CN118130646A - Prostaglandin intermediate detection method - Google Patents
Prostaglandin intermediate detection method Download PDFInfo
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- CN118130646A CN118130646A CN202410146831.3A CN202410146831A CN118130646A CN 118130646 A CN118130646 A CN 118130646A CN 202410146831 A CN202410146831 A CN 202410146831A CN 118130646 A CN118130646 A CN 118130646A
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- 150000003180 prostaglandins Chemical class 0.000 title claims abstract description 46
- 238000001514 detection method Methods 0.000 title abstract description 60
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical group OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 51
- 239000000543 intermediate Substances 0.000 claims abstract description 38
- 239000000243 solution Substances 0.000 claims abstract description 28
- 150000002596 lactones Chemical class 0.000 claims abstract description 23
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 16
- 150000007524 organic acids Chemical class 0.000 claims abstract description 12
- 239000000741 silica gel Substances 0.000 claims abstract description 10
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 10
- 239000007864 aqueous solution Substances 0.000 claims abstract description 9
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 7
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical group [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 claims abstract description 3
- 238000000034 method Methods 0.000 claims description 46
- 238000010828 elution Methods 0.000 claims description 27
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 16
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 12
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 12
- 239000011259 mixed solution Substances 0.000 claims description 9
- 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
- 238000001914 filtration Methods 0.000 claims description 6
- 235000019253 formic acid Nutrition 0.000 claims description 6
- 239000002245 particle Substances 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 5
- 239000007924 injection Substances 0.000 claims description 5
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical group CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 5
- FPLYNRPOIZEADP-UHFFFAOYSA-N octylsilane Chemical group CCCCCCCC[SiH3] FPLYNRPOIZEADP-UHFFFAOYSA-N 0.000 claims description 5
- 239000000945 filler Substances 0.000 claims description 2
- 238000000926 separation method Methods 0.000 abstract description 10
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 239000000523 sample Substances 0.000 description 31
- 239000012488 sample solution Substances 0.000 description 18
- 238000002360 preparation method Methods 0.000 description 14
- 239000012085 test solution Substances 0.000 description 11
- 238000007865 diluting Methods 0.000 description 9
- 238000005303 weighing Methods 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 8
- 239000012491 analyte Substances 0.000 description 7
- 238000011895 specific detection Methods 0.000 description 7
- -1 Prostanoid compounds Chemical class 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 5
- 238000000889 atomisation Methods 0.000 description 5
- 150000001875 compounds Chemical class 0.000 description 5
- 230000014759 maintenance of location Effects 0.000 description 5
- 238000000691 measurement method Methods 0.000 description 5
- 229940094443 oxytocics prostaglandins Drugs 0.000 description 5
- 238000003908 quality control method Methods 0.000 description 5
- 238000010998 test method Methods 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 4
- 239000012490 blank solution Substances 0.000 description 4
- 238000004587 chromatography analysis Methods 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 108010014431 prostalin Proteins 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000000377 silicon dioxide Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 2
- UMMADZJLZAPZAW-OVXHCKHTSA-N carboprost tromethamine Chemical compound OCC([NH3+])(CO)CO.CCCCC[C@](C)(O)\C=C\[C@H]1[C@@H](O)C[C@H](O)[C@@H]1C\C=C/CCCC([O-])=O UMMADZJLZAPZAW-OVXHCKHTSA-N 0.000 description 2
- 229960005296 carboprost tromethamine Drugs 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- GGXICVAJURFBLW-CEYXHVGTSA-N latanoprost Chemical compound CC(C)OC(=O)CCC\C=C/C[C@H]1[C@@H](O)C[C@@H](O)[C@@H]1CC[C@@H](O)CCC1=CC=CC=C1 GGXICVAJURFBLW-CEYXHVGTSA-N 0.000 description 2
