CN105891402A - Method for detecting whether raw materials of honeysuckle extracts and related products are doped with honeysuckle stems - Google Patents
Method for detecting whether raw materials of honeysuckle extracts and related products are doped with honeysuckle stems Download PDFInfo
- Publication number
- CN105891402A CN105891402A CN201610395041.4A CN201610395041A CN105891402A CN 105891402 A CN105891402 A CN 105891402A CN 201610395041 A CN201610395041 A CN 201610395041A CN 105891402 A CN105891402 A CN 105891402A
- Authority
- CN
- China
- Prior art keywords
- extract
- flos lonicerae
- honeysuckle
- water
- lonicerae
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
Abstract
The invention discloses a method for detecting whether raw materials of honeysuckle extracts and related products are doped with honeysuckle stems. Whether raw materials of honeysuckle extracts and related products are doped with honeysuckle stems during production can be judged. The method includes the following steps of evenly mixing to-be-detected samples for extraction; conducting HPLC chromatographic analysis, wherein an HPLC chromatogram is detected, a target component chromatographic peak is positioned, and the peak area ratio is calculated. The method is easy and convenient to operate and simple in step; no organic reagent is used in the preparation process of a test article solution and a comparison product solution, and analysis cost is low.
Description
Technical field
The present invention relates to Chinese medicine extract Quality Control Technology field, former particularly to a kind of Flos Lonicerae extract and Related product
Whether material adulterates the detection method of Caulis Lonicerae.
Background technology
Flower and the stem branch of caprifoliaceae plant Radix Ophiopogonis Lonicera japonica Thunb. all can be used as medicine, the flower that dry flower or band are just opened
I.e. Flos Lonicerae (early summer is gathered before opening), is dried stem branch i.e. Caulis Lonicerae (autumn, season in winter two tap).By going to medical material market, medicine
Shop is investigated, and finds that both price variances are huge, and the price of Flos Lonicerae is about about 10 times of Caulis Lonicerae.Flos Lonicerae extract and phase
Close product to feed intake with Flos Lonicerae, but in order to cost-effective, some lawless person may replace feeding intake with Caulis Lonicerae, or mixes
Enter part Caulis Lonicerae to feed intake.
From appearance character, Flos Lonicerae and Caulis Lonicerae crude drug are easily distinguishable, but owing to deriving from same plant, both extracts
Composition close, even if Flos Lonicerae extract and Related product feed intake with Caulis Lonicerae, prior art also is difficult to be distinguished.Most
Flos Lonicerae extract and Related product are only using chlorogenic acid as index components, and the true and false passing judgment on product is good and bad, but in Caulis Lonicerae also
Containing a large amount of chlorogenic acids, said method can not prevent the behavior fed intake in violation of rules and regulations with Caulis Lonicerae, and the most not to this in prior art
The record of scheme that kind of behavior exercises supervision and checks.
Summary of the invention
Inspection party for the Caulis Lonicerae that solves whether to adulterate when feeding intake without Flos Lonicerae extract and Related product in above prior art
Method, whether the raw material that this application provides a kind of Flos Lonicerae extract and Related product adulterates the detection method of Caulis Lonicerae.
Through studying for a long period of time, isolated one caffeoylquinic acids glucosides: 5-O-[4 '-O-(β-D-Glucopyranose. from Caulis Lonicerae
Base) coffee acyl] quininic acid, determine that this structural formula of compound is as follows through HR-ESI-MS, 2D NMR technology:
This composition is one of main component of Caulis Lonicerae, but in Flos Lonicerae, this component content is the lowest.Research finds, passes through HPLC
Chromatogram Content of Chlorogenic Acid and the ratio of 5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl] quininic acid peak area, it can be determined that gold
The Caulis Lonicerae that whether adulterates in Flos Lonicerae extract and Related product feeds intake.
