CN114544787A - Method for detecting related substances in NADP (nicotinamide adenine dinucleotide phosphate) - Google Patents
Method for detecting related substances in NADP (nicotinamide adenine dinucleotide phosphate) Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 20
- 239000000126 substance Substances 0.000 title claims abstract description 17
- XJLXINKUBYWONI-NNYOXOHSSA-N NADP zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-NNYOXOHSSA-N 0.000 title description 45
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 30
- 239000007864 aqueous solution Substances 0.000 claims abstract description 12
- 239000011259 mixed solution Substances 0.000 claims abstract description 6
- 238000010829 isocratic elution Methods 0.000 claims abstract description 3
- XJLXINKUBYWONI-NNYOXOHSSA-O NADP(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-NNYOXOHSSA-O 0.000 claims abstract 3
- 238000001514 detection method Methods 0.000 claims description 18
- SHFJWMWCIHQNCP-UHFFFAOYSA-M hydron;tetrabutylazanium;sulfate Chemical compound OS([O-])(=O)=O.CCCC[N+](CCCC)(CCCC)CCCC SHFJWMWCIHQNCP-UHFFFAOYSA-M 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 3
- 239000000243 solution Substances 0.000 claims description 3
- 230000005526 G1 to G0 transition Effects 0.000 claims description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 2
- 239000011159 matrix material Substances 0.000 claims description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 2
- 239000000741 silica gel Substances 0.000 claims description 2
- 229910002027 silica gel Inorganic materials 0.000 claims description 2
- 239000000523 sample Substances 0.000 claims 1
- 239000012488 sample solution Substances 0.000 claims 1
- 238000000926 separation method Methods 0.000 abstract description 14
- 150000002500 ions Chemical class 0.000 abstract description 11
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 6
- 239000007857 degradation product Substances 0.000 description 26
- 239000002994 raw material Substances 0.000 description 22
- 239000012071 phase Substances 0.000 description 16
- 239000007788 liquid Substances 0.000 description 13
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 description 12
- 238000010521 absorption reaction Methods 0.000 description 12
- 229950006238 nadide Drugs 0.000 description 10
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 10
- 238000004458 analytical method Methods 0.000 description 7
- 230000014759 maintenance of location Effects 0.000 description 7
- 239000012085 test solution Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 239000012535 impurity Substances 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 239000005515 coenzyme Substances 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- PQLVXDKIJBQVDF-UHFFFAOYSA-N acetic acid;hydrate Chemical compound O.CC(O)=O PQLVXDKIJBQVDF-UHFFFAOYSA-N 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000001195 anabolic effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-M hydrogensulfate Chemical compound OS([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-M 0.000 description 1
- KTVYGDJRKCXTRA-UHFFFAOYSA-N hydron;pyrrolidin-1-amine;chloride Chemical compound Cl.NN1CCCC1 KTVYGDJRKCXTRA-UHFFFAOYSA-N 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229940101270 nicotinamide adenine dinucleotide (nad) Drugs 0.000 description 1
- 238000001612 separation test Methods 0.000 description 1
Images
Classifications
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- 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
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
-
- 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
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
-
- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
-
- 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
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
- G01N2030/324—Control of physical parameters of the fluid carrier of pressure or speed speed, flow rate
Abstract
The invention provides a method for detecting related substances in NADP, which is a high performance liquid chromatography. The method adopts a C18 reversed phase chromatographic column, and uses a mixed solution of ion pair aqueous solution and methanol as a mobile phase to carry out isocratic elution. The method realizes the effective separation of the NADP and related substances thereof by selecting the chromatographic column and the mobile phase, thereby accurately detecting the related substances.
Description
The technical field is as follows:
the invention belongs to the technical field of drug analysis, and particularly relates to a detection method of NADP-related substances.
Background art:
oxidized beta-Nicotinamide adenine dinucleotide phosphate (coenzyme II, NADP) is an extremely important nucleotide coenzyme, and the structure of the coenzyme is shown as a formula I. It is a substance formed by combining Nicotinamide Adenine Dinucleotide (NAD) and a phosphate molecule by an ester bond, and participates in various anabolic reactions, such as the synthesis of lipids, fatty acids and nucleotides, and coenzyme II is used as a carrier of hydrogen transfer and a medium of phosphate transfer in organisms to participate in various synthetic reactions.
In addition, NADP is widely used as an additive for foods, cosmetics and the like in the fields of scientific research, foods, cosmetics and the like. NADP is produced from NAD, NAD which is not completely reacted may be produced during the reaction, and NADP may be degraded into NMN, NR, NA, NAM, etc. during the reaction. At present, no literature report exists on the NADP analysis method. In order to ensure the subsequent development and production quality of NADP, the quality of NADP needs to be controlled. Therefore, it is very urgent for pharmaceutical manufacturers to research and obtain a method for detecting NADP-related substances.
