated saffron yellow pigment B, ganoderic acid A, schisandrol A, astilbene glycoside, rhodioloside, and 5-hydroxymethylfurfural in Yanbenchu ​​antibacterial solution. A fingerprint chromatogram (316 nm) of Yanbenchu ​​antibacterial solution was also established simultaneously. The robustness and repeatability of this method were investigated to better facilitate quality control (see...). Figure 39).

[0010] The technical solution adopted by this invention to solve its technical problem is as follows: (1) A. Chromatographic conditions and system suitability test; octadecylsilane-bonded silica gel was used as the packing material; 0.1% phosphoric acid water was used as mobile phase A, and acetonitrile was used as mobile phase B; the detection wavelength of 316 nm was used for the establishment of characteristic chromatograms and the determination of the contents of cimicifugoside and glycyrrhizin; the detection wavelength of 402 nm was used for the determination of the contents of hydroxysafflower pigment A and dehydrated safflower yellow pigment B; the detection wavelength of 254 nm was used for the determination of the contents of ganoderic acid A and schisandrol A; and the detection wavelength of 275 nm was used for the determination of the contents of astilbin, rhodioloside, and 5-hydroxymethylfurfural; the column temperature was 30℃; and the flow rate was 0.7 mL·min. -1 Injection volume 10µL; The theoretical plate number calculated for the peaks of cimicifugoside, glycyrrhizin, hydroxysafflower pigment A, dehydrated safflower yellow pigment B, ganoderic acid A, schisandrin A, astilbene, rhodioloside, and 5-hydroxymethylfurfural is not less than 5000. B. Preparation of reference solution: Accurately weigh appropriate amounts of ganoderic acid A, schisandrin A, and astilbene reference standards, add methanol to prepare solutions containing 20 μg of each per ml, and shake well to obtain the reference solution. Accurately weigh appropriate amounts of hydroxysaffron A, dehydrated saffron B, and rhodioloside reference standards, and add methanol to prepare solutions containing 50 μg of each per ml. Shake well to obtain the final product. Accurately weigh appropriate amounts of cimicifugin glycoside, glycyrrhizin, and 5-hydroxymethylfurfural reference standards, and add methanol to prepare solutions containing 100 μg of each per ml. Shake well to obtain the final product. C. Preparation of the test solution: Take Yanbenchu ​​antibacterial solution, shake well, filter, and take the filtrate to obtain the test solution; D. Assay: Accurately pipette 10 μl each of the reference solution and the test solution into the liquid chromatograph, determine the content of each of the nine components, and calculate the content of each component.

[0011] The simultaneous determination refers to the simultaneous determination of the contents of cimicifugoside, glycyrrhizin, hydroxysafflower pigment A, dehydrated safflower yellow pigment B, ganoderic acid A, schisandrol A, astilbene, rhodioloside, and 5-hydroxymethylfurfural in Yanbenchu ​​antibacterial solution by switching different wavelengths under the same elution conditions, and the construction of characteristic chromatograms.

[0012] The present invention relates to a method for simultaneously determining nine index components and constructing characteristic chromatograms in Yanbenchu ​​antibacterial solution, and its application in constructing characteristic chromatograms of Yanbenchu ​​antibacterial solution.

[0013] The present invention relates to the application of a method for simultaneously determining nine index components and constructing characteristic chromatograms in Yanbenchu ​​antibacterial solution in the determination of the content of Yanbenchu ​​antibacterial solution.

[0014] Yan Benchu ​​Antibacterial Solution is an antibacterial solution with a disinfection license issued by Shijiazhuang Zangnuo Pharmaceutical Co., Ltd. It has inhibitory effects on Staphylococcus aureus, Escherichia coli, and Candida albicans.

