CN1320356C - Method for detecting Kidney tonifying mind easing extractive quality standard - Google Patents
Method for detecting Kidney tonifying mind easing extractive quality standard Download PDFInfo
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- CN1320356C CN1320356C CNB2004100893309A CN200410089330A CN1320356C CN 1320356 C CN1320356 C CN 1320356C CN B2004100893309 A CNB2004100893309 A CN B2004100893309A CN 200410089330 A CN200410089330 A CN 200410089330A CN 1320356 C CN1320356 C CN 1320356C
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Abstract
The present invention belongs to the technical field of traditional Chinese medicines, which discloses a detection method of the quality standard of a traditional Chinese medicine, namely a paste for kidney invigoration mental tranquilization. The present invention improves the original quality standard, increases the qualitative identification of folium eriobotryae, and establishes the content detection of red sage root protocatechuic aldehyde. The quality control of the present invention improves product stability, and is favorable to the quality control of industral production.
Description
Technical field:
The invention belongs to the medicine technical field.Be specifically related to method for detecting Kidney tonifying mind easing extractive quality standard.
Background technology:
The kidney-nourishing tranquilization paste is a traditional soft extract, form by 20 Six-element Chinese medicines such as genseng, pearl powder, adenophora tetraphylla, radix glehniae, the red sage root, the Radix Astragali, the fruit of Chinese magnoliavine (system), mulberry fruit, the fruit of glossy privet (system), Radix Angelicae Sinensis, barrenwort, rhizoma cyperi (system), glossy ganoderma, rhizoma nardostachyos, reticulate millettia, mussel, acorus graminens soland, polygala root (system), oyster, teasel root, giant knotweed, loguat leaf, dried orange peel, glutinous rehmannia, Fructus Hordei Germinatus, rice buds, has tonifying Qi and blood, the effect that kidney-nourishing is calmed the nerves.Be used for the Light-headedness due to the deficiency of qi and blood, soreness and weakness of waist and knees, night sweat insomnia; Also can be used for postpartum, aftercare.The red sage root is a ministerial drug in the side, has stasis-dispelling and pain-killing, activating blood to promote menstruation, the effect of the relieving restlessness that clears away heart-fire.
In the proper mass standard eight flavor medicines such as genseng have wherein been carried out qualitative identification, but do not reflected the assay item of the index components of medicine inherent quality, can't effectively guarantee the inherent quality of product aborning, and then pharmaceutical effectiveness can not get guaranteeing.
Summary of the invention:
Technical matters to be solved by this invention is to improve the initial quality standard for overcoming above-mentioned weak point, improves the quality of products, and has increased the wherein qualitative identification of loguat leaf, has set up protocatechualdehyde C in the red sage root
7H
6O
3Assay.
The invention provides method for detecting Kidney tonifying mind easing extractive quality standard, this method comprises the qualitative identification of loguat leaf wherein and has set up the assay of protocatechualdehyde in the red sage root.
The qualitative identification method of loguat leaf is the thin layer differential method in the kidney-nourishing tranquilization paste provided by the invention, and this method comprises the following steps: to measure according to thin-layered chromatography (Chinese Pharmacopoeia version appendix in 2000 VIB).
Get this product 100g, put in the conical flask, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 200ml, shake up, extracted (sonicated or hot reflux or cold soaking) 20 minutes, put cold after, filter, filtrate evaporate to dryness, residue add water 80ml makes dissolving, extracts 2 times with water saturated normal butyl alcohol jolting, each 50ml, merge normal butyl alcohol liquid, use the saturated water washing of normal butyl alcohol 2 times, each 80ml, discard water liquid, normal butyl alcohol is recycled to dried, and residue solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 1ml makes dissolving, as need testing solution; Other gets loguat leaf control medicinal material 2g, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 30ml extracted (sonicated or hot reflux or cold soaking) 20 minutes, filtered, the filtrate evaporate to dryness, residue extracts secondary with water-saturated n-butanol after adding water 20ml dissolving, each 20ml, merge normal butyl alcohol liquid, evaporate to dryness, residue solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 1ml makes dissolving, in contrast medicinal material solution; Test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same high-efficient silica gel G thin layer plate, with benzene-acetone (10～2): (5～0.2) are developping agent, launch, take out, dry, spray ethanol solution of sulfuric acid with 10%, 105 ℃ to be heated to spot colour developing clear, in the test sample chromatogram, with the corresponding position of loguat leaf control medicinal material chromatogram on, show the spot of same color.
