CN102297912A - Quality control method of licorice medicinal material whole-time multi-wavelength fusion fingerprint chromatogram - Google Patents
Quality control method of licorice medicinal material whole-time multi-wavelength fusion fingerprint chromatogram Download PDFInfo
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Abstract
The invention discloses a quality control method of a licorice medicinal material whole-time multi-wavelength fusion fingerprint chromatogram, which combines modern analysis detection means with information processing means, effectively fuses the chromatogram information of traditional Chinese medicine index components, can overcome the disadvantage of insufficient information content during single-wavelength single-index detection, and thus can perfectly reflect the internal quality of the traditional Chinese medicinal material. The quality control method of the invention has simple operation, is stable and reliable, has good reproducibility, and is applicable to quality evaluation and control of traditional Chinese medicinal materials and new medicine research and development of traditional Chinese medicine. Especially, the invention can be used as one of the indexes for licorice medicinal material quality control and identification.
Description
Invention field
The invention belongs to the technical field that adopts modern analysis detection means and information processing means that the quality of Chinese crude drug is detected, the control method that relates to a kind of Chinese crude drug quality, a kind of licorice medicinal materials all the period of time multi-wavelength of saying so more specifically merges fingerprint pattern quality control method.
Background technology
According to record of Pharmacopoeia of People's Republic of China version in 2010: this product is the dry root and the rhizome of glycyrrhizic legume, glycyrrhiza inflate bat or glycyrrhiza glabra.Beginning is stated from Sheng Nong's herbal classic, classifies as top grade.The gentle current twelve regular channels of this product property of medicine, traditional medicine are thought effects such as its useful gas bowl spares, clearing heat and detoxicating, expelling phlegm and arresting coughing, relieving spasm to stop pain, coordinating the drug actions of a prescription has saying of " ten sides, nine grass ".Clinically be mainly used in chronic hepatitis cirrhosis, canker, just send out treatment of diseases such as EA, pernicious vomiting severe, dysmenorrheal caused by qi stagnation and blood stasis, lumber ertebral disc be outstanding and the chronic liver disease liver histological improves, chronic liver disease liver cancer becomes inhibition etc.In China, Radix Glycyrrhizae mainly originates in areas such as the Inner Mongol, Gansu, Xinjiang, northeast, Shanxi.Mainly contain triterpenes and flavones ingredient in the Radix Glycyrrhizae, wherein triterpenes components mainly contains glycyrrhizic acid, enoxolone etc.; Flavones ingredient mainly contains liquiritin, Radix Glycyrrhizae aglycon, isoliquiritin etc.
Along with science and technology development, generally using and the information processing technology progressively perfect of modern advanced analysis detecting instrument had Chinese medicine analyzing and testing level and significantly improves, and the traditional Chinese medicine quality control method is updated.By setting up finger-print the Chinese crude drug index components is carried out a kind of development trend that assay becomes traditional Chinese medicine quality control and estimates in recent years.Traditional Chinese medicine fingerprint is meant some Chinese crude drug or Chinese patent drug through after suitably handling, the spectrogram or the chromatogram that can manifest Chinese crude drug or pcm chemical feature that utilize present information acquisition technique and quality analysis means to obtain.Since under two different chromatographic conditions, use different pre-treating methods to Radix Glycyrrhizae in liquiritin, isoliquiritin and mono-ammonium glycyrrhizinate detect respectively and can't realize the Radix Glycyrrhizae finger-print is carried out comprehensively and the overall evaluation, therefore, follow the information maximization principle and set up licorice medicinal materials all the period of time multi-wavelength and merge finger-print and its index components is carried out assay can obtain a kind of comparatively effective quality control method, thereby guarantee the safe, stable and reliable of licorice medicinal materials quality.
Licorice medicinal materials occupies crucial status in Chinese crude drug.At present, commercially available licorice medicinal materials quality is uneven, so the stability of licorice medicinal materials is difficult to guarantee.The assay of 2010 editions Chinese Pharmacopoeia regulations: this product is pressed dry product and is calculated, and contains liquiritin and must not be less than 0.50%; Glycyrrhizic acid must not be less than 2.0% in ammonium glycyrrhetate.
Summary of the invention
The object of the invention is to adopt modern analysis detection means and information processing means to set up the brand-new finger-print of licorice medicinal materials, reach content assaying method based on all the period of time multi-wavelength fusion of this finger-print, easier, reasonable, reliable with the method for quality control of guaranteeing licorice medicinal materials.