The present invention is obtained through the following steps:
Whether the raw material of a kind of Flos Lonicerae extract and Related product adulterates the detection method of Caulis Lonicerae, comprises the following steps:
(1) test sample Flos Lonicerae extract and Related product are taken, mixing, take appropriate, precision adds water 50mL, and close plug is weighed
Weight, ultrasonic power 500W, frequency 40kHz processes 30 minutes, takes out, lets cool, more weighed weight, supplies less loss with water
Weight, shake up, filter, collect subsequent filtrate；
(2) accurate subsequent filtrate of drawing, injection chromatograph of liquid, chromatographic condition: with octadecylsilane chemically bonded silica as filler；
With acetonitrile as mobile phase A, with the glacial acetic acid solution of Volume fraction 0.1%～1% as Mobile phase B, use gradient elution mode:
0min → 20min: acetonitrile 8% → 15%, the volume parts glacial acetic acid solution 92% → 85% than 0.1%～1%, detection wavelength is
327nm, positions with reference substance, measures sample Content of Chlorogenic Acid and 5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl] Kui
The ratio X of thujic acid peak area；
In step (2), ratio X is less than 69, it is judged that for being mixed with the Caulis Lonicerae medicine exceeding gross weight 5% in the Chinese medicine honeysuckle that feeds intake
Material.
In step (2), ratio X is less than 42, it is judged that for being mixed with the Caulis Lonicerae exceeding gross weight 10% in the Chinese medicine honeysuckle that feeds intake
Medical material.
In step (2), ratio X is less than 25, it is judged that for being mixed with the Caulis Lonicerae exceeding gross weight 20% in the Chinese medicine honeysuckle that feeds intake
Medical material.
Flos Lonicerae extract and Related product in above-mentioned steps (1), including with the methanol of water or low concentration or ethanol solution (methanol
Or ethanol percent by volume is less than 50%) as Extraction solvent, the single extract of Chinese medicine honeysuckle obtained, Flos Lonicerae and other
Chinese medicine jointly extracts and obtains compound extract, and with the addition of the medicine of said extracted thing, food, health food and cosmetics.
When being applied to compound extract and Related product thereof, this method is applicable to chlorogenic acid therein and 5-O-[4 '-O-(β-D-pyrans Portugal
Grape glycosyl) coffee acyl] quininic acid is derived only from feed intake Flos Lonicerae or the situation of Caulis Lonicerae medical material, say, that in other Chinese medicine not
Containing chlorogenic acid and 5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl] quininic acid.
In above-mentioned steps (2), the preparation of location reference substance solution, takes chlorogenic acid and 5-O-[4 '-O-(β-D-pyrans Portugal the most respectively
Grape glycosyl) coffee acyl] quininic acid reference substance is appropriate, accurately weighed, add water make mass concentration be respectively 20 μ g/mL and
The solution of 30 μ g/mL.
Described method, preferred orientation 5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl] quininic acid reference substance is by following
Step obtains:
(1) taking Caulis Lonicerae medical material 500g, be ground into coarse powder, adding 8 times amount volume fractions is the methanol solution of 50%, ultrasonic merit
Rate 500W, frequency 40kHz extraction 2 times, each 0.5 hour, united extraction liquid also filtered, and reclaims with Rotary Evaporators and extracts
Solvent, to dry, obtains dry extract a；
(2) dry extract a adds equivalent water and makes dissolving, is purified with D101 macroporous adsorptive resins, and column packing consumption is dry leaching
2 times of cream weight, with water as eluting solvent, 3 column volumes of eluting, collect eluent and merge, reclaiming with Rotary Evaporators
Solvent is to doing to obtain dry extract b；
(3) dry extract b adds equivalent water and makes dissolving, is purified with Sephadex LH-20 gel column, and column volume is sample water
15 times of liquor capacity, with water as eluting solvent, 3 column volumes of eluting, collect using 1/40th column volumes as portion and wash
De-liquid, detects with HPLC, merges the eluent containing target component, with Rotary Evaporators recycling design to dry, with
Purification 2 times repeatedly of Sephadex LH-20 gel column, obtain dry extract c；
(4) dry extract c adds water and makes dissolving, and 0.45 μm microporous filter membrane filters, and through half preparation HPLC purification, chromatographic column is ten
Eight alkyl silane bonded silica gels, flowing is acetonitrile-0.4% glacial acetic acid solution mutually, volume ratio 4:96, and 327nm detects, and collection contains
The eluent of target component, collects that liquid is concentrated can obtain 5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl] quinine after drying
Acid, calculates according to areas of peak normalization method, and purity is more than 98%.
After in step (1), Flos Lonicerae extract to be checked and Related product are dissolved in water, every 100mL is equivalent to containing Flos Lonicerae former
Medical material 0.5～5g.
