CN114685610A - Lactone hydrolysate of daptomycin RS-8a impurity and preparation method thereof - Google Patents
Lactone hydrolysate of daptomycin RS-8a impurity and preparation method thereof Download PDFInfo
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- 108010013198 Daptomycin Proteins 0.000 title claims abstract description 59
- 229960005484 daptomycin Drugs 0.000 title claims abstract description 59
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- 239000000413 hydrolysate Substances 0.000 title claims abstract description 55
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- 238000002360 preparation method Methods 0.000 title abstract description 15
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- 238000000034 method Methods 0.000 claims abstract description 13
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- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 8
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- 238000000855 fermentation Methods 0.000 claims description 15
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- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 12
- 239000000047 product Substances 0.000 claims description 10
- 238000010828 elution Methods 0.000 claims description 8
- 238000004108 freeze drying Methods 0.000 claims description 8
- 238000000746 purification Methods 0.000 claims description 8
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- 229920005989 resin Polymers 0.000 claims description 8
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 7
- 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 6
- 239000007864 aqueous solution Substances 0.000 claims description 6
- 235000019253 formic acid Nutrition 0.000 claims description 6
- 239000012043 crude product Substances 0.000 claims description 4
- 230000003301 hydrolyzing effect Effects 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
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- 238000001914 filtration Methods 0.000 claims description 3
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- 238000001514 detection method Methods 0.000 description 5
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- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 4
- 238000005481 NMR spectroscopy Methods 0.000 description 4
- 108010009736 Protein Hydrolysates Proteins 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
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- 238000005520 cutting process Methods 0.000 description 1
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- 125000003074 decanoyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C(*)=O 0.000 description 1
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- MYPYJXKWCTUITO-LYRMYLQWSA-N vancomycin Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-N 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
Abstract
The invention provides a lactone hydrolysate of daptomycin RS-8a impurity, a preparation method thereof and application of the lactone hydrolysate as a reference substance, wherein the daptomycin RS-8a impurity with higher purity is obtained by preparation and separation, sodium hydroxide is added, the hydrolysis time is controlled, the lactone hydrolysate of the RS-8a impurity is obtained by directional hydrolysis, and the chemical structure of the lactone hydrolysate is determined by mass spectrometry and nuclear magnetism. The method solves the problem that impurity standard substances are difficult to separate and obtain, provides technical support for optimizing the industrial production of daptomycin raw materials, and provides guarantee for the improvement of the subsequent daptomycin process quality standard and the optimization of an analysis method.
Description
Technical Field
The invention relates to the technical field of medicines, and in particular relates to a lactone hydrolysate impurity of daptomycin RS-8a impurities, and a preparation method and application thereof.
Background
Daptomycin (Daptomycin) is a monocyclolipopeptide antibiotic originally studied by Lilly, Cubist pharmaceutical company, and is an N-decanoyl derivative of a lipopeptide compound a21978C, which is a fermentation product of streptomyces roseosporus, and contains 13 amino acids, 10 of which form cyclododecopeptide, and the other 3 of which are connected with decanoyl to form a side chain, and is a second generation glycopeptide antibiotic drug following vancomycin. In 2003, the U.S. Food and Drug Administration (FDA) was passing quicklyDaptomycin (Daptomycin) (trade name) for injection approved by the Rapid Authority procedure) Can be used for treating complicated skin and skin structure infection caused by gram-positive sensitive strains, such as abscess, operation incision infection and skin ulcer.
Daptomycin is a fermentation product, is obtained by separating and purifying fermentation filtrate, can generate a large amount of pigment and byproducts with similar structures with the daptomycin during the fermentation process, and international application WO2001053330 discloses 14 impurities of daptomycin, such as dehydrated daptomycin, isomeric daptomycin, daptomycin lactone hydrolysate and the like.