[0015] The formulation and preparation process are as follows: Prescription: Rhodiola rosea 40-60 parts by weight, Phyllanthus emblica 40-60 parts by weight, Astragalus membranaceus 40-60 parts by weight, Angelica sinensis 30-50 parts by weight, Lonicera japonica 40-60 parts by weight, Glycyrrhiza uralensis 10-30 parts by weight, Prunus mume 5-10 parts by weight, Schisandra chinensis 5-10 parts by weight, Smilax glabra 10-30 parts by weight, Dictamnus dasycarpus 5-15 parts by weight, Ganoderma lucidum 10-30 parts by weight, Saposhnikovia divaricata 5-10 parts by weight, Lithospermum erythrorhizon 5-10 parts by weight, Curcuma zedoaria 5-10 parts by weight, Paeonia lactiflora 40-60 parts by weight, Bupleurum chinense 5-10 parts by weight, Carthamus tinctorius 5-10 parts by weight.

[0016] Preparation process: Soak the above-mentioned medicinal ingredients in 8 times the amount of water for 30 minutes, bring to a boil over high heat, then simmer over low heat for 1 hour, filter, add 6 times the amount of water, bring to a boil over high heat, then simmer over low heat for 30 minutes, filter, combine the filtrates, concentrate to an appropriate amount, centrifuge the decoction, and take the supernatant to obtain the final product.

[0017] Preferably, the concentration is reduced to 1 / 5 to 1 / 3 of the combined filtrate volume.

[0018] This formula contains seventeen medicinal ingredients. To ensure the high-quality therapeutic effect of Yan Benchu ​​antibacterial liquid, the quality of medicinal materials is strictly controlled. Simultaneous quantitative research was conducted on the following components in the antibacterial liquid: cimicifugin glycoside, glycyrrhizin, hydroxysafflower pigment A, dehydrated safflower yellow pigment B, ganoderic acid A, schisandrol A, astilbene glycoside, rhodioloside, and 5-hydroxymethylfurfural, and characteristic spectra were constructed.

[0019] The application of Yanbenchu ​​antibacterial liquid in the preparation of antibacterial drugs and / or sanitary disinfection products, the antibacterial effect including inhibition of Staphylococcus aureus, Escherichia coli and Candida albicans.

[0020] An antibacterial drug is prepared from a Yanbenchu ​​antibacterial solution of the present invention.

[0021] A hygiene and disinfection product is prepared from a Yanbenchu ​​antibacterial liquid of the present invention.

[0022] The principle of this invention is as follows:

[0023] Based on the principles of like dissolves like and the differences in dosage forms, gradient elution is employed, with polarity increasing from low to high, utilizing the maximum absorption of each substance (see...). Figures 10-18Using this wavelength as the detection wavelength not only ensures suitable peak size and area but also significantly reduces interference peaks, resulting in a stable baseline and good peak separation in the liquid chromatography spectrum, laying the foundation for simultaneous determination of multiple components. Furthermore, based on the fact that the injection volume of each component exhibits a good linear relationship with its peak area within a certain range, it can be used for quantitative determination.

[0024] The innovative points and beneficial effects of this invention are as follows:

[0025] (1) Gradient elution was adopted, with 0.1% phosphoric acid water as mobile phase A and acetonitrile as mobile phase B. Under the same elution conditions, the wavelength was switched to simultaneously determine nine index components and establish characteristic chromatograms. This allows several effective components of traditional Chinese medicine with completely different properties and structures to be quantitatively determined in a single step using the same mobile phase. The quantitative chromatograms of multiple components showed stable baselines and good peak separation. The method involves no extraction, concentration, or evaporation, and is simple, rapid, accurate, and reproducible. It is easy to popularize and master, effectively improving detection efficiency, reducing detection costs, and minimizing environmental pollution.