The determination method of red sage root flavour of a drug Central Plains catechu aldehyde is the high performance liquid chromatogram determination method in the kidney-nourishing tranquilization paste provided by the invention, and this method comprises the following steps: to measure according to high performance liquid chromatography (Chinese Pharmacopoeia version appendix in 2000 VI D).
Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica, with methyl alcohol-0.8% glacial acetic acid solution (15～2): (85～98) are moving phase, treat that the protocatechualdehyde chromatographic peak goes out behind the peak with methyl alcohol-0.8% glacial acetic acid solution (95～70): (5～30) wash-out 3～25 minutes; The detection wavelength is 280nm.Theoretical cam curve is calculated by the protocatechualdehyde peak should be not less than 6000～2000.
It is an amount of that the preparation precision of reference substance solution takes by weighing the protocatechualdehyde reference substance, and solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) is made the solution that every 1ml contains 6.8989 μ g, promptly.
This product 30.7712g is got in the preparation of need testing solution, put in the beaker, the accurate title, decide, add 25ml solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) stir and to make dissolving, be transferred in the 50ml measuring bottle, with solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) cyclic washing beaker repeatedly on a small quantity, washing lotion is incorporated in the same measuring bottle, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, precision is measured 5ml, puts in the 10ml measuring bottle solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 4ml, shake up, extracted (sonicated or hot reflux or cold soaking) 10 minutes, put cold after, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, filter, get subsequent filtrate, promptly.
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Calculate
The every g of this product contains the red sage root (with protocatechualdehyde (C as a result
7H
6O
3) meter) 22.42 μ g.
Protocatechualdehyde Determination on content during kidney-nourishing of the present invention is calmed the nerves in the red sage root flavour of a drug has been passed through campaign:
(1) instrument and reagent
Instrument: Agilent---1100 liquid chromatographs;
Reference substance: derive from Nat'l Pharmaceutical ﹠ Biological Products Control Institute's (lot number: 0844-200003 uses for assay);
Sample lot number: (1) 030901 (2) 030902 (3) 031001
Reagent: methyl alcohol is chromatogram alcohol.
(2) chromatographic condition
Through groping, adopt and detect wavelength: 280nm, flow velocity: 0.5～1.5ml/min, through test, determine to adopt moving phase: methyl alcohol-0.5% glacial acetic acid solution (15～2): (85～98), treat that the protocatechualdehyde chromatographic peak goes out usefulness methyl alcohol-0.8% glacial acetic acid solution (95～70) behind the peak: (5～30) wash-out 3～25 minutes.Chromatographic column: C18 (15cm * 5 μ m), column temperature: 45～20 ℃.
(3) test sample preparation method
Get this product 30g, put in the beaker, the accurate title, decide, add 25ml solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) stir and to make dissolving, be transferred in the 50ml measuring bottle, with solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) cyclic washing beaker repeatedly on a small quantity, washing lotion is incorporated in the same measuring bottle, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, precision is measured 5ml, puts in the 10ml measuring bottle solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 4ml, shake up, extracted (sonicated or hot reflux or cold soaking) 10 minutes, put cold after, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, filter, get subsequent filtrate, promptly.