- 229960001160 latanoprost Drugs 0.000 description 2
- WGFOBBZOWHGYQH-MXHNKVEKSA-N lubiprostone Chemical compound O1[C@](C(F)(F)CCCC)(O)CC[C@@H]2[C@@H](CCCCCCC(O)=O)C(=O)C[C@H]21 WGFOBBZOWHGYQH-MXHNKVEKSA-N 0.000 description 2
- 229960000345 lubiprostone Drugs 0.000 description 2
- QQCOAAFKJZXJFP-GANRVJIKSA-N methyl (Z)-7-[(1S,2S,3S,5R)-3,5-dihydroxy-2-[(E,3R)-3-hydroxy-3-methyloct-1-enyl]cyclopentyl]hept-5-enoate Chemical group CCCCC[C@@](C)(O)\C=C\[C@@H]1[C@@H](O)C[C@@H](O)[C@H]1C\C=C/CCCC(=O)OC QQCOAAFKJZXJFP-GANRVJIKSA-N 0.000 description 2
- OJLOPKGSLYJEMD-URPKTTJQSA-N methyl 7-[(1r,2r,3r)-3-hydroxy-2-[(1e)-4-hydroxy-4-methyloct-1-en-1-yl]-5-oxocyclopentyl]heptanoate Chemical compound CCCCC(C)(O)C\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1CCCCCCC(=O)OC OJLOPKGSLYJEMD-URPKTTJQSA-N 0.000 description 2
- 229960005249 misoprostol Drugs 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 229940127293 prostanoid Drugs 0.000 description 2
- 238000013441 quality evaluation Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 229960003400 sulprostone Drugs 0.000 description 2
- UQZVCDCIMBLVNR-TWYODKAFSA-N sulprostone Chemical compound O[C@@H]1CC(=O)[C@H](C\C=C/CCCC(=O)NS(=O)(=O)C)[C@H]1\C=C\[C@@H](O)COC1=CC=CC=C1 UQZVCDCIMBLVNR-TWYODKAFSA-N 0.000 description 2
- 229960004458 tafluprost Drugs 0.000 description 2
- WSNODXPBBALQOF-VEJSHDCNSA-N tafluprost Chemical compound CC(C)OC(=O)CCC\C=C/C[C@H]1[C@@H](O)C[C@@H](O)[C@@H]1\C=C\C(F)(F)COC1=CC=CC=C1 WSNODXPBBALQOF-VEJSHDCNSA-N 0.000 description 2
- MKPLKVHSHYCHOC-AHTXBMBWSA-N travoprost Chemical compound CC(C)OC(=O)CCC\C=C/C[C@H]1[C@@H](O)C[C@@H](O)[C@@H]1\C=C\[C@@H](O)COC1=CC=CC(C(F)(F)F)=C1 MKPLKVHSHYCHOC-AHTXBMBWSA-N 0.000 description 2
- 229960002368 travoprost Drugs 0.000 description 2
- TVHAZVBUYQMHBC-SNHXEXRGSA-N unoprostone Chemical compound CCCCCCCC(=O)CC[C@H]1[C@H](O)C[C@H](O)[C@@H]1C\C=C/CCCC(O)=O TVHAZVBUYQMHBC-SNHXEXRGSA-N 0.000 description 2
- 229960004317 unoprostone Drugs 0.000 description 2
- CORRAXPZMWTYIP-TWEVDUBQSA-N (3ar,4s,5r,6as)-4-(hydroxymethyl)-5-(oxan-2-yloxy)-3,3a,4,5,6,6a-hexahydrocyclopenta[b]furan-2-one Chemical compound O([C@H]1[C@@H]([C@H]2CC(=O)O[C@H]2C1)CO)C1CCCCO1 CORRAXPZMWTYIP-TWEVDUBQSA-N 0.000 description 1
- GMVPRGQOIOIIMI-UHFFFAOYSA-N (8R,11R,12R,13E,15S)-11,15-Dihydroxy-9-oxo-13-prostenoic acid Natural products CCCCCC(O)C=CC1C(O)CC(=O)C1CCCCCCC(O)=O GMVPRGQOIOIIMI-UHFFFAOYSA-N 0.000 description 1
- YRCZWPOLIAQXCV-NFBCFJMWSA-N 1-chloro-7-[(1S,2S)-2-octylcyclopentyl]heptan-1-ol Chemical compound ClC(CCCCCC[C@H]1CCC[C@@H]1CCCCCCCC)O YRCZWPOLIAQXCV-NFBCFJMWSA-N 0.000 description 1
- AXXJUGQWKHTJBS-OALUTQOASA-N ClC(=CCCCCC[C@H]1CCC[C@@H]1CCCCCCCC)O Chemical compound ClC(=CCCCCC[C@H]1CCC[C@@H]1CCCCCCCC)O AXXJUGQWKHTJBS-OALUTQOASA-N 0.000 description 1
- 229960000711 alprostadil Drugs 0.000 description 1
- 229940053200 antiepileptics fatty acid derivative Drugs 0.000 description 1
- 229940114079 arachidonic acid Drugs 0.000 description 1
- 235000021342 arachidonic acid Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 229960002986 dinoprostone Drugs 0.000 description 1
- XEYBRNLFEZDVAW-ARSRFYASSA-N dinoprostone Chemical compound CCCCC[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1C\C=C/CCCC(O)=O XEYBRNLFEZDVAW-ARSRFYASSA-N 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- OJZYRQPMEIEQFC-UAWLTFRCSA-N limaprost Chemical compound CCCC[C@H](C)C[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1CCCC\C=C\C(O)=O OJZYRQPMEIEQFC-UAWLTFRCSA-N 0.000 description 1
- 229950009365 limaprost Drugs 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 238000003541 multi-stage reaction Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- GMVPRGQOIOIIMI-DWKJAMRDSA-N prostaglandin E1 Chemical compound CCCCC[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1CCCCCCC(O)=O GMVPRGQOIOIIMI-DWKJAMRDSA-N 0.000 description 1
- XEYBRNLFEZDVAW-UHFFFAOYSA-N prostaglandin E2 Natural products CCCCCC(O)C=CC1C(O)CC(=O)C1CC=CCCCC(O)=O XEYBRNLFEZDVAW-UHFFFAOYSA-N 0.000 description 1
- 150000003814 prostanoids Chemical class 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000006257 total synthesis reaction Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
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- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Library & Information Science (AREA)
