CN104749259A - Method for determining olanzapine tablets and relative substance content by high performance liquid chromatography with gradient elution - Google Patents

Abstract

The invention discloses a method for determining olanzapine tablets and relative substance content by high performance liquid chromatography with gradient elution, which employs the following operation condition: a)an anti-phase C8 chromatographic column is employed as a chromatographic column; b)flow velocity is 1.5ml/min, detection wavelength is 220nm, column temperature is 35 DEG C, temperature of a sample to be measured is 5 DEG C; c)a sodium dodecyl sulfate buffer is prepared by the following steps: 1500ml water is taken, 5.0ml phosphoric acid is added, 50% of sodium hydroxide solution is used for adjusting pH value to 2.5, 13.0g sodium dodecyl sulfate is added and vibrated for dissolving and filtering; d)a mobile phase A is prepared by acetonitrile and the sodium dodecyl sulfate buffer according to volume ratio being 48: 52; and e)a mobile phase B is prepared by acetonitrile and the sodium dodecyl sulfate buffer according to volume ratio being 70: 30. The method has the advantages of good specialization and high accuracy, a main peak and impurity can be effectively separated, and relative substance of olanzapine tablets can be accurately detected.

Description

A kind of high effective liquid chromatography for measuring Olanzapine Tablets of gradient elution and the method for related substances content

Technical field

The present invention relates to a kind of Pharmaceutical Analysis field, the high performance liquid chromatography method for separating and analyzing being specifically related to a kind of gradient elution measures the method for Olanzapine Tablets and related substances content.

Background technology

This product Main Ingredients and Appearance is Olanzapine.Its chemical name is: 2-methyl isophthalic acid 0-(4-methyl isophthalic acid-piperazine)-4H-thieno [2,3-b] [1,5] benzodiazepine.General by name: Zyprexa, molecular formula: C 17 H 20 N 4 S molecular weight: 312.43; This product is Film coated tablets, aobvious faint yellow or yellow after removing dressing.

This product belongs to antipsychotic class medicine.

Related substances separation is the important indicator of Drug's control.The method of inspection setting up Accurate Determining impurity is even more important, and will carry out strict control to related substance, just be unable to do without strong, the highly sensitive analytical approach of specificity, and to the screening of analytical approach and checking.American Pharmacopeia 35 has recorded this kind, but still has problems when checked for impurities content, and chromatographic peak profile is poor, and magazins' layout degree is bad.

Summary of the invention

The object of the invention is the deficiency existed for prior art, provide a kind of high effective liquid chromatography for measuring Olanzapine of gradient elution and the method for related substances content.

The object of the invention is to be realized by following technological means:

Adopt following operating conditions:

A. chromatographic column adopts anti-phase C8 chromatographic column;

B. flow velocity is 1.5ml/min, and determined wavelength is 220nm, and column temperature is 35 ° of C, and testing sample temperature is 5 ° of C;

C. sodium lauryl sulfate buffer is prepared through following methods, water intaking 1500ml, and add phosphoric acid 5.0ml, by 50% sodium hydroxide solution adjust ph to 2.5, add lauryl sodium sulfate 13.0g, jolting makes dissolving, filters;

D. mobile phase A is to prepare at 48: 52 by the volume ratio of acetonitrile and sodium lauryl sulfate buffer;

E. Mobile phase B is to prepare at 70: 30 by the volume ratio of acetonitrile and sodium lauryl sulfate buffer.

Compared with prior art the present invention has following obvious advantage: specificity is good, accuracy is high, and main peak can effectively be separated with impurity, can detect Olanzapine Tablets related substance exactly.

Accompanying drawing explanation

Fig. 1 is that acid destroys chromatogram;

Fig. 2 is that alkali destroys chromatogram;

Fig. 3 is Oxidative demage chromatogram;

Fig. 4 is high temperature chromatogram;

Fig. 5 is that illumination destroys chromatogram;

Fig. 6 is detection limit chromatogram;

Fig. 7 is system suitability contrast colors spectrograms;

Fig. 8 is sample detection chromatogram;

Fig. 9 is Unite States Standard (USS) chromatogram.

Embodiment

The high effective liquid chromatography for measuring injection Olanzapine of gradient elution and a method for related substances content, adopt following operating conditions:

A. chromatographic column adopts anti-phase C8 chromatographic column;

B. flow velocity is 1.5ml/min, and determined wavelength is 220nm, and column temperature is 35 ° of C, and testing sample temperature is 5 ° of C;

C. sodium lauryl sulfate buffer is prepared through following methods, water intaking 1500ml, and add phosphoric acid 5.0ml, by 50% sodium hydroxide solution adjust ph to 2.5, add lauryl sodium sulfate 13.0g, jolting makes dissolving, filters;

D. mobile phase A is to prepare at 48: 52 by the volume ratio of acetonitrile and sodium lauryl sulfate buffer;

E. Mobile phase B is to prepare at 70: 30 by the volume ratio of acetonitrile and sodium lauryl sulfate buffer.

One, applicability contrast test

Solvent is to prepare at 60: 40 by the volume ratio of acetonitrile and sodium lauryl sulfate buffer, get respectively Olanzapine reference substance, impurity I, II, III, IV, V, VI, VII in right amount each, put in same measuring bottle, solubilizer is appropriate, ultrasonicly make dissolving and to make in every 1ml about containing the solution of Olanzapine 20 μ g, each 2 μ g of other impurity, as system suitability solution with solvent dilution.Precision measures 20 μ l injection liquid chromatographies, and record chromatogram, the degree of separation of Olanzapine and impurity IV (N-oxide) should be not less than 3.0, and the tailing factor of Olanzapine main peak should be not more than 1.5.

Two, sample determination

Get Olanzapine reference substance appropriate, accurately weighed, also dilute the solution made containing 2.5 μ g in every 1ml by dissolution with solvents, product solution in contrast; Get this product several pieces, porphyrize, gets fine powder appropriate, accurately weighed, add sample solvent appropriate, ultrasonic and jolting frequently 10 ~ 20 minutes, makes Olanzapine dissolve, lets cool, the solution about containing Olanzapine 0.5mg in every 1ml is made with solvent dilution, centrifugal, get supernatant as need testing solution.Precision measures each 20 μ l of above-mentioned two kinds of solution injection liquid chromatography respectively, record chromatogram.As aobvious impurity peaks in need testing solution chromatogram, measure each impurity peak area, its content is rolled over by the peak area of the Self-control method reference substance of the correction up factor, and the correction factor of impurity II is 0.4, and the correction factor of other impurity calculates by 1.0.Impurity I must not cross 0.1%, and impurity II must not cross 0.2%, and impurity III must not cross 0.15%, and impurity IV, V, VI all must not cross 0.5%, and impurity VII must not cross 0.1%, and single unknown impuritie must not cross 0.2%, and total impurities must not cross 1.5%.Should solvent peak be deducted during calculating, disregard the peak that gradient drift produces simultaneously.

Three, specificity test

Acid destroys: get this product 2, porphyrize, get fine powder 0.2g, accurately weighed, be placed in 10ml measuring bottle, the ultrasonic and jolting frequently of solubilizer 10 ~ 20 minutes, makes Olanzapine dissolve, let cool, add 1mol/l hydrochloric acid 2ml, 1h is heated in water-bath, add 2mol/l NaOH 2ml, neutralization, with solvent dilution to scale.

Alkali destroys: get this product 2, porphyrize, get fine powder 0.2g, accurately weighed, be placed in 10ml measuring bottle, the ultrasonic and jolting frequently of solubilizer 10 ~ 20 minutes, makes Olanzapine dissolve, let cool, add 1mol/l NaOH 2ml, 1h is heated in water-bath, add 2mol/l hydrochloric acid 2ml, neutralization, with solvent dilution to scale.

Oxidative demage: get this product 2, porphyrize, get fine powder 0.2g, accurately weighed, be placed in 10ml measuring bottle, the ultrasonic and jolting frequently of solubilizer 10 ~ 20 minutes, makes Olanzapine dissolve, lets cool, add 3,30% hydrogen peroxide, place 30 minutes, with solvent dilution to scale.

High temperature: get this product 2, porphyrize, get fine powder 0.2g, accurately weighed, be placed in 10ml measuring bottle, the ultrasonic and jolting frequently of solubilizer 10 ~ 20 minutes, makes Olanzapine dissolve, lets cool, and boiling water heating 4h, lets cool, with solvent dilution to scale.

Illumination destroys: get this product 2, porphyrize, get fine powder 0.2g, accurately weighed, is placed in 10ml measuring bottle, and the ultrasonic and jolting frequently of solubilizer 10 ~ 20 minutes, makes Olanzapine dissolve, let cool, with solvent dilution to scale.Put 4500lx according to 5 days.

Four, detectability

Get reference substance solution, adopt stepwise dilution method, sample introduction successively.When signal to noise ratio (S/N ratio) (S/N) is about 3, the Olanzapine Tablets of this correspondence is detectability.

Five, repeatability

Get and prepare 6 parts of need testing solutions with a collection of egg plain film difficult to understand, sample introduction measures and records chromatogram and peak area respectively, calculates relative standard deviation:

Six, testing result

Detect and be limited to 0.0023%, repeatability better RSD is 0.52%, and related substance testing result sees the following form:

Claims (1)

1. the high effective liquid chromatography for measuring Olanzapine of gradient elution and a method for related substances content, is characterized in that, adopt following operating conditions:

A. chromatographic column adopts anti-phase C8 chromatographic column;

B. flow velocity is 1.5ml/min, and determined wavelength is 220nm, and column temperature is 35 ° of C, and testing sample temperature is 5 ° of C;

C. sodium lauryl sulfate buffer is prepared through following methods, water intaking 1500ml, and add phosphoric acid 5.0ml, by 50% sodium hydroxide solution adjust ph to 2.5, add lauryl sodium sulfate 13.0g, jolting makes dissolving, filters;

D. mobile phase A is to prepare at 48: 52 by the volume ratio of acetonitrile and sodium lauryl sulfate buffer;

E. Mobile phase B is to prepare at 70: 30 by the volume ratio of acetonitrile and sodium lauryl sulfate buffer.