(4) selection of extraction time
Get this product 30g, put in the beaker, the accurate title, decide, add 25ml solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) stir and to make dissolving, be transferred in the 50ml measuring bottle, with solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) cyclic washing beaker repeatedly on a small quantity, washing lotion is incorporated in the same measuring bottle, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, precision is measured 5ml, put in the 10ml measuring bottle, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 4ml, shake up, (a) jolting respectively, (b) extracted (sonicated or hot reflux or cold soaking) 5 minutes, (c) extracted (sonicated or hot reflux or cold soaking) 10 minutes, (d) extracted (sonicated or hot reflux or cold soaking) 15 minutes, (f) extracted (sonicated or hot reflux or cold soaking) 20 minutes, put cold after, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, filter, get subsequent filtrate and measure, the results are shown in following table: selective extraction (sonicated or hot reflux or cold soaking) can propose the survey composition in 10 minutes fully.
Extraction time
| Extraction time (minute) | |
| 0 | 387.688 |
| 5 | 396.959 |
| 10 | 412.207 |
| 15 | 406.030 |
| 20 | 410.007 |
(5) typical curve
Getting the protocatechualdehyde reference substance solution difference sample introduction of variable concentrations, is ordinate Y with the peak area, and sample size is horizontal ordinate X, do return typical curve, the result shows that sample size and peak area are good linear relationship, the results are shown in following table in protocatechualdehyde sample size 8.536～1280.40ng scope.
Typical curve
| Sample size (ng) | 8.536 | 17.072 | 42.68 | 85.36 | 256.08 | 512.16 | 853.6 | 1280.40 |
| Peak area | 37.565 | 76.154 | 199.314 | 408.407 | 1261.499 | 2551.758 | 4270.248 | 6420.847 |
| Regression equation | Y＝5.0226X-15.3237 r＝0.99999 | |||||||
(6) precision test
Precision is measured reference substance solution 10 μ l, injects liquid chromatograph, the sample introduction analysis, and replication 5 times, the record peak area the results are shown in following table.
The precision test
| 1 | 2 | 3 | 4 | 5 | RSD(％) | |
| Peak area | 500.507 | 498.412 | 499.343 | 497.223 | 497.544 | 0.27 |
(7) stability test
Get same need testing solution and preparing back 0 hour, 3 hours, 8 hours, 15 hours, 21 hours injection liquid chromatographs respectively, the sample introduction measurement result, its peak area there is no significant change, illustrates that need testing solution is more stable, the results are shown in following table.
Stability test
| Standing time (hour) | 0 | 3 | 8 | 15 | 21 |
| Peak area | 369.909 | 360.525 | 366.677 | 363.186 | 363.457 |
(8) replica test
Sample thief (lot number: 030901) 30g, put in the beaker, the accurate title, decide, add 25ml solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) stir and to make dissolving, be transferred in the 50ml measuring bottle, with solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) cyclic washing beaker repeatedly on a small quantity, washing lotion is incorporated in the same measuring bottle, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, precision is measured 5ml, puts in the 10ml measuring bottle solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 4ml, shake up, extracted (sonicated or hot reflux or cold soaking) 10 minutes, put cold after, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, filter, get subsequent filtrate, promptly.Inject liquid chromatograph, measure respectively, the results are shown in following table.
Replica test
| Content (μ g/g) | |
| 1 | 21.826 |
| 2 | 22.947 |
| 3 | 22.716 |
| 4 | 22.671 |
| 5 | 21.964 |
| On average | 22.42 |
| RSD | 2.2％(n＝5) |
(9) average recovery is measured
Sample thief (lot number: 030901) 15g, put in the beaker, the accurate title, decide, the accurate protocatechualdehyde reference substance that adds is a certain amount of, add 25ml solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) stir and to make dissolving, be transferred in the 50ml measuring bottle, with solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) cyclic washing beaker repeatedly on a small quantity, washing lotion is incorporated in the same measuring bottle, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, precision is measured 5ml, put in the 10ml measuring bottle, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 4ml, shake up, extracted (sonicated or hot reflux or cold soaking) 10 minutes, put cold after, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, filter, get subsequent filtrate, promptly.Inject liquid chromatograph, measure respectively, the results are shown in following table.
