CN102323340A - Action of pyrro-quinoline quinone and amino acid neurotransmitters - Google Patents
Action of pyrro-quinoline quinone and amino acid neurotransmitters Download PDFInfo
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Abstract
The invention relates to a method for researching of action of pyrro-quinoline quinone and amino acid neurotransmitters, belonging to the technical field of bioanalysis. In the invention, the reaction between pyrro-quinoline quinine (PQQ) and the five neurotransmitters such as Gly (Glycine), Dser (D-serine), Glu (Glutamate), Asp (Aspartate) and GABA (Gamma-aminobutyric Acid) is researched by adopting an HPLC (High-performance Liquid Chromatography) method, the reaction kinetics, the rate constant and the activation energy data of the PQQ and the five neurotransmitters are researched through measurement of the concentration of residual PQQ in reactions at different temperatures and different time points, and the reactive capability of the PQQ and the five neurotransmitters is investigated. Through the methodology research, the HPLC method established in the invention can be used for research of the reaction between the PQQ and the amino acid neurotransmitter, and the detection limit is 0.05mu m. According to bibliographical information, in the reaction between the PQQ and glycine through capillary electrophoresis detection, the detection limit is 0.1-0.2mu m. The magnitude of reaction activation energy determines the level of difficulty, and the higher the activation energy is, the more difficult the reaction is conducted. EGABA>EGlu>EAsp>EDser>EGly, and the increase of temperature is favorable for quick reaction.
Description
Technical field
The research method of pyrrolo-quinoline purine quinone and amino acid neurotransmitter effect belongs to the bioassay technique field.
Background technology
Pyrrolo-quinoline purine quinone, chemical name is 4,5-dihydro-4; 5-titanium dioxide-1-hydrogen pyrroles (2,3f) quinone-2,7; The 9-tricarboxylic acids has another name called methaxatin (PQQ), is a kind of oxidoreducing enzyme prothetic group; Mainly be distributed among prokaryotes and part plant and the mammal, can participate in the electron transfer process of body redox reaction and respiratory chain as the donor of electronics.PQQ has stronger radicals scavenging function, and the excitatory neuron poison that N-methyl D-asparagine acid acceptor (NMDA) is mediated has neuroprotection; The redox site effect of PQQ and NMDA reduces and causes Ca in the cell by glutamic acid
2+Increase, reduce nerve cell death, promote the generation of nerve growth factor, in brain, promote neural factor to produce and neuroprotection; In recent years research shows that PQQ has the better prevention effect to Parkinson's disease; Thereby PQQ has more important research prospect aspect neuropsychiatric disease.
The variation of central nervous system disease and amino acid neurotransmitter has bigger relation.Glutamic acid (Glu) and asparatate (Asp) are the important excitatory neurotransmitters of brain; GABA (GABA) and glycocoll (Gly) are important inhibiting nerve mediators, and these amino acid neurotransmitters and multiple the nervous system disease have confidential relation.The nervous system disease that D conformation serine (Dser) is considered to be overexcited with nmda receptor as a kind of novel neurotransmitter or down-regulated expression is relevant is relevant.PQQ can react with various neurotransmitter amino acids and generate the oxazole compounds, and reaction equation is formula as follows.Its critical function aspect nervous system maybe be relevant with its effect to neurotransmitter amino acid, therefore study PQQ and neurotransmitter amino acid reaction pair research its very important researching value is being arranged at neuroprotection aspect machine-processed.The present invention adopts the reaction of HPLC method research PQQ and Gly, Ser, Glu, Asp and 5 kinds of neurotransmitters of GABA; Through measuring the concentration of different temperatures, different time points reaction residue PQQ; The data such as reaction kinetics, rate constant and energy of activation of research PQQ and 5 kinds of neurotransmitters; Investigate the reaction capacity of PQQ and 5 kinds of neurotransmitters; PQQ provides important researching value at the mechanism of action aspect the neuroprotective to research, and on assay method, improves, and the HPLC method of employing makes PQQ reach preferably with corresponding amino acid to separate.Correlative study does not appear in the newspapers.
Summary of the invention
The object of the invention is to propose the research method of a kind of pyrrolo-quinoline purine quinone and amino acid neurotransmitter effect, and it is having very important researching value aspect neuroprotection mechanism the reaction pair research of research PQQ and neurotransmitter amino acid.
