CN114814034B - Liquid chromatography method for simultaneously detecting contents of saponin and flavone in ophiopogon japonicus - Google Patents
Liquid chromatography method for simultaneously detecting contents of saponin and flavone in ophiopogon japonicus Download PDFInfo
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- 244000248557 Ophiopogon japonicus Species 0.000 title claims abstract description 75
- 238000000034 method Methods 0.000 title claims abstract description 28
- 229930182490 saponin Natural products 0.000 title claims abstract description 28
- 150000007949 saponins Chemical class 0.000 title claims abstract description 28
- 229930003944 flavone Natural products 0.000 title claims abstract description 25
- 235000011949 flavones Nutrition 0.000 title claims abstract description 25
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 title claims abstract description 16
- 150000002212 flavone derivatives Chemical class 0.000 title claims abstract description 16
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 title claims abstract description 16
- 238000004811 liquid chromatography Methods 0.000 title claims abstract description 11
- 239000001397 quillaja saponaria molina bark Substances 0.000 title claims description 16
- 239000000523 sample Substances 0.000 claims abstract description 37
- 229930195210 Ophiopogon Natural products 0.000 claims abstract description 27
- 235000017709 saponins Nutrition 0.000 claims abstract description 27
- 239000002608 ionic liquid Substances 0.000 claims abstract description 26
- 238000000105 evaporative light scattering detection Methods 0.000 claims abstract description 23
- 239000012488 sample solution Substances 0.000 claims abstract description 20
- 239000000126 substance Substances 0.000 claims abstract description 13
- 238000004458 analytical method Methods 0.000 claims abstract description 9
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- 239000013558 reference substance Substances 0.000 claims abstract description 8
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- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 126
- 239000000243 solution Substances 0.000 claims description 25
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- QRCTUFJCBXMBPP-UHFFFAOYSA-N Ovaliflavanone A Chemical compound O1C=2C(CC=C(C)C)=C(O)C(CC=C(C)C)=CC=2C(=O)CC1C1=CC=CC=C1 QRCTUFJCBXMBPP-UHFFFAOYSA-N 0.000 claims description 7
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- BMWPBKOFJSHJAW-UHFFFAOYSA-N Saponin B Natural products CC1(C)CCC2(CCC3(C)C(=CCC4C5(C)CCC(OC6OC(CO)C(O)C(OC7OC(CO)C(O)C(O)C7O)C6=O)C(C)(C)C5CCC34C)C2C1)C(=O)O BMWPBKOFJSHJAW-UHFFFAOYSA-N 0.000 claims description 5
- LVTJOONKWUXEFR-UEZXSUPNSA-N protodioscin Chemical compound O([C@@H]1[C@@H](CO)O[C@H]([C@@H]([C@H]1O)O[C@H]1[C@@H]([C@H](O)[C@@H](O)[C@H](C)O1)O)O[C@@H]1CC2=CC[C@H]3[C@@H]4C[C@@H]5O[C@]([C@H]([C@@H]5[C@@]4(C)CC[C@@H]3[C@@]2(C)CC1)C)(O)CC[C@@H](C)CO[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@@H](C)[C@H](O)[C@@H](O)[C@H]1O LVTJOONKWUXEFR-UEZXSUPNSA-N 0.000 claims description 5
- QDQWGYLCDZBAMD-UHFFFAOYSA-N saponin C Natural products CC1C2C3CCC4C5(C)CCC(O)C(C)(COC6OC(CO)C(O)C(O)C6O)C5CCC4(C)C3(C)CCC27C8OC8C1(C)OC7=O QDQWGYLCDZBAMD-UHFFFAOYSA-N 0.000 claims description 5
- 239000002904 solvent Substances 0.000 claims description 5
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 4
- ZONYXWQDUYMKFB-UHFFFAOYSA-N SJ000286395 Natural products O1C2=CC=CC=C2C(=O)CC1C1=CC=CC=C1 ZONYXWQDUYMKFB-UHFFFAOYSA-N 0.000 claims description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 238000010828 elution Methods 0.000 claims description 4
- 229930003949 flavanone Natural products 0.000 claims description 4
- 150000002208 flavanones Chemical class 0.000 claims description 4
- 235000011981 flavanones Nutrition 0.000 claims description 4
- 235000019253 formic acid Nutrition 0.000 claims description 4
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 4
- 239000002245 particle Substances 0.000 claims description 4
- 238000002137 ultrasound extraction Methods 0.000 claims description 4
- 239000000706 filtrate Substances 0.000 claims description 3
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- 239000000945 filler Substances 0.000 claims description 2
- 238000000227 grinding Methods 0.000 claims description 2
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 2
- 239000000377 silicon dioxide Substances 0.000 claims description 2
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- KAIPKTYOBMEXRR-UHFFFAOYSA-N 1-butyl-3-methyl-2h-imidazole Chemical compound CCCCN1CN(C)C=C1 KAIPKTYOBMEXRR-UHFFFAOYSA-N 0.000 claims 1