Average recovery
| Sample weighting amount (g) | Add reference substance amount (mg) | Record total amount (μ g) | Record content (μ g) in the sample | The recovery (%) | Mean value (%) | RSD |
| 15.0494 | 0.34144 | 682.269 | 337.407 | 101.002 | 100.86 | 1.4％ (n＝6) |
| 15.5466 | 0.34144 | 686.825 | 348.554 | 99.071 | ||
| 16.5872 | 0.4268 | 802.245 | 371.885 | 100.834 | ||
| 16.5152 | 0.4268 | 810.754 | 370.270 | 103.206 | ||
| 17.5172 | 0.51216 | 904.181 | 392.735 | 99.860 | ||
| 16.9478 | 0.51216 | 898.378 | 379.969 | 101.220 |
(10) sample determination
Sample thief (lot number: (1) 030901 (2) 030902 (3) 031001) 30g, put in the beaker, the accurate title, decide, add 25ml solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) stir and to make dissolving, be transferred in the 50ml measuring bottle, with solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) cyclic washing beaker repeatedly on a small quantity, washing lotion is incorporated in the same measuring bottle, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, precision is measured 5ml, puts in the 10ml measuring bottle solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 4ml, shake up, extracted (sonicated or hot reflux or cold soaking) 10 minutes, put cold after, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, filter, get subsequent filtrate, promptly.Inject liquid chromatograph, measure respectively, the results are shown in following table:
Sample determination
| The sample lot number | Average content (μ g/g) |
| 030901 | 22.42 |
| 030902 | 19.84 |
| 031001 | 21.35 |
(11) negative sample is measured
Get the sample 30g that does not contain the red sage root, put in the beaker, the accurate title, decide, add 25ml solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) stir and to make dissolving, be transferred in the 50ml measuring bottle, with solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) cyclic washing beaker repeatedly on a small quantity, washing lotion is incorporated in the same measuring bottle, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, precision is measured 5ml, puts in the 10ml measuring bottle solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 4ml, shake up, extracted (sonicated or hot reflux or cold soaking) 10 minutes, put cold after, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, filter, get subsequent filtrate, promptly.Inject liquid chromatograph, sample introduction 10 μ l analyze.The result is noiseless to sample determination.
In order to investigate the steadiness that quality standard improves back " kidney-nourishing tranquilization paste ", carried out long-term stable experiment:
1, test condition:
Commercially available back " kidney-nourishing tranquilization paste " is placed (25 ℃ ± 2 ℃) under the room temperature condition, and relative humidity 60% ± 10% is observed behind the placement certain hour, and sampling detects, high spot reviews: proterties, differentiate that content is checked the health examination project.
2, test result
" kidney-nourishing tranquilization paste " shows that through the test of product stability one year over high spot reviews projects such as the physicochemical property of product and health examination all meet " kidney-nourishing tranquilization paste " quality standard requirement, and product quality is comparatively stable.
Long-term stable experiment
Lot number: 030101 (RT:25 ℃ ± 2 ℃ and RH:60% ± 10%)
| 0 month | March | June | September | Dec | 16 months | ||
| Proterties | Qualified | Qualified | Qualified | Qualified | Qualified | Qualified | |
| Differentiate | 1 | + | + | + | + | + | + |
| 2 | + | + | + | + | + | + | |
| 3 | + | + | + | + | + | + | |
| 4 | + | + | + | + | + | + | |
| 5 | + | + | + | + | + | + | |
| 6 | + | + | + | + | + | + | |
| 7 | + | + | + | + | + | + | |
| 8 | + | + | + | + | + | + | |
| 9 | * | * | * | + | + | + | |
| Content (μ g/g) | * | * | * | 21.69 | 22.21 | 21.78 | |
| Check | Insolubles | Qualified | Qualified | Qualified | Qualified | Qualified | Qualified |
| Loading amount | Qualified | Qualified | Qualified | Qualified | Qualified | Qualified | |
| The hygiene standard | Bacterium (individual/gram) | ＜10 | ＜10 | ＜10 | ＜10 | ＜10 | ＜10 |
| Mould saccharomycete (individual/gram) | ＜10 | ＜10 | ＜10 | ＜10 | ＜10 | ＜10 | |
| Escherichia coli | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | |
| Mite lives | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | |
| Conclusion | Qualified | Qualified | Qualified | Qualified | Qualified | Qualified | |
Long-term stable experiment
Lot number: 030102 (RT:25 ℃ ± 2 ℃ and RH:60% ± 10%)
| 0 month | March | June | September | Dec | 16 months | ||
| Proterties | Qualified | Qualified | Qualified | Qualified | Qualified | Qualified | |
| Differentiate | 1 | + | + | + | + | + | + |
| 2 | + | + | + | + | + | + | |
| 3 | + | + | + | + | + | + | |
| 4 | + | + | + | + | + | + | |
| 5 | + | + | + | + | + | + | |
| 6 | + | + | + | + | + | + | |
| 7 | + | + | + | + | + | + | |
| 8 | + | + | + | + | + | + | |
| 9 | * | * | * | + | + | + | |
| Content (μ g/g) | * | * | * | 20.63 | 20.89 | 20.15 | |
| Check | Insolubles | Qualified | Qualified | Qualified | Qualified | Qualified | Qualified |
| Loading amount | Qualified | Qualified | Qualified | Qualified | Qualified | Qualified | |
| The hygiene standard | Bacterium (individual/gram) | ＜10 | ＜10 | ＜10 | ＜10 | ＜10 | ＜10 |
| Mould saccharomycete (individual/gram) | ＜10 | ＜10 | ＜10 | ＜10 | ＜10 | ＜10 | |
| Escherichia coli | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | |
| Mite lives | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | |
| Conclusion | Qualified | Qualified | Qualified | Qualified | Qualified | Qualified | |
Long-term stable experiment
Lot number: 030103 (RT:25 ℃ ± 2 ℃ and RH:60% ± 10%)
| 0 month | March | June | September | Dec | 16 months | ||
| Proterties | Qualified | Qualified | Qualified | Qualified | Qualified | Qualified | |
| Differentiate | 1 | + | + | + | + | + | + |
| 2 | + | + | + | + | + | + | |
| 3 | + | + | + | + | + | + | |
| 4 | + | + | + | + | + | + | |
| 5 | + | + | + | + | + | + | |
| 6 | + | + | + | + | + | + | |
| 7 | + | + | + | + | + | + | |
| 8 | + | + | + | + | + | + | |
| 9 | * | * | * | + | + | + | |
| Content (μ g/g) | * | * | * | 21.85 | 21.32 | 22.03 | |
| Check | Insolubles | Qualified | Qualified | Qualified | Qualified | Qualified | Qualified |
| Loading amount | Qualified | Qualified | Qualified | Qualified | Qualified | Qualified | |
| The hygiene standard | Bacterium (individual/gram) | ＜10 | ＜10 | ＜10 | ＜10 | ＜10 | ＜10 |
| Mould saccharomycete (individual/gram) | ＜10 | ＜10 | ＜10 | ＜10 | ＜10 | ＜10 | |
| Escherichia coli | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | |
| Mite lives | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect | |
| Conclusion | Qualified | Qualified | Qualified | Qualified | Qualified | Qualified | |
Description of drawings:
Fig. 1 loguat leaf thin-layer chromatogram
1, negative sample 2, sample 3, loguat leaf control medicinal material
Protocatechualdehyde reference substance chromatogram in Fig. 2 red sage root
Protocatechualdehyde sample chromatogram figure in Fig. 3 red sage root
Protocatechualdehyde negative sample chromatogram in Fig. 4 red sage root
Embodiment
Example one, lot number 030901
1, the loguat leaf thin layer is differentiated
Get this product 100g, put in the conical flask, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 200ml, shake up, extracted (sonicated or hot reflux or cold soaking) 20 minutes, put cold after, filter, filtrate evaporate to dryness, residue add water 80ml makes dissolving, extracts 2 times with water saturated normal butyl alcohol jolting, each 50ml, merge normal butyl alcohol liquid, use the saturated water washing of normal butyl alcohol 2 times, each 80ml, discard water liquid, normal butyl alcohol is recycled to dried, and residue solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 1ml makes dissolving, as need testing solution.Other gets loguat leaf control medicinal material 2g, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 30ml extracted (sonicated or hot reflux or cold soaking) 20 minutes, filtered, the filtrate evaporate to dryness, residue extracts secondary with water-saturated n-butanol after adding water 20ml dissolving, each 20ml, merge normal butyl alcohol liquid, evaporate to dryness, residue solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 1ml makes dissolving, in contrast medicinal material solution.Test according to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same high-efficient silica gel G thin layer plate, with benzene one acetone (10～2): (5～0.2) are developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ to be heated to the spot colour developing clear.In the test sample chromatogram, with the corresponding position of loguat leaf control medicinal material chromatogram on, show the spot of same color.