Technical scheme of the present invention: the research method of pyrrolo-quinoline purine quinone PQQ and amino acid neurotransmitter effect; Adopt the reaction of HPLC method research PQQ and Gly, Dser, Glu, Asp and 5 kinds of neurotransmitters of GABA; Through measuring the concentration of different temperatures, different time points reaction residue PQQ; Reaction kinetics, rate constant and the activation energy data of research PQQ and 5 kinds of neurotransmitters, the reaction capacity of investigation PQQ and 5 kinds of neurotransmitters;
PQQ with corresponding amino acid reaction equation is:
(1) HPLC analysis condition:
Adopt the C18 reverse-phase chromatographic column, model: Amethyst C18-P, 5 μ m, 4.6 * 250mm improves moving phase, A: acetonitrile; The B:20mM potassium dihydrogen phosphate, the 10mM TBAB contains 0.1%SDS (sodium dodecylsulphonate), uses phosphoric acid to transfer pH as between the 4.0-6.0; The v/v of A/B is 35/65, and flow velocity 0.8mL/min detects wavelength 249nm;
(2) wavelength determination
Take by weighing a certain amount of PQQ, Gly, Dser, Glu, Asp, GABA, with the dissolving of pH 7.0 phosphate buffers, being mixed with the PQQ solution concentration is 1mM respectively, each Freamine concentration is 25mM; By v/v is that 1:1 mixes, and lucifuge is hatched 24h in 37 ℃ of water-baths, carries out uv scan with the phosphate buffer of pH 7.0 as blank, and each sample uv absorption of ultraviolet spectrogram demonstration 249nm place is higher; Select 249nm as detecting wavelength;
(3) influence of pH value
Moving phase adopts different pH values (2.0-10.0) test, with PQQ and OXAZOLE degree of separation between the two pH is mapped, and pH is that each degree of separation of 4.0-6.0 is better, and PQQ reaches preferably with 5 kinds of OXAZOLE and separates;
Said 5 kinds of OXAZOLE:OX1 are PQQ-Gly, and OX2 is PQQ-Dser, and OX3 is PQQ-Glu, and OX4 is PQQ-Asp, and OX5 is PQQ-GABA;
(4) add the influence of SDS in the damping fluid to separating
In damping fluid, add 0.05%, 0.1%, 0.15% respectively, the 0.2%SDS test, when SDS content was 0.1%, each component peak type symmetry was better, PQQ and 5 kinds of OXAZOLE can both reach baseline separation;
(5) linearity, degree of accuracy and detectability are measured
Accurately take by weighing a certain amount of PQQ; Use pH 7.0 phosphate buffers to be mixed with the solution of concentration as 25mM; Be diluted to the standard solution of 0.25 ~ 2.5mM variable concentrations with same buffer, sample introduction is mapped to sample concentration with integral area respectively; Getting regression equation is A=87503294C+345390, R
2=0.9997, linear in 0.25 ~ 2.5mM scope; Under the chromatographic condition of confirming on the same day continuous sample introduction 6 times, be 2.4% (n=6) according to calculated by peak area gained relative standard deviation, the relative standard deviation of retention time is 0.39% (n=6), minimum detectable activity is 2.0ng (S/N>=3);
(6) mensuration of PQQ and 5 seed amino acids reaction residue PQQ concentration
With PQQ is the 1:1 hybrid reaction with 5 seed amino acids by v/v respectively; Lucifuge; Place 37,50 ℃ to hatch respectively, in 0.5,1,2.5,5,7.5,10, each time point abstraction reaction liquid 10 μ L of 24h, inject liquid chromatograph; HPLC analyzes, and calculates the concentration of the PQQ that dissociates according to peak area and the linear equation of PQQ;
(7) mensuration of PQQ and the 5 seed amino acid reaction rate constants and the order of reaction
Experimental data with each temperature, each time point determining is calculated reaction rate constants at different levels by following formula:
First order reaction, n=1:
TwoOrder reaction, n=2:a=b:
,
TwoOrder reaction, n=2:a ≠ b:
,
Third-order reaction; N=3:
Zero-order reaction, n=0:x=k
0T,
In the formula: a is the initial concentration of reactant A, and b is the initial concentration of reactant B, and k is a reaction rate constant, and t is the time, and x is a t product concentration constantly, and n is the order of reaction;
With the experimental data of each temperature, each time point press following formula calculating zero level, one-level,
TwoThe rate constant of level, third-order reaction confirms that with rate constant variance minimum method the order of reaction is n=1, i.e. PQQ and 5 seed amino acids reaction is first order reaction; Calculate the rate constants k that obtains at each temperature simultaneously, and calculate the mean speed constant;
(8) mensuration of reaction activity
Calculate the energy of activation of each reaction according to the Arrhenius formula:
In the formula: E
aBe reaction activity, T is an absolute temperature, and R is a Reed fort constant, R=8.314 J/molK,
Calculate reaction activity by following formula;
(9) half life determination
Half life period t
1/2Be half required time of reactant consumption, the half life period in reaction time at different levels can calculate by following formula:
First order reaction: t
1/2=ln2/k
1,
Second order reaction: a=b:t
1/2=1/ (k
2A),
Second order reaction: a ≠ b:t
1/2(A) ≠ t
1/2(B),
Third-order reaction: a=b=c:t
1/2=3/ (2k
3a
2),
Zero-order reaction: t
1/2=a/(2k
0),
According to reaction equation, calculate the half life period t under the differential responses temperature
1/2
Beneficial effect of the present invention:
1. the HPLC method of setting up can be used in the research of PQQ and neurotransmitter amino acid reaction through methodological study, detects and is limited to 0.05 μ M.The Capillary Electrophoresis of bibliographical information detects PQQ and glycocoll, detects to be limited to 0.1 –, 0.2 μ M.