- 238000002360 preparation method Methods 0.000 claims 1
- 239000012088 reference solution Substances 0.000 claims 1
- ITMCEJHCFYSIIV-UHFFFAOYSA-M triflate Chemical compound [O-]S(=O)(=O)C(F)(F)F ITMCEJHCFYSIIV-UHFFFAOYSA-M 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 24
- 238000000605 extraction Methods 0.000 abstract description 24
- 239000003960 organic solvent Substances 0.000 abstract description 5
- 238000000638 solvent extraction Methods 0.000 abstract description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 13
- 238000009210 therapy by ultrasound Methods 0.000 description 6
- PXKPKGHXANCVMC-UHFFFAOYSA-N 3-butyl-1-methyl-1,2-dihydroimidazol-1-ium;trifluoromethanesulfonate Chemical compound OS(=O)(=O)C(F)(F)F.CCCCN1CN(C)C=C1 PXKPKGHXANCVMC-UHFFFAOYSA-N 0.000 description 5
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- 235000017173 flavonoids Nutrition 0.000 description 4
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 description 3
- 235000008696 isoflavones Nutrition 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- 150000005856 steroid saponins Chemical class 0.000 description 3
- IAZSXUOKBPGUMV-UHFFFAOYSA-N 1-butyl-3-methyl-1,2-dihydroimidazol-1-ium;chloride Chemical compound [Cl-].CCCC[NH+]1CN(C)C=C1 IAZSXUOKBPGUMV-UHFFFAOYSA-N 0.000 description 2
- XGBLLQBZRQMYNV-UHFFFAOYSA-N 1-butyl-3-methyl-2H-imidazole nitric acid Chemical compound [N+](=O)(O)[O-].C(CCC)N1CN(C=C1)C XGBLLQBZRQMYNV-UHFFFAOYSA-N 0.000 description 2
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- GOMNOOKGLZYEJT-UHFFFAOYSA-N isoflavone Chemical compound C=1OC2=CC=CC=C2C(=O)C=1C1=CC=CC=C1 GOMNOOKGLZYEJT-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
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- OIWSIWZBQPTDKI-UHFFFAOYSA-N 1-butyl-3-methyl-2h-imidazole;hydrobromide Chemical compound [Br-].CCCC[NH+]1CN(C)C=C1 OIWSIWZBQPTDKI-UHFFFAOYSA-N 0.000 description 1
- ZNNXXAURXKYLQY-UHFFFAOYSA-N 1-butyl-3-methyl-2h-imidazole;sulfuric acid Chemical compound OS(O)(=O)=O.CCCCN1CN(C)C=C1 ZNNXXAURXKYLQY-UHFFFAOYSA-N 0.000 description 1
- QZKBXQUHXDHAOA-UHFFFAOYSA-N Br(=O)(=O)O.C(CCC)N1CN(C=C1)C Chemical compound Br(=O)(=O)O.C(CCC)N1CN(C=C1)C QZKBXQUHXDHAOA-UHFFFAOYSA-N 0.000 description 1
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- 206010011224 Cough Diseases 0.000 description 1
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- 241000234280 Liliaceae Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241001448421 Ophiopogon jaburan Species 0.000 description 1
- 206010068319 Oropharyngeal pain Diseases 0.000 description 1
- 201000007100 Pharyngitis Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- PYKYMHQGRFAEBM-UHFFFAOYSA-N anthraquinone Natural products CCC(=O)c1c(O)c2C(=O)C3C(C=CC=C3O)C(=O)c2cc1CC(=O)OC PYKYMHQGRFAEBM-UHFFFAOYSA-N 0.000 description 1
- 150000004056 anthraquinones Chemical class 0.000 description 1
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- 150000002515 isoflavone derivatives Chemical class 0.000 description 1
- 229920006008 lipopolysaccharide Polymers 0.000 description 1
- 239000011344 liquid material Substances 0.000 description 1
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- 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
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- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N2030/042—Standards
- G01N2030/047—Standards external
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- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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Abstract
The invention discloses a liquid chromatography method for simultaneously detecting the contents of saponins and flavones in Zhejiang dwarf lilyturf tuber, which comprises the following steps: 1) Respectively taking a Zhejiang ophiopogonin reference substance and a flavone reference substance, preparing a mixed standard substance solution, and detecting the obtained mixed standard substance solution under the HPLC-DAD-ELSD analysis condition to obtain a standard curve; 2) Taking a sample of Zhejiang ophiopogon japonicus to be detected, preparing a sample solution, and detecting by using an HPLC-DAD-ELSD combined method; 3) And calculating the content of saponins and flavones in the sample of the Zhejiang ophiopogon japonicus. The invention adopts ionic liquid to extract the characteristic components of the Zhejiang ophiopogon root, and compared with the traditional organic solvent extraction, the method is a more efficient and greener extraction method; the established HPLC-DAD-ELSD detection method has the advantages of rapidness, accuracy, good repeatability and the like.
