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Efficient liquid phase chromatographic analysis of compound bone peptide

A high-performance liquid chromatography, compound osteopeptide technology, applied in the field of drug analysis, can solve the problems of short retention time, difficult baseline separation, no ideal method for compound osteopeptide analysis, etc., and achieves simple operation, convenient use, and extended retention time. Effect

Inactive Publication Date: 2009-12-02
NANJING NORMAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because polypeptides, protein peptides, and amino acids have no UV absorption, it is often difficult to separate the baseline and have a short retention time using commonly used high-performance liquid chromatography analysis methods. Therefore, there is still no ideal method for the analysis of compound osteopeptides.

Method used

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  • Efficient liquid phase chromatographic analysis of compound bone peptide
  • Efficient liquid phase chromatographic analysis of compound bone peptide
  • Efficient liquid phase chromatographic analysis of compound bone peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 ion-pair reagent---the chromatographic investigation of benzyltrimethylammonium chloride

[0034] Chromatographic conditions: mobile phase: pH=4.5 acetic acid-sodium acetate buffer solution (0.015mol / L) containing benzyltrimethylammonium chloride 5mmol / L; flow rate 1ml / min; UV detection wavelength: 240nm; column temperature: room temperature .

[0035] Samples: scorpion extract, bone peptide injection (provided by Nanjing Jianlida Technology Development Co., Ltd., batch number: 050603).

[0036] The reverse phase column is a C8 column.

[0037] Take an appropriate amount of scorpion extract and bone peptide injection, dilute with mobile phase, and inject 20 μl into the chromatograph respectively. The obtained chromatograms can be found in figure 1 , figure 2 .

[0038] Conclusion: Osteopeptide and scorpion were clearly distinguished under chromatographic conditions. After repeated repetitions, the retention time T of the characteristic peak of osteopep...

Embodiment 2

[0039] Comparison of several chromatographic columns of embodiment 2

[0040] Chromatographic conditions: Lichrospher C8, Lichrospher C18, and LichrospherPhenyl chromatographic columns are used respectively, and the specifications are all 4.6mmx300mm, 5μm; mobile phase: acetic acid-sodium acetate buffer solution (0.015mol / L) with pH=4.5 containing benzyltrimethyl Ammonium chloride 5mmol / L; flow rate 1ml / min; UV detection wavelength: 240nm; column temperature: room temperature.

[0041] Sample: Scorpion extract (batch number: 050603) and bone peptide injection

[0042] After diluting the two samples by a certain number of times with the mobile phase, 20 μl each was injected into three different chromatographic systems, and the obtained chromatograms are shown in the attached figure 1 , figure 2 , image 3 , Figure 4 , Figure 5 , Figure 6 . Analyze the chromatograms, see Table 1.

[0043] Table 1

[0044]

[0045] Conclusion: All kinds of reversed-phase columns c...

Embodiment 3

[0046] Embodiment 3 investigates the mobile phase of different proportions of organic solvents

[0047] Chromatographic conditions: mobile phase: C% (C = 10, 5 or 0) methanol acetic acid-sodium acetate buffer solution (pH = 4.5) containing benzyltrimethylammonium chloride 5mmol / L; flow rate 1ml / min; chromatographic column : LichrospherPhenyl column (4.6mmx300mm, 5μm), ultraviolet detection wavelength: 262nm; column temperature: room temperature.

[0048] Sample: Scorpion extract (batch number: 050603) and bone peptide injection

[0049] After the samples were diluted with the mobile phase, 20 μl was injected into the chromatograph, and the difference between the two samples in the resulting chromatogram was compared. The analysis results are shown in Table 2.

[0050] Table 2

[0051]

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Abstract

The high-efficient liquid phase chromatogram method for compound bone peptide comprises: selecting a chromatographic column and a phase-inversion column; setting condition as room temperature, 1ml / min flow rate, and 220-310nm UV wavelength; adding pH stable system as acetic acid-acetate or phosphoric acid-phosphate buffer solution system; selecting the ion couple agent as the cation group and anion group; detecting sample curve of liquid-phase chromatogram; determining the compound content by the feature peak to classify the bone peptide and compound peptide. This invention can obtain the corresponding fingerprint graph for efficient classification.

Description

technical field [0001] The invention relates to a drug analysis method, in particular to a method for analyzing raw materials and preparations of bone peptide, compound bone peptide and scorpion extract by using reversed-phase ion-pair high-performance liquid chromatography. Background technique [0002] The main components of osseopeptides are water-soluble substances extracted from limb bones of healthy animals, such as: polypeptides, protein peptides, amino acids, organic calcium, phosphorus, inorganic calcium, inorganic salts, trace elements, etc. Scorpion extract is a water-soluble substance extracted from scorpion, and its main component type is similar to bone peptide. Compound bone peptide is a compound preparation obtained by adding a certain amount of scorpion extract to bone peptide. Common bone peptide injections (formerly known as bone peptide injections) and compound bone peptide injections are now on the market. Osteopeptide injection is a sterilized aqueous...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02B01J20/281
Inventor 任勇王晓洁马学琴朱贯臣吉爱宁
Owner NANJING NORMAL UNIVERSITY
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