Described Flos Lonicerae is dry flower or the first flower opened of band of caprifoliaceae plant Radix Ophiopogonis Lonicera japonica Thunb..
Described Caulis Lonicerae is the dry stem branch of caprifoliaceae plant Radix Ophiopogonis Lonicera japonica Thunb..
Described gradient elution mode be gradient change component (composition, ionic strength etc.) or the pH of eluent, to by layer
The method that on analysis post, different components was eluted out within the rational time.
Beneficial effect:
1) the inventive method solves in prior art smoothly without throwing with Caulis Lonicerae the most in violation of rules and regulations in Flos Lonicerae extract and Related product
The detection method of material；
2) method is easy and simple to handle, and step is simple；The preparation process of need testing solution and reference substance solution does not uses organic reagent,
Reduce analysis cost.
Accompanying drawing explanation
Fig. 1 is 2.3 lower 5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl] quininic acid reference substance solution HPLC in embodiment 1
Chromatogram；
Fig. 2 is 2.3 lower chlorogenic acid reference substance solution HPLC chromatogram in embodiment 1；
Fig. 3 is the HPLC chromatogram of 2.3 lower Flos Lonicerae extract need testing solutions in embodiment 1；
Fig. 4 is the HPLC chromatogram of 3.1 lower No. 8 Chinese medicine honeysuckles self-control Flos Lonicerae extract need testing solutions in embodiment 1；
Fig. 5 is the HPLC chromatogram of 3.2 lower No. 2 Caulis Lonicerae medical materials self-control Caulis Lonicerae extract need testing solutions in embodiment 1；
Fig. 6 is the HPLC chromatogram making the Flos Lonicerae extract need testing solution that 5% Caulis Lonicerae feeds intake in embodiment 1 under 3.3 by oneself；
Detailed description of the invention
Below by way of specific embodiment, the present invention is further illustrated, not limitation of the invention, according to known in this field
Prior art, embodiments of the present invention are not limited to specific embodiment.
Embodiment 1
1 instrument and reagent
Instrument: Sartorius CP225D electronic balance；Agilent 1200 high performance liquid chromatograph.
Reference substance and source: 5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl] quininic acid (composition A), self-control, with peak
Area normalization method measures, and content is more than 98%；Chlorogenic acid (composition B), National Institute for Food and Drugs Control provides, lot number:
110753-201314, content: 96.6%.
Reagent: acetonitrile, methanol are chromatographically pure, water is purified water prepared by Millipore, and other reagent are analytical pure.
2 chromatographic conditions
The preparation of 2.1 reference substance solution
5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl] quininic acid reference substance solution: precision weighs 5-O-[4 '-O-(β-D-pyrrole
Glucopyranoside base) coffee acyl] quininic acid reference substance 15.36mg, put in 100mL measuring bottle, be dissolved in water and be diluted to scale,
Shake up, as reference substance stock solution A (0.1536mg/mL).Precision measures above-mentioned reference substance stock solution A 10mL, puts 50mL
In measuring bottle, it is diluted with water to scale, shakes up, i.e. obtain reference substance solution A that concentration is 30.72 μ g/mL.
Chlorogenic acid reference substance solution: precision weighs chlorogenic acid reference substance 11.00mg, puts in 100mL measuring bottle, adds 50% methanol molten
Solve and be diluted to scale, shaking up, as reference substance stock solution B (0.1063mg/mL).Precision measures above-mentioned reference substance stock solution
B 5mL, puts in 25mL measuring bottle, adds 50% methanol dilution to scale, shakes up, i.e. obtain the comparison that concentration is 21.25 μ g/mL
Product solution B.
The preparation of 2.2 Flos Lonicerae extract need testing solutions takes Flos Lonicerae extract to be checked (market purchase), pulverizes, mixing,
Take about 0.1g (calculating, be equivalent to Flos Lonicerae crude drug 0.5g) by extraction ratio 20%, accurately weighed, put in tool plug conical flask, essence
The close 50mL that adds water, close plug, weighed weight, ultrasonic power 500W, frequency 40kHz processes 30 minutes, takes out, puts
Cold, more weighed weight, supply the weight of less loss with water, shake up, filter, take subsequent filtrate, to obtain final product.