Description
Technical Field
The invention belongs to the technical field of natural pharmacy and analysis and detection of traditional Chinese medicines, and particularly relates to a liquid chromatography method for simultaneously detecting contents of saponins and flavones in Zhejiang dwarf lilyturf tuber.
Background
Zhejiang dwarf lilyturf tuber is one of the most well-known genuine medicinal materials of Zhejiang province, namely Zhejiang eight ingredients, and is a dried root tuber of lilyturf turber Ophiopogon japonicus (L.f.) ker-Gawl. Of the family Liliaceae, and has been used for thousands of years in the treatment of acute cough, sore throat, cardiovascular and cerebrovascular diseases and other diseases. Because of the huge differences of the Zhejiang dwarf lilyturf tuber in the aspects of cultivation conditions, regional environment, growth period and the like, the Zhejiang dwarf lilyturf tuber and the Sichuan dwarf lilyturf tuber are different in chemical composition and content and have various characteristics. Pharmacological research shows that the Zhejiang ophiopogon root extract has the same effect on H 2 O 2 The induced myocardial cell injury and lipopolysaccharide-induced macrophage inflammation of mice have stronger inhibition effect, while Zhejiang and Sichuan dwarf lilyturf tuber show different selective cytotoxicity to different tumor cell strains. Studies show that steroid saponin and high isoflavone components are the main drug effect substances, and the research results further show that the ophiopogon japonicus saponin B and ophiopogon japonicus saponin C, the methylophiopogon japonicus flavanone A and methylophiopogon japonicus flavanone B are the main characteristic steroid saponin and high isoflavone components in the ophiopogon japonicus. Because the component difference exists between the Zhejiang dwarf lilyturf tuber and the Sichuan dwarf lilyturf tuber at present, the market price of the Zhejiang dwarf lilyturf tuber is far higher than that of the Sichuan dwarf lilyturf tuber, and the establishment of the detection method for detecting the characteristic components of the Zhejiang dwarf lilyturf tuber has very important significance for the identification and quality control of the Zhejiang dwarf lilyturf tuber medicinal material.
At present, the literature reports that HPLC-MS and HPLC-UV-ELSD are used for identifying different components in the ophiopogon japonicus and analyzing fingerprints, and HPLC-UV, HPLC-ELSD and HPLC-MS are used for respectively measuring partial characteristic flavone or saponin of the ophiopogon japonicus. However, due to the lack of the characteristic component reference substance, the detection sensitivity of the saponin is low, and the like, the method for quantitatively analyzing the flavonoid and the saponin in the Zhejiang ophiopogon japonicus is necessary to be explored and established conveniently and accurately. In addition, in the prior pretreatment of the analysis and detection of the components of the Zhejiang ophiopogon japonicus, organic solvents such as methanol and the like are mainly adopted for extraction, so that the defects of poor extraction selectivity, more impurities, certain safety risks, environmental pollution and the like exist. As a new green solvent, the ionic liquid has the excellent characteristics of low vapor pressure, liquid Cheng Kuan, high thermal stability, good dissolution capacity and the like compared with the traditional organic solvent; the structure of the ionic liquid has designability, and the physical and chemical properties of the ionic liquid can be adjusted by changing or modifying the ionic liquid so as to meet application requirements. In recent years, the unique advantages of different structural combinations of anions and cations of ionic liquids are gradually revealed in the pretreatment of extraction of natural active substances of different classes. In addition, the HPLC-DAD-ELSD combined technology is a liquid chromatography method capable of simultaneously detecting and analyzing the ophiopogon flavonoids and saponins, and has the advantages of time saving and simplicity.