Laboratory personnel find that a large amount of unknown impurities exist in direct fermentation liquor of daptomycin and fermentation liquor treated by a subsequent extraction and purification process according to the existing extraction process, but the compound of the formula (I) is listed in the quality standard of daptomycin without the import medicine standard, and the compound is generally treated as the unknown impurities in actual work, so that huge clinical risks exist.
Therefore, the analysis confirms the compound shown in the formula (I) and provides a reference substance, which has a remarkable significance on the quality control of the production of the daptomycin bulk drug and the preparation production, and in addition, in the process of carrying out drug quality detection, high-purity impurities are required to be used as the reference substance of corresponding impurities for the quality detection of the daptomycin bulk drug and the preparation.
Disclosure of Invention
In order to solve the technical problems, the invention provides a novel lactone hydrolysate of daptomycin RS-8a impurity, which has a structure shown in a formula (I):
the preparation method of the lactone hydrolysate is characterized in that a NaOH solution is added into a solution containing RS-8a for hydrolysis to obtain the RS-8a lactone hydrolysate.
Further, the solution containing daptomycin RS-8a comprises direct fermentation broth of daptomycin or fermentation broth treated by a subsequent extraction and purification process.
Further, the fermentation liquid after the subsequent extraction and purification process can be a daptomycin resin 818 analysis combined liquid.
Further, 0.1mol/LNaOH solution is added into the solution containing the daptomycin RS-8a impurity, after hydrolysis is carried out for 25-35 minutes, 0.1mol/LHCl solution with the same volume as NaOH solution is added for neutralization, so as to obtain the daptomycin RS-8a lactone hydrolysate, and the volume ratio of the solution containing RS-8a to the 0.1mol/LNaOH solution is 1: 2.
Further, the hydrolysis time was 30 minutes.
The preparation method of the lactone hydrolysate is characterized by comprising the following steps:
1) crude RS-8 a: taking daptomycin fermentation liquor or daptomycin resin 818, resolving the combined solution, filtering, eluting the filtrate by a chromatographic column, collecting the eluent, and concentrating under reduced pressure to obtain a crude product of RS-8 a;
2) purification of RS-8 a: dissolving the RS-8a crude product prepared in the step 1), adsorbing the dissolved product on a column, eluting the dissolved product by using eluent, collecting the eluent in sections, and combining the eluent with high purity to obtain RS-8a combined solution;
3) crude RS-8a lactone hydrolysate: adding 0.1mol/LNaOH solution into the RS-8a combined solution, hydrolyzing for 30 minutes, adding an isovolumetric 0.1mol/LHCl solution for neutralization to obtain a crude RS-8a lactone hydrolysate solution;
4) purification of RS-8a lactone hydrolysate: and (3) adsorbing the crude RS-8a lactone hydrolysate solution obtained in the step 3) by using a column, eluting by using eluent, collecting by sections, combining the eluent with higher purity, concentrating under reduced pressure to obtain a concentrated RS-8a lactone hydrolysate solution, freeze-drying and freeze-drying, wherein the freeze-dried solid is the RS-8a lactone hydrolysate.
Further, the volume ratio of the RS-8a combined solution to the 0.1mol/LNaOH solution is 1: 2.
Furthermore, the daptomycin fermentation broth used in step 1) is a fermentation broth which is not treated after fermentation culture, usually, the first step in the daptomycin fermentation broth treatment process is adsorption and desorption treatment by using an HZ818 resin, and the product obtained by the treatment is a daptomycin resin 818 desorption combined solution.
Further, the chromatographic column used in the step 1) is a C18 column, and the elution is carried out by using water after the chromatographic column is loaded, wherein the elution amount is 2 times of the column volume; then eluting with 30% ethanol water solution with the elution amount of 1.6 times of the column volume; then eluting with 50% ethanol water solution with the elution amount being 2 times of the column volume, and collecting the eluate in the stage; finally, eluting with ethanol, wherein the elution amount is 2 times of the column volume.