- Engineering & Computer Science (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a detection method of prostaglandin intermediate, which comprises the following steps: detecting prostaglandin intermediates in the test object by high performance liquid chromatography; the prostaglandin intermediate is selected from the group consisting of a coriolis lactone; wherein, in the high performance liquid chromatography, the detector is an electric fog detector; the chromatographic column is a silicane bonded silica gel chromatographic column; the mobile phase A is water or an aqueous solution containing organic acid; the mobile phase B is methanol or a methanol solution containing organic acid; the volume ratio of the mobile phase A to the mobile phase B is 100 (0.01-100). The detection method provided by the invention has the characteristics of high sensitivity, good separation degree, strong specificity, good repeatability, simplicity, rapidness and the like.
Description
Technical Field
The invention belongs to the field of analysis and detection, and particularly relates to a detection method of a prostaglandin intermediate.
Background
Prostaglandins (prostaglandin, PG) are a class of fatty acid derivatives produced by the arachidonic acid metabolic pathway, mainly comprising PGA, PGB, PGC, PGD, PGE, PGF and other series of compounds, which exist in various tissues in the body and have various physiological activities (Korbecki J et al.2014, acta biochem Pol). Prostanoid compounds have very high drug-properties, and up to now, more than 20 prostanoid drugs are marketed in bulk worldwide (Peng H, et al 2017,Org Biomol Chem).
The first report in 1969 by Corey et al was the realization of the chemical total synthesis of prostaglandins by means of a Kelly lactone as key intermediate (Corey EJ,1969,J Am Chem Soc). Although a number of process routes have been developed for prostaglandin synthesis, the current industrial synthesis of PG class of drugs still relies heavily on the class of Kerilides, from which multi-step reactions are still required to obtain prostaglandins (Zhang F, et al 2021,Nat Chem;Cunningham L,et al,2022,Org Lett).
The quality control of the coriolis lactone as a key intermediate for synthesizing prostaglandin raw material is closely related to the quality of the final product and related preparations, so that it is necessary to establish a simple and efficient detection and analysis method for quality control of the prostaglandin synthesis key intermediate, coriolis lactone. Since the ultraviolet absorption of the coriolis lactone is very low, the conventional ultraviolet detector is difficult to detect and analyze, and no related analysis method is reported at present.
Disclosure of Invention
In order to overcome at least one of the problems of the prior art, it is an object of the present invention to provide a method for detecting prostaglandin intermediates.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
In a first aspect, the present invention provides a method for detecting a prostaglandin intermediate, comprising the steps of:
Detecting prostaglandin intermediates in the test object by high performance liquid chromatography;
the prostaglandin intermediate is selected from the group consisting of a coriolis lactone;
Wherein, in the high performance liquid chromatography,
The detector is an electric fog detector;
the chromatographic column is a silicane bonded silica gel chromatographic column;
the mobile phase A is water or an aqueous solution containing organic acid; the mobile phase B is methanol or a methanol solution containing organic acid; the volume ratio of the mobile phase A to the mobile phase B is 100 (0.01-100).
The invention adopts an electric fog type detector to detect prostaglandin intermediate, the analyte is atomized and dried into particles, the particles of the analyte collide with charged nitrogen and are charged, the charged particles of the analyte enter a collector, the charge quantity of the particles of the analyte is measured by an electrometer, and the generated signal current is in direct proportion to the content of the analyte. The detection principle is not dependent on the chromophores (while ultraviolet detectors need to detect them), nor does it need to form gas-phase ions (mass spectrometric detectors need to form gas-phase ions for detection). Thus, the present invention allows for the selection of an electrospray detector as a detector-established assay for quality control of the coriolis lactones and related materials.