The present invention is achieved by the following technical solutions:
Licorice medicinal materials all the period of time multi-wavelength merges the method for quality control of finger-print, it is characterized in that this method carries out as follows:
(1) preparation of need testing solution: the about 5g of extracting liquorice medicinal powder, the accurate title, decide, and puts in the round-bottomed flask, precision adds entry 100mL, claims to decide weight, reflux 2 hours, put cold, claim to decide weight, water is supplied the weight that subtracts mistake, shakes up again, filter, the accurate subsequent filtrate 20mL that draws is added on the AB-8 resin column, with distilled water 120mL washing, use the 100mL95% ethanol elution again, collect ethanol eluate, evaporate to dryness, residue add 20% acetonitrile to be made dissolving and is transferred in the 10mL measuring bottle, add 20% acetonitrile and be settled to scale, shake up, filter, get subsequent filtrate promptly;
(2) preparation of reference substance solution: it is an amount of that precision takes by weighing mono-ammonium glycyrrhizinate reference substance, liquiritin reference substance and isoliquiritin reference substance, add methyl alcohol and make the solution that every 1mL contains mono-ammonium glycyrrhizinate reference substance 0.154mg, liquiritin reference substance 0.587mg and isoliquiritin reference substance 0.362mg respectively, promptly;
(3) chromatographic condition and system suitability test: with octadecyl silane is the Agilent TC-C of filling agent
18Chromatographic column; With 0.2 ‰ formic acid water is mobile phase A, is Mobile phase B with the acetonitrile, and gradient elution: 0 ~ 10min is 5% ~ 10%B, and 10 ~ 61min is 10% ~ 27%B, and 61 ~ 70min is 27% ~ 40%B, and 70 ~ 90min is 40% ~ 50%B; Flow velocity 1.0mL/min; 30 ℃ of column temperatures; The DAD detecting device detects under wavelength 254nm, 279nm and 362 nm simultaneously; Sample size 10 μ L; Theoretical cam curve is calculated, and should be not less than 3000;
(4) all the period of time multi-wavelength fusion: adopt multi-wavelength to merge fingerprint pattern technology the spectral data under 254nm, 279nm and three wavelength of 362 nm is carried out all the period of time fusion, obtain a chromatogram that can comprise three wavelength information simultaneously.
(5) sample size is measured: use the finger-print data after merging, the content of index components in the calculation sample.
(6) above-mentioned definite content assaying method is carried out efficiency evaluation, comprising specificity, accuracy, precision, repeatability, stability, linearity, scope and durability.
The method of quality control of licorice medicinal materials of the present invention, the preparation of preferred need testing solution: the about 5g of extracting liquorice medicinal powder, the accurate title, decide, put in the round-bottomed flask, precision adds entry 100mL, claims to decide weight, reflux 2 hours is put coldly, claims to decide weight again, water is supplied the weight that subtracts mistake, shake up, filter, the accurate subsequent filtrate 20mL that draws, be added on the AB-8 resin column, with distilled water 120mL washing, use the 100mL95% ethanol elution again, collect ethanol eluate, evaporate to dryness, residue add 20% acetonitrile make the dissolving and be transferred in the 10mL measuring bottle, add 20% acetonitrile and be settled to scale, shake up, filter, get subsequent filtrate promptly;
Licorice medicinal materials method of quality control of the present invention, preferred detection method: with octadecyl silane is the Agilent TC-C of filling agent
18Chromatographic column; With 0.2 ‰ formic acid water is mobile phase A, is Mobile phase B with the acetonitrile, and gradient elution: 0 ~ 10min is 5% ~ 10%B, and 10 ~ 61min is 10% ~ 27%B, and 61 ~ 70min is 27% ~ 40%B, and 70 ~ 90min is 40% ~ 50%B; Flow velocity 1.0mL/min; 30 ℃ of column temperatures; The DAD detecting device detects under wavelength 254nm, 279nm and 362 nm simultaneously; Sample size 10 μ L; Theoretical cam curve is calculated, and should be not less than 3000;
Licorice medicinal materials method of quality control of the present invention, obtain spectral data under 254nm, 279nm and three wavelength of 362 nm by analysis and detecting instrument, exploit information is handled the spectral data under three wavelength of means all the period of time fusion, obtains a chromatogram that can comprise three wavelength information simultaneously.
