CN104614475A - Thirst quenching clearing particle content detection method - Google Patents
Thirst quenching clearing particle content detection method Download PDFInfo
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- CN104614475A CN104614475A CN201310538597.0A CN201310538597A CN104614475A CN 104614475 A CN104614475 A CN 104614475A CN 201310538597 A CN201310538597 A CN 201310538597A CN 104614475 A CN104614475 A CN 104614475A
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Abstract
The invention relates to a thirst quenching clearing particle content detection method, the method comprises the following steps: preparation of a to-be-tested sample solution, to be more specific, taking a thirst quenching clearing preparation, adding proper amount of extraction solvent methanol or ethanol, ultrasonically dissolving, adding a solvent to adjust the solution concentration, filtering with a microfiltration membrane to obtain filtrate; preparation of a control stock solution, to be more specific, taking neomangiferin, mangiferin, timosaponin BII, palmatine and berberine controls, adding methanol for dissolving to obtain the control stock solution; determination of content, to be more specific, respectively injecting the to-be-tested sample solution and the control stock solution into UPLC-PDA-ELSD chromatograph to obtain a chromatogram, and according to the chromatogram, using the peak area method for calculation of content of the neomangiferin, mangiferin, timosaponin BII, palmatine and berberine.
Description
Technical field:
The present invention relates to a kind of effective constituent determination method of medicine, particularly one is quenched one's thirst particle Chinese traditional medicine active constituent content detection method clearly
Background technology:
Particle clearly of quenching one's thirst is a kind of Chinese patent drug for the treatment of diabetes of having gone on the market, and its formula is composed as follows: the wind-weed, rhizoma atractylodis, the coptis, cattail pollen, wolf's milk and appropriate amount of auxiliary materials, and its preparation method comprises the following steps:
A () gets the wind-weed, rhizoma atractylodis, the coptis, cattail pollen, wolf's milk, for subsequent use;
(b) rhizoma atractylodis extraction by steam distillation volatile oil 5 hours, by Rhzoma Atractylodis Lanceae volatile oil beta-cyclodextrin inclusion compound, Atractylis oil and beta-schardinger dextrin-feed intake by 1: 6, and 50 DEG C of dryings, obtained Benexate Hydrochloride is for subsequent use, and extract filters, and obtains filtrate 1;
C the residue after () wind-weed, the coptis, cattail pollen, wolf's milk and rhizoma atractylodis water extraction adds 8 times amount soak by water twice, first time decocts 2 hours, second time decocts 1 hour, merge twice decocting liquid, filter to get filtrate 2, merging filtrate 1 and 2, concentrated filtrate become proportion be 1.10 ~ 1.15 clear cream, dry dry extract;
D (): above-mentioned dry extract and volatile oil clathrate compound, fully mix, and obtains pharmaceutically active substance, acceptable auxiliary material is mixed and made into the dosage forms such as particle with medicine further.
Quench one's thirst clearly in particle, each taste medicine plays different effects, wherein, the arduous temperature of rhizoma atractylodis, enter taste two warp, fragrant odour, benefaction and not keeping, both can be drying damp and strengthening spleen, again can activating QI to alleviate the depression, therefore ZHU Dan-xi calls it: " rhizoma atractylodis are Foot yangming Channel medicine, and smell is pungent strong; Gu Zhiqi is sent out in strong stomach invigorating the spleen ", " can separate all strongly fragrant ".The wind-weed is sweet cold, and enter lung, stomach, kidney channel, have nourishing Yin and falling fire, the merit of clearing heat and moistening dryness, Compendium of Material Medica is thought, " then moisten the dry and enriching yin of stomach under the wind-weed, on then clearing lung-heat gold and purging intense heat.", " herbal classic " meaning " hanker, except perverse trend by main quenching one's thirst.", two medicines are all monarch, the mutual compatibility of two medicines, concurrence nourishing kidney-yin, and invigorating the spleen is transported, and change Yu Zhigong; Coptis smell bitter cold, has heat-clearing and damp-drying drug, purging intense heat and detonicating, with for minister, the auxiliary wind-weed, to protect kidney yin, helps rhizoma atractylodis turbid with dampness elimination; Pollen Typhae taste cool in nature is sweet pungent, enters the heart, liver two warp, cooling blood and removing stasis, blood Tianjin cloth and scorchingly to separate, and the stasis of blood is degassed row, and cloudy Tianjin is spontaneous, and can improve diabetes and merge cardio cerebrovascular affection, with for assistant; Wolf's milk is natural hypoglycemic quality goods among the people, and mildly bitter flavor is pungent and property is put down, clearing heat and detoxicating, and blood circulation and channel invigorating is not only kind hypoglycemic, and also have good effect to diabetes complicated ulcer sore furuncle, with for making, all medicines share, then play nourishing YIN and clearing away fire altogether, the merit that invigorating the spleen is invigorated blood circulation, all diseases can alleviate and heal it.All medicines above, according to the basic theory of science of TCM formulas, ingenious compatibility, tightly combines, and plays nourishing Yin and falling fire, clearing away heat and eliminating dampeness, merit promoting blood circulation and removing blood stasis altogether, thus reaches the object to healing.