Disclosure of Invention
Aiming at the defects of the existing method for detecting the components of the Zhejiang dwarf lilyturf tuber, the invention establishes an analysis and detection method for analyzing the characteristic components of the Zhejiang dwarf lilyturf tuber based on ionic liquid extraction and liquid chromatography. Aims at solving the problems that the conventional detection method has less consideration on saponin and flavone components and the organic solvent is used in the extraction to cause safety pollution, and establishing a method for extracting and simultaneously detecting characteristic saponin and flavone components in the Zhejiang ophiopogon japonicus with higher efficiency and greener color.
The method is characterized in that firstly, proper ionic liquid solution is selected for ultrasonic-assisted extraction of characteristic components of the Zhejiang dwarf lilyturf tuber sample, then, the characteristic components of the Zhejiang dwarf lilyturf tuber are further enriched by a solid phase extraction method, and finally, the detection is carried out by an HPLC-DAD-ELSD method.
The technical scheme of the invention is as follows:
1) Sample extraction
Firstly, grinding radix ophiopogonis tuberous root or fibrous root medicinal material powder into fine powder, accurately weighing a proper amount of radix ophiopogonis sample powder in a triangular flask with a plug, adding an ionic liquid solution into an ultrasonic cleaning instrument, ultrasonically extracting for a plurality of minutes, centrifuging, taking clear liquid, filtering, taking a proper amount of filtrate, adsorbing the filtrate on a solid-phase extraction column, eluting with water, 25% methanol and 100% methanol in sequence, collecting methanol eluent, rotationally evaporating and concentrating, accurately fixing the volume to 2ml by using methanol, and filtering by using a microporous filter membrane to obtain a sample solution to be detected.
The ionic liquid is any one of 1-butyl-3-methylimidazole nitrate, 1-butyl-3-methylimidazole triflate, 1-butyl-3-methylimidazole chloride and 1-butyl-3-methylimidazole bromate, and preferably 1-butyl-3-methylimidazole triflate.
The concentration of the ionic liquid aqueous solution is 0.7-1.50 mol/L, preferably 1mol/L;
the liquid-solid ratio of the ionic liquid to the ophiopogon root sample or fibrous root medicinal material powder is 10-50 ml/g, preferably 40ml/g;
the ultrasonic extraction time of the dwarf lilyturf tuber sample or fibrous root medicinal material is 30-120 min, preferably 60min.
2) HPLC-DAD-ELSD chromatography condition column: a chromatographic column with octadecylsilane chemically bonded silica as filler. The column size is 4.6mm by 100mm particle size 2.7 μm, preferably Welch BoltimateTM-C18 column (4.6 mm by 100mm,2.7 μm);
mobile phase: acetonitrile is used as phase A, and 0.1% formic acid is used as phase B; the gradient elution procedure was: 0-15 min, 30-40% of A; 15-25 min, 40-80% A; the sample injection amount is 5-40 mu L; column temperature: 30-40 ℃, preferably 35 ℃;
the flow rate is 0.8-1.2 mL/min, preferably 1mL/min;
a DAD detector with a detection wavelength of 270-300 nm, preferably 296nm;
ELSD detector, atomizer mode heating of 0-40%, drift tube temperature of 50-90 ℃ and carrier gas pressure of 20-40 psi, preferably atomizer mode heating of 20%, drift tube temperature of 70 ℃ and carrier gas pressure of 25psi.
3) Standard Curve establishment
The characteristic components of the Zhejiang dwarf lilyturf tuber in the technical scheme are ophiopogon root saponin B (S1), ophiopogon root saponin C (S2), methyl ophiopogon root flavanone A (H1) and methyl ophiopogon root flavanone B (H2) respectively. The standard substances of the 4 compounds are respectively weighed, methanol is used as a solvent to prepare mixed standard substance solutions with different gradient concentrations, and the obtained mixed standard solutions are detected under the HPLC-DAD-ELSD analysis condition to obtain a detection HPLC spectrogram. The saponins S1 and S2 respectively draw standard curves by taking the logarithmic value of the peak area as an ordinate and the logarithmic value of the sample injection amount concentration as an abscissa; and respectively drawing standard curves of the flavones H1 and H2 by taking the peak area as an ordinate and the sample injection amount concentration as an abscissa.
4) Sample analysis assay
Comparing the sample spectrogram obtained in the step 1) with the standard substance spectrogram obtained in the step 3) to obtain qualitative results of saponin and flavonoid components in the sample; substituting the peak area values of the anthraquinone components in the spectrogram of the sample obtained in the step 1) into the corresponding standard curves obtained in the step 3), and calculating to obtain the contents of the saponin and flavonoid components in the sample.