[0026] (2) This method for simultaneously determining these nine components (cicadaein glycoside, glycyrrhizin, hydroxysaffron pigment A, dehydrated saffron yellow pigment B, ganoderic acid A, schisandrol A, astilbene glycoside, rhodioloside, and 5-hydroxymethylfurfural) is reported for the first time. Furthermore, the characteristic chromatogram contains over 40 peaks. In addition to the components measured above, ferulic acid, gallic acid, caffeic acid, and rhodioloside were identified as characteristic components of the chromatogram. The detection efficiency is significantly improved, and the instrument cost is significantly reduced. Moreover, and more importantly, it upgrades the process from one-to-one measurement and evaluation to one-to-multiple evaluations, achieving twice the result with half the effort.

[0027] (3) The advent of the method of the present invention not only provides a method for the simultaneous detection of multiple medicinal materials (Saposhnikovia divaricata, Glycyrrhiza uralensis, Carthamus tinctorius, Ganoderma lucidum, Schisandra chinensis, Smilax glabra, Rhodiola rosea) in the Yan Benchu ​​antibacterial liquid prescription, but also provides a reference and research ideas for the simultaneous determination of these components in other compound preparations.

[0028] (4) The key technology of this invention is that acetonitrile in the mobile phase has the characteristic of pre-retention time of various chemical components. When combined with phosphoric acid solution, it can make acidic components exist as molecules and eliminate the tailing of ions. Through infinitely repeated gradient ratio studies between acetonitrile and phosphoric acid of multiple concentrations, an elution condition was found that can make various chemical components with large polarity differences appear in the same mobile phase, and the peak retention time is appropriate, the baseline is stable, and the reproducibility is good. Attached Figure Description

[0029] Figure 1 Linear relationship graph of cimicifugoside; Figure 2Linear relationship graph of glycyrrhizin; Figure 3 Linear relationship graph of hydroxysaffron A; Figure 4 Linear relationship graph of dehydrated safflower yellow pigment B; Figure 5 Linear relationship graph of ganoderic acid A; Figure 6 Linear relationship graph of schisandrin A; Figure 7 Linear relationship graph of astilbin; Figure 8 Linear relationship diagram of rhodioloside; Figure 9 Linear relationship graph of 5-hydroxymethylfurfural; Figure 10 Ultraviolet absorption spectrum of cimicifugoside; Figure 11 Ultraviolet absorption spectrum of glycyrrhizin; Figure 12 Ultraviolet absorption spectrum of hydroxysaffron A; Figure 13 UV absorption spectrum of dehydrated safflower yellow pigment B Figure 14 UV absorption spectrum of ganoderic acid A Figure 15 UV absorption spectrum of schisandrin A Figure 16 UV absorption spectrum of astilbin Figure 17 UV absorption spectrum of rhodioloside Figure 18 UV absorption spectrum of 5-hydroxymethylfurfural Figure 19 HPLC chromatogram of cimicifugoside; Figure 20 HPLC chromatogram of glycyrrhizin; Figure 21 HPLC chromatogram of hydroxysaffron A; Figure 22 HPLC chromatogram of dehydrated safflower yellow pigment B; Figure 23 HPLC chromatogram of ganoderic acid A; Figure 24 HPLC chromatogram of schisandrin A; Figure 25 HPLC chromatogram of astilbin; Figure 26 HPLC chromatogram of rhodioloside; Figure 27HPLC chromatogram of 5-hydroxymethylfurfural; Figure 28 HPLC chromatogram of Yan Benchu ​​antibacterial solution sample (316nm); 1 is cimicifugoside, 2 is glycyrrhizin; Figure 29 HPLC chromatogram of Yanbenchu ​​antibacterial solution sample (402nm); 1 is labeled as hydroxysafflower pigment A, and 2 is labeled as dehydrated safflower yellow pigment B; Figure 30 HPLC chromatogram of Yan Benchu ​​antibacterial solution sample (254nm); 1 is ganoderic acid A, 2 is schisandrol A; Figure 31 HPLC chromatogram of Yan Benchu ​​antibacterial solution sample (275nm); 1 is 5-hydroxymethylfurfural, 2 is astilbin, and 3 is rhodioloside; Figure 32 HPLC chromatogram of a blank sample without Saposhnikovia divaricata Figure 33 The HPLC chromatogram of a blank sample without licorice is shown. Figure 34 The HPLC chromatogram of a blank sample without safflower is shown. Figure 35 The HPLC chromatogram of a blank sample without Ganoderma lucidum is shown. Figure 36 The HPLC chromatogram of a blank sample without Schisandra chinensis; Figure 37 The HPLC chromatogram of a blank sample without Smilax glabra (Tufuling). Figure 38 This is the HPLC chromatogram of a blank sample that does not contain Rhodiola rosea. Figure 39 Characteristic spectrum of Yan Benchu ​​antibacterial liquid; 1 is gallic acid, 2 is 5-hydroxymethylfurfural, 3 is caffeic acid, 4 is hydroxysafflower pigment A, 5 is ferulic acid, 6 is cimicifugin glycoside, 7 is glycyrrhizin, 8 is astilbene glycoside, 9 is dehydrated safflower yellow pigment B, 10 is rhodioloside, 11 is rhodioloside, 12 is ganoderic acid A, and 13 is schisandrol A.