Licorice medicinal materials method of quality control of the present invention will can obtain one group of spectral data after the spectral data all the period of time fusion under 254nm, 279nm and three wavelength of 362 nm.
Licorice medicinal materials method of quality control of the present invention, the finger-print data that obtain can be used for the fingerprint qualitative analysis of licorice medicinal materials.
Licorice medicinal materials method of quality control of the present invention, the finger-print data that obtain can be used for the quantitative test of licorice medicinal materials index components.
Licorice medicinal materials all the period of time multi-wavelength fusion fingerprint pattern quality control method of the present invention is compared the good effect that is had with the assay method of existing Chinese Pharmacopoeia regulation and is:
(1) the present invention is open first by setting up the method that licorice medicinal materials all the period of time multi-wavelength merges finger-print and its index components carried out assay, the method is to same sample heterogeneity multi-wavelength qualitative and quantitative analysis simultaneously, thus perfect licorice medicinal materials method of quality control.
(2) the present invention adopts modern advanced analysis detection technique, and a plurality of index components of licorice medicinal materials are carried out analyzing and testing, thereby obtains extremely abundant quantity of information.
(3) the present invention carries out all the period of time fusion by information processing means with the chromatogram under the licorice medicinal materials multi-wavelength, is guaranteeing to obtain a chromatogram that reacts a plurality of wavelength information simultaneously under the minimum prerequisite of data message loss.
(4) method of quality control of the present invention, its advantage is simple to operate, and is reliable and stable, precision, repeatability, the recovery all reach the quantitative test requirement, can comprehensively analyze the quality of licorice medicinal materials on a macro scale.
(5) the present invention is merged finger-print by setting up licorice medicinal materials all the period of time multi-wavelength, each constituents in the licorice medicinal materials is carried out the fingerprint qualitative and quantitative analysis, disclose the inherent quality feature of licorice medicinal materials truly comprehensively, thereby guaranteed the safe, effective, reliable, stable of licorice medicinal materials, guaranteed the equalization stable of its quality and the standardization of technology.
Description of drawings
Fig. 1 is reference substance liquiritin under the 254nm wavelength, isoliquiritin, mono-ammonium glycyrrhizinate chromatogram.
Fig. 2 is reference substance liquiritin under the 279nm wavelength, isoliquiritin, mono-ammonium glycyrrhizinate chromatogram.
Fig. 3 is reference substance liquiritin under the 362nm wavelength, isoliquiritin, mono-ammonium glycyrrhizinate chromatogram.
Fig. 4 is the UV scanning figure of liquiritin.
Fig. 5 is the UV scanning figure of isoliquiritin.
Fig. 6 is the UV scanning figure of mono-ammonium glycyrrhizinate.
Fig. 7 is a HPLC finger-print under the licorice medicinal materials 254nm wavelength.
Fig. 8 is a HPLC finger-print under the licorice medicinal materials 279nm wavelength.
Fig. 9 is a HPLC finger-print under the licorice medicinal materials 362nm wavelength.
Figure 10 is chromatogram and the fusion peak that licorice medicinal materials all the period of time multi-wavelength merges.