Wherein, the coriolis lactone is also named: the chemical name of the colestolide is: (3 aR,4S,5R,6 aS) -4- (hydroxymethyl) -5- ((tetrahydro-2H-pyran-2-yl) oxy) hexahydro-2H-cyclopenta [ b ] furan-2-one having the structural formula:
preferably, the organic acid is at least one selected from formic acid, acetic acid and trifluoroacetic acid.
Preferably, the volume ratio of the organic acid to the water in the mobile phase A is (0.01-0.5): 100; and/or, in the mobile phase B, the volume ratio of the organic acid to the methanol is (0.01-0.5): 100.
Preferably, the chromatographic column is an octyl silane bonded silica gel chromatographic column or an octadecyl silane bonded silica gel chromatographic column.
Preferably, the chromatographic column has a length of 150-250 mm, an inner diameter of 2-5 mm, and a filler particle size of 3-5 μm.
Preferably, the column temperature of the chromatographic column is 25-40 ℃.
Preferably, the temperature of an atomization chamber of the electric fog type detector is 35-50 ℃, the filtering constant is 0.1-5, and the data acquisition frequency is 2-50 Hz.
Preferably, the flow rate of the mobile phase A and/or the mobile phase B is 0.5-1.5 mL/min.
Preferably, the sample injection concentration of the object to be detected is 0.01-100 mg/mL; the sample injection volume is 5-20 mu L.
Preferably, the analyte is formulated into an analyte solution and then detected by liquid chromatography.
Preferably, the solvent in the solution of the object to be detected is at least one selected from water, methanol and acetonitrile.
Preferably, in the high performance liquid chromatography, the elution method of the chromatographic column is gradient elution; the gradient elution is as follows: firstly, the volume ratio is (9-19): eluting the mixed solution of the mobile phase A and the mobile phase B in the (0-1) for 1-55 min; then the volume ratio is (0-1): eluting the mixed solution of the mobile phase A and the mobile phase B in the steps of (9-19) for 0.1-10 min; and then the volume ratio is (9-19): eluting the mixed solution of the mobile phase A and the mobile phase B in the (0-1) for 3-8 min; further preferably, in the high performance liquid chromatography, the elution method of the chromatographic column is gradient elution; the gradient elution is as follows: firstly, the volume ratio is (9-19): 1 and a mobile phase A and a mobile phase B for eluting for 1-55 min; then the volume ratio is 1: eluting the mixed solution of the mobile phase A and the mobile phase B in the steps of (9-19) for 0.1-10 min; and then the volume ratio is (9-19): 1 and the mixed solution of the mobile phase A and the mobile phase B is eluted for 3 to 8 minutes.
Preferably, the prostaglandins include PGF 2α series compounds and derivatives thereof, PGE series compounds and derivatives thereof.
Preferably, the PGF 2α series of compounds and derivatives thereof are selected from the group consisting of bemeprostil, latanoprost, travoprost, tafluprost, carboprost tromethamine, carboprost methyl, chloroprostaenol, lubiprostone, unoprostone, fluprostol, phenol prostalin, and prostalin.
Preferably, the PGE series compound and its derivatives are selected from the group consisting of dienoprost, alprostadil, misoprostol, rimalaprost, rioprost, enprost, enoprost, gemfibroprost, sulprostone.
The method of the present invention can be used for detecting intermediate products of the aforementioned prostaglandins and derivatives thereof.
The beneficial effects of the invention are as follows: the detection method provided by the invention can be used for efficiently detecting the coriolis lactone, has the characteristics of high sensitivity, good separation degree, strong specificity, good repeatability, simpleness, rapidness and the like, and can be suitable for quality control and evaluation of prostaglandin bulk drugs and preparations thereof. Specifically, the invention adopts an electric fog type detector to effectively analyze and detect the compound, has the advantages of strong specificity, high sensitivity, good repeatability, rapidness and simplicity, and can be suitable for intermediate quality control and quality evaluation of final products and preparations of prostaglandin raw material medicine synthesis processes of PGF 2α series compounds and derivatives thereof (such as bemeprostil, latanoprost, travoprost, tafluprost, carboprost tromethamine, carboprost methyl, chlorprostanol, lubiprostone, unoprostone, fluprostol, phenolic prostalin and prostalin), PGE series compounds and derivatives thereof (such as dinoprostone, prostadil, misoprostol, limaprost, rioprost, enoprost, gemfibroprost and sulprostone).