Particle (XiaoKeQing granules, XKQ) clearly of quenching one's thirst is made up of the wind-weed, rhizoma atractylodis, the coptis, cattail pollen and wolf's milk 5 taste medicine.Chinese patent: 200310107286,200410048002.4 disclose relevant formula and preparation.At present, granular mass standard content clearly of quenching one's thirst only has carried out assay to the sarsasapogenin in wind-weed hydrolysate under measuring item, globality and the complicacy of Chinese medicine preparation cannot be characterized comprehensively, owing to quenching one's thirst, particle is clearly prepared from by 5 kinds of Chinese crude drugs, its effective constituent complexity is various, be necessary to find a kind of method that can detect wherein Multiple components, with the content of product in effective production control process and quality, inventors performed the experiment of multiple detection method for this reason, final acquisition is a kind of to be applicable to quenching one's thirst the method for particle clearly, and 5 the composition Neomangiferins higher to wherein content, mangiferin, timosaponin BII, palmatine and jamaicin have carried out assay.
Summary of the invention:
The present invention adopt UPLC-PDA-ELSD chromatography to quench one's thirst clearly in preparation Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin carried out assay, for the quality of quenching one's thirst clearly provides safety, effectively, accurately controls.
The invention provides a kind of detection method of the preparation clearly of quenching one's thirst, particularly including the detection method of the particle clearly of quenching one's thirst.
Said method comprising the steps of:
Prepared by need testing solution:
Cancel thirsty clear preparation, add Extraction solvent methyl alcohol or appropriate amount of ethanol, ultrasonic dissolution, then add solvent adjustment solution concentration, miillpore filter filters, and gets filtrate, both.
The preparation of reference substance storing solution:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance, add methyl alcohol and dissolve, obtain reference substance storing solution.
Assay:
Above-mentioned need testing solution and reference substance storing solution are injected UPLC-PDA-ELSD chromatograph respectively, obtain chromatogram, according to chromatogram, the content of Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin in need testing solution is calculated by peak area method, and the content of these compositions in the preparation clearly of obtaining quenching one's thirst.
Wherein, described UPLC-PDA-ELSD chromatograph, its chromatographic condition is as follows:
Chromatographic column: be selected from: UPLC C18 or T3 post;
Mobile phase: be selected from: solvent orange 2 A (acetate buffer solution (containing 0.3-0.5% acetic acid and 0.05-0.1% triethylamine)) – solvent B(acetonitrile); Solvent orange 2 A (0.1% aqueous formic acid) – solvent B(acetonitrile); One in solvent orange 2 A (0.1% aqueous acetic acid) – solvent B(acetonitrile).
The present invention adopts gradient elution to carry out wash-out, described gradient elution: be selected from the one in following four different elution protocol:
①0～4min，5%–10%B，4～10min，10%–15%B，10～13min，15%–22%B，13～15min，22%–30%B，15～18min，30%B，18～20min，30%–80%B，20～23min，80%–5%B；
②0～4min，5%B，4～5min，5%–10%B，5～8min，10%B，8～10min，10-15%B，10～11min，15-12%B，11～13min，12%–22%B，13～15min，22%–30%B，15～18min，30%B，18～20min，30%–80%B，20～23min，80%–5%B；
③0～4min，5%B，4～5min，5%–10%B，5～8min，10%B，8～10min，10-15%B，10～11min，15-12%B，11～13min，12%–20%B，13～14min，20%–25%B，14～18min，25–30%B，20～23min，80%–5%B；
④0～4min，5%B，4～5min，5%–10%B，5～8min，10%B，8～10min，10-20%B，10～13min，20%B，13～14min，20%–25%B，14～18min，25–30%B，20～22min，80%–5%B；
Column temperature: 25-35 DEG C
Flow velocity: 0.15-0.25ml/min
ELSD air velocity: 2.0-3.0Lmin
-1;
Drift tube temperature: 100-110 DEG C;
Sample size is 2-4 μ L.
Determined wavelength: 240-340nm
Preferably, need testing solution preparation method of the present invention is as follows:
Cancel thirsty clear preparation 1-7g, with 50-80% ethanol or the ultrasonic extraction of 25-80% methyl alcohol, ultrasonic time is 20-40min;
Preferred, need testing solution preparation method of the present invention is as follows: Extraction solvent is the ethanol of 75%, or the methyl alcohol of 70-80%.
Most preferred need testing solution preparation method of the present invention is as follows:
Cancel thirsty clear preparation (particle clearly of preferably quenching one's thirst) 6g, accurately weighed, put in 25ml measuring bottle, add 75% methyl alcohol appropriate, ultrasonic (power 500w) 30min, is placed to room temperature, adds 75% methanol constant volume to scale, and 0.22 μm of miillpore filter filters, and gets subsequent filtrate, both.
Preferably, the preparation preparation method of reference substance storing solution of the present invention is as follows:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance, add the methyl alcohol of 70-80%.Dissolve, obtain reference substance storing solution.
Preferred, the preparation preparation method of reference substance storing solution of the present invention is as follows:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance, add the methyl alcohol of 75%.
Dissolve, obtain reference substance storing solution.
Most preferred, the preparation preparation method of reference substance storing solution of the present invention is as follows:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance appropriate, accurately weighed, add 75% methyl alcohol and dissolve, obtain mass concentration and be respectively 0.244,0.784,0.772,0.070 and 0.224 μ g μ L
-1reference substance storing solution.