Further, the chromatographic column used in the step 2) is a C18 column, the eluent a used is 0.1% formic acid aqueous solution, the eluent B is ethanol, and the volume ratio of the two is eluent a: eluent B60: and 40, combining the eluates with the purity of more than 70 percent.
Further, the volume ratio of the RS-8a combined solution to the 0.1mol/LNaOH solution in the step 3) is 1: 2.
Further, the chromatographic column used in the step 4) is a C18 column, the eluent A used is 0.1% formic acid aqueous solution, the eluent B is ethanol, and the volume ratio of the two is eluent A: eluent B ═ 60: 40.
the daptomycin RS-8a lactone hydrolysate shown in the formula (I) is applied as an impurity reference substance in the quality control of daptomycin.
Furthermore, the daptomycin RS-8a lactone hydrolysate can be used as a reference substance for detecting the content of impurities in daptomycin medicines, or can be used as a reference substance for detecting the content of impurities in daptomycin in the daptomycin extraction and separation process.
Compared with the prior art, the invention has the beneficial effects that:
(1) the invention provides a novel daptomycin lactone hydrolysate, a preparation and separation method thereof, and application of the daptomycin lactone hydrolysate as a standard reference substance;
(2) according to the invention, the impurity RS-8a with higher purity is obtained by preparation and separation, sodium hydroxide is added, the hydrolysis time is controlled, the RS-8a lactone hydrolysate is obtained by directional hydrolysis, the structure of the lactone hydrolysate is determined by mass spectrometry and nuclear magnetism, and the difficulty that a standard product is difficult to separate and obtain is solved;
(3) the method provides technical support for optimizing the industrial production of the daptomycin raw material, and guarantees the improvement of the subsequent daptomycin process quality standard and the optimization of an analysis method.
Drawings
FIG. 1 is a liquid phase detection profile of the original daptomycin product;
FIG. 2 is a liquid phase detection profile of the Memanticin product;
FIG. 3 is a liquid phase detection spectrum of daptomycin resin 818;
FIG. 4 is a high resolution first order mass spectrum of the RS-8a lactone hydrolysate prepared in example two;
FIG. 5 is a high resolution secondary mass spectrum of the RS-8a lactone hydrolysate prepared in example two;
FIG. 6 is a hydrogen spectrum of the RS-8a lactone hydrolysate prepared in example two;
FIG. 7 is a carbon spectrum of the RS-8a lactone hydrolysate prepared in example two.
Detailed Description
Example one
Determination of a preparation method of lactone hydrolysate of daptomycin RS-8a impurity.
The liquid phase analysis of the daptomycin resin 818 resolved pool showed that the sample contained an impurity peak with a retention time of 1.05(RRT1.05) as shown in FIG. 3.
The mass spectrum analysis (instrument: Agilent HPLC 1290-QTOF 6545A) is carried out on the impurities of the combined solution by adopting a two-dimensional desalting multi-center cutting technology to obtain the mass-to-charge ratio of an RRT1.05 impurity peak of 826.8774[ M +2H]2+And secondary fragment analysis is carried out by taking 826.8774 as parent ion, the secondary fragment and the secondary fragment of daptomycin lactone hydrolysate only have difference on capric acid fragment, and the mass-to-charge ratio of RRT1.05 impurity peak to RRT1.55 impurity peak (RS-8a) is 817.8707[ M +2H ]]2+The difference is one molecule of water, so the lactone hydrolysate with the RRT1.05 impurity peak as the RRT1.55 impurity peak is supposed to have the RRT1.55 impurity peak with normalized content of about 5.5% in the merged liquid and higher than the RRT1.05 impurity peak (1.0%),and the separation distance from the main peak is far, so that the separation and the preparation are more convenient, and therefore, the RRT1.55 impurity peak is prepared and then hydrolyzed into the RRT1.05 impurity peak.
Example two
Preparation of lactone hydrolysate of daptomycin RS-8a impurity.