2. the size of reaction activity has determined the complexity of reaction, and energy of activation is high more, reacts more difficulty and carries out.
E
GABA＞E
Glu＞E
Asp＞E
Dser＞E
Gly。What the raising temperature helped reacting carries out fast.
3. calculate the rate constant of zero level, one-level, secondary, third-order reaction, software confirms that with rate constant variance minimum method the order of reaction is n=1, and promptly this reacts and is first order reaction.In reaction system, amino acid whose molal quantity is far longer than the molal quantity of PQQ so that the reaction before and after amino acid whose concentration can be similar to be regarded as constant.Therefore, in the case, the apparent order of reaction is that 1 prompting actual response comes down to second order reaction, that is reaction is a bimolecular elementary reaction, and reaction in other words is the amino acid of PQQ chelating a part of a part.
4. along with temperature of reaction raises, rate constant increases, and shows that improving temperature of reaction helps strengthening reaction rate.Visible by table 2, K
Gly＞K
Dser＞K
Asp＞K
Glu＞K
GABA
5. from reaction rate, reaction activity, the reaction of all visible PQQ of chromatogram and 5 kinds of neurotransmitters, wherein reacting the fastest with PQQ is Gly, 24h all is converted into product.PQQ and Dser also are easier to reaction, and Dser is as the endogenic ligand of nmda receptor, for nmda receptor be overexcited or a spirit type disease that function suppresses to cause provides new treatment target spot.This result provides extremely valuable foundation for the applied research of PQQ in spirit type disease.Reacting the slowest is GABA, and PQQ and its react hardly.
Description of drawings
The ultraviolet scanning spectrum figure of Fig. 1 PQQ-Gly and PQQ contrast.A、PQQ-Gly，O、PQQ。
The ultraviolet scanning spectrum figure of Fig. 2 PQQ-Dser and PQQ contrast.B、PQQ-Dser，O、PQQ。
The ultraviolet scanning spectrum figure of Fig. 3 PQQ-Glu and PQQ contrast.C、PQQ-Glu，O、PQQ。
The ultraviolet scanning spectrum figure of Fig. 4 PQQ-Asp and PQQ contrast.D、PQQ-Asp，O、PQQ。
The ultraviolet scanning spectrum figure of Fig. 5 PQQ-GABA and PQQ contrast.E、PQQ-GABA，O、PQQ。
Fig. 6 pH is to the influence of degree of separation.
The HPLC chromatogram of 37 ℃ of reactions of Fig. 7 PQQ and Gly 24h.PQQ-Gly(OX1)。
The HPLC chromatogram of 37 ℃ of reactions of Fig. 8 PQQ and Dser 24h.PQQ-Dser(OX2)。
The HPLC chromatogram of 37 ℃ of reactions of Fig. 9 PQQ and Glu 24h.PQQ-Glu(OX3)。
The HPLC chromatogram of 37 ℃ of reactions of Figure 10 PQQ and Asp 24h.PQQ-Asp(OX4)。
The HPLC chromatogram of 37 ℃ of reactions of Figure 11 PQQ and GABA 24h.PQQ-GABA(OX5)。
Embodiment
One, HPLC analysis condition:
Employing C18 reverse-phase chromatographic column (model: Amethyst C18-P, 5 μ m, 4.6 * 250mm), moving phase is improved A: acetonitrile; The B:20mM potassium dihydrogen phosphate, the 10mM TBAB contains 0.1%SDS, uses phosphoric acid to transfer pH as between the 4.0-6.0; A/B is 35/65 (v/v), and flow velocity 0.8mL/min detects wavelength 249nm;
Two, wavelength determination
Take by weighing a certain amount of PQQ, Gly, Dser, Glu, Asp and GABA, with the dissolving of pH 7.0 phosphate buffers, being mixed with PQQ concentration is 1mM respectively, amino acid concentration is 25mM; Press 1:1 and mix, lucifuge is hatched 24h in 37 ℃ of water-baths, carries out uv scan with the phosphate buffer of pH 7.0 as blank, and ultraviolet spectrogram is seen Fig. 1-Fig. 5, and each sample uv absorption of 249nm place is higher; Select 249nm as detecting wavelength.