Preferably, chromatographic condition of the present invention is as follows:
Chromatographic column: Waters Acquity UPLC HSS C
18(2.1 × 100mm, 1.8 μm);
Mobile phase: solvent orange 2 A (acetate buffer solution (containing 0.5% acetic acid and 0.1% triethylamine)) – solvent B(acetonitrile),
Gradient elution (0 ~ 4min, 5% – 10%B, 4 ~ 8min, 10% – 15%B, 8 ~ 13min, 15% – 22%B, 13 ~ 15min, 22% – 30%B, 15 ~ 18min, 30%B, 18 ~ 20min, 30% – 80%B, 20 ~ 23min, 80% – 5%B; Gradient elution process only illustrates the content of solvent B, and remainder is solvent orange 2 A); Column temperature: 30 DEG C;
Flow velocity: 0.2mLmin
-1;
Sample size: 3 μ L;
Determined wavelength: 258nm;
ELSD air velocity: 2.5Lmin
-1;
Drift tube temperature: 105 DEG C,
Yield value: 1.
Relational language related to the present invention is explained
UPLC-PDA-ELSD: the visible ultraviolet of ultra high efficiency liquid phase multi-wavelength and evaporative light detection method.
To quench one's thirst preparation clearly, comprise particle clearly of quenching one's thirst, tablet clearly of quenching one's thirst, gel resin wafer of quenching one's thirst, clearing dropping pill agent of quenching one's thirst, suppository clearly of quenching one's thirst, oral solutions clearly of quenching one's thirst, injection clearly of quenching one's thirst.Can according to the method preparation in known patent.Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin are known drug composition, can commercially obtain.
The present invention may be used for above-mentioned five kinds of compositions in detection of drugs preparation and content thereof, and therefore the present invention is also applicable to the other drug preparation simultaneously containing above-mentioned five kinds of compositions.
The preferred method of the present invention is through experiment screening acquisition, and the selection result is as follows:
System suitability test
Get test sample (lot number 130301) solution, reference substance solution (0.50mL reference substance storing solution adds 75% methanol constant volume to 2mL) and negative control solution, detect according to chromatographic process in embodiment 1, record chromatogram compares that ultraviolet and evaporative light to detect in spectrogram determination test sample ultraviolet chromatogram that No. 1 peak is Neomangiferin, No. 2 peaks are mangiferin, No. 4 peaks are palmatine, No. 5 peaks are jamaicin, and in ELSD chromatogram, No. 3 peaks are timosaponin BII.These theoretical cam curve >=120000, system condition No. 2 peaks.(indicate 1 in literary composition: Neomangiferin 2 mangiferin; 3 timosaponin BII; 4. palmatine; 5. jamaicin)
Linear relationship is investigated
Precision measures reference substance storing solution each 0.10 prepared by embodiment 1 method, 0.25, 0.50, 0.60 and 1.00mL put 2mL volumetric flask, add 75% methyl alcohol precision and be diluted to scale, obtain the reference substance solution of 5 different quality concentration, measure according to chromatographic condition sample introduction in embodiment 1, Neomangiferin, mangiferin, palmatine and jamaicin with reference substance concentration X for horizontal ordinate, peak area Y is that ordinate carries out linear regression, timosaponin BII with the logarithm value X of reference substance concentration for horizontal ordinate, the logarithm value Y of peak area is that ordinate carries out linear regression, obtain regression equation, the results are shown in Table 1, show that each reference substance solution is linearly good within the scope of respective concentration.
The each component linear equation of table 1 and concentration range thereof
Precision test
The above-mentioned linearity control product solution of accurate absorption (0.50mL reference substance storing solution adds 75% methanol constant volume to 2mL), continuous sample introduction 5 times, the RSD value recording Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin retention time is respectively 0.3%, 0.3%, 0.1%, 0.1% and 0.1%, the RSD of peak area is respectively 2.5%, and 2.3%, 2.7%, 2.4% and 2.5%, show that instrument precision is good.
Precision data:
Replica test
Sample thief (lot number 130301) 5 parts, prepare need testing solution by embodiment 1 method, detect by embodiment 1 method, the RSD value of the retention time of result Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin peak area is respectively 0.1%, 0.2%, 0.1%, 0.1% and 0.1%, RSD is respectively 2.9%, 2.6%, 2.9%, 2.7% and 2.9%, show that the method repeatability is good.
Repeatability data:
Stability test
The above-mentioned linearity control product solution of accurate absorption (0.50mL reference substance storing solution adds 75% methanol constant volume to 2mL), respectively at 0,2,4,8,12,24h sample introduction is analyzed, and the RSD value of the retention time of result Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin is respectively 0.3%, 0.3%, 0.1%, 0.1% and 0.1%, RSD is respectively 3.0%, 2.6%, 2.5%, 2.8% and 3.0%, show that this solution is good at 24h internal stability.
Average recovery is tested
Get the particle (lot number 130301) clearly of quenching one's thirst of 9 parts of known content, every part of 3.0g, the reference substance solution of suitable concentration is quantitatively added according to 80%, 100%, 120% of often kind of component amount to be measured in sample, need testing solution is prepared by embodiment 1 method, obtain the solution of three kinds of variable concentrations, each 3 parts, measure by the chromatographic condition of embodiment 1, calculate the recovery and relative standard deviation value.The average recovery rate that result records Neomangiferin is 98.5%, RSD value is 1.1%; The average recovery rate of mangiferin is 98.8%, RSD value is 0.7%; The average recovery rate of timosaponin BII is 99.2%, RSD value is 0.6%; The average recovery rate of palmatine is 98.0%, RSD value is 1.1%; The average recovery rate of jamaicin is 98.0%, RSD value is 1.2%.
Sample size measures
Precision takes 11 batch samples, prepares need testing solution respectively by embodiment 1 method, measures by embodiment 1 condition.The results are shown in Table 2.