2.3 chromatographic condition
Dikma Technologies PLATISILTMODS C18(250mm×4.6mm,5mm)；With acetonitrile as mobile phase A,
With 0.4% glacial acetic acid solution as Mobile phase B, the regulation according to the form below carries out gradient elution；Detection wavelength is 327nm.Flow velocity
1.0mL/min, column temperature 35 DEG C.Number of theoretical plate presses the calculating of 5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl] quininic acid peak should
It is not less than 8000.
Precision draws 2.1 lower reference substance solution A and reference substance solution B, 2.2 lower Flos Lonicerae extract need testing solutions respectively
Each 10 μ L, inject chromatograph of liquid, are measured.5-O-[4 '-O-(β-D-glucopyranosyl) in result reference substance and test sample
Coffee acyl] the peak type of quininic acid and chlorogenic acid chromatographic peak is preferable, and number of theoretical plate, separating degree, symmetrical factor all meet related request,
Prove that this chromatographic condition is feasible.Chromatogram is shown in Fig. 1～3 respectively.
3 methodological studies
The ratio of two kinds of compositions in 3.1 Flos Lonicerae extracts
Scheme: collect many batches of Chinese medicine honeysuckles, makes standard Flos Lonicerae extract by oneself, investigates wherein composition B and the ratio of composition A
Relation.Flos Lonicerae extract typically carries out reflux, extract, with water for solvent, therefore simulates above-mentioned carrying during preparation standard Flos Lonicerae extract
Take process.
The preparation of self-control Flos Lonicerae extract need testing solution: extracting honeysuckle medical material, pulverizes, and mixing takes about 1g, accurately weighed,
Precision adds water 100mL, weighs, and reflux, extract, 1h lets cool, more weighed weight, supplies the weight of less loss with water, shakes up, filter
Cross, take subsequent filtrate, to obtain final product.
Respectively precision is drawn 2.1 lower reference substance solution A and reference substance solution B, is made Flos Lonicerae extract test sample by oneself under 3.1
The each 10 μ L of solution, inject chromatograph of liquid, measure, and calculate.The results are shown in Table 1.
Table 1 is made Flos Lonicerae extract by oneself and is investigated result
The ratio of two kinds of compositions in 3.2 Caulis Lonicerae extracts
Scheme: collect many batches of Caulis Lonicerae medical materials, with the extraction process of 3.1 lower Flos Lonicerae extracts, makes Caulis Lonicerae extract by oneself,
Investigate wherein composition B and the proportionate relationship of composition A.
The preparation of self-control Caulis Lonicerae extract need testing solution: take Caulis Lonicerae medical material, pulverizes, and mixing takes about 1g, accurately weighed,
Precision adds water 100mL, weighs, and reflux, extract, 1h lets cool, more weighed weight, supplies the weight of less loss with water, shakes up, filter
Cross, take subsequent filtrate, to obtain final product.
Respectively precision is drawn 2.1 lower reference substance solution A and reference substance solution B, is made Caulis Lonicerae extract by oneself under 3.2 and supply
The each 10 μ L of test sample solution, inject chromatograph of liquid, measure, and calculate.The results are shown in Table 2.
Table 2 is made Caulis Lonicerae extract by oneself and is investigated result
According to table 1 measurement result, in Flos Lonicerae extract, the maximum of composition B and composition A peak area ratio is 236.70,
Little value is 118.98, and meansigma methods is 186.76；According to table 2 measurement result, two kinds of Component peak area ratios of Caulis Lonicerae extract are
Big value is only 1.41, and minima is 0.53, and meansigma methods is 1.05.Flos Lonicerae extract is compared with Caulis Lonicerae extract, above-mentioned
Peak area ratio difference huge (average value 177.87 times), thus can be by measuring this ratio reflection Flos Lonicerae extract
Whether feed intake with Caulis Lonicerae.
The ratio of two kinds of compositions in 3.3 Flos Lonicerae extracts that Caulis Lonicerae feeds intake in varing proportions
Scheme: extracting honeysuckle medical material, add different proportion Caulis Lonicerae medical material, prepare extract, investigate wherein composition B with become
Divide the proportionate relationship of A.