Compared with the prior art, the method adopts the ionic liquid to extract the characteristic components of the Zhejiang dwarf lilyturf tuber, and is a more efficient and more green extraction method compared with the traditional organic solvent extraction method; the established HPLC-DAD-ELSD detection method has the advantages of rapidness, accuracy, good repeatability and the like, so the invention provides a method suitable for simultaneously detecting the qualitative and quantitative detection of the saponin and flavone components in the Zhejiang ophiopogon root.
Drawings
The structural formula of the characteristic components of the Zhejiang ophiopogon root in the figure 1 is shown in the specification, wherein S1 is ophiopogon root saponin B, S, ophiopogon root saponin C, H1 is ophiopogon root flavanone A, H2 and ophiopogon root flavanone B (the components marked by S1, S2, H1 and H2 in the following figures are the same as the present figure);
FIG. 2 shows HPLC detection chromatogram of mixed standard solution of characteristic components of radix Ophiopogonis, wherein (A) is HPLC-ELSD chromatogram; (B) HPLC-DAD detection wavelength 296nm chromatogram;
FIG. 3 influence of different ionic liquid types on extraction rate of characteristic components of Zhejiang dwarf lilyturf tuber;
FIG. 4 is the effect of different ionic liquid concentrations on extraction rate of characteristic components of Zhejiang dwarf lilyturf tuber;
FIG. 5 is the influence of different liquid-material ratios on the extraction rate of the characteristic components of Zhejiang dwarf lilyturf tuber;
FIG. 6 influence of different ultrasonic times on extraction rate of characteristic components of Zhejiang radix Ophiopogonis;
FIG. 7 HPLC detection chromatogram of radix Ophiopogonis Thunberg root tuber sample, (A) is HPLC-ELSD chromatogram; (B) HPLC-DAD detection wavelength 296nm chromatogram;
FIG. 8 HPLC detection chromatogram of radix Ophiopogonis fiber root sample, (A) is HPLC-ELSD chromatogram; (B) HPLC-DAD detection wavelength 296nm chromatogram.
Detailed Description
The present invention is described in further detail below with reference to specific examples, and specific examples are given.
The characteristic components of the Zhejiang ophiopogon saponin and the flavone are ophiopogon saponin B (S1), ophiopogon saponin C (S2), ophiopogon flavone A (H1) and ophiopogon flavone B (H2), and the structural formulas of the characteristic components are shown in figure 1. The standard substances of the 4 compounds are respectively weighed, methanol is used as a solvent to prepare mixed standard substance solutions with different gradient concentrations, the obtained mixed standard solutions are detected under the HPLC-DAD-ELSD analysis condition, and the detection HPLC spectrogram is obtained, and the HPLC chromatograph of the mixed standard substance is shown in figure 2.
Example 1: sample extraction and pretreatment method
Taking about 0.1-1 g of ophiopogon japonicus sample powder, precisely weighing, placing the sample powder into a conical bottle with a plug, precisely adding 10ml of ionic liquid solution with a certain concentration, sealing, and performing ultrasonic treatment for 30-120 minutes to obtain an extracting solution. After the extracting solution is centrifugated, 5ml of supernatant is added into a C18 solid phase extraction column pretreated by methanol and water, then 10ml of water, 10ml of 25% methanol and 5ml of methanol are sequentially added for eluting, the methanol eluent is collected, distilled and concentrated, and then the methanol is accurately fixed to volume to 2ml, and filtered by an organic microporous filter membrane with the thickness of 0.45 mu m, thus obtaining a sample solution. Sucking 5-40 mu L of sample solution, and injecting the sample solution into high performance liquid chromatography for measurement.
Example 2: optimization of HPLC-DAD-ELSD detection conditions
By mixing different particle sizes (2.7 μm and5.0 μm) shows that the smaller 4.6X100 mm chromatographic column with 2.7 μm particle size does not significantly increase column pressure, is suitable for common liquid chromatography, but has greatly shortened detection time, better separation effect and greatly reduced solvent cost, wherein Welch BoltimateTM-C is preferred 18 And (5) a column. Acetonitrile is used as a mobile phase of A phase, and 0.1% formic acid is used as a B phase; the gradient elution procedure was: 0-15 min, 30-40% of A; 15-25 min, 40-80% A; the sample injection amount is 5-40 mu L. The column temperature is optionally 30-40 ℃, preferably 35 ℃. The flow rate is optionally 0.8-1.2 mL/min, preferably 1mL/min. The DAD detector can optionally detect wavelengths of 270-300 nm, preferably 296nm, based on the maximum absorption of ophiopogon japonicus homoisoflavones H1 and H2. The ELSD detector can be heated in a sprayer mode of 0-40%, the temperature of the drift tube is 50-90 ℃ and the carrier gas pressure is 20-40 psi, and according to the detected signal intensity of the ophiopogon saponin S1 and S2 under the above conditions, the sprayer mode is preferred to be heated in 20%, the temperature of the drift tube is 70 ℃ and the carrier gas pressure is 25psi.