Filtering the combined solution after the daptomycin resin 818 is resolved, taking 250ml of the combined solution to be adsorbed by a peristaltic pump, loading the mixture on a C18 fast liquid phase preparative chromatographic column (chromatographic column parameters are C18, 330g, 40-60 mu m,) Eluting with water (500ml), 30% ethanol water solution (400ml), 50% ethanol water solution (500ml) and ethanol (500ml) in sequence, collecting eluate by stages, analyzing the eluate by HPLC, collecting the fraction (50% ethanol water solution) with high RS-8a purity, concentrating under reduced pressure to remove most ethanol to obtain RS-8a crude purification combined concentrated solution for further refining.
Adsorbing the refined upper column liquid in the previous step by using a C18 column with the diameter of 10 mu m, and then eluting by using an eluent, wherein the eluent A is 0.1% formic acid aqueous solution, the eluent B is ethanol, and the volume ratio of the eluent A to the eluent B is: eluent B60: 40, collecting the eluent in sections at the flow rate of 25ml/min, merging the eluent with higher purity (more than 70%), adding 0.1mol/l NaOH solution into the fraction merging solution with the RS-8a purity of more than 70% in proportion (merging solution: 0.1mol/l NaOH solution is 1:2), hydrolyzing for 30 minutes, adding equal volume of 0.1mol/l HCl solution for neutralizing to obtain a preparation stock solution of the RS-8a lactone hydrolysate, wherein the RRT of the RS-8a lactone hydrolysate is 1.05 according to liquid phase analysis.
Adsorbing the stock solution of the RS-8a lactone hydrolysate in the previous step by using a C18 column with the diameter of 10 mu m, and then eluting by using an eluent, wherein the eluent A: b is about 60: 40, the eluent A is 0.1% formic acid aqueous solution, the eluent B is ethanol, and the volume ratio of the eluent A to the eluent B is that: eluent B60: 40, collecting the eluent in sections at the flow rate of 25ml/min, collecting and merging the eluent, decompressing and concentrating to remove ethanol to obtain a concentrated solution of the RS-8a lactone hydrolysate, freeze-drying and freeze-drying, wherein the solid after freeze-drying is the RRT1.05 impurity.
EXAMPLE III
And (3) identifying the structure of the lactone hydrolysate of the daptomycin RS-8a impurity.
The daptomycin RS-8a lactone hydrolysate prepared in the second example is subjected to high-resolution primary mass spectrometry, and M/z is 845.8518[ M + K + H ]]2+,826.8774[M+2H]2+,837.8678[M+Na+H]2+The peak of the double charge excimer ion is consistent with the theoretical value, and the spectrum is shown in figure 4.
The daptomycin RS-8a lactone hydrolysate prepared in example two is subjected to high resolution secondary mass spectrometry, the spectrum is shown in figure 5, and the main mass spectrometry fragmentation fragments and fragment peak analysis are shown in figure 5 and table 1.