TABLE 1 basic information on NADP and its raw materials, degradation products
The invention content is as follows:
the invention aims to provide a rapid, simple and efficient detection method for detecting related substances in NADP aiming at the defects of the prior art.
The technical scheme adopted by the invention is as follows: selecting a chromatographic column as a reversed phase C18 chromatographic column, taking a mixed solution of ion pair aqueous solution and methanol as a mobile phase, wherein the volume ratio of the mobile phase is the ion pair aqueous solution: 65-75% of methanol: 35-25, and performing isocratic elution at a flow rate of 0.6-1.2 mL/min.
Further, the stationary phase of the reversed phase C18 chromatographic column adopts ultra-pure spherical silica gel matrix bonded with C18 group, preferably an chromatogram C18 chromatographic column (CNW Athena C18-WP).
Furthermore, the temperature of the chromatographic column in the column incubator is 20-35 ℃, preferably 28 ℃.
Further, the mobile phase is a mixed solution of an ion pair aqueous solution and methanol.
Further, the mobile phase has a volume ratio of ion to aqueous solution: 65-75% of methanol: 35-25, preferably the volume ratio of ion to water solution: 70 parts of methanol: 30.
furthermore, the ion pair aqueous solution is potassium dihydrogen phosphate aqueous solution added with ion pair reagent tetrabutylammonium hydrogen sulfate.
Furthermore, the flow rate of the mobile phase is 0.6-1.2 mL/min, preferably 1.0 mL/min.
Furthermore, the concentration of the test solution is 0.5-2 mg/mL, preferably 1 mg/mL.
The amount of the sample is further 5-20 uL, preferably 10 uL.
Further, the detection wavelength is 260 nm.
Further, the running time is not less than 12min, preferably 16 min.
The method has the advantages that the reversed phase C18 chromatographic column and the special mobile phase are selected, the NADP and the initial main raw materials and the degradation products in the synthesis of the NADP are effectively separated, and the related substances are accurately tested.
Drawings
FIG. 1 high performance liquid chromatogram of example 1 (main partial NADP UV absorption chart)
FIG. 2 high performance liquid chromatogram of example 1 (absorption chart of NAD as a main raw material in ultraviolet)
FIG. 3 high performance liquid chromatogram of example 1 (degradation product NMN UV absorption chart)
FIG. 4 high performance liquid chromatogram of example 1 (degradation product NR UV absorption chart)
FIG. 5 high performance liquid chromatogram of example 1 (degradation product NA UV absorption graph)
FIG. 6 high performance liquid chromatogram of example 1 (degradation product NAM ultraviolet absorption chart)
FIG. 7 high performance liquid chromatogram of example 1 (method resolution chromatogram)
FIG. 8 high performance liquid chromatogram of example 1 (method sensitivity spectrum)
FIG. 9 high performance liquid chromatogram of example 1 (test sample detection spectrum)
FIG. 10 high performance liquid chromatogram of example 2
FIG. 11 high performance liquid chromatogram of example 3
FIG. 12 high performance liquid chromatogram of example 4
FIG. 13 high performance liquid chromatogram of example 5
Detailed Description
The technical content of the present invention is further described below with reference to specific examples for better understanding of the content of the present invention, but the scope of the present invention is not limited thereto.
Example 1 high performance liquid chromatography analysis of related substances in NADP
1. Selection of detection wavelength
The invention considers the ultraviolet absorption of the initial main raw material (NAD) and degradation products (NMN, NR, NAM and NA) for the synthesis of the NADP, and the specific method is as follows: taking appropriate amount of NADP and the raw materials and degradation products thereof, respectively adding water to dissolve the NADP and the raw materials and the degradation products into 1mg/mL solution, introducing 10uL of sample, using two-way pipelines, uniformly adopting flow rate according to 0.2mL/min, uniformly adopting mixed solution (30:70:0.1, V/V/V) of water/acetonitrile/trifluoroacetic acid as a mobile phase, and determining the maximum absorption wavelength of the NADP and the raw materials and the degradation products thereof by adopting the full-wavelength scanning function of a diode array detector of a high-efficiency liquid phase, wherein the ultraviolet absorption diagram is shown in attached figures 1-6, and the results are shown in Table 2.
TABLE 22 maximum absorption wavelength of Aminopyrrolidine hydrochloride and its specific impurities
| Compound abbreviation (description) | Wavelength of maximum absorption (nm) |
| NADP (Main branch) | 259 |
| NAD (raw material) | 258 |
| NMN (degradation products) | 263 |
| NR (degradation product) | 265 |
| NA (degradation product) | 261 |
| NAM (degradation product) | 260 |
With the general attached figures 1-6, all compounds have ultraviolet absorption at 260nm, so the selected detection wavelength is 260 nm.