[0030] Figures 1-9 In the figure, the vertical axis represents the peak area; the horizontal axis represents the injection volume (ng). Figures 10-18 In the figure, the vertical axis represents the absorption intensity (mAU); the horizontal axis represents the absorption wavelength (nm). Figures 19-39 In the graph, the horizontal axis represents the retention time (minutes), and the vertical axis represents the response value (mAU). Detailed Implementation

[0031] Example 1

[0032] (1) A. Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material; 0.1% phosphoric acid water was used as mobile phase A, and acetonitrile was used as mobile phase B; the detection wavelength of 316 nm was used for the establishment of characteristic chromatograms and the determination of the contents of cimicifugoside and glycyrrhizin; the detection wavelength of 402 nm was used for the determination of the contents of hydroxysafflower pigment A and dehydrated safflower yellow pigment B; the detection wavelength of 254 nm was used for the determination of the contents of ganoderic acid A and schisandrol A; and the detection wavelength of 275 nm was used for the determination of the contents of astilbin, rhodioloside, and 5-hydroxymethylfurfural; the column temperature was 30℃; and the flow rate was 0.7 mL·min. -1 Injection volume 10µL; The calculated peak values ​​of the theoretical plate for cimicifugin, glycyrrhizin, hydroxysafflower pigment A, dehydrated safflower yellow pigment B, ganoderic acid A, schisandrol A, astilbene, rhodioloside, and 5-hydroxymethylfurfural are all not less than 5000. B. Preparation of reference solution: Accurately weigh appropriate amounts of ganoderic acid A, schisandrin A, and astilbene reference standards, add methanol to prepare solutions containing 20 μg of each per ml, and shake well to obtain the reference solution. Accurately weigh appropriate amounts of hydroxysaffron A, dehydrated saffron B, and rhodioloside reference standards, and add methanol to prepare solutions containing 50 μg of each per ml. Shake well to obtain the final product. Accurately weigh appropriate amounts of cimicifugin glycoside, glycyrrhizin, and 5-hydroxymethylfurfural reference standards, and add methanol to prepare solutions containing 100 μg of each per ml. Shake well to obtain the final product. C. Preparation of the test solution: Take Yanbenchu ​​antibacterial solution, shake well, filter, and take the filtrate to obtain the test solution; For the assay, accurately pipette 10 μl each of the reference solution and the test solution and inject them into the liquid chromatograph for determination. Calculate the content of each of the nine components.