The influence of embodiment 2 pH values
Because PQQ and amino acid reaction have generated new material, analyze the changes of contents of PQQ and must select suitable separation condition, make PQQ reach effectively with the product oxazole and separate; We adopt ion-pairing agent, and in moving phase, add an amount of modifying agent SDS, make peak type symmetry better, and moving phase pH value is very big to the retention time influence, must select suitable pH value to make PQQ and each separating substances; Select
PH 2.0-10.0 makes an experiment, and investigates the separation case of PQQ and product, and resolution representes separating effect with degree of separation; (degree of separation computing formula: R=2 (2-tR1)/(W1+W2); TR is a retention time, and W is a half-peak breadth), and with degree of separation pH is mapped.PH is that the 4.0-6.0 degree of separation is higher.PQQ reaches preferably with each amino acid reactants and separates; PQQ reference substance retention time is 8.55min, and OX1 (PQQ-Gly) retention time is 9.47min, and OX2 (PQQ-Dser) retention time is 9.37 min; OX3 (PQQ-Glu) retention time is 7.29 min; OX4 (PQQ-Asp) retention time is 7.37 min, and OX5 (PQQ-GABA) retention time is 11.12 min, and the HPLC chromatogram is seen Fig. 7-Figure 11.
Add the influence of SDS in embodiment 3 damping fluids to separating
In damping fluid, add 0.05%, 0.1%, 0.15% respectively, the 0.2%SDS test, when SDS content was 0.1%, PQQ and 5 kinds of OXAZOLE can both reach baseline separation.
Embodiment 4
Linearity, degree of accuracy and detectability are measured
Accurately take by weighing a certain amount of PQQ; Use pH 7.0 phosphate buffers to be mixed with the solution of concentration as 25mM; Be diluted to the standard solution of 0.25 ~ 2.5mM variable concentrations with same buffer, sample introduction is mapped to sample concentration with integral area respectively; Getting regression equation is A=87503294C+345390, R
2=0.9997, linear in 0.25 ~ 2.5mM scope.Under the chromatographic condition of confirming on the same day continuous sample introduction 6 times, be about 2.4% (n=6) according to calculated by peak area gained relative standard deviation, the relative standard deviation of retention time is about 0.39% (n=6).Minimum detectable activity is 2.0ng (S/N >=3).
The mensuration of PQQ and 5 seed amino acids reaction residue PQQ concentration
PQQ is pressed the 1:1 hybrid reaction with 5 seed amino acids respectively; Lucifuge; Place 37,50 ℃ to hatch respectively, in 0.5,1,2.5,5,7.5,10, each time point abstraction reaction liquid 10 μ L of 24h, inject liquid chromatograph; HPLC analyzes, and calculates the concentration of the PQQ that dissociates according to peak area and the linear equation of PQQ.
The concentration of each reaction time point PQQ of table 1 and the free PQQ of amino acid reaction
PQQ respectively with the mensuration of the 5 seed amino acid reaction rate constants and the order of reaction
Experimental data with each temperature, each time point determining is calculated reaction rate constants at different levels by following formula.
First order reaction (n=1):
TwoOrder reaction (n=2): a=b,
TwoOrder reaction (n=2): a ≠ b,
Third-order reaction (n=3):
Zero-order reaction (n=0): x=k
0t
In the formula: a is the initial concentration of reactant A, and b is the initial concentration of reactant B, and k is a reaction rate constant, and t is the time, and x is a t product concentration constantly, and n is the order of reaction.
With the experimental data of each temperature, each time point press following formula calculating zero level, one-level,
TwoThe rate constant of level, third-order reaction, software confirms that with rate constant variance minimum method the order of reaction is n=1, promptly this reacts and is first order reaction.