Assay result (the mgg of 5 kinds of chemical compositions in table 211 batch sample
-1)
Beneficial effect of the present invention
At present, granular mass standard content clearly of quenching one's thirst only has carried out assay to the sarsasapogenin in wind-weed hydrolysate under measuring item, and detection method is HPLC-ELSD method, particle clearly of quenching one's thirst is made up of the wind-weed, rhizoma atractylodis, the coptis, cattail pollen and wolf's milk 5 taste medicine, only carry out measuring the quality effectively cannot learning the particle clearly of quenching one's thirst to a kind of composition of wind-weed hydrolysate, therefore, we have carried out abundant extraction to granular preparation clearly of quenching one's thirst and have carried out chromatographic resolution to ingredient in laboratory.In extraction for the purpose of Overall Acquisition formulation chemist composition, and with UPLC-PDA-ELSD method, chromatographic resolution is carried out to composition, more original quicker, accurate containing the survey method method of inspection, more fully can analyze the quality of the pharmaceutical preparations.
Accompanying drawing explanation
Quench one's thirst particle (A), reference substance (B) and negative control clearly of Fig. 1 (lacks wind-weed C; Lack coptis D; Lack rhizoma atractylodis E; Lack cattail pollen F; Lack wolf's milk G) UPLC-PDA chromatogram
Quench one's thirst particle (A), reference substance (B) and negative control clearly of Fig. 2 (lacks wind-weed C; Lack coptis D; Lack rhizoma atractylodis E; Lack cattail pollen F; Lack wolf's milk G) UPLC-ELSD chromatogram
Embodiment:
Further illustrate the present invention by the following examples.
Embodiment 1
Instrument and reagent
ACQUITY UPLC(Waters, US, comprise quaternary geopressure gradient pump, vacuum degassing machine, automatic sampler, column oven, PDA detecting device, Empower3 chromatographic work station), evaporative light-scattering detector (ELSD) (Alltech ELSD2000), KQ-500DE type numerical control ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd.), analytical balance (XS105 type electronic analytical balance, Mettler Toledo), MilliporeSimplicity ultrapure water system.
Acetonitrile, methyl alcohol (chromatographic grade, Merck); Glacial acetic acid (chromatographic grade, Concord, Tianjin company limited); Triethylamine (chromatographically pure, Tianjin Yuan Li Chemical Co., Ltd.); Absolute ethyl alcohol (analyzing pure, Li Anlongbohua Tianjin medical chemistry company limited); Mangiferin (lot number 111607-200402), timosaponin BII(lot number 222839-201102) and jamaicin (lot number 110713-201212) reference substance purchased from National Institute for Food and Drugs Control; Neomangiferin and palmatine reference substance are purchased from Phytomarker Co., Ltd..
Control medicinal material: the wind-weed (Rhizoma Anemarrhenae), the coptis (Coptis chinensis Franch), Atractylis lancea (Atractylodes lancea (Thunb.) DC.), cattail pollen (Typha angustifolia L.) and wolf's milk (Herba Euphorbiae Humifusae) provide by Tianjin Tasly Modern Chinese Medicine Resource Co., Ltd.;
Particulate samples clearly of quenching one's thirst to be detected is provided by Tianjin Tasly Pharmaceutical Group Co., Ltd..
Chromatographic condition
Chromatographic column: Waters Acquity UPLC HSS C
18(2.1 × 100mm, 1.8 μm); Mobile phase: solvent orange 2 A (acetate buffer solution (containing 0.5% acetic acid and 0.1% triethylamine)) – solvent B(acetonitrile), gradient elution (0 ~ 4min, 5% – 10%B, 4 ~ 8min, 10% – 15%B, 8 ~ 13min, 15% – 22%B, 13 ~ 15min, 22% – 30%B, 15 ~ 18min, 30%B, 18 ~ 20min, 30% – 80%B, 20 ~ 23min, 80% – 5%B; Gradient elution process only illustrates the content of solvent B, and remainder is solvent orange 2 A); Column temperature: 30 DEG C; Flow velocity: 0.2mLmin
-1; Sample size: 3 μ L; Determined wavelength: 258nm; ELSD air velocity: 2.5Lmin
-1; Drift tube temperature: 105 DEG C, yield value: 1.
The preparation of need testing solution and reference substance storing solution
The preparation of need testing solution
Cancel thirsty clear particle and be about 6.0g, accurately weighed, put in 25ml measuring bottle, add 75% methyl alcohol appropriate, ultrasonic (power 500w) 30min, is placed to room temperature, adds 75% methanol constant volume to scale, and 0.22 μm of miillpore filter filters, and gets subsequent filtrate, both.
The preparation of reference substance storing solution
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance appropriate, accurately weighed, add 75% methyl alcohol and dissolve, obtain mass concentration and be respectively 0.244,0.784,0.772,0.070 and 0.224 μ g μ L
-1reference substance storing solution.
Embodiment 2
1, to quench one's thirst the detection method of preparation clearly, comprise the following steps:
Prepared by need testing solution:
Cancel thirsty clear preparation 1g, extract with 50% EtOH Sonicate, ultrasonic time is 20min.
Add solvent adjustment solution concentration again, miillpore filter filters, and gets filtrate, both,
The preparation of reference substance storing solution:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance appropriate, accurately weighed, add 75% methyl alcohol and dissolve, obtain mass concentration and be respectively 0.244,0.784,0.772,0.070 and 0.224 μ g μ L
-1reference substance storing solution.