The detection method of the invention can be also suitable for detecting the related substances of the coriolis lactone, and has the advantages of high sensitivity, good separation degree, strong specificity, good repeatability and the like.
Drawings
FIG. 1 is a chromatographic test chart of a blank solution in the present invention.
FIG. 2 is a chromatographic test chart of the sample solution in example 5.
Detailed Description
Specific embodiments of the present invention will be described in further detail below with reference to the drawings and examples, but the practice and protection of the present invention are not limited thereto. It should be noted that the following processes, if not specifically described in detail, can be realized or understood by those skilled in the art with reference to the prior art. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
Example 1:
The embodiment provides a prostaglandin intermediate detection method, which comprises the following specific detection steps and detection conditions:
Preparation of test solution: and (3) taking a proper amount of a sample to be measured, precisely weighing, adding a water-methanol (volume ratio is 50:50) solution for dissolving and diluting to prepare a solution containing about 1mg of the sample to be measured in 1 mL.
The measuring method comprises the following steps: precisely measuring 10 mu L of the sample solution, and injecting into a liquid chromatograph.
Chromatographic conditions:
The chromatographic column is filled with octyl silane bonded silica gel (4.6 mm. Times.150 mm,5 μm); the flow rate is 0.8mL/min; mobile phase a was an aqueous solution containing 0.1% formic acid, mobile phase B was a methanol solution containing 0.1% formic acid, and gradient elution was performed according to the elution procedure in table 1 below; column temperature is 40 ℃; the detector is an electric fog detector (the temperature of an atomization chamber is 50 ℃, the filtering constant is 5, and the data acquisition frequency is 10 Hz); the sample volume was 10. Mu.L.
Table 1 elution procedure for chromatographic column
The detection results of the coriolis lactones in the test samples according to the above test methods are shown in table 2 below:
TABLE 2 detection results of Kerilactone in samples to be measured
| Chromatographic peak | Retention time/min | Peak area/pA min | Peak height/pA | Degree of separation |
| Kerilactone peak A | 9.72 | 12.56 | 113.53 | 1.40 |
| Kerilactone peak B | 10.05 | 11.72 | 113.14 | 2.97 |
As can be seen from Table 2, there are two detection peaks, peak A and peak B, respectively, in the detection method in this example, the detection of the Coriolis lactone in the sample to be detected can be performed rapidly and efficiently, and the effective separation can be performed.
Example 2:
The embodiment provides a prostaglandin intermediate detection method, which comprises the following specific detection steps and detection conditions:
Preparation of test solution: and (3) taking a proper amount of a sample to be measured, precisely weighing, adding methanol for dissolving and diluting to prepare a solution containing about 10mg of the sample to be measured in 1 mL.
The measuring method comprises the following steps: precisely measuring 10 mu L of the sample solution, and injecting into a liquid chromatograph.
Chromatographic conditions:
The chromatographic column is a chromatographic column (4.6 mm multiplied by 250mm,5 μm) filled with octyl silane bonded silica gel; the flow rate is 1mL/min; mobile phase a was an aqueous solution containing 0.2% acetic acid, mobile phase B was a methanol solution containing 0.2% acetic acid, and gradient elution was performed according to the elution procedure in table 3 below; the column temperature is 25 ℃; the detector is an electric fog detector (the temperature of an atomization chamber is 35 ℃, the filtering constant is 3.6, and the data acquisition frequency is 10 Hz); the sample volume was 10. Mu.L.
TABLE 3 elution procedure for chromatography columns
The results of the detection of the coriolis lactones in the test samples according to the above test methods are shown in table 4 below:
TABLE 4 detection results of Kerilactones in samples to be tested
| Chromatographic peak | Retention time/min | Peak area/pA min | Peak height/pA | Degree of separation |
| Kerilactone peak A | 21.33 | 86.34 | 890.76 | 0.85 |
| Kerilactone peak B | 21.56 | 86.13 | 889.37 | 1.57 |
As can be seen from Table 4, the detection method in this example can rapidly and efficiently detect the Coriolis lactone in the sample to be detected, and can perform effective separation.
Example 3:
The embodiment provides a prostaglandin intermediate detection method, which comprises the following specific detection steps and detection conditions:
Preparation of test solution: and (3) taking a proper amount of a sample to be measured, precisely weighing, adding a water-methanol (volume ratio is 50:50) solution for dissolving and diluting to prepare a solution containing about 1mg of the sample to be measured in 1 mL.
The measuring method comprises the following steps: precisely measuring 10 mu L of the sample solution, and injecting into a liquid chromatograph.