Calculate the rate constants k that obtains at each temperature simultaneously by last, and calculate the mean speed constant.
Mean speed constant, half life period, the energy of activation of table 2 PQQ and the reaction of 5 seed amino acids
The mensuration of embodiment 7 reaction activities
Calculate the energy of activation of each reaction according to the Arrhenius formula:
ln(k
2?/?k
1)?=?(E
a/R)(1/T
1?–?1/T
2)（2）
In the formula: E
aBe reaction activity, T is an absolute temperature, and R is Reed fort constant (R=8.314 J/molK).
Calculate reaction activity by formula (2), the result sees table 2.
Half life period t
1/2Be half required time of reactant consumption, the half life period in reaction time at different levels can calculate by following formula:
First order reaction: t
1/2=
,
Second order reaction: a=b:t
1/2=
,
Second order reaction: a ≠ b:t
1/2(A) ≠ t
1/2(B),
Third-order reaction: a=b=c:t
1/2=
,
Zero-order reaction: t
1/2=
,
According to following formula, calculate the half life period t under the differential responses temperature
1/2, the result sees table 2.
Claims (1)
1. the research method of pyrrolo-quinoline purine quinone PQQ and amino acid neurotransmitter effect; It is characterized in that adopting the reaction of HPLC method research PQQ and Gly, Dser, Glu, Asp and 5 kinds of neurotransmitters of GABA; Through measuring the concentration of different temperatures, different time points reaction residue PQQ; Reaction kinetics, rate constant and the activation energy data of research PQQ and 5 kinds of neurotransmitters, the reaction capacity of investigation PQQ and 5 kinds of neurotransmitters;
(1) HPLC analysis condition
Adopt the C18 reverse-phase chromatographic column, model: Amethyst C18-P, 5 μ m, 4.6 * 250mm improves moving phase, A: acetonitrile; B:20mM potassium dihydrogen phosphate, 10mM TBAB, contain 0.1% sodium dodecylsulphonate SDS, use phosphoric acid to transfer pH to be 4.0-6.0; The v/v of A/B is 35/65, and flow velocity 0.8mL/min detects wavelength 249nm;
(2) wavelength determination
Take by weighing a certain amount of PQQ, Gly, Dser, Glu, Asp, GABA, with the dissolving of pH 7.0 phosphate buffers, being mixed with the PQQ solution concentration is 1mM respectively, and each Freamine concentration is 25mM; PQQ solution is that 1:1 mixes by v/v respectively with each Freamine, and lucifuge is hatched 24h in 37 ℃ of water-baths, carries out uv scan with the phosphate buffer of pH 7.0 as blank, and ultraviolet spectrogram shows that each sample uv absorption of 249nm place is higher; Select 249nm as detecting wavelength;
(3) influence of pH value
Selection is mapped to pH with degree of separation, and pH is that the 4.0-6.0 degree of separation is higher, and PQQ reaches preferably with 5 kinds of OXAZOLE reactants and separates;
Said 5 kinds of OXAZOLE:OX1 are PQQ-Gly, and OX2 is PQQ-Dser, and OX3 is PQQ-Glu, and OX4 is PQQ-Asp, and OX5 is PQQ-GABA;
(4) add the influence of SDS in the damping fluid to separating
In damping fluid, add 0.05%, 0.1%, 0.15% respectively, the 0.2%SDS test, when SDS content was 0.1%, PQQ and 5 kinds of OXAZOLE can both reach baseline separation;
(5) linearity, degree of accuracy and detectability are measured
Accurately take by weighing a certain amount of PQQ; Use pH 7.0 phosphate buffers to be mixed with the solution of concentration as 25mM; Be diluted to the standard solution of 0.25 ~ 2.5mM variable concentrations with same buffer, sample introduction is mapped to sample concentration with integral area respectively; Getting regression equation is A=87503294C+345390, R
2=0.9997, linear in 0.25 ~ 2.5mM scope; Under the chromatographic condition of confirming on the same day continuous sample introduction 6 times, be 2.4% according to calculated by peak area gained relative standard deviation, the relative standard deviation of retention time is 0.39%, minimum detectable activity is 2.0ng;
(6) mensuration of PQQ and 5 seed amino acids reaction residue PQQ concentration
With PQQ solution is the 1:1 hybrid reaction with 5 seed amino acid solution by v/v respectively; Lucifuge; Place 37,50 ℃ to hatch respectively, in 0.5,1,2.5,5,7.5,10, each time point abstraction reaction liquid 10 μ L of 24h, inject liquid chromatograph; HPLC analyzes, and calculates the concentration of the PQQ that dissociates according to peak area and the linear equation of PQQ;
