CN104698022A - Method for measuring purity of aryltetralin lactone lignans on basis of quantitative nuclear magnetic resonance hydrogen spectrum method - Google Patents
Method for measuring purity of aryltetralin lactone lignans on basis of quantitative nuclear magnetic resonance hydrogen spectrum method Download PDFInfo
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Abstract
The invention relates to a method for measuring the purity of aryltetralin lactone lignans on the basis of a quantitative nuclear magnetic resonance hydrogen spectrum method. The problems that an existing technology for measuring the purity of the aryltetralin lactone lignans is high in cost, complex in operation method and high in time consumption and reference substances can not be easily obtained can be effectively solved. The method includes the steps of weighing 10 mg to 14 mg of internal standard substances, dissolving the internal standard substances in 0.8 mL to 1.2 mL of DMSO-d6 to prepare an internal standard substance solution, weighing 10 mg to 50 mg of aryltetralin lactone lignans, dissolving the aryltetralin lactone lignans in 0.8 mL to 1.2 mL of DMSO-d6 to prepare a target analyte solution, weighing 80 microliters to 120 microliters of internal standard substance solution, 180 microliters to 220 microliters of target analyte solution and 180 microliters to 220 microliters of DMSO-d6 to prepare a sample solution, suspending the sample solution for 25 s to 35 s to carry out a quantitative nuclear magnetic test, measuring the peak area of aromatic protons of the internal standard substances and H-2,6 of the aryltetralin lactone lignans to obtain the molar mass ratio of target analyte to the internal standard substances, and then measuring the purity through the mass ratio of the target analyte to the internal standard substances. The method is low in cost, can rapidly and accurately evaluate the purity of the aryltetralin lactone lignans and has high economic and social benefits.
Description
Technical field
The present invention relates to field of medicine and chemical technology, particularly a kind of method measuring arylnaphthalene lactone type lignanoid purity based on quantitative hydrogen nuclear magnetic resonance spectroscopy.
Background technology
Due to biosynthetic complicacy, the structure of natural products is also complicated various.Often be difficult to Separation of Natural Products closely similar for structure to come based on the separation and purification means of the chromatographic processes of routine, and the highly active impurity being mixed with trace in natural products can cause bioactive wrong conclusion.According to relevant report, natural products ursolic acid has 120 kinds of biologically actives, and some biologically active is antipodal in different documents, as antibacterial activity, and cytotoxic activity.Occur that the reason of such conclusion is, the active ursolic acid sample used of test is not single ursolic acid chemical entities, and the impurity coexisted with it has had a strong impact on Activity Results.Therefore the natural products Purity assessment method of Erecting and improving is the necessary condition of its evaluated biological activity accurately.Highly purified natural products is also that relevant evaluation of medical materials' quality is necessary simultaneously.Therefore the Purity assessment of natural products is very important.
Arylnaphthalene lactone type lignanoid is the more lignans of the number that finds from plant, in more than 40 Plants, has report at present.This compounds has the effects such as antitumor, antiviral, antibacterial, desinsection.Arylnaphthalene lactone type Lignanoids compounds take podophyllotoxin as representative, its derivant Etoposide (etoposide, and Teniposide (teniposide VP-16), VM-26) be applied to clinical the eighties in eighties of last century, good result for the treatment of is all had to kinds cancers such as breast cancer, carcinoma of testis, small-cell carcinoma of the lung, lymph cancer, leukemia of children, take podophyllotoxin as the arylnaphthalene lactone type lignanoid of representative be one of class natural products that research is the most deep at present, enjoy the extensive concern of Chinese scholars always.The content of this compounds in relevant medicinal material is mainly through thin-layer chromatography, high performance liquid chromatography, Capillary Electrophoresis chromatogram, wherein general with the efficient liquid-phase chromatography method with UV-detector, this method does not have the compound of uv absorption to detect for those, high-purity standard items must be had to compare, these standard items are expensive, not easily obtain, and this method of operating complicated and time consumption is long.Quantitative hydrogen nuclear magnetic resonance spectroscopy, because its operation is fast and convenient, sample consumption is few, provide structural information and quantitative analysis results qualitatively simultaneously, particularly quantitatively hydrogen nuclear magnetic resonance spectroscopy does not rely on the high-purity standard items of measured object, and the cost of therefore quantitative test reduces greatly.Utilize quantitative hydrogen nuclear magnetic resonance spectroscopy, purity testing that outer target high performance liquid chromatography carries out arylnaphthalene lactone type lignanoid is a very important research work to select internal standard compound to substitute to use the standard items of target detection thing to do.But it is not yet seen the open report adopting quantitative hydrogen nuclear magnetic resonance spectroscopy to measure the purity of arylnaphthalene lactone type lignanoid.
Summary of the invention
For above-mentioned situation, in order to overcome the defect of prior art, the invention provides a kind of method measuring arylnaphthalene lactone type lignanoid purity based on quantitative hydrogen nuclear magnetic resonance spectroscopy, effectively can solve that existing arylnaphthalene lactone type lignanoid purity detecting technical costs is high, reference substance not easily obtains, method of operating is complicated, the problem of length consuming time.
The technical scheme that the present invention solves is that the method comprises the following steps: the internal standard compound taking 10-14mg, is dissolved in the DMSO-d6 of 0.8-1.2mL, is prepared into internal standard substance solution; Take the arylnaphthalene lactone type lignanoid of 10-50mg, be dissolved in the DMSO-d6 of 0.8-1.2mL, be prepared into target analytes solution; Measure the internal standard substance solution of 80-120 μ L, the target analytes solution of 180-220 μ L, the DMSO-d6 of 180-220 μ L, transfer in centrifuge tube, make sample solution, by this sample solution suspendible 25-35s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of quantitative nuclear-magnetism test: scanning times (NS), 8 or 16; Spectral width (SW), 15-25ppm; Relaxation time (D1), 16-35s; Probe temperature, 20-30 DEG C; Pulse angle, 30 °, or 45 °, or 90 °; Acquisition time (AQ), 2.5-3.5s; Measure aromatic and arylnaphthalene lactone type lignanoid H-2 on internal standard compound, the peak area of 6, draws the molal weight ratio of target analytes relative to internal standard compound, then by the mass ratio foundation formula of target analytes with internal standard compound:
Calculate the purity P of target analytes
x, wherein:
Ix is the peak area of target analytes; I
stdfor interior target peak area; Nx is the nuclear number of target analytes; N
stdfor the nuclear number of interior target; Mx is the molal weight of target analytes; M
stdfor interior target molal weight; Mx is the quality of target analytes; m
stdfor interior target quality; Px is the purity of target analytes; P
stdfor the purity of internal standard compound;
Described arylnaphthalene lactone type lignanoid is deoxypodophyllotoxin, podophyllotoxin, 4-demethylpodophyllotoxin, podophyllotoxin-7 '-O-β-D-glucopyranoside, any one in 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside;
Described internal standard compound is Isosorbide-5-Nitrae-dinitro benzene, 3,4,5-trichloropyridine, any one in the chloro-2-nitrobenzene of 1,3,5-tri-.
Cost of the present invention is low, can evaluate the purity of arylnaphthalene lactone type lignanoid fast and accurately, have good economic and social benefit.
Accompanying drawing explanation
Fig. 1 is the quantitative nuclear magnetic spectrogram of the present invention.
Fig. 2 is the mol ratio linear regression result figure that the 4-demethylpodophyllotoxin of the quantitative nmr for the determination of the present invention and Isosorbide-5-Nitrae-dinitro benzene mol ratio and gravimetry measure.
Embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is described in further detail.
The present invention can be provided by following examples in concrete enforcement.
The inventive method comprises the following steps: the internal standard compound taking 10-14mg, is dissolved in the DMSO-d of 0.8-1.2mL
6in, be prepared into internal standard substance solution; Take the arylnaphthalene lactone type lignanoid of 10-50mg, be dissolved in the DMSO-d of 0.8-1.2mL
6in, be prepared into target analytes solution; Measure the internal standard substance solution of 80-120 μ L, the target analytes solution of 180-220 μ L, the DMSO-d of 180-220 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 25-35s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times (NS), 8 or 16; Spectral width (SW), 15-25ppm; Relaxation time (D1), 16-35s; Probe temperature, 20-30 DEG C; Pulse angle, 30 °, or 45 °, or 90 °; Acquisition time (AQ), 2.5-3.5s; Measure aromatic and arylnaphthalene lactone type lignanoid H-2 on internal standard compound, the peak area of 6, draws the molal weight ratio of target analytes relative to internal standard compound, then goes out the purity of target analytes by target analytes and the mass ratio foundation formulae discovery of internal standard compound;
Described arylnaphthalene lactone type lignanoid is deoxypodophyllotoxin, podophyllotoxin, 4-demethylpodophyllotoxin, podophyllotoxin-7 '-O-β-D-glucopyranoside, 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside, any one in sinolignan A;
R
1=CH
3or H; R
2=H, OH, glc or 6-acetyl-glc
Described internal standard compound is Isosorbide-5-Nitrae-dinitro benzene, 3,4,5-trichloropyridine, any one in the chloro-2-nitrobenzene of 1,3,5-tri-;
Described arylnaphthalene lactone type lignanoid purity obtains according to following formula
Ix is the peak area of target analytes; I
stdfor interior target peak area; Nx is the nuclear number of target analytes; N
stdfor the nuclear number of interior target; Mx is the molal weight of target analytes; M
stdfor interior target molal weight; Mx is the quality of target analytes; m
stdfor interior target quality; Px is the purity of target analytes; P
stdfor the purity of internal standard compound.
Embodiment 1
The inventive method comprises the following steps: the Isosorbide-5-Nitrae-dinitro benzene taking 12.47mg, is dissolved in the DMSO-d of 1.0mL
6in, be prepared into Isosorbide-5-Nitrae-dinitro benzene internal standard substance solution; Take the deoxypodophyllotoxin of 28.61mg, be dissolved in the DMSO-d of 1.0mL
6in, be prepared into deoxypodophyllotoxin target analytes solution; Measure the Isosorbide-5-Nitrae-dinitro benzene internal standard substance solution of 100 μ L, the deoxypodophyllotoxin target analytes solution of 200 μ L, the DMSO-d of 200 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 30s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times (NS), 16; Spectral width (SW), 20ppm; Relaxation time (D1), 16s; Probe temperature, 25 DEG C; Pulse angle, 30 °; Acquisition time (AQ), 3.2s; Measure aromatic and deoxypodophyllotoxin H-2 on Isosorbide-5-Nitrae-dinitro benzene, the peak area of 6, show that deoxypodophyllotoxin is relative to 1, the molal weight ratio of 4-dinitro benzene, then the purity being gone out deoxypodophyllotoxin by the mass ratio of deoxypodophyllotoxin and Isosorbide-5-Nitrae-dinitro benzene according to formulae discovery.
Embodiment 2
The inventive method comprises the following steps: the Isosorbide-5-Nitrae-dinitro benzene taking 12.85mg, is dissolved in the DMSO-d of 1.0mL
6in, be prepared into Isosorbide-5-Nitrae-dinitro benzene internal standard substance solution; Take the podophyllotoxin of 29.95mg, be dissolved in the DMSO-d of 1.0mL
6in, be prepared into podophyllotoxin target analytes solution; Measure the Isosorbide-5-Nitrae-dinitro benzene internal standard substance solution of 100 μ L, the podophyllotoxin target analytes solution of 200 μ L, the DMSO-d of 200 μ L
6, transfer in centrifuge tube, obtain sample solution; By this sample solution suspendible 30s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times (NS), 16; Spectral width (SW), 20ppm; Relaxation time (D1), 16s; Probe temperature, 25 DEG C; Pulse angle, 30 °; Acquisition time (AQ), 3.2s; Measure aromatic and podophyllotoxin H-2 on Isosorbide-5-Nitrae-dinitro benzene, the peak area of 6, draws the molal weight ratio of podophyllotoxin relative to Isosorbide-5-Nitrae-dinitro benzene, then goes out the purity of podophyllotoxin by podophyllotoxin and the mass ratio foundation formulae discovery of Isosorbide-5-Nitrae-dinitro benzene.
Embodiment 3
The inventive method comprises the following steps: the Isosorbide-5-Nitrae-dinitro benzene taking 12.90mg, is dissolved in the DMSO-d of 1.0mL
6in, be prepared into Isosorbide-5-Nitrae-dinitro benzene internal standard substance solution; Take the 4-demethylpodophyllotoxin of 29.35mg, be dissolved in the DMSO-d of 1.0mL
6in, be prepared into 4-demethylpodophyllotoxin target analytes solution; Measure the Isosorbide-5-Nitrae-dinitro benzene internal standard substance solution of 100 μ L, the 4-demethylpodophyllotoxin target analytes solution of 200 μ L, the DMSO-d of 200 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 30s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times (NS), 8; Spectral width (SW), 25ppm; Relaxation time (D1), 35s; Probe temperature, 30 DEG C; Pulse angle, 90 °; Acquisition time (AQ), 3.5s; Measure 1, aromatic and 4-demethylpodophyllotoxin H-2 on 4-dinitro benzene, the peak area of 6, show that 4-demethylpodophyllotoxin is relative to 1, the molal weight ratio of 4-dinitro benzene, gone out the purity of 4-demethylpodophyllotoxin again according to formulae discovery by the mass ratio of 4-demethylpodophyllotoxin and Isosorbide-5-Nitrae-dinitro benzene.
Embodiment 4
The inventive method comprises the following steps: the Isosorbide-5-Nitrae-dinitro benzene taking 11.87mg, is dissolved in the DMSO-d of 1.0mL
6in, be prepared into Isosorbide-5-Nitrae-dinitro benzene internal standard substance solution; Take 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside of 39.84mg, be dissolved in the DMSO-d of 1.0mL
6in, be prepared into 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside target analytes solution; Measure the Isosorbide-5-Nitrae-dinitro benzene internal standard substance solution of 100 μ L, 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside target analytes solution of 200 μ L, the DMSO-d of 200 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 30s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times (NS), 8; Spectral width (SW), 15ppm; Relaxation time (D1), 16s; Probe temperature, 20 DEG C; Pulse angle, 30 °; Acquisition time (AQ), 3.2s; Measure 1, aromatic and 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside H-2 on 4-dinitro benzene, the peak area of 6, show that 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside is relative to 1, the molal weight ratio of 4-dinitro benzene, gone out the purity of 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside again according to formulae discovery by the mass ratio of 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside and Isosorbide-5-Nitrae-dinitro benzene.
Embodiment 5
The inventive method comprises the following steps: the Isosorbide-5-Nitrae-dinitro benzene taking 11.80mg, is dissolved in the DMSO-d of 1.0mL
6in, be prepared into Isosorbide-5-Nitrae-dinitro benzene internal standard substance solution; Take podophyllotoxin-7 '-O-β-D-glucopyranoside of 40.24mg, be dissolved in the DMSO-d of 1.0mL
6in, be prepared into podophyllotoxin-7 '-O-β-D-glucopyranoside target analytes solution; Measure the Isosorbide-5-Nitrae-dinitro benzene internal standard substance solution of 100 μ L, podophyllotoxin-7 '-O-β-D-glucopyranoside target analytes solution of 200 μ L, the DMSO-d of 200 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 30s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times (NS), 16; Spectral width (SW), 20ppm; Relaxation time (D1), 16s; Probe temperature, 25 DEG C; Pulse angle, 30 °; Acquisition time (AQ), 3.2s; Measure 1, aromatic and podophyllotoxin-7 '-O-β-D-glucopyranoside H-2 on 4-dinitro benzene, the peak area of 6, show that podophyllotoxin-7 '-O-β-D-glucopyranoside is relative to 1, the molal weight ratio of 4-dinitro benzene, gone out the purity of podophyllotoxin-7 '-O-β-D-glucopyranoside again according to formulae discovery by the mass ratio of podophyllotoxin-7 '-O-β-D-glucopyranoside and Isosorbide-5-Nitrae-dinitro benzene.
Embodiment 6
The inventive method comprises the following steps: the method comprises the following steps: the Isosorbide-5-Nitrae-dinitro benzene taking 12.83mg, is dissolved in the DMSO-d of 1.0mL
6in, be prepared into Isosorbide-5-Nitrae-dinitro benzene internal standard substance solution; Take the sinolignan A of 47.08mg, be dissolved in the DMSO-d of 1.0mL
6in, be prepared into sinolignan A target analytes solution; Measure the Isosorbide-5-Nitrae-dinitro benzene internal standard substance solution of 100 μ L, the sinolignan A target analytes solution of 200 μ L, the DMSO-d of 200 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 30s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times (NS), 16; Spectral width (SW), 20ppm; Relaxation time (D1), 35s; Probe temperature, 25 DEG C; Pulse angle, 90 °; Acquisition time (AQ), 3.5s; Measure the H-2 of aromatic and sinolignan A on Isosorbide-5-Nitrae-dinitro benzene, the peak area of 6, show that sinolignan A is relative to 1, the molal weight ratio of 4-dinitro benzene, then the purity being gone out sinolignan A by the mass ratio of sinolignan A and Isosorbide-5-Nitrae-dinitro benzene according to formulae discovery.
Embodiment 7
The inventive method comprises the following steps: 3,4, the 5-trichloropyridines taking 10.91mg, are dissolved in the DMSO-d of 0.8mL
6in, be prepared into 3,4,5-trichloropyridine internal standard substance solution; Take the podophyllotoxin of 12.11mg, be dissolved in the DMSO-d of 0.8mL
6in, be prepared into podophyllotoxin target analytes solution; Measure 3,4, the 5-trichloropyridine internal standard substance solution of 80 μ L, the podophyllotoxin target analytes solution of 220 μ L, the DMSO-d of 220 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 25s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times (NS), 16; Spectral width (SW), 20ppm; Relaxation time (D1), 16s; Probe temperature, 25 DEG C; Pulse angle, 30 °; Acquisition time (AQ), 3.2s; Measure aromatic and podophyllotoxin H-2 on Isosorbide-5-Nitrae-dinitro benzene, the peak area of 6, draw the molal weight ratio of podophyllotoxin relative to 3,4,5-trichloropyridine, gone out the purity of podophyllotoxin again according to formulae discovery by the mass ratio of podophyllotoxin and 3,4,5-trichloropyridine.
Embodiment 8
The inventive method comprises the following steps: 1,3, the 5-tri-chloro-2-nitrobenzene taking 11.52mg, is dissolved in the DMSO-d of 1.2mL
6in, be prepared into 1,3,5-tri-chloro-2-nitrobenzene internal standard substance solution; Take the sinolignan A of 15.86mg, be dissolved in the DMSO-d of 1.2mL
6in, be prepared into sinolignan A target analytes solution; Measure 1,3, the 5-tri-chloro-2-nitrobenzene internal standard substance solution of 120 μ L, the sinolignan A target analytes solution of 180 μ L, the DMSO-d of 180 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 35s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times (NS), 16; Spectral width (SW), 20ppm; Relaxation time (D1), 16s; Probe temperature, 25 DEG C; Pulse angle, 30 °; Acquisition time (AQ), 3.2s; Measure 1,3, the H-2 of aromatic and sinolignan A on the chloro-2-nitrobenzene of 5-tri-, the peak area of 6, show that sinolignan A is relative to 1,3, the molal weight ratio of the chloro-2-nitrobenzene of 5-tri-, the purity of sinolignan A is gone out again by the mass ratio foundation formulae discovery of the chloro-2-nitrobenzene of sinolignan A and 1,3,5-tri-.
The parameter of above-described embodiment 1-8 and the result of arylnaphthalene lactone type lignanoid purity are as table 1
The inventive method is reliable and stable, and can measure the purity of arylnaphthalene lactone type lignanoid exactly, regarding assay data is as follows:
1. experiment material
4-demethylpodophyllotoxin is bought in Nanjing Qing Ze Pharmaceutical Technology Co., Ltd (purity is 98.0%).Isosorbide-5-Nitrae-dinitro benzene is bought and is changed into Industrial Co., Ltd in Tokyo.Chromatogram methyl alcohol is bought in α Cygni friend fine chemicals company limited, and DMSO-d6 buys in Cambridge Isotope Laboratories.Quantitative nuclear magnetic spectrogram adopts Bruker Avance 500MHz nmr determination.High performance liquid chromatography adopts Waters Alliance 2489 piece-rate system and Waters 2695 UV detect system to measure.
2. methodology confirmation
2.1. linearly investigate
Accurately take a certain amount of 4-demethylpodophyllotoxin and 1,4-dinitro benzene is in the sample hose of 5ml, the DMSO-d6 adding 0.5ml dissolves, obtain the 4-demethylpodophyllotoxin of different mol ratio example (0.946,0.6291,0.3078,1.516,2.947) and the hybrid standard sample solution of Isosorbide-5-Nitrae-dinitro benzene.Carry out linear regression by the mol ratio measured the 4-demethylpodophyllotoxin of quantitative nmr for the determination and Isosorbide-5-Nitrae-dinitro benzene mol ratio and gravimetry, obtain linear equation, Y=1.002X – 0.001, R2=1.Show that the method has good linear.The preparation of linear sample and test result are in table 1 and Fig. 1.
The preparation of table 1 linear sample and test result
2.2. specificity
The specificity investigating the method is investigated by the nuclear-magnetism hydrogen signal of the interference that may exist in bioassay standard sample solution.Test result shows, chemical shift is from δ 6.2 to 9.0 within the scope of this, and baseline is flat, H-2 in target detection thing, 6 and Isosorbide-5-Nitrae-dinitro benzene on fragrant Hydrogen Proton signal do not receive the interference of any hydrogen signal, peak shape is symmetrical, and the signal to noise ratio (S/N ratio) of these two signals is greater than 1000.Therefore the specificity of the method is stronger.
2.3. accuracy
The accuracy of the method is investigated by the average recovery rate and relative standard deviation (RSD) measuring three groups of samples.The results are shown in Table 2, three groups of samples be on average recovered as 100.2%, relative standard deviation (RSD) is 0.51%.Illustrate that the method can measure the purity of arylnaphthalene lactone type lignanoid exactly.
The accuracy measurement result of table 2.4-demethylpodophyllotoxin,
| Group number | 4-demethylpodophyllotoxin (mg) | 1.4-dinitro benzene (mg) | Purity (%) | The recovery (%) |
| Set-1 | 5.94 | 1.29 | 98.9 | 100.9 |
| Set-1 | 5.77 | 1.25 | 97.3 | 99.3 |
| Set-1 | 6.67 | 1.38 | 98.7 | 100.7 |
| Set-2 | 9.85 | 1.07 | 97.8 | 99.8 |
| Set-2 | 10.00 | 1.09 | 98.3 | 100.3 |
| Set-2 | 11.13 | 1.16 | 98.6 | 100.6 |
| Set-3 | 15.03 | 1.06 | 97.9 | 99.9 |
| Set-3 | 17.10 | 1.18 | 98.1 | 100.1 |
| Set-3 | 15.64 | 1.09 | 98.3 | 100.3 |
| Mean | 98.2 | 100.2 | ||
| RSD(％) | 0.51 |
2.4. precision
The result of withinday precision and day to day precision is respectively in table 3 and table 4.The relative standard deviation (RSD) of withinday precision and day to day precision is respectively 0.32% and 0.36%, shows that the method precision is good.
The withinday precision measurement result of table 3 4-demethylpodophyllotoxin
| Numbering | 4-demethylpodophyllotoxin (mg) | 1.4-dinitro benzene (mg) | Purity (%) |
| 1 | 5.94 | 1.29 | 98.3 |
| 2 | 6.02 | 1.34 | 98.2 |
| 3 | 5.87 | 1.30 | 97.6 |
| 4 | 5.96 | 1.33 | 98.0 |
| 5 | 6.07 | 1.34 | 98.0 |
| 6 | 6.01 | 1.30 | 97.5 |
| Mean | 97.9 | ||
| RSD(％) | 0.32 |
The day to day precision measurement result of table 4 4-demethylpodophyllotoxin
| Numbering | 4-demethylpodophyllotoxin (mg) | 1.4-dinitro benzene (mg) | Purity (%) |
| 1 | 6.04 | 1.32 | 98.2 |
| 2 | 6.01 | 1.30 | 98.3 |
| 3 | 5.85 | 1.29 | 97.6 |
| 4 | 5.86 | 1.31 | 98.1 |
| 5 | 6.07 | 1.36 | 98.0 |
| 6 | 6.15 | 1.34 | 97.4 |
| Mean | 97.9 | ||
| RSD(％) | 0.36 |
2.5. stability
The standard model solution prepared measured purity respectively after 0,8,16 and 24 hour, and RSD is 0.34%, showed that sample solution was stable existence in 24 hours.The results are shown in Table 5.
The Stability Determination result of table 5 4-demethylpodophyllotoxin
2.6. quantitative nuclear-magnetism determination method and high performance liquid chromatography are to the comparative studies of arylnaphthalene lactone type lignanoid purity testing
For confirming the accuracy of the method further, the purity of high performance liquid chromatography to arylnaphthalene lactone type lignanoid is adopted to measure.Liquid phase chromatogram condition is, methanol-water gradient elution (35-65%, 0-35min; 65-100%, 35-50min); Determined wavelength 210nm; Flow velocity 1.0ml/ml; Column temperature 25 DEG C.Statistics shows, quantitative nuclear-magnetism determination method and the measurement result of high performance liquid chromatography to arylnaphthalene lactone type lignanoid purity are basically identical.Specifically in table 6.
The quantitative nuclear-magnetism determination method of table 6 and high performance liquid chromatography are to the measurement result of arylnaphthalene lactone type lignanoid purity
According to the analysis result of this experiment, compared with traditional efficient liquid-phase chromatography method, quantitative hydrogen nuclear magnetic resonance spectroscopy has the following advantages:
1. quick.15min is needed for the pre-treatment of each quantitative nuclear magnetic resonance sample and analysis.And the pre-treatment of each sample of high performance liquid chromatography and analysis amount to 65min (sample pre-treatments 5min; Chromatographic column balance 10min; Stratographic analysis 50min).Each sample amounts hydrogen nuclear magnetic resonance spectroscopy can save 50min.
2. accurate.For the compound lacking conjugated system, the high performance liquid chromatography with UV-detector does not respond.Therefore, when being mixed with the impurity of this kind of shortage conjugated system in target analytes, analysis result is inaccurate.What quantitative hydrogen nuclear magnetic resonance spectroscopy detected is proton, is ubiquitous for proton organic compound, and whether target analytes and the organic impurities that coexists with it be containing conjugated system, and quantitative hydrogen nuclear magnetic resonance spectroscopy can detect accurately to it.
3. cost-saving.The quantitative hydrogen nuclear magnetic resonance spectroscopy organic molecule that structure simple molecules amount cheap and easy to get is moderate does interior mark, and this test adopts Isosorbide-5-Nitrae-dinitro benzene to do interior mark, and requirement is about 1mg, and cost is 15 yuan; The DMSO-d6 of 0.5ml, cost is 20 yuan.For quantitative hydrogen nuclear magnetic resonance spectroscopy, each sample needs about 35 yuan.High performance liquid chromatography, chromatogram methyl alcohol 60ml, cost about 2 yuan; The standard items of object are needed to be about 10mg, the cost of different arylnaphthalene lactone type lignanoids is also not quite similar, deoxypodophyllotoxin (335 yuan), podophyllotoxin (190 yuan), 4-demethylpodophyllotoxin (225 yuan), for high performance liquid chromatography, each sample needs 192-337 unit, cost-saving 157-302 unit.And arylnaphthalene lactone type lignanoid glucosides is as podophyllotoxin-7 '-O-β-D-Glucose glycosides, 4-demethylpodophyllotoxin-7 '-O-β-D-Glucose glycosides, sinolignan A is commercially difficult to buy, and the absolute purity therefore for these three arylnaphthalene lactone type lignanoid glucosides is measured and cannot be measured by high performance liquid chromatography.
Above-mentioned experiment can clearly show, the invention discloses arylnaphthalene lactone type lignanoid method for detecting purity, quantitative hydrogen nuclear magnetic resonance spectroscopy can measure the purity of arylnaphthalene lactone type lignanoid rapidly and accurately, cost is low, for arylnaphthalene lactone type lignanoid further the quality control of evaluated biological activity and relevant medicinal material accurately provide early stage basis, for the Purity assessment of natural products makes creative contribution, there is good economic and social benefit.
Claims (9)
1. measure a method for arylnaphthalene lactone type lignanoid purity based on quantitative hydrogen nuclear magnetic resonance spectroscopy, it is characterized in that, the method comprises the following steps: the internal standard compound taking 10-14mg, is dissolved in the DMSO-d of 0.8-1.2mL
6in, be prepared into inner mark solution; Take the arylnaphthalene lactone type lignanoid of 10-50mg, be dissolved in the DMSO-d of 0.8-1.2mL
6in, be prepared into target analytes solution; Measure the inner mark solution of 80-120 μ L, the target analytes solution of 180-220 μ L, the DMSO-d of 180-220 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 25-35s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times, 8 or 16; Spectral width, 15-25ppm; Relaxation time, 16-35s; Probe temperature, 20-30 DEG C; Pulse angle, 30 °, or 45 °, or 35 °; Acquisition time, 2.5-3.5s; Measure aromatic and arylnaphthalene lactone type lignanoid H-2 on internal standard compound, the peak area of 6, draws the molal weight ratio of target analytes relative to internal standard compound, then by the mass ratio foundation formula of target analytes with internal standard compound:
Calculate the purity P of target analytes
x, wherein:
Ix is the peak area of target analytes; I
stdfor interior target peak area; Nx is the nuclear number of target analytes; N
stdfor the nuclear number of interior target; Mx is the molal weight of target analytes; M
stdfor interior target molal weight; Mx is the quality of target analytes; m
stdfor interior target quality; Px is the purity of target analytes; P
stdfor the purity of internal standard compound;
Described arylnaphthalene lactone type lignanoid is deoxypodophyllotoxin, podophyllotoxin, 4-demethylpodophyllotoxin, podophyllotoxin-7 '-O-β-D-glucopyranoside, 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside, any one in sinolignanA;
Described internal standard compound is Isosorbide-5-Nitrae-dinitro benzene, 3,4,5-trichloropyridine, any one in the chloro-2-nitrobenzene of 1,3,5-tri-.
2. a kind of method measuring arylnaphthalene lactone type lignanoid purity based on quantitative hydrogen nuclear magnetic resonance spectroscopy according to claim 1, it is characterized in that, the method comprises the following steps: the Isosorbide-5-Nitrae-dinitro benzene taking 12.47mg, is dissolved in the DMSO-d of 1.0mL
6in, be prepared into Isosorbide-5-Nitrae-dinitro benzene inner mark solution; Take the deoxypodophyllotoxin of 28.61mg, be dissolved in the DMSO-d of 1.0mL
6in, be prepared into deoxypodophyllotoxin target analytes solution; Measure the Isosorbide-5-Nitrae-dinitro benzene inner mark solution of 100 μ L, the deoxypodophyllotoxin target analytes solution of 200 μ L, the DMSO-d of 200 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 30s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times 16; Spectral width, 20ppm; Relaxation time, 16s; Probe temperature, 25 DEG C; Pulse angle, 30 °; Acquisition time, 3.2s; Measure aromatic and deoxypodophyllotoxin H-2 on Isosorbide-5-Nitrae-dinitro benzene, the peak area of 6, show that deoxypodophyllotoxin is relative to 1, the molal weight ratio of 4-dinitro benzene, then the purity being gone out deoxypodophyllotoxin by the mass ratio of deoxypodophyllotoxin and Isosorbide-5-Nitrae-dinitro benzene according to formulae discovery.
3. a kind of method measuring arylnaphthalene lactone type lignanoid purity based on quantitative hydrogen nuclear magnetic resonance spectroscopy according to claim 1, it is characterized in that, the method comprises the following steps: the Isosorbide-5-Nitrae-dinitro benzene taking 12.85mg, is dissolved in the DMSO-d of 1.0mL
6in, be prepared into Isosorbide-5-Nitrae-dinitro benzene inner mark solution; Take the podophyllotoxin of 29.95mg, be dissolved in the DMSO-d of 1.0mL
6in, be prepared into podophyllotoxin target analytes solution; Measure the Isosorbide-5-Nitrae-dinitro benzene inner mark solution of 100 μ L, the podophyllotoxin target analytes solution of 200 μ L, the DMSO-d of 200 μ L
6, transfer in centrifuge tube, obtain sample solution; By this sample solution suspendible 30s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times, 16; Spectral width, 20ppm; Relaxation time, 16s; Probe temperature, 25 DEG C; Pulse angle, 30 °; Acquisition time, 3.2s; Measure aromatic and podophyllotoxin H-2 on Isosorbide-5-Nitrae-dinitro benzene, the peak area of 6, draws the molal weight ratio of podophyllotoxin relative to Isosorbide-5-Nitrae-dinitro benzene, then goes out the purity of podophyllotoxin by podophyllotoxin and the mass ratio foundation formulae discovery of Isosorbide-5-Nitrae-dinitro benzene.
4. a kind of method measuring arylnaphthalene lactone type lignanoid purity based on quantitative hydrogen nuclear magnetic resonance spectroscopy according to claim 1, it is characterized in that, the method comprises the following steps: the Isosorbide-5-Nitrae-dinitro benzene taking 12.90mg, is dissolved in the DMSO-d of 1.0mL
6in, be prepared into Isosorbide-5-Nitrae-dinitro benzene inner mark solution; Take the 4-demethylpodophyllotoxin of 29.35mg, be dissolved in the DMSO-d of 1.0mL
6in, be prepared into 4-demethylpodophyllotoxin target analytes solution; Measure the Isosorbide-5-Nitrae-dinitro benzene inner mark solution of 100 μ L, the 4-demethylpodophyllotoxin target analytes solution of 200 μ L, the DMSO-d of 200 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 30s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times, 8; Spectral width, 25ppm; Relaxation time, 35s; Probe temperature, 30 DEG C; Pulse angle, 90 °; Acquisition time, 3.5s; Measure 1, aromatic and 4-demethylpodophyllotoxin H-2 on 4-dinitro benzene, the peak area of 6, show that 4-demethylpodophyllotoxin is relative to 1, the molal weight ratio of 4-dinitro benzene, gone out the purity of 4-demethylpodophyllotoxin again according to formulae discovery by the mass ratio of 4-demethylpodophyllotoxin and Isosorbide-5-Nitrae-dinitro benzene.
5. a kind of method measuring arylnaphthalene lactone type lignanoid purity based on quantitative hydrogen nuclear magnetic resonance spectroscopy according to claim 1, it is characterized in that, the method comprises the following steps: the Isosorbide-5-Nitrae-dinitro benzene taking 11.87mg, is dissolved in the DMSO-d of 1.0mL
6in, be prepared into Isosorbide-5-Nitrae-dinitro benzene inner mark solution; Take 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside of 39.84mg, be dissolved in the DMSO-d of 1.0mL
6in, be prepared into 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside target analytes solution; Measure the Isosorbide-5-Nitrae-dinitro benzene inner mark solution of 100 μ L, 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside target analytes solution of 200 μ L, the DMSO-d of 200 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 30s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times, 8; Spectral width, 15ppm; Relaxation time, 16s; Probe temperature, 20 DEG C; Pulse angle, 30 °; Acquisition time, 3.2s; Measure 1, aromatic and 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside H-2 on 4-dinitro benzene, the peak area of 6, show that 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside is relative to 1, the molal weight ratio of 4-dinitro benzene, gone out the purity of 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside again according to formulae discovery by the mass ratio of 4-demethylpodophyllotoxin-7 '-O-β-D-glucopyranoside and Isosorbide-5-Nitrae-dinitro benzene.
6. a kind of method measuring arylnaphthalene lactone type lignanoid purity based on quantitative hydrogen nuclear magnetic resonance spectroscopy according to claim 1, it is characterized in that, the method comprises the following steps: the Isosorbide-5-Nitrae-dinitro benzene taking 11.80mg, is dissolved in the DMSO-d of 1.0mL
6in, be prepared into Isosorbide-5-Nitrae-dinitro benzene inner mark solution; Take podophyllotoxin-7 '-O-β-D-glucopyranoside of 40.24mg, be dissolved in the DMSO-d of 1.0mL
6in, be prepared into podophyllotoxin-7 '-O-β-D-glucopyranoside target analytes solution; Measure the Isosorbide-5-Nitrae-dinitro benzene inner mark solution of 100 μ L, podophyllotoxin-7 '-O-β-D-glucopyranoside target analytes solution of 200 μ L, the DMSO-d of 200 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 30s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times, 16; Spectral width, 20ppm; Relaxation time, 16s; Probe temperature, 25 DEG C; Pulse angle, 30 °; Acquisition time, 3.2s; Measure 1, aromatic and podophyllotoxin-7 '-O-β-D-glucopyranoside H-2 on 4-dinitro benzene, the peak area of 6, show that podophyllotoxin-7 '-O-β-D-glucopyranoside is relative to 1, the molal weight ratio of 4-dinitro benzene, gone out the purity of podophyllotoxin-7 '-O-β-D-glucopyranoside again according to formulae discovery by the mass ratio of podophyllotoxin-7 '-O-β-D-glucopyranoside and Isosorbide-5-Nitrae-dinitro benzene.
7. a kind of method measuring arylnaphthalene lactone type lignanoid purity based on quantitative hydrogen nuclear magnetic resonance spectroscopy according to claim 1, it is characterized in that, the method comprises the following steps: the Isosorbide-5-Nitrae-dinitro benzene taking 12.83mg, is dissolved in the DMSO-d of 1.0mL
6in, be prepared into Isosorbide-5-Nitrae-dinitro benzene inner mark solution; Take the sinolignan A of 47.08mg, be dissolved in the DMSO-d of 1.0mL
6in, be prepared into sinolignan A target analytes solution; Measure the Isosorbide-5-Nitrae-dinitro benzene inner mark solution of 100 μ L, the sinolignan A target analytes solution of 200 μ L, the DMSO-d of 200 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 30s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times, 8; Spectral width, 20ppm; Relaxation time, 35s; Probe temperature, 25 DEG C; Pulse angle, 90 °; Acquisition time, 3.5s; Measure the H-2 of aromatic and sinolignan A on Isosorbide-5-Nitrae-dinitro benzene, the peak area of 6, show that sinolignanA is relative to 1, the molal weight ratio of 4-dinitro benzene, then the purity being gone out sinolignan A by the mass ratio of sinolignan A and Isosorbide-5-Nitrae-dinitro benzene according to formulae discovery.
8. a kind of method measuring arylnaphthalene lactone type lignanoid purity based on quantitative hydrogen nuclear magnetic resonance spectroscopy according to claim 1, it is characterized in that, the method comprises the following steps: 3,4, the 5-trichloropyridines taking 10.91mg, are dissolved in the DMSO-d of 0.8mL
6in, be prepared into 3,4,5-trichloropyridine inner mark solution; Take the podophyllotoxin of 12.11mg, be dissolved in the DMSO-d of 0.8mL
6in, be prepared into podophyllotoxin target analytes solution; Measure 3,4, the 5-trichloropyridine inner mark solutions of 80 μ L, the podophyllotoxin target analytes solution of 220 μ L, the DMSO-d of 220 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 25s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times, 16; Spectral width, 20ppm; Relaxation time, 16s; Probe temperature, 25 DEG C; Pulse angle, 30 °; Acquisition time, 3.2s; Measure aromatic and podophyllotoxin H-2 on Isosorbide-5-Nitrae-dinitro benzene, the peak area of 6, draw the molal weight ratio of podophyllotoxin relative to 3,4,5-trichloropyridine, gone out the purity of podophyllotoxin again according to formulae discovery by the mass ratio of podophyllotoxin and 3,4,5-trichloropyridine.
9. a kind of method measuring arylnaphthalene lactone type lignanoid purity based on quantitative hydrogen nuclear magnetic resonance spectroscopy according to claim 1, it is characterized in that, the method comprises the following steps: take 1 of 11.52mg, 3, the chloro-2-nitrobenzene of 5-tri-, is dissolved in the DMSO-d of 1.2mL
6in, be prepared into 1,3,5-tri-chloro-2-nitrobenzene inner mark solution; Take the sinolignan A of 15.86mg, be dissolved in the DMSO-d of 1.2mL
6in, be prepared into sinolignan A target analytes solution; Measure 1,3, the 5-tri-chloro-2-nitrobenzene inner mark solution of 120 μ L, the sinolignan A target analytes solution of 180 μ L, the DMSO-d of 180 μ L
6, transfer in centrifuge tube, obtain sample solution, by this sample solution suspendible 35s, be transferred in nuclear magnetic tube, carry out quantitative nuclear-magnetism test, the condition of described quantitative nuclear-magnetism test: scanning times, 16; Spectral width, 20ppm; Relaxation time, 16s; Probe temperature, 25 DEG C; Pulse angle, 30 °; Acquisition time, 3.2s; Measure 1,3, the H-2 of aromatic and sinolignan A on the chloro-2-nitrobenzene of 5-tri-, the peak area of 6, show that sinolignan A is relative to 1,3, the molal weight ratio of the chloro-2-nitrobenzene of 5-tri-, the purity of sinolignan A is gone out again by the mass ratio foundation formulae discovery of the chloro-2-nitrobenzene of sinolignan A and 1,3,5-tri-.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106018456A (en) * | 2016-05-16 | 2016-10-12 | 山东省分析测试中心 | Method for determining purity of daptomycin on basis of hydrogen nuclear magnetic resonance |
| CN111855727A (en) * | 2020-07-23 | 2020-10-30 | 淮北师范大学 | Method for detecting purity and impurity content of foaming agent product |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH11118739A (en) * | 1997-10-08 | 1999-04-30 | Ihm:Kk | Computerized magnetic resonance analyzer, and measurement method by using analytical instrument having equal efficiency to the same for specifying material, and blend ratio of essential oil blend for specific use, and its application |
| JP2006194798A (en) * | 2005-01-14 | 2006-07-27 | Sumitomo Electric Ind Ltd | Analytical method for polyolefin resin composition |
| CN101429107A (en) * | 2008-11-18 | 2009-05-13 | 云南民族大学 | Lignanoid compounds, producing method, identification method and uses thereof |
| US20090311349A1 (en) * | 2008-06-05 | 2009-12-17 | Bionovo, Inc., A Delaware Corporation | Method of quantification of multiple bioactives from botanical compositions |
| CN103808749A (en) * | 2013-01-30 | 2014-05-21 | 河南省科高植物天然产物开发工程技术有限公司 | Method for identifying forsythia suspensa derivatives |
| CN104069348A (en) * | 2014-06-30 | 2014-10-01 | 天津中医药大学 | Sealwort extract as well as preparation method and use thereof |
-
2015
- 2015-04-02 CN CN201510156068.3A patent/CN104698022B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH11118739A (en) * | 1997-10-08 | 1999-04-30 | Ihm:Kk | Computerized magnetic resonance analyzer, and measurement method by using analytical instrument having equal efficiency to the same for specifying material, and blend ratio of essential oil blend for specific use, and its application |
| JP2006194798A (en) * | 2005-01-14 | 2006-07-27 | Sumitomo Electric Ind Ltd | Analytical method for polyolefin resin composition |
| US20090311349A1 (en) * | 2008-06-05 | 2009-12-17 | Bionovo, Inc., A Delaware Corporation | Method of quantification of multiple bioactives from botanical compositions |
| CN101429107A (en) * | 2008-11-18 | 2009-05-13 | 云南民族大学 | Lignanoid compounds, producing method, identification method and uses thereof |
| CN103808749A (en) * | 2013-01-30 | 2014-05-21 | 河南省科高植物天然产物开发工程技术有限公司 | Method for identifying forsythia suspensa derivatives |
| CN104069348A (en) * | 2014-06-30 | 2014-10-01 | 天津中医药大学 | Sealwort extract as well as preparation method and use thereof |
Non-Patent Citations (2)
| Title |
|---|
| 张炳火,李汉全: "鬼臼毒素在两种溶剂中核磁共振谱图数据的比较", 《安徽农业科学》 * |
| 李前荣 等: "4-叠氮-4-脱氧-4"-去甲基表鬼臼毒素的1H和13C核磁共振分析", 《分析化学研究简报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106018456A (en) * | 2016-05-16 | 2016-10-12 | 山东省分析测试中心 | Method for determining purity of daptomycin on basis of hydrogen nuclear magnetic resonance |
| CN111855727A (en) * | 2020-07-23 | 2020-10-30 | 淮北师范大学 | Method for detecting purity and impurity content of foaming agent product |
| CN111855727B (en) * | 2020-07-23 | 2023-12-19 | 淮北师范大学 | Method for detecting purity and impurity content of foaming agent product |
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