In table 1, in No. 8 Chinese medicine honeysuckle extracts, composition A and composition B each peak area, the two ratio are in meansigma methods water
Flat (No. 8 Chinese medicine honeysuckle extractive HPLC chromatograms are shown in Fig. 4), thus select this batch of medical material to carry out investigation and can reflect gold silver
The average level of flower pesticide material.In like manner, No. 2 Caulis Lonicerae medical material (No. 2 Caulis Lonicerae medicinal substances extract HPLC chromatogram in table 2 are selected
Figure is shown in Fig. 5) test.
Make the preparation of the Flos Lonicerae extract need testing solution that 1% Caulis Lonicerae feeds intake by oneself: take Caulis Lonicerae (Caulis Lonicerae medical material 2) and
Flos Lonicerae (Chinese medicine honeysuckle 8) medicinal powder mixes, and makes Caulis Lonicerae account for the 1% of medical material gross weight, takes about 1g, accurate title
Fixed, precision adds water 100mL, weighs, and reflux, extract, 1h lets cool, more weighed weight, supplies the weight of less loss with water, shakes up,
Filter, take subsequent filtrate, to obtain final product.
According to the method described above, the Flos Lonicerae extract need testing solution that 5%, 10%, 20% Caulis Lonicerae feeds intake is prepared respectively.
Accurate respectively draw 2.1 lower reference substance solution A and reference substance solution B, make Radix Ophiopogonis in varing proportions by oneself under 3.3
The each 10 μ L of Flos Lonicerae extract need testing solution that rattan feeds intake, inject chromatograph of liquid, measure, and calculate.The results are shown in Table 3.
The Flos Lonicerae extract investigation result that different proportion Caulis Lonicerae feeds intake made by oneself by table 3
According to table 3 measurement result, in the medical material that feeds intake, Caulis Lonicerae ratio is the biggest, and composition B and composition A peak area ratio are the least.With
5% Caulis Lonicerae carries out feed intake (HPLC chromatogram is shown in Fig. 6), and this ratio is 69.97, the gold the most substantially less than measured in table 1
Flos Lonicerae extract minimum ratio (118.98).
To sum up, when in Flos Lonicerae extract, composition B and composition A peak area ratio are less than 69, Flos Lonicerae extract crude drug
Material Flos Lonicerae may be mixed with the Caulis Lonicerae more than 5%；When in Flos Lonicerae extract, composition B and composition A peak area ratio are little
In 42 time, Flos Lonicerae extract raw medicinal material Flos Lonicerae may be mixed with the Caulis Lonicerae more than 10%；When in Flos Lonicerae extract
When composition B and composition A peak area ratio are less than 25, Flos Lonicerae extract raw medicinal material Flos Lonicerae may be mixed with more than 20%
Caulis Lonicerae.
4 sample determinations
Extract with the Chinese medicine honeysuckle containing different proportion Caulis Lonicerae, make checking Flos Lonicerae extract by oneself, to the side set up
Method is verified.Never buy many batches of Flos Lonicerae extracts (being shown in Table 4) with manufacturing enterprise, check for the method set up
Mix the situation that Caulis Lonicerae carries out feeding intake.
The preparation of need testing solution: take Flos Lonicerae extract to be measured and pulverize, mixing, take about 0.1g, accurately weighed, put tool plug cone
In shape bottle, precision adds water 50mL, close plug, weighed weight, ultrasonic power 500W, and frequency 40kHz processes 30 minutes, takes
Go out, let cool, more weighed weight, supply the weight of less loss with water, shake up, filter, take subsequent filtrate, to obtain final product.
Precision draws 2.1 lower reference substance solution A and reference substance solution B, each 10 μ L of above-mentioned need testing solution respectively, injects liquid
Chromatography, measures, and calculates.The results are shown in Table 4.
Table 4 commercially available Flos Lonicerae extract measurement result
According to upper table measurement result, self-control checking 1. number extract, B Yu the A peak area ratio (54.17) fed intake with 8% Caulis Lonicerae
2. number falling between the ratio that feeds intake of 5% and 10% Caulis Lonicerae measured under 3.3, the self-control checking fed intake with 15% Caulis Lonicerae is with
Extract, B and A peak area ratio (32.05) falls between the ratio that 10% and 20% Caulis Lonicerae of mensuration feeds intake under 3.3,
Prove that the method set up is accurate, reliable.