Following the chromatographic condition fumbling and condition optimization above, the following examples employed HPLC-DAD-ELSD conditions specifically were: waters e2695 liquid phase system with chromatographic column Welch BoltimateTM-C 18 Columns (4.6 mm. Times.100 mm,2.7 μm); the mobile phase takes acetonitrile as a phase A and 0.1% formic acid as a phase B, and the gradient elution procedure is as follows: 0-15 min, 30-40% of A; 15-25 min, 40-80% A; the sample injection amount is 5-40 mu L. The column temperature was 35℃and the flow rate was 1ml/min. Waters2996DAD detector, wavelength 296nm; the Waters2424ELSD detector was heated in the nebulizer mode to 20% and the drift tube temperature was 70 ℃ and carrier gas pressure was 25psi.
Example 3: selection of ionic liquids
1g of 5 parts of Zhejiang ophiopogon root tuber sample powder is weighed in parallel, precisely weighed, placed in a conical bottle with a plug, respectively and precisely added with 10ml of 1mol/L of 5 ionic liquid solutions such as 1-butyl-3-methylimidazole nitrate, 1-butyl-3-methylimidazole triflate, 1-butyl-3-methylimidazole bisulfate, 1-butyl-3-methylimidazole chloride, 1-butyl-3-methylimidazole bromide and the like, and subjected to sealing and ultrasonic treatment for 60 minutes to obtain an extracting solution. Centrifuging the extracting solution, taking 5ml of supernatant, adding a C18 solid phase extraction column pretreated by methanol and water, sequentially adding 10ml of water, 10ml of 25% methanol and 5ml of methanol for eluting, collecting methanol eluent, performing rotary evaporation and concentration, accurately fixing the volume to 2ml by methanol, filtering by an organic microporous filter membrane, obtaining a sample solution, sucking 10 mu L of the sample solution, and injecting the sample solution into a high performance liquid chromatography for determination. The above experiments were repeated 3 times.
As can be seen from FIG. 3, the extraction rates of two steroid saponins from the ionic liquid solutions of 5 identical butyl cations are not very different, while the extraction rate of two high isoflavones from the 1-butyl-3-methylimidazole triflate solution is highest and far higher than that of the other 4 ionic liquids, so that 1-butyl-3-methylimidazole triflate ([ Bmim ] OTf) is preferred.
Example 4: selection of ionic liquid concentration
1g of 6 parts of Zhejiang ophiopogon root tuber sample powder is weighed in parallel, precisely weighed, placed in a conical flask with a plug, respectively and precisely added with 10ml of [ Bmim ] OTf solution of 0, 0.50, 0.75, 1.0, 1.25 and 1.50mol/L, sealed, and subjected to ultrasonic treatment for 60 minutes to obtain an extracting solution. After the extracting solution is centrifugated, 5ml of supernatant is added into a C18 solid phase extraction column pretreated by methanol and water, then 10ml of water, 10ml of 25% methanol and 5ml of methanol are sequentially added for eluting, the methanol eluent is collected, the methanol is concentrated by rotary evaporation, the volume is accurately fixed to 2ml, an organic microporous filter membrane is used for filtering, a sample solution is obtained, and 10 mu L of sample solution is sucked and injected into high performance liquid chromatography for determination. The above experiments were repeated 3 times.
As can be seen from FIG. 4, the extraction yield of the two flavones increased significantly with increasing IL concentration, but the change was insignificant when the IL concentration was greater than 1 mol/L. In contrast, when IL was increased from 0mol/L to 1mol/L, the extraction rates of the two saponins did not change much, but as the concentration of IL increased, the extraction rates of the two saponins decreased significantly. Thus, a [ Bmim ] OTf solution concentration of 1mol/L is preferred.
Example 5: selection of liquid-to-material ratio
Respectively weighing 0.2 g, 0.25g, 0.333 g, 0.5 g and 1g of sample powder of the root tuber of Zhejiang dwarf lilyturf, precisely weighing, placing into a conical flask with a plug, precisely adding 10ml of 1mol/L [ Bmim ] OTf solution, sealing, and performing ultrasonic treatment for 60 minutes to obtain an extracting solution. After the extracting solution is centrifugated, 5ml of supernatant is added into a C18 solid phase extraction column pretreated by methanol and water, then 10ml of water, 10ml of 25% methanol and 5ml of methanol are sequentially added for eluting, the methanol eluent is collected, the methanol is concentrated by rotary evaporation, the volume is accurately fixed to 2ml, an organic microporous filter membrane is used for filtering, a sample solution is obtained, and 10 mu L of sample solution is sucked and injected into high performance liquid chromatography for determination. The above experiments were repeated 3 times.