Chromatographic conditions:
the chromatographic column is filled with octadecylsilane chemically bonded silica (4.6mm×150mm,5 μm); the flow rate is 0.7mL/min; mobile phase a was an aqueous solution containing 0.2% trifluoroacetic acid, mobile phase B was a methanol solution containing 0.2% trifluoroacetic acid, and gradient elution was performed according to the elution procedure in table 5 below; column temperature is 40 ℃; the detector is an electric fog detector (the temperature of an atomization chamber is 35 ℃, the filtering constant is 3.6, and the data acquisition frequency is 10 Hz); the sample volume was 10. Mu.L.
TABLE 5 elution procedure for chromatography columns
The results of the detection of the coriolis lactones in the test samples according to the above test methods are shown in table 6 below:
TABLE 6 detection results of Kerilactones in samples to be tested
| Chromatographic peak | Retention time/min | Peak area/pA min | Peak height/pA | Degree of separation |
| Kerilactone peak A | 14.87 | 9.39 | 89.19 | 1.04 |
| Kerilactone peak B | 15.02 | 9.14 | 89.62 | 2.53 |
As can be seen from Table 6, the detection method in this example can rapidly and efficiently detect the Coriolis lactone in the sample to be detected, and can perform effective separation.
Example 4:
The embodiment provides a prostaglandin intermediate detection method, which comprises the following specific detection steps and detection conditions:
Preparation of test solution: and (3) taking a proper amount of a sample to be measured, precisely weighing, adding a water-methanol (volume ratio is 50:50) solution for dissolving and diluting to prepare a solution containing about 10mg of the sample to be measured in 1 mL.
The measuring method comprises the following steps: precisely measuring 10 mu L of the sample solution, and injecting into a liquid chromatograph.
Chromatographic conditions:
The chromatographic column is a chromatographic column (4.6 mm×250mm,5 μm) filled with octadecylsilane chemically bonded silica; the flow rate is 1mL/min; mobile phase a was an aqueous solution containing 0.1% trifluoroacetic acid, and mobile phase B was a methanol solution containing 0.1% trifluoroacetic acid, and gradient elution was performed according to the elution procedure in table 7 below; the column temperature is 30 ℃; the detector is an electric fog detector (the temperature of an atomization chamber is 50 ℃, the filtering constant is 3.6, and the data acquisition frequency is 10 Hz); the sample volume was 10. Mu.L.
TABLE 7 elution procedure for chromatography columns
The results of the detection of the coriolis lactones in the test samples according to the above test methods are shown in table 8 below:
TABLE 8 detection results of Kerilactones in samples to be tested
| Chromatographic peak | Retention time/min | Peak area/pA min | Peak height/pA | Degree of separation |
| Kerilactone peak A | 13.68 | 104.18 | 765.08 | 0.95 |
| Kerilactone peak B | 13.53 | 105.33 | 788.35 | 1.74 |
As can be seen from Table 8, the detection method in this example can rapidly and efficiently detect the Coriolis lactone in the sample to be detected, and can effectively separate the same.
Example 5:
This example provides a method for detecting prostaglandin intermediate, wherein chromatographic conditions and test sample solutions are prepared in the same manner as in example 1, and the difference between this example and example 1 is that: the measurement method is different, and in this example, the measurement method is as follows: precisely measuring 10 μl of each of the blank solution (i.e. mixed solution of water and methanol with volume ratio of 50:50) and the sample solution, injecting into a liquid chromatograph, and recording the chromatogram, wherein the chromatogram of the blank solution is shown in figure 1, and the chromatogram of the sample solution is shown in figure 2. As can be seen by comparing FIGS. 1 to 2, the blank solution does not interfere with the detection of the chromatographic peaks A and B of the Coriolis lactone, and further shows that the determination method has good specificity.
Example 6:
the present example provides a method for detecting a prostaglandin intermediate, and the chromatographic conditions in the detection of this example are the same as those in example 1, and the difference between this example and example 1 is that:
Preparation of test solution: and (3) taking a proper amount of a sample to be measured, precisely weighing, adding a water-methanol (volume ratio is 50:50) solution for dissolving and diluting to prepare a solution containing about 0.01mg of the sample to be measured in 1 mL.
The measuring method comprises the following steps: precisely measuring 10 mu L of the sample solution, and injecting into a liquid chromatograph. The results measured according to the above test methods are shown in table 9 below.