Collect 18 batches of Flos Lonicerae extract samples of 11 manufacturing enterprises altogether, be directed to 9 batch sample compositions of 5 enterprises
B Yu A peak area ratio is less than 69, may be mixed with the Caulis Lonicerae more than 5%, account for the 50% of total batch；Wherein face, 8 batches of peaks
Long-pending ratio is even less than 25, more serious with the situation ratio that Caulis Lonicerae feeds intake.According to the above results, Flos Lonicerae extract manufacturing enterprise
The situation that incorporation Caulis Lonicerae feeds intake is commonplace, and inspection method disclosed by the invention will be sent out in terms of specification Flos Lonicerae extract market
Wave huge effect.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention should not be limited by the examples, other
The change made under any spirit without departing from the present invention and principle, modify, combine, substitute, simplify and all should be equivalence and replace
Change mode, within being included in protection scope of the present invention.
Claims (9)
1. whether the raw material of a Flos Lonicerae extract and Related product adulterates the detection method of Caulis Lonicerae, it is characterised in that include following step
Rapid:
(1) taking Flos Lonicerae extract to be checked and Related product, add water, supersound process makes dissolving, takes out, lets cool, shake up, filter
Cross, collect subsequent filtrate；
(2) subsequent filtrate is injected chromatograph of liquid, calculate chlorogenic acid and 5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl]
The ratio X of quininic acid peak area,
Ratio X is less than 69, it is judged that for being mixed with the Caulis Lonicerae medical material exceeding gross weight 5% in the Chinese medicine honeysuckle that feeds intake,
Ratio X is less than 42, it is judged that for being mixed with the Caulis Lonicerae medical material exceeding gross weight 10% in the Chinese medicine honeysuckle that feeds intake,
Ratio X is less than 25, it is judged that for being mixed with the Caulis Lonicerae medical material exceeding gross weight 20% in the Chinese medicine honeysuckle that feeds intake.
Method the most according to claim 1, it is characterised in that chromatographic condition in step (2): with octadecylsilane bonded silica
Glue is filler；With acetonitrile as mobile phase A, with the glacial acetic acid solution of Volume fraction 0.1%～1% as Mobile phase B, use ladder
Degree type of elution: 0min → 20min: acetonitrile 8% → 15%, the volume parts glacial acetic acid solution 92% → 85% than 0.1%～1%,
Detection wavelength is 327nm.
Method the most according to claim 1, it is characterised in that respectively with chlorogenic acid and 5-O-[4 '-O-(β-D-pyrrole in step (2)
Glucopyranoside base) coffee acyl] corresponding chromatographic peak positions by quininic acid reference substance.
Method the most according to claim 1, it is characterised in that ultrasonic power 500W in step (1), the process of frequency 40kHz
30 minutes.
Method the most according to claim 1, it is characterised in that Flos Lonicerae extract product, is to be less than with water or percent by volume
The methanol of 50% or ethanol solution are as Extraction solvent, the single extract of Chinese medicine honeysuckle obtained or Flos Lonicerae with other Chinese medicine altogether
Extract together the compound extract obtained, and with the addition of above-mentioned single extract or the medicine of compound extract, food, health care food
Product or cosmetics, do not contain chlorogenic acid and 5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl] quinine in other Chinese medicine described
Acid.
Method the most according to claim 3, it is characterised in that chlorogenic acid and 5-O-[4 '-O-(β-D-glucopyranosyl) caffeoyl
Base] quininic acid reference substance mass concentration is respectively 20 ± 2 μ g/mL and 30 ± 3 μ g/mL.