Assay:
Above-mentioned need testing solution and reference substance storing solution are injected UPLC-PDA-ELSD chromatograph respectively, obtain chromatogram, according to chromatogram, the content of Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin in need testing solution is calculated by peak area method, and the content of these compositions in the preparation clearly of obtaining quenching one's thirst
Wherein, described UPLC-PDA-ELSD chromatograph, its chromatographic condition is as follows:
Chromatographic column: be selected from: UPLC C18 post;
Mobile phase: be selected from: solvent orange 2 A (acetate buffer solution (containing 0.3-0.5% acetic acid and 0.05-0.1% triethylamine)) – solvent B(acetonitrile);
Gradient elution is adopted to carry out wash-out, described gradient elution: be selected from:
①0～4min，5%–10%B，4～10min，10%–15%B，10～13min，15%–22%B，13～15min，22%–30%B，15～18min，30%B，18～20min，30%–80%B，20～23min，80%–5%B；
Column temperature: 25 DEG C
Flow velocity: 0.15ml/min
ELSD air velocity: 2.0Lmin
-1;
Drift tube temperature: 100 DEG C;
Sample size is 2 μ L,
Determined wavelength: 240nm.
Embodiment 3
1, to quench one's thirst the detection method of preparation clearly, comprise the following steps:
Prepared by need testing solution:
Cancel thirsty clear preparation 7g, extract with 80% EtOH Sonicate, ultrasonic time is 40min.
The preparation of reference substance storing solution:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance appropriate, accurately weighed, add 75% methyl alcohol and dissolve, obtain mass concentration and be respectively 0.244,0.784,0.772,0.070 and 0.224 μ g μ L
-1reference substance storing solution.
Assay:
Above-mentioned need testing solution and reference substance storing solution are injected UPLC-PDA-ELSD chromatograph respectively, obtain chromatogram, according to chromatogram, the content of Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin in need testing solution is calculated by peak area method, and the content of these compositions in the preparation clearly of obtaining quenching one's thirst
Wherein, described UPLC-PDA-ELSD chromatograph, its chromatographic condition is as follows:
Chromatographic column: be selected from: T3 post;
Mobile phase: be selected from: solvent orange 2 A (0.1% aqueous formic acid) – solvent B(acetonitrile);
Gradient elution is adopted to carry out wash-out, described gradient elution: be selected from
0～4min，5%B，4～5min，5%–10%B，5～8min，10%B，8～10min，10-15%B，10～11min，15-12%B，11～13min，12%–22%B，13～15min，22%–30%B，15～18min，30%B，18～20min，30%–80%B，20～23min，80%–5%B；
Column temperature: 35 DEG C
Flow velocity: 0.25ml/min
ELSD air velocity: 3.0Lmin
-1;
Drift tube temperature: 110 DEG C;
Sample size is 4 μ L,
Determined wavelength: 340nm.
Embodiment 4
1, to quench one's thirst the detection method of preparation clearly, comprise the following steps:
Prepared by need testing solution:
Cancel thirsty clear preparation 3.5g, with the ultrasonic extraction of 25% methyl alcohol, then add solvent adjustment solution concentration, miillpore filter filters, and gets filtrate, both,
The preparation of reference substance storing solution:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance appropriate, accurately weighed, add 75% methyl alcohol and dissolve, obtain mass concentration and be respectively 0.244,0.784,0.772,0.070 and 0.224 μ g μ L
-1reference substance storing solution.
Assay:
Above-mentioned need testing solution and reference substance storing solution are injected UPLC-PDA-ELSD chromatograph respectively, obtain chromatogram, according to chromatogram, the content of Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin in need testing solution is calculated by peak area method, and the content of these compositions in the preparation clearly of obtaining quenching one's thirst
Wherein, described UPLC-PDA-ELSD chromatograph, its chromatographic condition is as follows:
Chromatographic column: be selected from: UPLC C18 post;
Mobile phase: be selected from: solvent orange 2 A (0.1% aqueous acetic acid) – solvent B(acetonitrile),
Gradient elution is adopted to carry out wash-out, described gradient elution: be selected from
0～4min，5%B，4～5min，5%–10%B，5～8min，10%B，8～10min，10-15%B，10～11min，15-12%B，11～13min，12%–20%B，13～14min，20%–25%B，14～18min，25–30%B，20～23min，80%–5%B；
Column temperature: 25-35 DEG C
Flow velocity: 0.15-0.25ml/min
ELSD air velocity: 2.0-3.0Lmin
-1;
Drift tube temperature: 100-110 DEG C;
Sample size is 2-4 μ L,
Determined wavelength: 240-340nm.
Embodiment 5
1, to quench one's thirst the detection method of preparation clearly, comprise the following steps:
Prepared by need testing solution:
Cancel thirsty clear preparation 5g, with the ultrasonic extraction of 80% methyl alcohol, then add solvent adjustment solution concentration, miillpore filter filters, and gets filtrate, both,
The preparation of reference substance storing solution:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance appropriate, accurately weighed, add 75% methyl alcohol and dissolve, obtain mass concentration and be respectively 0.244,0.784,0.772,0.070 and 0.224 μ g μ L
-1reference substance storing solution.