[0033] A Yanbenchu ​​antibacterial liquid comprises the following drugs: 40 parts by weight of Rhodiola rosea, 40 parts by weight of Phyllanthus emblica, 40 parts by weight of Astragalus membranaceus, 30 parts by weight of Angelica sinensis, 40 parts by weight of Lonicera japonica, 10 parts by weight of Glycyrrhiza uralensis, 5 parts by weight of Prunus mume, 5 parts by weight of Schisandra chinensis, 10 parts by weight of Smilax glabra, 5 parts by weight of Dictamnus dasycarpus, 10 parts by weight of Ganoderma lucidum, 5 parts by weight of Saposhnikovia divaricata, 5 parts by weight of Lithospermum erythrorhizon, 5 parts by weight of Curcuma zedoaria, 40 parts by weight of Paeonia lactiflora, 5 parts by weight of Bupleurum chinense, and 5 parts by weight of Carthamus tinctorius.

[0034] A method for preparing Yan Benchu ​​antibacterial liquid includes the following steps: soaking the above-mentioned medicinal ingredients in 8 times the amount of water for 30 minutes, boiling over high heat and then simmering over low heat for 1 hour, filtering, adding 6 times the amount of water, boiling over high heat and then simmering over low heat for 30 minutes, filtering, combining the filtrates, concentrating to obtain a decoction, concentrating to 1 / 5 of the volume of the combined filtrate, centrifuging the decoction, and taking the supernatant to obtain the liquid.

[0035] The application of Yanbenchu ​​antibacterial liquid in the preparation of antibacterial drugs and / or sanitary disinfection products, the antibacterial effect including inhibition of Staphylococcus aureus, Escherichia coli and Candida albicans.

[0036] An antibacterial drug was prepared from a Yanbenchu ​​antibacterial solution according to this embodiment.

[0037] A hygiene and disinfection product is prepared from a Yanbenchu ​​antibacterial liquid according to this embodiment.

[0038] Table 1. Injection volume and peak area of ​​cimicifugoside

[0039] Table 2. Glycyrrhizin injection amount and peak area

[0040] Table 3. Injection volume and peak area of ​​hydroxysafflower pigment A

[0041] Table 4. Injection amount and peak area of ​​dehydrated safflower yellow pigment B

[0042] Table 5. Injection amount and peak area of ​​ganoderic acid A

[0043] Table 6. Schisandrin A Injection Amount and Peak Area

[0044] Table 7. Astilbin Injection Amount and Peak Area

[0045] Table 8. Rhodioloside Injection Amount and Peak Area

[0046] Table 9. 5-Hydroxymethylfurfural injection amount and peak area

[0047] Table 10 Results of the recovery rate test of anchietin glycosides in the samples

[0048] Table 11 Results of glycyrrhizin recovery test in samples

[0049] Table 12 Results of the recovery rate test of hydroxysaffron A in the samples

[0050] Table 13 Results of the recovery rate test of dehydrated safflower yellow pigment B in the samples

[0051] Table 14 Results of Ganoderic Acid A Recovery in Samples

[0052] Table 15 Results of Schisandrin A Recovery in Samples

[0053] Table 16 Results of the recovery rate test of astilbin in samples

[0054] Table 17 Results of the recovery rate test of rhodioloside in the samples

[0055] Table 18 Results of 5-hydroxymethylfurfural recovery tests in samples

[0056] Table 19 Precision Experiment Results

[0057] Table 20 Stability test results

[0058] Table 21 Repeatability test results (ug / ml)

[0059] Example 2 The only difference between this embodiment and Embodiment 1 is that: A Yanbenchu ​​antibacterial liquid comprises the following drugs: 60 parts by weight of Rhodiola rosea, 60 parts by weight of Phyllanthus emblica, 60 parts by weight of Astragalus membranaceus, 50 parts by weight of Angelica sinensis, 60 parts by weight of Lonicera japonica, 30 parts by weight of Glycyrrhiza uralensis, 10 parts by weight of Prunus mume, 10 parts by weight of Schisandra chinensis, 30 parts by weight of Smilax glabra, 15 parts by weight of Dictamnus dasycarpus, 30 parts by weight of Ganoderma lucidum, 10 parts by weight of Saposhnikovia divaricata, 10 parts by weight of Lithospermum erythrorhizon, 10 parts by weight of Curcuma zedoaria, 60 parts by weight of Paeonia lactiflora, 10 parts by weight of Bupleurum chinense, and 10 parts by weight of Carthamus tinctorius.