Method the most according to claim 3, it is characterised in that location 5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl]
Quininic acid reference substance is obtained through the following steps:
(1) taking Caulis Lonicerae medical material 500g, be ground into coarse powder, adding 8 times amount volume fractions is the methanol solution of 50%, ultrasonic merit
Rate 500W, frequency 40kHz extraction 2 times, each 0.5 hour, united extraction liquid also filtered, and reclaims with Rotary Evaporators and extracts
Solvent, to dry, obtains dry extract a；
(2) dry extract a adds equivalent water and makes dissolving, is purified with D101 macroporous adsorptive resins, and column packing consumption is dry leaching
2 times of cream weight, with water as eluting solvent, 3 column volumes of eluting, collect eluent and merge, reclaiming with Rotary Evaporators
Solvent is to doing to obtain dry extract b；
(3) dry extract b adds equivalent water and makes dissolving, is purified with Sephadex LH-20 gel column, and column volume is sample water
15 times of liquor capacity, with water as eluting solvent, 3 column volumes of eluting, collect using 1/40th column volumes as portion and wash
De-liquid, detects with HPLC, merges the eluent containing target component, with Rotary Evaporators recycling design to dry, with
Purification 2 times repeatedly of Sephadex LH-20 gel column, obtain dry extract c；
(4) dry extract c adds water and makes dissolving, and 0.45 μm microporous filter membrane filters, and through half preparation HPLC purification, chromatographic column is ten
Eight alkyl silane bonded silica gels, flowing is acetonitrile-0.4% glacial acetic acid solution mutually, volume ratio 4:96, and 327nm detects, and collection contains
The eluent of target component, collects that liquid is concentrated can obtain 5-O-[4 '-O-(β-D-glucopyranosyl) coffee acyl] quinine after drying
Acid, calculates according to areas of peak normalization method, and purity is more than 98%.
Method the most according to claim 2, it is characterised in that with the glacial acetic acid solution of Volume fraction 0.4% as Mobile phase B.
Method the most according to claim 1, it is characterised in that in step (1), Flos Lonicerae extract to be checked and Related product add water
After dissolving, every 100mL is equivalent to containing Flos Lonicerae crude drug 0.5～5g.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610395041.4A CN105891402B (en) | 2016-06-03 | 2016-06-03 | Whether the raw material of Honegsukle flower P.E and Related product adulterates the detection method of caulis lonicerae |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610395041.4A CN105891402B (en) | 2016-06-03 | 2016-06-03 | Whether the raw material of Honegsukle flower P.E and Related product adulterates the detection method of caulis lonicerae |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105891402A true CN105891402A (en) | 2016-08-24 |
| CN105891402B CN105891402B (en) | 2017-06-06 |
Family
ID=56710738
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610395041.4A Active CN105891402B (en) | 2016-06-03 | 2016-06-03 | Whether the raw material of Honegsukle flower P.E and Related product adulterates the detection method of caulis lonicerae |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105891402B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109541099A (en) * | 2018-11-27 | 2019-03-29 | 山东省食品药品检验研究院 | A kind of discrimination method of Radix Paeoniae Alba rhizome or its extract |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101269182A (en) * | 2007-03-23 | 2008-09-24 | 北京亚东生物制药有限公司 | Pharmaceutical composition for treating children's wind-heat cold, preparation method and quality control method |
| US20110126313A1 (en) * | 2008-02-18 | 2011-05-26 | Norfolk Plant Sciences Limited | Methods and compositions for modifying plant flavonoid composition and disease resistance |
| CN103524348A (en) * | 2013-09-29 | 2014-01-22 | 湖南金地生物科技有限公司 | Process for extracting chlorogenic acid from honeysuckle and dry leaves of honeysucklestem through biological enzyme |
| CN105606734A (en) * | 2016-01-05 | 2016-05-25 | 华中科技大学 | Method for detecting honeysuckle flower and lonicerae flos medicinal materials through rapid resolution liquid chromatography |
-
2016
- 2016-06-03 CN CN201610395041.4A patent/CN105891402B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101269182A (en) * | 2007-03-23 | 2008-09-24 | 北京亚东生物制药有限公司 | Pharmaceutical composition for treating children's wind-heat cold, preparation method and quality control method |
| US20110126313A1 (en) * | 2008-02-18 | 2011-05-26 | Norfolk Plant Sciences Limited | Methods and compositions for modifying plant flavonoid composition and disease resistance |