Assay:
Above-mentioned need testing solution and reference substance storing solution are injected UPLC-PDA-ELSD chromatograph respectively, obtain chromatogram, according to chromatogram, the content of Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin in need testing solution is calculated by peak area method, and the content of these compositions in the preparation clearly of obtaining quenching one's thirst
Wherein, described UPLC-PDA-ELSD chromatograph, its chromatographic condition is as follows:
Chromatographic column: be selected from: T3 post;
Mobile phase: be selected from: solvent orange 2 A (acetate buffer solution (containing 0.3-0.5% acetic acid and 0.05-0.1% triethylamine)) – solvent B(acetonitrile);
Gradient elution is adopted to carry out wash-out, described gradient elution: be selected from
0～4min，5%B，4～5min，5%–10%B，5～8min，10%B，8～10min，10-20%B，10～13min，20%B，13～14min，20%–25%B，14～18min，25–30%B，20～22min，80%–5%B；
Column temperature: 25-35 DEG C
Flow velocity: 0.15-0.25ml/min
ELSD air velocity: 2.0-3.0Lmin
-1;
Drift tube temperature: 100-110 DEG C;
Sample size is 2-4 μ L,
Determined wavelength: 240-340nm.
Embodiment 6
To quench one's thirst the detection method of preparation clearly, comprise the following steps:
Prepared by need testing solution:
Cancel thirsty clear preparation 3g, extract with 75% EtOH Sonicate, then add solvent adjustment solution concentration, miillpore filter filters, and gets filtrate, both,
The preparation of reference substance storing solution:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance, add methyl alcohol and dissolve, obtain reference substance storing solution,
Assay:
Above-mentioned need testing solution and reference substance storing solution are injected UPLC-PDA-ELSD chromatograph respectively, obtain chromatogram, according to chromatogram, the content of Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin in need testing solution is calculated by peak area method, and the content of these compositions in the preparation clearly of obtaining quenching one's thirst
Wherein, described UPLC-PDA-ELSD chromatograph, its chromatographic condition is as follows:
Chromatographic column: be selected from: UPLC C18 or T3 post;
Mobile phase: be selected from: solvent orange 2 A (acetate buffer solution (containing 0.3-0.5% acetic acid and 0.05-0.1% triethylamine)) – solvent B(acetonitrile)
Gradient elution is adopted to carry out wash-out, described gradient elution: 0 ~ 4min, 5% – 10%B, 4 ~ 10min, 10% – 15%B, 10 ~ 13min, 15% – 22%B, 13 ~ 15min, 22% – 30%B, 15 ~ 18min, 30%B, 18 ~ 20min, 30% – 80%B, 20 ~ 23min, 80% – 5%B;
Column temperature: 35 DEG C
Flow velocity: 0.15ml/min
ELSD air velocity: 3.0Lmin
-1;
Drift tube temperature: 100 DEG C;
Sample size is 2 μ L,
Determined wavelength: 240nm.
Embodiment 7
To quench one's thirst the detection method of preparation clearly, comprise the following steps:
Prepared by need testing solution:
Cancel thirsty clear preparation 3g, with the ultrasonic extraction of 75% methyl alcohol, then add solvent adjustment solution concentration, miillpore filter filters, and gets filtrate, both,
The preparation of reference substance storing solution:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance, add methyl alcohol and dissolve, obtain reference substance storing solution,
Assay:
Above-mentioned need testing solution and reference substance storing solution are injected UPLC-PDA-ELSD chromatograph respectively, obtain chromatogram, according to chromatogram, the content of Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin in need testing solution is calculated by peak area method, and the content of these compositions in the preparation clearly of obtaining quenching one's thirst
Wherein, described UPLC-PDA-ELSD chromatograph, its chromatographic condition is as follows:
Chromatographic column: be selected from: UPLC C18 or T3 post;
Mobile phase: be selected from: solvent orange 2 A (0.1% aqueous formic acid) – solvent B(acetonitrile);
Gradient elution is adopted to carry out wash-out, described gradient elution:
0～4min，5%B，4～5min，5%–10%B，5～8min，10%B，8～10min，10-15%B，10～11min，15-12%B，11～13min，12%–22%B，13～15min，22%–30%B，15～18min，30%B，18～20min，30%–80%B，20～23min，80%–5%B；
Column temperature: 25 DEG C
Flow velocity: 0.25ml/min
ELSD air velocity: 2.0Lmin
-1;
Drift tube temperature: 100 DEG C;
Sample size is 4 μ L,
Determined wavelength: 340nm.
Embodiment 8
To quench one's thirst the detection method of preparation clearly, comprise the following steps:
Prepared by need testing solution:
Cancel thirsty clear preparation 3g, with the ultrasonic extraction of 50% methyl alcohol alcohol, then add solvent adjustment solution concentration, miillpore filter filters, and gets filtrate, both,
The preparation of reference substance storing solution:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance, add methyl alcohol and dissolve, obtain reference substance storing solution,
Assay:
Above-mentioned need testing solution and reference substance storing solution are injected UPLC-PDA-ELSD chromatograph respectively, obtain chromatogram, according to chromatogram, the content of Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin in need testing solution is calculated by peak area method, and the content of these compositions in the preparation clearly of obtaining quenching one's thirst
Wherein, described UPLC-PDA-ELSD chromatograph, its chromatographic condition is as follows:
Chromatographic column: be selected from: UPLC C18 or T3 post;
Mobile phase: be selected from: solvent orange 2 A (0.1% aqueous acetic acid) – solvent B(acetonitrile),
Gradient elution is adopted to carry out wash-out, described gradient elution:
0～4min，5%B，4～5min，5%–10%B，5～8min，10%B，8～10min，10-15%B，10～11min，15-12%B，11～13min，12%–20%B，13～14min，20%–25%B，14～18min，25–30%B，20～23min，80%–5%B；
Column temperature: 30 DEG C
Flow velocity: 0.2ml/min
ELSD air velocity: 2.5Lmin
-1;
Drift tube temperature: 105 DEG C;
Sample size is 3 μ L,
Determined wavelength: 258nm.
