TWI777327B - A distinguishing method for measuring the dissolution profile of megestrol acetate suspension - Google Patents

Abstract

The present invention provides a distinguishing method for measuring the dissolution profile of megestrol acetate suspension. The method includes the following steps: dissolving a surfactant in water to prepare a dissolving medium, wherein the surfactant includes sodium lauryl sulfate or polysorbate 80. The mass concentration of sodium lauryl sulfate ranges is from 0.35% to 0.45% while the surfactant is sodium lauryl sulfate; the mass concentration of polysorbate 80 ranges is from 0.8% to 1.0% while the surfactant is polysorbate 80. Then, shake the megestrol acetate oral suspension well, and prepare the samples after decompression and degassing. The samples are added to the dissolving medium through the automatic sample adding device, and the measurement is carried out according to the dissolution device stirring paddle method. Finally, take samples at sampling time points to determine the amount of dissolution at different time points.

Description

A discriminative method for measuring the dissolution curve of megestrol acetate suspension

The invention relates to the field of medicine, in particular to a method for determining the dissolution curve of a megestrol acetate suspension with discriminating power.

Dissolution: refers to the speed and degree of dissociation of active drugs from common preparations such as tablets, capsules or granules under specified conditions (that is, the amount of drug released when the drug reaches a certain time point in the dissolution medium). It is to examine whether the release of the drug at the specified time point can reach the limit specified in the standard.

Dissolution Profile: It is a dissolution-time curve drawn from the dissolution obtained by the formulation at different time points. The dissolution curve reflects the in vitro dissolution and dissolution process of the preparation, and the quality of the generic drug can be evaluated by comparing the similarity of multiple dissolution curves in vitro between the generic preparation and the original preparation.

Since 2012, the National Medical Products Administration (NMPA) has launched the quality consistency evaluation of generic drugs. The generic drugs that have been approved for the market will be consistent with the quality and efficacy of the original drug, so that the quality and efficacy of the generic drugs will be consistent with that of the original drugs. It is consistent with the original research drug. Among them, formulating a scientific and reasonable solid preparation dissolution curve is an important step to improve the success rate of in vivo bioequivalence test. It also provides a basis for including the characteristic dissolution curve of the drug in the corresponding quality standard, and provides a guarantee for the consistency of the quality of the drug between batches and the consistency of the drug quality before and after the process change.

At present, the various suspensions contained in domestic and foreign pharmacopeias require less measurement of solubility, only the megestrol acetate oral suspension, felbamate oral suspension, Indomethacin Oral Suspension specifies the requirements for the determination of solubility (measured by the second method of dissolution determination).

In the current market, the first/second method dissociation device is mainly designed to measure the solubility of solid preparations. When using this instrument to measure the dissolubility of liquid preparations, a syringe is generally used to manually add samples. Dissolution curves of suspensions are sensitive to changes in sample state, sample addition position, sample addition speed and sample injection needle diameter, which affect the precision of parallel detection of sample dissolution results.

The current commercially available megestrol acetate oral suspension is Megace®, specification: 40mg/ml (original preparation), in the prior art, the method for measuring the solubility of megestrol acetate oral suspension is in the US Pharmacopoeia and the Korean Pharmacopoeia All are recorded; the Korean Pharmacopoeia recording method is the dissolution method using 5% SDS aqueous solution as the dissolution medium. The dissolution rate of this method is too fast, and the difference between the prescription process of the generic drug and the original drug cannot be distinguished. Quality between generic drug companies; the US Pharmacopoeia contains three methods for determination of solubility, as shown in the following table:

When the commercially available preparation of megestrol acetate suspension, Megace®, adopts method 1 as the dissolution curve method, the dissolution rate of this method is faster (the dissolution rate has reached 85% in 15 minutes), and the obtained dissolution curve cannot effectively distinguish megestrol acetate. In vitro release of suspensions and effects on in vitro release.

When the commercially available preparation Megace® (specification: 40mg/ml) is completely dissolved under the conditions of method 2 and method 3, the solubility of megestrol acetate is 0.444mg/ml. It is determined that the raw material of megestrol acetate is in 0.5% SDS aqueous solution The medium solubility is 0.527mg/ml, and the conditions of method 2 and method 3 have seriously failed to meet the sink conditions. The dissolution curve is determined by this method, the dissolution rate is slow, and the distinction between the dissolution curves is not obvious. And there is no guarantee that the dissociation process can reach a dissociation amount of 85% or more.