Claims (4)
1. licorice medicinal materials all the period of time multi-wavelength merges the method for quality control of finger-print, it is characterized in that this method carries out as follows:
(1) preparation of need testing solution: the about 5g of extracting liquorice medicinal powder, the accurate title, decide, and puts in the round-bottomed flask, precision adds entry 100mL, claims to decide weight, reflux 2 hours, put cold, claim to decide weight, water is supplied the weight that subtracts mistake, shakes up again, filter, the accurate subsequent filtrate 20mL that draws is added on the AB-8 resin column, with distilled water 120mL washing, use the 100mL95% ethanol elution again, collect ethanol eluate, evaporate to dryness, residue add 20% acetonitrile to be made dissolving and is transferred in the 10mL measuring bottle, add 20% acetonitrile and be settled to scale, shake up, filter, get subsequent filtrate promptly;
(2) preparation of reference substance solution: it is an amount of that precision takes by weighing mono-ammonium glycyrrhizinate reference substance, liquiritin reference substance and isoliquiritin reference substance, add methyl alcohol and make the solution that every 1mL contains mono-ammonium glycyrrhizinate reference substance 0.154mg, liquiritin reference substance 0.587mg and isoliquiritin reference substance 0.362mg respectively, promptly;
(3) chromatographic condition and system suitability test: with octadecyl silane is filling agent Agilent TC-C
18Chromatographic column; With 0.2 ‰ formic acid water is mobile phase A, is Mobile phase B with the acetonitrile, and gradient elution: 0 ~ 10min is 5% ~ 10%B, and 10 ~ 61min is 10% ~ 27%B, and 61 ~ 70min is 27% ~ 40%B, and 70 ~ 90min is 40% ~ 50%B; Flow velocity 1.0mL/min; 30 ℃ of column temperatures; The DAD detecting device detects under wavelength 254nm, 279nm and 362 nm simultaneously; Sample size 10 μ L; Theoretical cam curve is calculated, and should be not less than 3000;
(4) all the period of time multi-wavelength fusion: adopt multi-wavelength to merge fingerprint pattern technology the spectral data under 254nm, 279nm and three wavelength of 362 nm is carried out all the period of time fusion, obtain a chromatogram that can comprise three wavelength information simultaneously;
(5) sample size is measured: use the finger-print data after merging, the content of index components in the calculation sample;
(6) above-mentioned definite content assaying method is carried out efficiency evaluation, comprising specificity, accuracy, precision, repeatability, stability, linearity, scope and durability.
2. licorice medicinal materials all the period of time multi-wavelength as claimed in claim 1 merges the method for quality control of finger-print, it is characterized in that the preparation of this method need testing solution: the about 5g of extracting liquorice medicinal powder, the accurate title, decide, put in the round-bottomed flask, precision adds entry 100mL, claims to decide weight, reflux 2 hours, put cold, claim to decide weight, water is supplied the weight that subtracts mistake, shakes up again, filter, the accurate subsequent filtrate 20mL that draws is added on the AB-8 resin column, with distilled water 120mL washing, use the 100mL95% ethanol elution again, collect ethanol eluate, evaporate to dryness, residue add 20% acetonitrile to be made dissolving and is transferred in the 10mL measuring bottle, add 20% acetonitrile and be settled to scale, shake up, filter, get subsequent filtrate promptly.
3. licorice medicinal materials all the period of time multi-wavelength as claimed in claim 1 merges the method for quality control of finger-print, it is characterized in that, this method chromatographic condition and system suitability test: with octadecyl silane is the Agilent TC-C of filling agent
18Chromatographic column; With 0.2 ‰ formic acid water is mobile phase A, is Mobile phase B with the acetonitrile, and gradient elution: 0 ~ 10min is 5% ~ 10%B, and 10 ~ 61min is 10% ~ 27%B, and 61 ~ 70min is 27% ~ 40%B, and 70 ~ 90min is 40% ~ 50%B; Flow velocity 1.0mL/min; 30 ℃ of column temperatures; The DAD detecting device detects under wavelength 254nm, 279nm and 362 nm simultaneously; Sample size 10 μ L; Theoretical cam curve is calculated, and should be not less than 3000.
4. licorice medicinal materials all the period of time multi-wavelength as claimed in claim 1 merges the method for quality control of finger-print, it is characterized in that, all the period of time multi-wavelength fusion: adopt multi-wavelength to merge fingerprint pattern technology the spectral data under 254nm, 279nm and three wavelength of 362 nm is carried out all the period of time fusion, obtain a chromatogram that can comprise three wavelength information simultaneously.
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Cited By (10)
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| CN102539576A (en) * | 2012-01-30 | 2012-07-04 | 辽宁中医药大学 | Control method for simultaneously detecting content of nine components through five wavelength merging method for licorice medicinal materials |
| CN102680629A (en) * | 2012-05-18 | 2012-09-19 | 新疆全安药业有限公司 | Method for establishing Anweiyang capsule fingerprint spectrum |
| CN102680628A (en) * | 2012-05-18 | 2012-09-19 | 新疆全安药业有限公司 | Method for establishing fingerprint of An Weiyang |
| CN102692461A (en) * | 2012-05-22 | 2012-09-26 | 辽宁中医药大学 | Whole-time three-wavelength fusion method for simultaneously determining contents of four ingredients in Flos Carthami |
| CN102692460A (en) * | 2012-05-22 | 2012-09-26 | 辽宁中医药大学 | Process for determining content of four components of honeysuckle during full-time-interval three-wavelength fusion |
| CN102759585A (en) * | 2012-07-02 | 2012-10-31 | 辽宁华润本溪三药有限公司 | Method for controlling full-time interval multi-wavelength fused fingerprint quality of qi stagnation and stomachache granules |
| CN105467051A (en) * | 2014-09-11 | 2016-04-06 | 富力 | Dachuanxiong tablet whole-time multi-wavelength fusion fingerprint quality control method |
| CN109932432A (en) * | 2017-12-15 | 2019-06-25 | 富力 | The method of quality control of Shenyi capsule bulk pharmaceutical chemicals all the period of time multi-wavelength fusion finger-print |
| CN110274970A (en) * | 2018-12-28 | 2019-09-24 | 吉林三九金复康药业有限公司 | The method for building up for melting poor finger-print and its application in Yixiesheng capsule Quality Control |
| CN111443154A (en) * | 2020-05-21 | 2020-07-24 | 贵州中医药大学 | Research method of medicinal genetic relationship of glycyrrhiza |
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| CN102539576A (en) * | 2012-01-30 | 2012-07-04 | 辽宁中医药大学 | Control method for simultaneously detecting content of nine components through five wavelength merging method for licorice medicinal materials |
| CN102680629B (en) * | 2012-05-18 | 2014-08-06 | 新疆全安药业有限公司 | Method for establishing Anweiyang capsule fingerprint spectrum |
| CN102680629A (en) * | 2012-05-18 | 2012-09-19 | 新疆全安药业有限公司 | Method for establishing Anweiyang capsule fingerprint spectrum |
| CN102680628A (en) * | 2012-05-18 | 2012-09-19 | 新疆全安药业有限公司 | Method for establishing fingerprint of An Weiyang |
| CN102680628B (en) * | 2012-05-18 | 2015-01-14 | 新疆全安药业有限公司 | Method for establishing fingerprint of An Weiyang |
| CN102692460A (en) * | 2012-05-22 | 2012-09-26 | 辽宁中医药大学 | Process for determining content of four components of honeysuckle during full-time-interval three-wavelength fusion |
| CN102692461B (en) * | 2012-05-22 | 2013-12-18 | 辽宁中医药大学 | Whole-time three-wavelength fusion method for simultaneously determining contents of four ingredients in Flos Carthami |
| CN102692461A (en) * | 2012-05-22 | 2012-09-26 | 辽宁中医药大学 | Whole-time three-wavelength fusion method for simultaneously determining contents of four ingredients in Flos Carthami |
| CN102759585B (en) * | 2012-07-02 | 2013-12-04 | 辽宁华润本溪三药有限公司 | Method for controlling full-time interval multi-wavelength fused fingerprint quality of qi stagnation and stomachache granules |
| CN102759585A (en) * | 2012-07-02 | 2012-10-31 | 辽宁华润本溪三药有限公司 | Method for controlling full-time interval multi-wavelength fused fingerprint quality of qi stagnation and stomachache granules |
| CN105467051A (en) * | 2014-09-11 | 2016-04-06 | 富力 | Dachuanxiong tablet whole-time multi-wavelength fusion fingerprint quality control method |
| CN105467051B (en) * | 2014-09-11 | 2017-07-28 | 富力 | The method of quality control of big Ligusticum wallichii piece all the period of time multi-wavelength fusion finger-print |
| CN109932432A (en) * | 2017-12-15 | 2019-06-25 | 富力 | The method of quality control of Shenyi capsule bulk pharmaceutical chemicals all the period of time multi-wavelength fusion finger-print |
| CN110274970A (en) * | 2018-12-28 | 2019-09-24 | 吉林三九金复康药业有限公司 | The method for building up for melting poor finger-print and its application in Yixiesheng capsule Quality Control |
| CN110274970B (en) * | 2018-12-28 | 2021-11-05 | 吉林三九金复康药业有限公司 | Method for establishing melting difference fingerprint spectrum and application of melting difference fingerprint spectrum in quality control of Yixuesheng capsules |
| CN111443154A (en) * | 2020-05-21 | 2020-07-24 | 贵州中医药大学 | Research method of medicinal genetic relationship of glycyrrhiza |
| CN111443154B (en) * | 2020-05-21 | 2023-03-07 | 贵州中医药大学 | Research method of medicinal genetic relationship of glycyrrhiza |
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