Embodiment 9
To quench one's thirst the detection method of preparation clearly, comprise the following steps:
Prepared by need testing solution:
Cancel thirsty clear preparation 3g, with the ultrasonic extraction of 50% methyl alcohol alcohol, then add solvent adjustment solution concentration, miillpore filter filters, and gets filtrate, both,
The preparation of reference substance storing solution:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance, add methyl alcohol and dissolve, obtain reference substance storing solution,
Assay:
Above-mentioned need testing solution and reference substance storing solution are injected UPLC-PDA-ELSD chromatograph respectively, obtain chromatogram, according to chromatogram, the content of Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin in need testing solution is calculated by peak area method, and the content of these compositions in the preparation clearly of obtaining quenching one's thirst
Wherein, described UPLC-PDA-ELSD chromatograph, its chromatographic condition is as follows:
Chromatographic column: be selected from: UPLC C18 or T3 post;
Mobile phase: be selected from: solvent orange 2 A (acetate buffer solution (containing 0.3-0.5% acetic acid and 0.05-0.1% triethylamine)) – solvent B(acetonitrile); Gradient elution is adopted to carry out wash-out, described gradient elution:
0～4min，5%B，4～5min，5%–10%B，5～8min，10%B，8～10min，10-20%B，10～13min，20%B，13～14min，20%–25%B，14～18min，25–30%B，20～22min，80%–5%B；
Column temperature: 30 DEG C
Flow velocity: 0.2ml/min
ELSD air velocity: 2.5Lmin
-1;
Drift tube temperature: 105 DEG C;
Sample size is 3 μ L,
Determined wavelength: 258nm.
Claims (10)
1. to quench one's thirst the detection method of preparation clearly, comprise the following steps:
Prepared by need testing solution:
Cancel thirsty clear preparation, add Extraction solvent methyl alcohol or appropriate amount of ethanol, ultrasonic dissolution, then add solvent adjustment solution concentration, miillpore filter filters, and gets filtrate, both,
The preparation of reference substance storing solution:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance, add methyl alcohol and dissolve, obtain reference substance storing solution,
Assay:
Above-mentioned need testing solution and reference substance storing solution are injected UPLC-PDA-ELSD chromatograph respectively, obtain chromatogram, according to chromatogram, the content of Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin in need testing solution is calculated by peak area method, and the content of these compositions in the preparation clearly of obtaining quenching one's thirst
Wherein, described UPLC-PDA-ELSD chromatograph, its chromatographic condition is as follows:
Chromatographic column: UPLC C18 or T3 post;
Mobile phase: be selected from: solvent orange 2 A acetate buffer solution is wherein containing 0.3-0.5% acetic acid and 0.05-0.1% tri-Yi An – solvent B acetonitrile; Solvent orange 2 A 0.1% first acid aqueous solution – solvent B acetonitrile; Solvent orange 2 A 0.1% vinegar acid aqueous solution – solvent B acetonitrile, in one;
Gradient elution is adopted to carry out wash-out, described gradient elution: to be selected from the one in following four different elution protocol:
①0～4min，5%–10%B，4～10min，10%–15%B，10～13min，15%–22%B，13～15min，22%–30%B，15～18min，30%B，18～20min，30%–80%B，20～23min，80%–5%B；
②0～4min，5%B，4～5min，5%–10%B，5～8min，10%B，8～10min，10-15%B，10～11min，15-12%B，11～13min，12%–22%B，13～15min，22%–30%B，15～18min，30%B，18～20min，30%–80%B，20～23min，80%–5%B；
③0～4min，5%B，4～5min，5%–10%B，5～8min，10%B，8～10min，10-15%B，10～11min，15-12%B，11～13min，12%–20%B，13～14min，20%–25%B，14～18min，25–30%B，20～23min，80%–5%B；
④0～4min，5%B，4～5min，5%–10%B，5～8min，10%B，8～10min，10-20%B，10～13min，20%B，13～14min，20%–25%B，14～18min，25–30%B，20～22min，80%–5%B；
Column temperature: 25-35 DEG C
Flow velocity: 0.15-0.25ml/min
ELSD air velocity: 2.0-3.0Lmin
-1;
Drift tube temperature: 100-110 DEG C;
Sample size is 2-4 μ L,
UV detect wavelength: 240-340nm.
2. method according to claim 1, is characterized in that, described need testing solution preparation method is as follows:
Cancel thirsty clear preparation 1-7g, with 50-80% ethanol or the ultrasonic extraction of 25-80% methyl alcohol, ultrasonic time is 20-40min.
3. method according to claim 1, is characterized in that, described need testing solution preparation method is as follows:
Extraction solvent is the ethanol of 75%, or the methyl alcohol of 70-80%.
4. method according to claim 1, is characterized in that, described need testing solution preparation method is as follows:
Cancel thirsty clear preparation 6g, accurately weighed, put in 25ml measuring bottle, add 75% methyl alcohol appropriate, the ultrasonic 30min of power 500w, is placed to room temperature, adds 75% methanol constant volume to scale, and 0.22 μm of miillpore filter filters, and gets subsequent filtrate, both.
5. method according to claim 1, is characterized in that, the preparation method of described reference substance storing solution is as follows:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance, add the methyl alcohol of 70-80%, dissolve, obtain reference substance storing solution.
6. method according to claim 1, is characterized in that, the preparation method of described reference substance storing solution is as follows:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance, add the methyl alcohol of 75%, dissolve, obtain reference substance storing solution.