2, catechu aldehyde in red sage root Central Plains is measured
Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version appendix in 2000 VI D).
Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica, with methyl alcohol-0.8% glacial acetic acid solution (15～2): (85～98) are moving phase, treat that the protocatechualdehyde chromatographic peak goes out behind the peak with methyl alcohol-0.8% glacial acetic acid solution (95～70): (5～30) wash-out 3～25 minutes; The detection wavelength is 280nm.Theoretical cam curve is calculated by the protocatechualdehyde peak should be not less than 6000～2000
It is an amount of that the preparation precision of reference substance solution takes by weighing the protocatechualdehyde reference substance, and solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) is made the solution that every 1ml contains 6.8989 μ g, promptly.
This product 30.7712g is got in the preparation of need testing solution, put in the beaker, the accurate title, decide, add 25ml solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) stir and to make dissolving, be transferred in the 50ml measuring bottle, with solvent (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) cyclic washing beaker repeatedly on a small quantity, washing lotion is incorporated in the same measuring bottle, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, precision is measured 5ml, puts in the 10ml measuring bottle solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) 4ml, shake up, extracted (sonicated or hot reflux or cold soaking) 10 minutes, put cold after, solubilizer (methyl alcohol, ethanol, water, normal butyl alcohol or chloroform) be diluted to scale, shake up, filter, get subsequent filtrate, promptly.
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, promptly.
Calculate
The every g of this product contains the red sage root (with protocatechualdehyde (C as a result
7H
6O
3) meter) 22.42 μ g.
Example two,
Lot number: 030902
By 030901 batch of detection, testing result: (1) differentiates and is positive that (2) red sage root is (with protocatechualdehyde (C
7H
6O
3) meter) 19.84 μ g.
Example three,
Lot number: 031001
By 030901 batch of detection, testing result: (1) differentiates and is positive that (2) red sage root is (with protocatechualdehyde (C
7H
6O
3) meter) 21.35 μ g.
Claims (3)
1, a kind of method for detecting Kidney tonifying mind easing extractive quality standard is characterized in that utilizing thin-layer chromatography that loguat leaf is carried out qualitative identification and with the HPLC method protocatechualdehyde in the red sage root carried out assay.
2, a kind of method for detecting Kidney tonifying mind easing extractive quality standard according to claim 1, the qualitative identification that it is characterized in that wherein said loguat leaf comprises the following steps: to get this product 100g, put in the conical flask, solubilizer methyl alcohol, ethanol, water, normal butyl alcohol or chloroform 200ml, shake up, extract and use sonicated, hot reflux or cold soaking 20 minutes, put cold after, filter the filtrate evaporate to dryness, residue adds water 80ml makes dissolving, extract 2 times with water saturated normal butyl alcohol jolting, each 50ml merges normal butyl alcohol liquid, with the saturated water washing of normal butyl alcohol 2 times, each 80ml discards water liquid, and normal butyl alcohol is recycled to dried, residue solubilizer methyl alcohol, ethanol, water, normal butyl alcohol or chloroform 1ml make dissolving, as need testing solution; Other gets loguat leaf control medicinal material 2g, solubilizer methyl alcohol, ethanol, water, normal butyl alcohol or chloroform 30ml extract with sonicated, hot reflux or cold soaking 20 minutes, filter, the filtrate evaporate to dryness, residue extracts secondary with water-saturated n-butanol after adding water 20ml dissolving, each 20ml, merge normal butyl alcohol liquid, evaporate to dryness, residue solubilizer methyl alcohol, ethanol, water, normal butyl alcohol or chloroform 1ml make dissolving, in contrast medicinal material solution; Test according to thin-layered chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively on same high-efficient silica gel G thin layer plate, with benzene-acetone 10～2: 5～0.2 is developping agent, launch, take out, dry, spray ethanol solution of sulfuric acid with 10%, 105 ℃ to be heated to spot colour developing clear, in the test sample chromatogram, with the corresponding position of loguat leaf control medicinal material chromatogram on, show the spot of same color.