TABLE 9 detection results of Kerilactones in samples to be tested
| Chromatographic peak | Retention time/min | Peak area/pA min | Peak height/pA | Signal to noise ratio |
| Kerilactone peak A | 9.52 | 0.1361 | 14.63 | 3.83 |
| Kerilactone peak B | 9.85 | 0.1285 | 14.32 | 3.61 |
As is clear from Table 9, when the concentration of the sample solution was 0.01mg/mL, the signal to noise ratio of each of the Kerilactone peaks A and B was more than 3, further indicating that the detection limit of the analysis method of the present invention was 0.01mg/mL.
Example 7:
The present example provides a method for detecting a prostaglandin intermediate, wherein the chromatographic conditions and the measurement method are the same as those in example 1, and the difference between the present example and example 1 is that:
6 parts of the test solutions were prepared in parallel according to the preparation method of the test solution in example 1, and then the 6 parts of the test solutions were tested for the coriolis lactones, and specific test results are shown in the following table 10.
TABLE 10 detection results of Kerilactones in samples to be tested
As is clear from Table 10, in the sample solution, the peak area RSD of the Kerilactone peak A was 2.91% and the peak area RSD of the Kerilactone peak B was 2.85%, and it was further revealed that the measurement method of the present invention has good reproducibility.
Example 8:
The present example provides a method for detecting prostaglandin intermediate, wherein the chromatographic conditions, the preparation method of the sample solution and the measurement method are the same as those of example 1, and the difference between this example and example 1 is that: the sample solutions were allowed to stand for 0 hours, 24 hours, and 48 hours, respectively, and then were subjected to sample injection detection, and specific detection results are shown in table 11 below.
TABLE 11 detection results of Kerilactones in samples to be tested
As is clear from Table 11, after the sample solutions were left for 0 hours, 24 hours and 48 hours, the peak areas RSD of the Kerilactone peaks A and B in the sample solutions were 2.94% and 2.63%, respectively, further showing that the sample solutions were stable well within 48 hours at room temperature.
Comparative example 1
The embodiment provides a prostaglandin intermediate detection method, which comprises the following specific detection steps and detection conditions:
preparation of test solution: and (3) taking a proper amount of a sample to be measured, precisely weighing, adding a water-methanol (volume ratio is 50:50) solution for dissolving and diluting to prepare a solution containing about 10mg of the sample to be measured in 1 mL.
The measuring method comprises the following steps: precisely measuring 10 mu L of the sample solution, and injecting into a liquid chromatograph.
Chromatographic conditions:
The chromatographic column is filled with octyl silane bonded silica gel (4.6 mm. Times.150 mm,5 μm); the flow rate is 0.8mL/min; mobile phase a was an aqueous solution containing 0.1% formic acid and mobile phase B was a methanol solution containing 0.1% formic acid, followed by gradient elution according to the elution procedure in table 12 below; column temperature is 40 ℃; the detector is an ultraviolet detector, and the detection wavelength is 210nm; the sample volume was 10. Mu.L.
Table 12 elution procedure for chromatographic column
The results obtained by the above-described detection method are shown in Table 13.
TABLE 13 detection results of Kerilactones in samples to be tested
As is clear from Table 13, the detection method in this example did not detect the chromatographic peak of the Kerill, indicating that the UV absorption of the Kerill was very small and that it was difficult to detect the analysis by a UV detector.
Comparative example 2
The present example provides a method for detecting a prostaglandin intermediate, which differs from comparative example 1 in that: the detection wavelength of the ultraviolet detector is 254nm; preparation of test solution: and (3) taking a proper amount of a sample to be measured, precisely weighing, adding a water-methanol (volume ratio is 50:50) solution for dissolving and diluting to prepare a solution containing about 1mg of the sample to be measured in 1 mL. The results obtained by the detection according to the detection method in this example are shown in Table 14.
TABLE 14 detection results of Kerilactones in samples to be tested
As is clear from Table 14, the detection method in this example did not detect the chromatographic peak of the Kerill, indicating that the UV absorption of the Kerill was very small and that it was difficult to detect the analysis by a UV detector.
Comparative example 3
The embodiment provides a prostaglandin intermediate detection method, which comprises the following specific detection steps and detection conditions:
Preparation of test article: and (3) taking a proper amount of a sample to be measured, precisely weighing, adding a water-methanol (volume ratio is 50:50) solution for dissolving and diluting to prepare a solution containing about 10mg of the sample to be measured in 1 mL.
The measuring method comprises the following steps: precisely measuring 10 mu L of the sample solution, and injecting into a liquid chromatograph.