As can be seen from fig. 5, the extraction rate of the characteristic component of ophiopogon root gradually increases with an increase in the liquid-to-material ratio, but when the liquid-to-material ratio is increased to 50ml/g, the liquid-to-material ratio of 50ml/g is preferable.
Example 6: selection of ultrasound time
Weighing 0.25g of sample powder of 5 parts of radix ophiopogonis root tuber in parallel, precisely weighing, placing into a conical bottle with a plug, precisely adding 10mL of 1mol/L [ Bmim ] OTf solution, sealing, performing ultrasonic treatment for 30, 45, 60, 90 and 120 minutes, centrifuging the extracting solution, taking 5mL of supernatant, adding a C18 solid phase extraction column pretreated by methanol and water, sequentially adding 10mL of water, 10mL of 25% methanol and 5mL of methanol for eluting, collecting methanol eluent, performing rotary evaporation and concentration, accurately fixing the volume of the methanol to 2mL, filtering by an organic microporous filter membrane, obtaining a sample solution, and sucking 10 mu L of the sample solution for high performance liquid chromatography determination. The above experiments were repeated 3 times.
As can be seen from fig. 6, when the ultrasonic time is controlled to be 60 minutes or less, the extraction amount of flavone increases with time, and the extraction rate of specific components increases, and further increases with time, the total extraction rate of characteristic components decreases due to the decrease in the extraction rate of saponins. Thus, a preferred 60min ultrasound time is considered in combination.
Example 7: methodological verification
Under the preferable conditions of extraction and chromatography, the invention provides a method for analyzing the characteristic components of the Zhejiang dwarf lilyturf tuber by combining ionic liquid ultrasonic extraction with HPLC-DAD-ELSD through experimental study, wherein the linear range, limit of restriction (LOD) and quantitative Limit (LOQ) results are shown in table 1, the repeatability, precision and stability are shown in table 2, and the sample recovery rate results are shown in table 3. The results show that the method has good repeatability, precision, stability and accuracy.
TABLE 1 Standard Curve equation, linear correlation coefficient, linear Range, detection Limit (LOD), quantitative Limit (LOQ)
TABLE 2 repeatability, precision and stability (RSD/%)
TABLE 3 sample recovery rate
Example 8: determination of different radix Ophiopogonis tuberous root and fibrous root samples
Respectively weighing 0.25g of 6 different Zhejiang dwarf lilyturf tuber sample powders, precisely weighing, placing into a conical flask with a plug, precisely adding 10ml of 1mol/L, sealing, and performing ultrasonic treatment for 60 minutes to obtain an extracting solution. Centrifuging the extracting solution, taking 5ml of supernatant, adding a C18 solid phase extraction column pretreated by methanol and water, sequentially adding 10ml of water, 10ml of 25% methanol and 5ml of methanol for eluting, collecting methanol eluent, concentrating by rotary evaporation, accurately fixing the volume of the methanol to 2ml, filtering by an organic microporous filter membrane to obtain a sample solution, sucking 10 mu L of the sample solution, and injecting the sample solution into a high performance liquid chromatography for determination. The above experiments were repeated 3 times.
The detection results are shown in Table 4, and the HPLC chromatograms of representative radix Ophiopogonis tuberous root and fibrous root samples are shown in FIG. 7 and FIG. 8. The result shows that the method is suitable for detecting the root tuber and fibrous root samples of the ophiopogon japonicus.