TABLE 1 analysis of the fragments of the RS-8a lactone hydrolysate
The lactone hydrolysate of the daptomycin RS-8a impurity prepared in example two was subjected to nuclear magnetic resonance, and the obtained nuclear magnetic resonance hydrogen spectrum and nuclear magnetic resonance carbon spectrum are shown in FIGS. 6 and 7, and the data are as follows:
1H NMR:(600MHz,DMSO-d6)δ12.28(br.s,5H),10.79(s,1H),8.39(d,J=6.6Hz,H),8.38(d,J=6.6Hz,1H), 8.33(d,J=6.6Hz,1H),8.27(d,J=6.6Hz,1H),8.19(d,J=7.2Hz,1H),8.12(br.s,1H),8.07(br.s,1H),8.02(d, J=6.6Hz,1H),7.97(d,J=8.4Hz,1H),7.96(d,J=8.4Hz,1H),7.93(d,J=6.0Hz,1H),7.85(d,J=7.8Hz,1H), 7.74(d,J=7.6Hz,1H),7.71(overlap,2H),7.66(br.s,1H),7.57(d,J=7.8Hz,1H),7.39(br.s,1H),7.31(d,J=7.8Hz, 1H),7.30(br.s,1H),7.25(d,J=7.2Hz,1H),7.19(br.s,1H),7.15(br.s,1H),7.04(t,J=7.8Hz,1H),6.97(overlap, 1H),6.96(overlap,1H),6.76(t,J=7.8Hz,1H),6.54(t,J=7.2Hz,1H),4.95(br.s,1H),4.72(m,1H),4.63(m,1H), 4.55(overlap,1H),4.55(overlap,1H),4.55(overlap,1H),4.43(overlap,1H),4.43(overlap,1H),4.43(overlap, 1H),4.33(m,1H),4.24(t,J=6.6Hz,1H),4.14(m,1H),4.02(m,1H),3.76(s,2H),3.76(s,2H),3.59(m,1H), 3.54(m,1H),3.41(overlap,1H),3.37(overlap,1H),3.09(overlap,1H),2.92(dd,J=13.8,4.2Hz,1H), 2.78(overlap,1H),2.77(overlap,2H),2.72(overlap,1H),2.68(overlap,1H),2.55(overlap,1H),2.52(overlap, 1H),2.50(overlap,1H),2.47(overlap,2H),2.39(overlap,1H),2.34(overlap,1H),2.05(t,J=7.8Hz,2H),1.98(dd, J=13.6,11.2Hz,1H)),1.72(m,1H),1.55(overlap,1H),1.55(overlap,2H),1.38(m,1H),1.26(overlap,1H), 1.25(overlap,2H),1.19(overlap,1H),1.18(t,J=7.2Hz,3H),1.12(overlap,2H),1.12(overlap,2H),1.10(overlap, 2H),1.06(d,J=6.0Hz,3H),1.01(overlap,1H),0.83(d,J=6.6Hz,3H),0.82(t,J=6.6Hz,3H),0.81(d,J=5.4Hz, 3H).
13C,NMR(600MHz,DMSO-d6)δ197.88,173.73,173.00,172.92,172.22,172.10,172.04,171.95,171.71, 171.68,171.33,171.25,170.93,170.73,170.43,170.06,169.75,168.87,168.75,151.19,136.04,134.34, 131.14,127.28,123.73,120.81,118.39,118.15,116.98,116.16,114.47,111.26,109.91,66.57,61.94,58.87, 55.42,54.94,54.12,51.59,49.86,49.71,49.63,48.48,48.03,42.23,42.21,40.09,38.54,37.81,36.97,36.48, 36.12,36.01,35.67,35.10,33.72,32.92,29.13,28.93,28.65,27.25,26.35,25.02,23.32,19.54,19.10,18.07, 14.55,11.24。
the invention determines the RRT1.05 impurity structure as shown in formula (I) through mass spectrum, hydrogen spectrum and carbon spectrum:
Claims (10)
2. the method for preparing lactone hydrolysate of claim 1, wherein the lactone hydrolysate containing daptomycin RS-8a impurity is obtained by adding NaOH solution to a solution containing daptomycin RS-8a impurity for hydrolysis.
3. The method for preparing lactone hydrolysate of claim 2, wherein the lactone hydrolysate containing daptomycin RS-8a impurity is obtained by adding 0.1mol/l NaOH solution to a solution containing daptomycin RS-8a impurity, hydrolyzing for 25-35 minutes, and then adding 0.1mol/l LHCl solution having the same volume as that of NaOH solution to neutralize.