7. method according to claim 1, is characterized in that, the preparation method of described reference substance storing solution is as follows:
Get Neomangiferin, mangiferin, timosaponin BII, palmatine and jamaicin reference substance appropriate, accurately weighed, add 75% methyl alcohol and dissolve, obtain mass concentration and be respectively 0.244,0.784,0.772,0.070 and 0.224 μ g μ L
-1reference substance storing solution.
8. method according to claim 1, is characterized in that, described chromatographic condition is as follows:
Chromatographic column: Waters Acquity UPLC HSS C
18;
Mobile phase: solvent orange 2 A is that acetate buffer solution wherein contains 0.5% acetic acid and 0.1% San Yi An – solvent B is acetonitrile,
Condition of gradient elution is:
0 ~ 4min, 5% – 10%B, 4 ~ 8min, 10% – 15%B, 8 ~ 13min, 15% – 22%B, 13 ~ 15min, 22% – 30%B, 15 ~ 18min, 30%B, 18 ~ 20min, 30% – 80%B, 20 ~ 23min, 80% – 5%B; Gradient elution process only illustrates the content of solvent B, and remainder is solvent orange 2 A.
9. method according to claim 1, is characterized in that, described chromatographic condition is as follows:
Column temperature: 30 DEG C;
Flow velocity: 0.2mLmin
-1;
Sample size: 3 μ L;
Determined wavelength: 258nm;
ELSD air velocity: 2.5Lmin
-1;
Drift tube temperature: 105 DEG C,
Yield value: 1.
10. preparation clearly of quenching one's thirst as claimed in claim 1 is particle clearly of quenching one's thirst.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107991416A (en) * | 2018-01-26 | 2018-05-04 | 江苏省中医院 | There is the quality determining method for the Chinese medicine compound prescription for treating urgent, chronic nephropyeltis and hyperplasia of prostate |
| CN109187820A (en) * | 2018-11-09 | 2019-01-11 | 湖南国华制药有限公司 | The method for building up and its finger-print of a kind of compound formula chinaroot greenbrier granules UPLC finger-print and application |
| CN110274963A (en) * | 2018-03-13 | 2019-09-24 | 天士力医药集团股份有限公司 | One kind is quenched one's thirst clear fingerprint atlas detection method |
| CN110618231A (en) * | 2019-10-23 | 2019-12-27 | 山东省中医药研究院 | Preparation process and quality standard research of Baihu decoction granules |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030102265A1 (en) * | 2001-12-03 | 2003-06-05 | Thierry Gandelheid | Method of separating compound(s) from mixture(s) |
| EP1380329A1 (en) * | 2002-07-12 | 2004-01-14 | Shimadzu Corporation | Preparative liquid chromatograph using plural detectors |
| US20110300547A1 (en) * | 2010-06-08 | 2011-12-08 | Tang ke xuan | Method of utilizing the pts gene and anti-sense ads to increase patchouli alcohol content in artemisia annua l. |
-
2013
- 2013-11-01 CN CN201310538597.0A patent/CN104614475B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030102265A1 (en) * | 2001-12-03 | 2003-06-05 | Thierry Gandelheid | Method of separating compound(s) from mixture(s) |
| EP1380329A1 (en) * | 2002-07-12 | 2004-01-14 | Shimadzu Corporation | Preparative liquid chromatograph using plural detectors |
| US20110300547A1 (en) * | 2010-06-08 | 2011-12-08 | Tang ke xuan | Method of utilizing the pts gene and anti-sense ads to increase patchouli alcohol content in artemisia annua l. |
Non-Patent Citations (2)
| Title |
|---|
| 孙海胜等: "HPLC法测定消渴清颗粒中菝葜皂苷元的含量", 《西北药学杂志》 * |
| 韩健等: "用HPLC-ELSD法同时测定不同产地知母饮片中芒果苷、新芒果苷和知母皂苷BII的含量", 《药学服务与研究》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107991416A (en) * | 2018-01-26 | 2018-05-04 | 江苏省中医院 | There is the quality determining method for the Chinese medicine compound prescription for treating urgent, chronic nephropyeltis and hyperplasia of prostate |
| CN107991416B (en) * | 2018-01-26 | 2020-10-09 | 江苏省中医院 | Quality detection method of traditional Chinese medicine compound with effects of treating acute and chronic pyelonephritis and prostatic hyperplasia |
| CN110274963A (en) * | 2018-03-13 | 2019-09-24 | 天士力医药集团股份有限公司 | One kind is quenched one's thirst clear fingerprint atlas detection method |
| CN109187820A (en) * | 2018-11-09 | 2019-01-11 | 湖南国华制药有限公司 | The method for building up and its finger-print of a kind of compound formula chinaroot greenbrier granules UPLC finger-print and application |
| CN109187820B (en) * | 2018-11-09 | 2021-07-23 | 湖南国华制药有限公司 | Method for establishing UPLC fingerprint of compound formula chinaroot greenbrier granules |
| CN110618231A (en) * | 2019-10-23 | 2019-12-27 | 山东省中医药研究院 | Preparation process and quality standard research of Baihu decoction granules |
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Address after: 300410 Tianjin city Beichen District Huaihe road and road intersection Dingjiang tianzhijiao Park forensic Center for Intellectual Property Department Patentee after: Tasly Pharmaceutical Group Limited by Share Ltd Address before: 300410 Tianjin city Beichen District Huaihe road and road intersection Dingjiang tianzhijiao Park forensic Center for Intellectual Property Department Patentee before: Tasly Pharmaceutical Group Co., Ltd. |