3, a kind of method for detecting Kidney tonifying mind easing extractive quality standard according to claim 1 is characterized in that the content assaying method of protocatechualdehyde in the wherein said red sage root comprises the following steps:
According to high effective liquid chromatography for measuring
(1) chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filling agent, with methyl alcohol-0.8% glacial acetic acid solution 15～2: 85～98 is moving phase, treats to use methyl alcohol-0.8% glacial acetic acid solution 95～70: 5～30 wash-outs 3～25 minutes after the protocatechualdehyde chromatographic peak goes out the peak; The detection wavelength is 280nm.Theoretical cam curve is pressed the calculating of protocatechualdehyde peak should be between 6000～2000;
(2) preparation of reference substance solution: it is an amount of that precision takes by weighing the protocatechualdehyde reference substance, and solubilizer methyl alcohol, ethanol, water, normal butyl alcohol or chloroform are made the solution that every 1ml contains 8 μ g, promptly;
(3) preparation of need testing solution: get this product 30g, put in the beaker, the accurate title, decide, add the 25ml solvent methanol, ethanol, water, normal butyl alcohol or chloroform stir and make dissolving, be transferred in the 50ml measuring bottle, use solvent methanol, ethanol, water, normal butyl alcohol or chloroform be cyclic washing beaker repeatedly on a small quantity, and washing lotion is incorporated in the same measuring bottle, solubilizer methyl alcohol, ethanol, water, normal butyl alcohol or chloroform are diluted to scale, shake up, precision is measured 5ml, puts in the 10ml measuring bottle solubilizer methyl alcohol, ethanol, water, normal butyl alcohol or chloroform 4ml, shake up, extracted sonicated or hot reflux or cold soaking 10 minutes, put cold after, solubilizer methyl alcohol, ethanol, water, normal butyl alcohol or chloroform are diluted to scale, shake up, filter, get subsequent filtrate, promptly;
(4) determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, to measure, the every gram of this product contains the red sage root with protocatechualdehyde C
7H
6O
3Meter must not be less than 16 μ g.
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| CN106442834A (en) * | 2016-08-31 | 2017-02-22 | 吉林修正药业新药开发有限公司 | Method for constructing HPLC characteristic spectra of kidney-tonifying nerve-calming drug |
| CN109668851A (en) * | 2019-03-04 | 2019-04-23 | 安徽协和成制药有限公司 | The quality standard detecting method of monarch drug in a prescription glossy ganoderma extract in a kind of pearl ganoderma lucidum slice |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR960004026B1 (en) * | 1992-11-11 | 1996-03-25 | 김규원 | Angiogenesis inhibitor composition comprising ursolic acid |
| CN1264040A (en) * | 1999-08-11 | 2000-08-23 | 河北省药品检验所 | Method for detecting protocatechuic aldehyde in red sage root |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR960004026B1 (en) * | 1992-11-11 | 1996-03-25 | 김규원 | Angiogenesis inhibitor composition comprising ursolic acid |
| CN1264040A (en) * | 1999-08-11 | 2000-08-23 | 河北省药品检验所 | Method for detecting protocatechuic aldehyde in red sage root |
Non-Patent Citations (1)
| Title |
|---|
| 含枇杷叶的中成药薄层色谱鉴别探讨 李茂森，中国药品标准，第4卷第2期 2003 * |
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