2. Selection of other chromatographic conditions
The present invention finally employs chromatographic conditions as shown in Table 3.
TABLE 3 chromatographic conditions
The experimental procedure was as follows:
(1) separation degree test procedure of method
100mg of NADP, NAD, NR, NAM, NMN and NA are taken out of 10mg to 100mL volumetric flasks, dissolved and diluted to the scale by the diluent, and shaken up to be used as a resolution test solution. And (3) introducing a sample of the resolution test solution according to the chromatographic conditions, recording a chromatogram, and referring to an attached figure 7, wherein the NADP retention time Rt is 6.356min, the NAD retention time Rt is 9.504min, the NMN retention time Rt is 4.975min, the NR retention time Rt is 5.413min, the NA retention time Rt is 7.740min, the NAM retention time Rt is 10.691min, the resolution R between adjacent compounds is more than 1.5, and the resolution test solution completely meets the base line separation requirement (the base line separation requirement R is more than or equal to 1.5). The results are summarized in Table 4 below.
TABLE 4 degrees of separation test results
| Abbreviation of Compound | Retention time Rt, min | Degree of separation |
| NMN | 4.975 | — |
| NR | 5.413 | 1.622 |
| NADP | 6.356 | 2.715 |
| NA | 7.740 | 3.835 |
| NAD | 9.504 | 4.495 |
| NAM | 10.691 | 2.808 |
(2) Sensitivity test procedure for the method
Weighing raw materials NAD and degradation products NMN, NR, NA and NAM respectively in 10mg to 100mL volumetric flasks, dissolving and uniformly mixing the raw materials NAD and the degradation products NMN, NR, NA and NAM with water, precisely transferring the raw materials into 1mL to 100mL volumetric flasks, weighing 100mg NADP and adding the raw materials, dissolving and filtering the raw materials with water to prepare a sensitivity test solution containing 0.1% of the raw materials and the degradation products, injecting samples according to the chromatographic conditions, recording a chromatogram, and recording the signal-to-noise ratio S/N of chromatographic peaks corresponding to the raw materials and the degradation products in the chromatogram, wherein the signal-to-noise ratio S/N is more than 3, so that the signal-to-noise ratio requirement of the detection limit is met (the signal-to-noise ratio is more than or equal to 3 as the detection limit requirement). The results are summarized in Table 5 below.
TABLE 5 summary of sensitivity test results
(3) Test step for detecting test article
Weighing 100mg of NADP into a 100mL volumetric flask, dissolving in water, mixing uniformly, filtering, introducing sample according to the chromatographic conditions to detect the purity, and recording a chromatogram, wherein the detection result of the degradation product impurity NR in the chromatogram is 1.489%, the detection result of the degradation product impurity NR in the chromatogram is 0.945%, the detection result of the initial main raw material NAD residue is 0.219%, and the maximum single impurity of other unknown structures is 0.424%. The method can realize detection of related substances in NADP test sample.
Example 2 high Performance liquid chromatography analysis of related substances in NADP
In this example, the separation degree of NADP and its raw materials and degradation products in a conventional mobile phase was examined, and only the mobile phase was replaced with a mixed solution of water, acetonitrile and trifluoroacetic acid (70/30/0.1, V/V) under the same conditions and by the same method as in example 1, and the separation degree test solution was injected, so that peaks of NADP and its raw materials and degradation products were too early, and baseline separation of NADP and its raw materials and degradation products was not achieved, as shown in fig. 10.
Example 3 high Performance liquid chromatography analysis of related substances in NADP
In this example, the influence of ions added to the mobile phase on the chromatographic behavior of the reagent was examined, and the same conditions and methods as in example 1 were used, only ammonium tetrabutyl hydrogen sulfate was not added to the mobile phase, and in the sample separation degree experiment, the NADP peak profile was poor, and baseline separation of NADP and its raw materials and degradation products was not achieved, as shown in fig. 11.
Example 4 high Performance liquid chromatography analysis of related substances in NADP
This example examined the effect of the mobile phase flow rate varying beyond the range on the resolution of the process, using the same conditions and method as in example 1, only replacing the flow rate from 1.0mL/min to 1.3mL/min, feeding the resolution test solution, feeding the resolution test, was able to separate NADP and its raw materials, degradation products, but where the resolution between NR and NMN was only 1.422, failing to meet the baseline resolution requirement of 1.5, see figure 12.