| CN103524348A (en) * | 2013-09-29 | 2014-01-22 | 湖南金地生物科技有限公司 | Process for extracting chlorogenic acid from honeysuckle and dry leaves of honeysucklestem through biological enzyme |
| CN105606734A (en) * | 2016-01-05 | 2016-05-25 | 华中科技大学 | Method for detecting honeysuckle flower and lonicerae flos medicinal materials through rapid resolution liquid chromatography |
Non-Patent Citations (5)
| Title |
|---|
| EFFICIENT N NCUBE ET AL: "Analyses of chlorogenic acids and related cinnamic acid derivatives from Nicotiana tabacumtissues with the aid of UPLC-QTOF-MS MS based on the in-source collision-induced dissociation method", 《CHEMISTRY CENTRAL JOURNAL》 * |
| JIA-YU ZHANG ET AL: "Diagnostic fragment-ion-based and extension strategy coupled to DFIs intensity analysis for identification of chlorogenic acids isomers in Flos Lonicerae Japonicae by HPLC-ESI-MSn", 《TALANTA》 * |
| RAKESH JAISWAL ET AL: "Identification and characterization of chlorogenic acids, chlorogenic acid glycosides and flavonoids from Lonicera henryi L. (Caprifoliaceae) leaves by LC–MSn", 《PHYTOCHEMISTRY》 * |
| SHUI-HAN ZHANG ET AL: "Typical ultraviolet spectra in combination with diagnostic mass fragmentation analysis for the rapid and comprehensive profiling of chlorogenic acids in the buds of Lonicera macranthoides", 《ANAL BIOANAL CHEM》 * |
| 赵昱 等: "咖啡酰奎尼酸类化合物研究进展", 《中国中药杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109541099A (en) * | 2018-11-27 | 2019-03-29 | 山东省食品药品检验研究院 | A kind of discrimination method of Radix Paeoniae Alba rhizome or its extract |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105891402B (en) | 2017-06-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104215698B (en) | The fingerprint atlas detection method of Callicarpa nudiflora medical material, intermediate and preparation thereof and standard finger-print | |
| CN105259295B (en) | Quality detection method for ginseng, cassia twig and poria cocos oral solution | |
| CN105606734A (en) | Method for detecting honeysuckle flower and lonicerae flos medicinal materials through rapid resolution liquid chromatography | |
| CN102928523B (en) | Wild chrysanthemum flower fingerprint determination method, its application, and wild chrysanthemum flower quality detection method | |
| CN104297026A (en) | Method for extracting effective flavonoid components in traditional Chinese medicine pericarpium citri reticulatae | |
| CN104020235A (en) | Method for simultaneously determining content of chlorogenic acid and galuteolin in lonicera japonica | |
| CN106706786A (en) | Method for determining content of six ginsenoside ingredients of folium ginseng | |
| CN103808835A (en) | Method for simultaneously measuring contents of 10 chemical components in four-substance soup decoction by HPLC-DAD (high performance liquid chromatography-diode array detection) method | |
| CN109668970A (en) | A kind of ultra performance liquid chromatography detection method of Chinese medicine composition | |
| CN110243988A (en) | The method for building up of strong medicine white flower nine inner bright HPLC finger-print | |
| CN102520103A (en) | Contrast Chinese patent medicine extract of pills of six ingredients with rehmannia, and preparation method and application thereof | |
| CN104007198B (en) | A kind of glossy ganoderma emperor's preparation HPLC standard finger-print and construction method thereof and application | |
| CN104155383B (en) | The detection method of blue or green Pu granule | |
| CN103575821A (en) | Detection method of 14 chemical components in Tangminling preparation | |
| CN102998383A (en) | Method for testing content of main components in Rhodiola rosea extracts | |
| CN105891402B (en) | Whether the raw material of Honegsukle flower P.E and Related product adulterates the detection method of caulis lonicerae | |
| CN104730158A (en) | Content measurement method for modified huoxiangzhengqi soft capsule | |
| CN106290645A (en) | The construction method of a kind of Lhasa rhubarb finger printing and standard finger-print thereof | |
| CN102008541B (en) | Method for simultaneously detecting three main active ingredients in sugar-free type compound wintercreeper preparation | |
| CN104237441A (en) | Method for simultaneous detection of iridoid glycoside, phenylethanoid glycoside, flavone and dicaffeoyl ingredients in lamiophlomis rotata | |
| CN100432670C (en) | Method for inspecting Chinese-medicinal preparation Kaiyinwan | |
| CN104597168B (en) | Ramulus Mori refines decoction pieces content assaying method | |
| CN104483411B (en) | A kind of capsule of weeping forsythia and the detection method containing capsule of weeping forsythia product | |
| CN103340406A (en) | Preparation method of cordyceps sinensis granules | |
| CN104458954B (en) | A kind of dodder formulation granule finger printing and method for building up thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |