Novel method for high-sensitivity and wide detection range fluorescence detection of alendronate sodium (ALDS)

A technology of sodium alendronate and sodium alendronate, which is applied in the field of fluorescence detection of sodium alendronate, can solve the problems of large measurement error, complicated operation, and poor sensitivity, and achieve high selectivity detection and low background Signal, practical effect

Inactive Publication Date: 2018-02-02
CHONGQING MEDICAL UNIVERSITY
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Since the molecular structure of alendronate sodium contains two phosphonic acid groups, it has strong polarity, is easily ionized, and has no chromophoric group, so the content of the drug cannot be directly determined by conventional ultraviolet detectors or fluorescence detectors.
The current national standard (WS1-(X-312)-2004Z[18]) adopts the molybdenum blue colorimetric method with large measurement error for the content of alendronate sodium tablets, which is not conducive to the quality control of drugs
Li Yun and others reported Cu 2+ Complex reversed-phase ion-pair chromatography-ultraviolet detection method to determine the content of alendronate sodium tablets, but this method needs to combine alendronate sodium with Cu 2+ After vigorously shaking for 40 minutes, a good complex is formed, and the sensitivity is poor, and the detection linear range is narrow, so it is not suitable for the detection of biological samples
Another literature report uses high performance liquid chromatography to detect the content of alendronate sodium in pharmaceutical preparations or biological samples. Since alendronate sodium does not have a chromogenic group, it is necessary to use o-phthalaldehyde, diazomethane , Chloroformic acid fluorene and other derivatives are derivatized to generate ultraviolet absorbing groups. This method needs to go through a tedious solid phase extraction process, and the operation is complicated. At the same time, diazomethane is explosive and toxic, and there are potential safety hazards.

Method used

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  • Novel method for high-sensitivity and wide detection range fluorescence detection of alendronate sodium (ALDS)
  • Novel method for high-sensitivity and wide detection range fluorescence detection of alendronate sodium (ALDS)
  • Novel method for high-sensitivity and wide detection range fluorescence detection of alendronate sodium (ALDS)

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Experimental program
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Effect test

Embodiment 1

[0040] Example 1 Synthesis and spectroscopic determination of DNA-AgNCs

[0041] Preparation of DNA stock solution

[0042] The lyophilized DNA powder was centrifuged at 6000rpm for 10min, and 31μL of PB buffer (pH 6.6) was slowly added to prepare a 100μmol / L stock solution, which was stored in a refrigerator at 4°C for later use.

[0043] Synthesis of DNA-AgNCs

[0044] Add AgNO to the prepared DNA solution at a molar ratio of 1:6 3 solution (18.6μmol / L), vortexed in 20mmol / L PB buffer (pH 6.6) for 5min, incubated in the dark for 20min, and then quickly added AgNO 3 Equimolar concentration of NaBH 4 The solution (18.6mol / L) was stored in a refrigerator at 4°C for future use.

[0045] The synthesized DNA-AgNCs were imaged by high-resolution transmission electron microscope, transmission electron microscope model: Tecnai G2F20S-TWIN; measurement conditions: accelerating voltage 300kV. see results figure 1 , figure 1 Middle a and b are high-resolution transmission electro...

Embodiment 2

[0048] The preparation and detection method of embodiment 2 solution

[0049] Preparation of Alendronate Sodium Standard Stock Solution

[0050] Accurately weigh 0.0162g of alendronate sodium in a brown vial, dissolve and dilute to 5mL with ultrapure water, and prepare 10 - 2 mol / L standard stock solution, stored in a refrigerator at 4°C. Before use, dilute to concentrations of: 10 -3 , 5×10 -4 、10 -4 , 5×10 -5 、10 -5 , 5×10 -6 、10 -6 、10 -7 、10 -8 mol / L series of standard solutions.

[0051] Preparation of alendronate sodium test solution

[0052] Take 20 tablets of Alendronate Sodium Tablets from 3 batches respectively, accurately weigh them, grind them finely, and accurately weigh an appropriate amount of fine powder (approximately equivalent to 1 mg of Alendronic Acid), put them in an 8mL measuring bottle, add an appropriate amount of water and ultrasonically Dissolve alendronate sodium, dilute to the mark with water, shake well, centrifuge at 3000rpm for 3min...

Embodiment 3

[0058] Example 3 Investigation of Influencing Factors

[0059] The inventors studied Cu 2+ The influence of concentration, pH of the system and reagent addition sequence on the detection results is as follows.

[0060] Cu 2+ The influence of concentration on detection results: a certain concentration of Cu 2+ The solution was added to the mixed solution of PB buffer (pH 7.4) and DNA-AgNCs (310nmol / L) and vortexed to mix well, and stood at room temperature for 15min, and finally made up to 400μL with ultrapure water, and stood at room temperature for 20min. Measure the fluorescence parameters according to Example 2. Cu 2+ When the concentration is 0-1200nmol / L, the fluorescence intensity of DNA-AgNCs in the system is as follows: image 3 shown. Depend on image 3 It can be seen that when Cu 2+ After adding, the fluorescence of DNA-AgNCs was rapidly quenched, with Cu 2+ As the concentration increases, the fluorescence intensity gradually decreases until it is completely...

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Abstract

The invention provides a novel method for high-sensitivity and wide detection range fluorescence detection of alendronate sodium (ALDS). A background signal is lowered by the aid of quenching effect of copper ions on silver nanoclusters in oligonucleotide-fluorescent silver nanoclusters, the ALDS in samples is combined with the copper ions to form a compound, the copper ions are enabled to be awayfrom the oligonucleotide-fluorescent silver nanoclusters to have fluorescence in the siliver nanoclusters to be restored, and content of the ALDS is calculated by monitoring the change of fluorescence intensity of the DNA-AgNCs compound probe by the concentration of the ALDS and drawing a standard curve of fluorescence recovery degree and content of the ALDS. The method is applicable to detectionof content of the ALDS in the samples (pharmaceutic preparations or biological samples such as urine samples and serum samples), and has the advantages of wide linear range, high flexibility, less interference, time saving, green and safety and the like, and the method is suitable for large-scale promotion.

Description

technical field [0001] The invention relates to the fluorescence detection of alendronate sodium, in particular to a new method for fluorescence detection of alendronate sodium with high sensitivity and wide detection range. Background technique [0002] Alendronate sodium (Alendronate sodium, ALDS), chemically named (4-amino-1-hydroxybutylene) diphosphonic acid monosodium salt trihydrate, is the third generation of aminobisphosphonate bone resorption inhibitors . Alendronate sodium has a strong affinity with intraosseous hydroxyapatite, can enter bone matrix hydroxyapatite crystals to inhibit osteoclast activity, and indirectly inhibit bone resorption through osteoblasts. It is mainly used clinically to treat bone osteoporosis and osteitis deformans. Since the molecular structure of alendronate sodium contains two phosphonic acid groups, it has strong polarity, is easily ionized, and has no chromophoric group, so the content of the drug cannot be directly determined by a ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432
Inventor 张普贾春燕尚京川
Owner CHONGQING MEDICAL UNIVERSITY
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