TABLE 4 detection and analysis results (mg/g) of characteristic Components in samples of different sources of Zhejiang radix Ophiopogonis
。
Claims (4)
1. The liquid chromatography method for simultaneously detecting the contents of saponin and flavone in the ophiopogon japonicus is characterized by comprising the following steps of:
1) Respectively taking a Zhejiang ophiopogonin reference substance and a flavone reference substance, preparing a mixed standard substance solution, and detecting the obtained mixed standard substance solution under the HPLC-DAD-ELSD analysis condition to obtain a standard curve;
2) Taking a sample of Zhejiang ophiopogon japonicus to be detected, preparing a sample solution, and detecting by using an HPLC-DAD-ELSD combined method; the preparation method of the sample solution comprises the following steps: grinding the root tuber or fibrous root medicinal material powder of the radix ophiopogonis sample to be detected into fine powder, accurately weighing a proper amount of the radix ophiopogonis sample powder in a triangular flask with a plug, adding an ionic liquid aqueous solution into an ultrasonic cleaning instrument, performing ultrasonic extraction for 60min with the ionic liquid aqueous solution having a liquid-solid ratio of 40ml/g and the radix ophiopogonis sample powder, centrifuging, taking supernatant, filtering, adsorbing filtrate on a solid-phase extraction column, eluting sequentially with water, 25% methanol and 100% methanol, collecting methanol eluent, performing rotary evaporation concentration, accurately fixing the volume to 2ml with methanol, and filtering with a microporous filter membrane to obtain a sample solution; the ionic liquid is 1-butyl-3-methylimidazole trifluoro methanesulfonate; the concentration of the ionic liquid aqueous solution is 1mol/L;
3) Calculating the content of saponins and flavones in a Zhejiang ophiopogon sample;
wherein, the HPLC-DAD-ELSD has the following chromatographic conditions:
chromatographic column: selecting octadecylsilane chemically bonded silica as a chromatographic column filled with the filler, wherein the specification of the chromatographic column is 4.6mm multiplied by 100mm, and the particle size of the chromatographic column is 2.7 mu m;
mobile phase: acetonitrile is used as phase A, and 0.1% formic acid is used as phase B; the gradient elution procedure was: 0-15 min, 30-40% of A; 15-25 min, 40-80% A; the sample injection amount is 5-40 mu L;
column temperature: 30-40 ℃;
the flow rate is 0.8-1.2 mL/min;
a DAD detector for detecting 270 to 300nm wavelengths;
ELSD detector, atomizer mode heating of 0-40%, drift tube temperature of 50-90 ℃ and carrier gas pressure of 20-40 psi.
2. The liquid chromatography method for simultaneously detecting the contents of saponins and flavones in Zhejiang ophiopogon japonicus according to claim 1, wherein the saponin reference substance in the step 1) is selected from the group consisting of ophiopogon japonicus saponin B, denoted as S1, ophiopogon japonicus saponin C, denoted as S2; the flavone reference substance is selected from methyl ophiopogon root flavanone A, and is marked as H1, methyl ophiopogon root flavanone B, and is marked as H2.
3. The liquid chromatography method for simultaneously detecting the contents of saponins and flavones in Zhejiang dwarf lilyturf tuber according to claim 1, wherein the solvent for preparing the reference solution in the step 1) is methanol.
4. The liquid chromatography method for simultaneously detecting the contents of saponins and flavones in Zhejiang ophiopogon root according to claim 1, wherein in the step 1), the standard curves comprise a saponin reference standard curve and a flavone reference standard curve, wherein the saponin reference standard curve is respectively drawn by taking the logarithmic value of peak area as an ordinate and the logarithmic value of sample injection concentration as an abscissa; and (3) respectively drawing a standard curve of the flavone reference substance by taking the peak area as an ordinate and the sample injection amount concentration as an abscissa.
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Non-Patent Citations (7)
| Title |
|---|
| HPLC-DAD-ELSD 同时测定知母中黄酮和 皂苷类成分的含量;苏晓楠 等;《中国中药杂志》;第40卷(第1期);第108-111页 * |
| HPLC-DAD-ELSD法同时测定黄芪中5个成分的含量;梁瑾 等;《药物分析杂志》(第02期);全文 * |
| 乌饭树树叶总黄酮的超声提取工艺研究;周一峰;《中国药师》;第18卷(第1期);全文 * |
| 甲醇溶液直接吸收-离子液体富集-气相色谱-质谱法测定卷烟主流烟气中吡啶、喹啉和苯乙烯;庹苏行;杨华武;吴名剑;戴云辉;;分析化学(第07期);全文 * |
| 直接衍生-离子液体富集-高效液相色谱法分析水基胶中脂肪族醛酮;万小红;吴名剑;蒋新宇;戴云辉;李绍晔;龚淑果;;色谱(第12期);全文 * |
| 离子液体- 超声辅助- 固相萃取法提取 富集急性子药材中的4 种凤仙萜四醇皂苷;吴谦 等;《分析化学( FENXI HUAXUE)》;第46卷(第6期);第910-916页 * |
| 苏晓楠 等.HPLC-DAD-ELSD 同时测定知母中黄酮和 皂苷类成分的含量.《中国中药杂志》.2015,第40卷(第1期),第108-111页. * |
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