[0060] A method for preparing Yan Benchu ​​antibacterial liquid includes the following steps: soaking the above-mentioned medicinal ingredients in 8 times the amount of water for 30 minutes, boiling over high heat and then simmering over low heat for 1 hour, filtering, adding 6 times the amount of water, boiling over high heat and then simmering over low heat for 30 minutes, filtering, combining the filtrates, concentrating to obtain a decoction, concentrating to 1 / 3 of the volume of the combined filtrate, centrifuging the decoction, and taking the supernatant to obtain the liquid.

[0061] The application of Yanbenchu ​​antibacterial liquid in the preparation of antibacterial drugs and / or sanitary disinfection products, the antibacterial effect including inhibition of Staphylococcus aureus, Escherichia coli and Candida albicans.

[0062] An antibacterial drug was prepared from a Yanbenchu ​​antibacterial solution according to this embodiment.

[0063] A hygiene and disinfection product is prepared from a Yanbenchu ​​antibacterial liquid according to this embodiment.

Claims

1. A method for simultaneously determining nine indicator components and constructing characteristic chromatograms in Yanbenchu ​​antibacterial solution, characterized in that: Includes the following steps: A. Chromatographic conditions and system suitability test: The column was filled with octadecylsilane-bonded silica gel; mobile phase A was 0.1% phosphoric acid water, and mobile phase B was acetonitrile; the detection wavelength was 316 nm for establishing characteristic chromatograms and determining the content of cimicifugoside and glycyrrhizin; the detection wavelength was 402 nm for determining the content of hydroxysafflower pigment A and dehydrated safflower yellow pigment B; the detection wavelength was 254 nm for determining the content of ganoderic acid A and schisandrol A; and the detection wavelength was 275 nm for determining the content of astilbin, rhodioloside, and 5-hydroxymethylfurfural; the column temperature was 30℃; and the flow rate was 0.7 mL·min. -1 Injection volume 10µL; The theoretical plate number calculated for the peaks of cimicifugoside, glycyrrhizin, hydroxysafflower pigment A, dehydrated safflower yellow pigment B, ganoderic acid A, schisandrin A, astilbene, rhodioloside, and 5-hydroxymethylfurfural is not less than 5000. B. Preparation of reference solutions: Accurately weigh appropriate amounts of ganoderic acid A, schisandrin A, and astilbene reference standards, add methanol to prepare solutions containing 20 μg of each per ml, shake well, and the solution is obtained. Accurately weigh appropriate amounts of hydroxysafflower pigment A, dehydrated safflower pigment B, and rhodioloside reference standard, and add methanol to prepare solutions containing 50 μg of each per ml. Shake well to obtain the final product. Accurately weigh appropriate amounts of cimicifugin glycoside, glycyrrhizin, and 5-hydroxymethylfurfural reference standards, and add methanol to prepare solutions containing 100 μg of each per ml. Shake well to obtain the final product. C. Preparation of test solution: Take Yanbenchu ​​antibacterial solution, shake well, filter, and take the filtrate to obtain the test solution; D. Determination method: Accurately pipette 10 μl each of the reference solution and the test solution into the liquid chromatograph, determine the content of each of the nine components, and calculate the content of each component.

2. The method according to claim 1, characterized in that, The aforementioned "one test, multiple evaluations" refers to the simultaneous determination of the contents of nine components in Yanbenchu ​​antibacterial solution—citricaside, glycyrrhizin, hydroxysaffron pigment A, dehydrated saffron yellow pigment B, ganoderic acid A, schisandrol A, astilbene, rhodioloside, and 5-hydroxymethylfurfural—using the same elution conditions and switching different wavelengths, and the construction of characteristic chromatograms. In addition to the components measured above, ferulic acid, gallic acid, caffeic acid, and rhodioloside were also identified as characteristic components of the chromatograms.