Example 5 high performance liquid chromatography analysis of related substances in NADP
In this example, the influence of the variation of the volume ratio of the mobile phase components beyond the range on the separation degree of the method is examined, and the sample injection separation degree experiment can separate NADP and its raw materials and degradation products by only mixing the ion pair aqueous solution and methanol at a ratio of 70:30 into the ion pair aqueous solution and methanol at a ratio of 60:40 under the same conditions and method as in example 1, but the separation degree between NADP and NR is only 0.553, which cannot meet the requirement of baseline separation 1.5, and is shown in FIG. 13.
Claims (6)
1. A method for detecting related substances in NADP is characterized in that a chromatographic column is selected as a reversed-phase C18 chromatographic column, a mixed solution of an ion-pair aqueous solution and methanol is used as a mobile phase, and the volume ratio of the mobile phase is that of the ion-pair aqueous solution: 65-75% of methanol: 35-25, and performing isocratic elution at a flow rate of 0.6-1.2 mL/min.
2. The detection method as claimed in claim 1, wherein the stationary phase of the C18 chromatographic column is bonded with C18 group by using ultra-pure spherical silica gel matrix.
3. The detection method according to claim 1, wherein the ion-pair aqueous solution is an aqueous potassium dihydrogen phosphate solution to which an ion-pair reagent tetrabutylammonium hydrogen sulfate is added.
4. The detection method according to claim 1, wherein the concentration of the sample solution is 0.5 to 2mg/mL, and the amount of the sample is 5 to 20 uL.
5. The detection method according to claim 1, wherein the detection wavelength is 260 nm.
6. The detection method according to claim 1, wherein the operation time is 12-80 min.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105866270A (en) * | 2016-03-30 | 2016-08-17 | 中山百灵生物技术有限公司 | Method for determining content of beta-NADP disodium salt and relevant substances |
| CN108303475A (en) * | 2017-12-25 | 2018-07-20 | 新绎健康科技有限公司 | A kind of detection method of energy matter |
| CN108431015A (en) * | 2016-12-14 | 2018-08-21 | 邦泰生物工程(深圳)有限公司 | A kind of purifying process of NADPH |
| US20190064126A1 (en) * | 2017-08-31 | 2019-02-28 | Agilent Technologies, Inc. | Methods of liquid chromatography for anionic compounds |
| CN111351877A (en) * | 2020-04-01 | 2020-06-30 | 上海中科新生命生物科技有限公司 | Tissue energy metabolism substance analysis method based on UPLC-MSMS |
| CN111380978A (en) * | 2020-03-16 | 2020-07-07 | 重庆市食品药品检验检测研究院 | Method for simultaneously determining contents of coenzymes NADP and FAD in medicine |
-
2020
- 2020-11-25 CN CN202011336198.2A patent/CN114544787A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105866270A (en) * | 2016-03-30 | 2016-08-17 | 中山百灵生物技术有限公司 | Method for determining content of beta-NADP disodium salt and relevant substances |
| CN108431015A (en) * | 2016-12-14 | 2018-08-21 | 邦泰生物工程(深圳)有限公司 | A kind of purifying process of NADPH |
| US20190064126A1 (en) * | 2017-08-31 | 2019-02-28 | Agilent Technologies, Inc. | Methods of liquid chromatography for anionic compounds |
| CN108303475A (en) * | 2017-12-25 | 2018-07-20 | 新绎健康科技有限公司 | A kind of detection method of energy matter |
| CN111380978A (en) * | 2020-03-16 | 2020-07-07 | 重庆市食品药品检验检测研究院 | Method for simultaneously determining contents of coenzymes NADP and FAD in medicine |
| CN111351877A (en) * | 2020-04-01 | 2020-06-30 | 上海中科新生命生物科技有限公司 | Tissue energy metabolism substance analysis method based on UPLC-MSMS |
Non-Patent Citations (3)
| Title |
|---|
| OGASAWARA YUKI 等: "Determination of reduced nicotinamide adenine dinucleotide phosphate concentration using high-performance liquid chromatography with fluorescence detection: ratio of the reduced form as a biomarker of oxidative stress", BIOLOGICAL AND PHARMACEUTICAL BULLETIN, vol. 32, no. 11, 1 November 2009 (2009-11-01) * |
| 俞海伦 等: "高效液相色谱法同时测定大鼠心肌组织中磷酸肌酸和5种磷酸腺苷的含量", 应用技术学报, no. 01, 30 March 2017 (2017-03-30) * |
| 洪平等: "高效液相色谱法测定骨骼肌ATP、ADP、AMP、NAD+、NADH含量", 中国运动医学杂志, vol. 21, no. 1, 5 January 2002 (2002-01-05), pages 57 - 60 * |
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