The chromatographic column is a chromatographic column (4.6 mm×250mm,5 μm) filled with octadecylsilane chemically bonded silica; the flow rate is 1mL/min; mobile phase a was an aqueous solution containing 0.1% trifluoroacetic acid and mobile phase B was a methanol solution containing 0.1% trifluoroacetic acid, followed by gradient elution according to the elution procedure shown in table 15 below; the column temperature is 30 ℃; the detector is an ultraviolet detector, and the detection wavelength is 210nm; the sample volume was 10. Mu.L.
TABLE 15 elution procedure for chromatography columns
TABLE 16 detection results of Coriolis lactone in samples to be tested
As can be seen from Table 16, the detection method in this example did not detect the chromatographic peak of the Kerill, indicating that the UV absorption of the Kerill was very small and that it was difficult to detect the analysis by a UV detector.
Comparative example 4
This example provides a method for detecting a prostaglandin intermediate, which differs from comparative example 3 in that: the detection wavelength of the ultraviolet detector is 254nm. Preparation of test solution: and (3) taking a proper amount of a sample to be measured, precisely weighing, adding a water-methanol (volume ratio is 50:50) solution for dissolving and diluting to prepare a solution containing about 1mg of the sample to be measured in 1 mL. The results of measurement according to the detection method in this example are shown in Table 17 below.
TABLE 17 detection results of Coriolis lactone in samples to be tested
As is clear from Table 17, the detection method in this example did not detect the chromatographic peak of the Kerill, indicating that the UV absorption of the Kerill was very small and that it was difficult to detect the analysis by a UV detector.
While the embodiments of the present invention have been described in detail, the present invention is not limited to the above embodiments, and various changes may be made without departing from the spirit of the present invention within the knowledge of those skilled in the art. Furthermore, embodiments of the invention and features of the embodiments may be combined with each other without conflict.
Claims (10)
1. A method for detecting a prostaglandin intermediate, characterized by: the method comprises the following steps:
Detecting prostaglandin intermediates in the test object by high performance liquid chromatography;
the prostaglandin intermediate is selected from the group consisting of a coriolis lactone;
Wherein, in the high performance liquid chromatography,
The detector is an electric fog detector;
the chromatographic column is a silicane bonded silica gel chromatographic column;
The mobile phase A is water or an aqueous solution containing organic acid; the mobile phase B is methanol or a methanol solution containing organic acid;
the volume ratio of the mobile phase A to the mobile phase B is 100 (0.01-100).
2. The method for detecting a prostaglandin intermediate as defined in claim 1, wherein: the organic acid is at least one selected from formic acid, acetic acid and trifluoroacetic acid.
3. The method for detecting a prostaglandin intermediate as defined in claim 1 or 2, wherein: the volume ratio of the organic acid to the water in the mobile phase A is (0.01-0.5): 100; and/or, in the mobile phase B, the volume ratio of the organic acid to the methanol is (0.01-0.5): 100.
4. The method for detecting a prostaglandin intermediate as defined in claim 1, wherein: the chromatographic column is octyl silane bonded silica gel chromatographic column or octadecyl silane bonded silica gel chromatographic column.
5. The method for detecting a prostaglandin intermediate as defined in claim 1 or 4, wherein: the length of the chromatographic column is 150-250 mm, the inner diameter is 2-5 mm, and the particle size of the filling agent is 3-5 mu m.
6. The method for detecting a prostaglandin intermediate as defined in claim 1 or 4, wherein: the column temperature of the chromatographic column is 25-40 ℃.
7. The method for detecting a prostaglandin intermediate as defined in claim 1, wherein: the temperature of an atomizing chamber of the electric fog type detector is 35-50 ℃, the filtering constant is 0.1-5, and the data acquisition frequency is 2-50 Hz.
8. The method for detecting a prostaglandin intermediate as defined in claim 1, wherein: the flow rate of the mobile phase A and/or the mobile phase B is 0.5-1.5 mL/min.
9. The method for detecting a prostaglandin intermediate as defined in claim 1, wherein: the sample injection concentration of the object to be detected is 0.01-100 mg/mL; the sample injection volume is 5-20 mu L.
10. The method for detecting a prostaglandin intermediate as defined in claim 1, wherein: in the high performance liquid chromatography, the elution method of the chromatographic column is gradient elution; the gradient elution is as follows: firstly, the volume ratio is (9-19): eluting the mixed solution of the mobile phase A and the mobile phase B in the (0-1) for 1-55 min; then the volume ratio is (0-1): eluting the mixed solution of the mobile phase A and the mobile phase B in the steps of (9-19) for 0.1-10 min; and then the volume ratio is (9-19): eluting the mixed solution of the mobile phase A and the mobile phase B in the (0-1) for 3-8 min.
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