(7) mensuration of PQQ and the 5 seed amino acid reaction rate constants and the order of reaction
Experimental data with each temperature, each time point determining is calculated reaction rate constants at different levels by following formula:
First order reaction; N=1:
TwoOrder reaction, n=2:a=b:
,
TwoOrder reaction, n=2:a ≠ b:
,
Third-order reaction; N=3:
Zero-order reaction, n=0:x=k
0T,
In the formula: a is the initial concentration of reactant A, and b is the initial concentration of reactant B, and k is a reaction rate constant, and t is the time, and x is a t product concentration constantly, and n is the order of reaction;
With the experimental data of each temperature, each time point press following formula calculating zero level, one-level,
TwoThe rate constant of level, third-order reaction confirms that with rate constant variance minimum method the order of reaction is n=1, i.e. PQQ and 5 seed amino acids reaction is first order reaction; Calculate the rate constants k that obtains at each temperature simultaneously, and calculate the mean speed constant;
(8) mensuration of reaction activity
Calculate the energy of activation of each reaction according to the Arrhenius formula:
In the formula: E
aBe reaction activity, T is an absolute temperature, and R is a Reed fort constant, R=8.314 J/molK,
Calculate reaction activity by following formula;
(9) half life determination
Half life period t
1/2Be half required time of reactant consumption, the half life period in reaction time at different levels can calculate by following formula:
First order reaction: t
1/2=
,
Second order reaction: a=b:t
1/2=
,
Second order reaction: a ≠ b:t
1/2(A) ≠ t
1/2(B),
Third-order reaction: a=b=c:t
1/2=
,
Zero-order reaction: t
1/2=
,
According to formula, calculate the half life period t under the differential responses temperature
1/2
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103163200A (en) * | 2013-03-25 | 2013-06-19 | 江苏省原子医学研究所 | Method for measuring content of pyrroloquinoline quinone dimethyl ester by micellar electrokinetic chromatography |
| CN106841452A (en) * | 2017-02-28 | 2017-06-13 | 江苏省原子医学研究所 | A kind of method that HPLC combinations UPLC MS detections PQQ is acted on lactic acid |
| CN109085269A (en) * | 2018-08-15 | 2018-12-25 | 江苏省原子医学研究所 | The method of PQQ and Lys and Arg reflex action research |
| CN112461975A (en) * | 2020-09-11 | 2021-03-09 | 福建省产品质量检验研究院 | Method for detecting coenzyme content in feed additive |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3739334B1 (en) * | 2018-01-12 | 2022-07-27 | Mitsubishi Gas Chemical Company, Inc. | A method for quantitative analysis of pyrroloquinoline quinone or a salt thereof |
-
2011
- 2011-06-09 CN CN 201110152463 patent/CN102323340B/en not_active Expired - Fee Related
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| ZDENĚK GLATZ等: "Determination of pyrroloquinoline quinone by capillary zone electrophoresis", 《JOURNAL OF CHROMATOGRAPHY B》 * |
| ZHOUPENG ZHANG等: "High performance liquid chromatographyic separation and PH-dependent electrochemical properties of pyrroloquinoline quinone and three closely related isomeric analogues", 《BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS》 * |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103163200A (en) * | 2013-03-25 | 2013-06-19 | 江苏省原子医学研究所 | Method for measuring content of pyrroloquinoline quinone dimethyl ester by micellar electrokinetic chromatography |
| CN106841452A (en) * | 2017-02-28 | 2017-06-13 | 江苏省原子医学研究所 | A kind of method that HPLC combinations UPLC MS detections PQQ is acted on lactic acid |
| CN106841452B (en) * | 2017-02-28 | 2019-02-22 | 江苏省原子医学研究所 | A kind of method that HPLC combination UPLC-MS detection PQQ acts on lactic acid |
| CN109085269A (en) * | 2018-08-15 | 2018-12-25 | 江苏省原子医学研究所 | The method of PQQ and Lys and Arg reflex action research |
| CN109085269B (en) * | 2018-08-15 | 2021-03-09 | 江苏省原子医学研究所 | Method for studying reaction behavior of PQQ with Lys and Arg |
| CN112461975A (en) * | 2020-09-11 | 2021-03-09 | 福建省产品质量检验研究院 | Method for detecting coenzyme content in feed additive |
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|---|---|
| CN102323340B (en) | 2013-02-27 |
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