3. The method of claim 1, wherein, Yan Benchu ​​antibacterial liquid contains the following ingredients: Rhodiola rosea 40-60 parts by weight, Phyllanthus emblica 40-60 parts by weight, Astragalus membranaceus 40-60 parts by weight, Angelica sinensis 30-50 parts by weight, Lonicera japonica 40-60 parts by weight, Glycyrrhiza uralensis 10-30 parts by weight, Prunus mume 5-10 parts by weight, Schisandra chinensis 5-10 parts by weight, Smilax glabra 10-30 parts by weight, Dictamnus dasycarpus 5-15 parts by weight, Ganoderma lucidum 10-30 parts by weight, Saposhnikovia divaricata 5-10 parts by weight, Lithospermum erythrorhizon 5-10 parts by weight, Curcuma zedoaria 5-10 parts by weight, Paeonia lactiflora 40-60 parts by weight, Bupleurum chinense 5-10 parts by weight, and Carthamus tinctorius 5-10 parts by weight.

4. The method of claim 3, wherein, The preparation method of Yan Benchu ​​antibacterial liquid is as follows: Soak the above-mentioned medicinal ingredients in 8 times the amount of water for 30 minutes, bring to a boil over high heat, then simmer over low heat for 1 hour, filter, add 6 times the amount of water, bring to a boil over high heat, then simmer over low heat for 30 minutes, filter, combine the filtrates, concentrate to obtain the decoction, centrifuge the decoction, and take the supernatant to obtain the liquid.

5. The method of claim 4, wherein, Concentrate to 1 / 5 to 1 / 3 of the combined filtrate volume.

6. A color original bacteriostatic solution, characterized by comprising: The formula includes the following herbs: Rhodiola rosea 40-60 parts by weight, Phyllanthus emblica 40-60 parts by weight, Astragalus membranaceus 40-60 parts by weight, Angelica sinensis 30-50 parts by weight, Lonicera japonica 40-60 parts by weight, Glycyrrhiza uralensis 10-30 parts by weight, Prunus mume 5-10 parts by weight, Schisandra chinensis 5-10 parts by weight, Smilax glabra 10-30 parts by weight, Dictamnus dasycarpus 5-15 parts by weight, Ganoderma lucidum 10-30 parts by weight, Saposhnikovia divaricata 5-10 parts by weight, Lithospermum erythrorhizon 5-10 parts by weight, Curcuma zedoaria 5-10 parts by weight, Paeonia lactiflora 40-60 parts by weight, Bupleurum chinense 5-10 parts by weight, and Carthamus tinctorius 5-10 parts by weight.

7. The method for preparing a paper initial bacteriostatic solution according to claim 6, characterized in that, Includes the following steps: Soak the above-mentioned medicinal ingredients in 8 times the amount of water for 30 minutes, bring to a boil over high heat, then simmer over low heat for 1 hour. Filter, add 6 times the amount of water, bring to a boil over high heat, then simmer over low heat for 30 minutes. Filter, combine the filtrates, and concentrate to obtain the decoction. Concentrate to 1 / 5 to 1 / 3 of the volume of the combined filtrate. Centrifuge the decoction and take the supernatant.

8. The application of the Yanbenchu ​​antibacterial liquid according to claim 6 in the preparation of antibacterial drugs and / or sanitary disinfection products, characterized in that, Antibacterial activity includes inhibition of Staphylococcus aureus, Escherichia coli, and Candida albicans.

9. A bacteriostatic pharmaceutical composition, characterized by comprising: It is prepared from the antibacterial solution of Yanbenchu ​​as described in claim 6.

10. A sanitary disinfecting article, characterized by It is prepared from the antibacterial solution of Yanbenchu ​​as described in claim 6.

Images