4. The method for producing a lactone hydrolysate of claim 1, comprising the steps of:
1) crude RS-8a impurity: taking daptomycin fermentation liquor or resin 818 to resolve the combined solution, filtering, eluting the filtrate by a chromatographic column, collecting eluent, and concentrating under reduced pressure to obtain a crude product of RS-8 a;
2) purification of RS-8a impurities: dissolving the RS-8a crude product prepared in the step 1), adsorbing the dissolved product on a column, eluting the dissolved product by using eluent, collecting the eluent in sections, and combining the eluent with high purity to obtain RS-8a combined solution;
3) crude RS-8a lactone hydrolysate: adding 0.1mol/LNaOH solution into the RS-8a combined solution, hydrolyzing for 30 minutes, adding an isovolumetric 0.1mol/LHCl solution for neutralization to obtain a crude RS-8a lactone hydrolysate solution;
4) purification of RS-8a lactone hydrolysate: and (3) adsorbing the crude RS-8a lactone hydrolysate solution obtained in the step 3) by using a column, eluting by using eluent, collecting by sections, combining the eluent with higher purity, concentrating under reduced pressure to obtain a concentrated RS-8a lactone hydrolysate solution, freeze-drying and freeze-drying the concentrated RS-8a lactone hydrolysate solution, and freeze-drying the solid RS-8a lactone hydrolysate.
5. The method for preparing lactone hydrolysate of claim 4, wherein the volume ratio of the RS-8a combined solution to the 0.1mol/LNaOH solution is 1: 2.
6. The method for producing a lactone hydrolysate of claim 4, wherein the column used in step 1) is a C18 column, and the column is eluted with water in an amount of 2 column volumes; then eluting with 30% ethanol water solution, wherein the elution amount is 1.6 times of the column volume; then eluting with 50% ethanol water solution with the elution amount being 2 times of the column volume, and collecting the eluate in the stage; finally, eluting with ethanol, wherein the elution amount is 2 times of the column volume.
7. The method according to claim 4, wherein the column used in step 2) is a C18 column, eluent A is 0.1% formic acid aqueous solution, eluent B is ethanol, and the volume ratio of eluent A to eluent B is: eluent B60: and 40, combining the eluates with the purity of more than 70%.
8. The method for preparing lactone hydrolysate of claim 4, wherein the volume ratio of the RS-8a combined solution to the 0.1mol/LNaOH solution in the step 3) is 1: 2.
9. The method according to claim 4, wherein the column used in step 4) is a C18 column, eluent A is 0.1% formic acid aqueous solution, eluent B is ethanol, and the volume ratio of eluent A to eluent B is: eluent B60: 40.
10. use of a daptomycin RS-8a lactone hydrolysate of formula (I) as defined in claim 1 as an impurity control in daptomycin quality control.
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CN102675426A (en) * | 2012-04-26 | 2012-09-19 | 杭州华东医药集团生物工程研究所有限公司 | Extraction and purification method of daptomycin |
CN105699554A (en) * | 2016-03-10 | 2016-06-22 | 杭州华东医药集团新药研究院有限公司 | High-purity daptomycin lactone hydrolysate and application thereof |
CN106866789A (en) * | 2015-12-11 | 2017-06-20 | 北大方正集团有限公司 | A kind of method for isolating and purifying Daptomycin RS-8 impurity |
CN106866791A (en) * | 2015-12-11 | 2017-06-20 | 北大方正集团有限公司 | A kind of preparation method of high-purity daptomycin lactone hydrolysate |
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CN102675426A (en) * | 2012-04-26 | 2012-09-19 | 杭州华东医药集团生物工程研究所有限公司 | Extraction and purification method of daptomycin |
CN106866789A (en) * | 2015-12-11 | 2017-06-20 | 北大方正集团有限公司 | A kind of method for isolating and purifying Daptomycin RS-8 impurity |
CN106866791A (en) * | 2015-12-11 | 2017-06-20 | 北大方正集团有限公司 | A kind of preparation method of high-purity daptomycin lactone hydrolysate |
CN105699554A (en) * | 2016-03-10 | 2016-06-22 | 杭州华东医药集团新药研究院有限公司 | High-purity daptomycin lactone hydrolysate and application thereof |
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