The object of the present invention is to overcome the deficiencies of existing pharmacopoeia data, provide a kind of assay method that can effectively distinguish prescription and process variables to megestrol acetate suspension in vitro release dissolution curve, and improve the precision and accuracy of the method, for The consistency of the batch-to-batch quality of the drug product is guaranteed.

In order to achieve the above object, the present invention provides a discriminative method for measuring the dissolution curve of megestrol acetate suspension, wherein the method comprises the following steps:

(1) preparation of dissolving medium: comprising the step of dissolving surfactant in water to prepare dissolving medium, wherein the surfactant comprises sodium dodecyl sulfate (SDS) or polysorbate 80 (tween 80), When the surfactant is sodium lauryl sulfate, the mass concentration range of sodium lauryl sulfate is 0.35% to 0.45%, and when the surfactant is polysorbate 80, the mass concentration range of polysorbate 80 is 0.8%~1.0%;

(2) Preparation of the sample to be tested: including taking the megestrol acetate oral suspension to be tested and shaking well, and degassing the sample under reduced pressure to obtain the sample to be tested;

(3) Determination of the dissociation amount: including adding the sample to be tested into the dissociation medium through the automatic sample adding device, measuring according to the stirring paddle method of the dissociation device, sampling at different sampling time points, and measuring the dissociation amount at different time points.

According to some specific embodiments of the present invention, wherein, step (2) includes vigorously shaking the megestrol acetate suspension to be tested, so that the suspension is fully mixed, and then degassed under reduced pressure for 30-60 minutes minutes, the vacuum degree is maintained at -50~-80KPa, and the sample to be tested is obtained.

The vigorous shaking is vigorous shaking as generally understood in the art, and according to some embodiments of the present invention, the vigorous shaking is shaking the sample vial up and down at a frequency of at least 80 times per minute.

According to some specific embodiments of the present invention, wherein, step (3) comprises using an automatic sample adding device to add the sample to be tested into the dissolving medium, wherein the automatic sample adding device (as shown in FIG. 9 ) at least consists of a syringe 2, an injection It consists of a pump 1, a sample needle 4, and a connecting hose 3 (preferably a silicone connecting hose).

Wherein, the syringe pump can be a conventional syringe pump in the field, and according to some specific embodiments of the present invention, wherein, the syringe pump in step (3) includes a single-channel or multi-channel syringe pump.

According to some specific embodiments of the present invention, wherein, the step (3) includes a sample addition amount ranging from 0.5ml to 10ml.

According to some specific embodiments of the present invention, wherein, the step (3) comprises that the sample adding position is 0.5-1.5 cm below the liquid surface of the dissolving medium, preferably 1 cm.

According to some specific embodiments of the present invention, wherein, step (3) comprises that the sample adding needle is a flat needle, and the inner diameter of the needle is 0.5mm~1.54mm (the corresponding international standard specification is 21G~14G).

According to some specific embodiments of the present invention, wherein, the sample adding time of step (3) is 10-15 seconds.

According to some specific embodiments of the present invention, wherein, the sampling time points of step (3) include 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, and 60 minutes.

According to some specific embodiments of the present invention, wherein, step (3) comprises a method for determining the dissociation amount including high performance liquid chromatography or ultraviolet spectrophotometry.

According to some specific embodiments of the present invention, wherein, the chromatographic conditions of the high performance liquid chromatography in step (3) include:

Detector: UV detector;

Chromatographic column: take ODS-3, 4.6mm×150mm, 5μm as the chromatographic column;

Detection wavelength: 292nm;

Column temperature: 35℃;

Mobile phase: take acetonitrile-water (65:35) as mobile phase;

Flow rate: 1.2ml/min.

According to some specific embodiments of the present invention, wherein, the detection wavelength of the ultraviolet spectrophotometry in step (3) is 292 nm.

According to some specific embodiments of the present invention, wherein, the method comprises the following steps:

(1) Preparation of dissolving medium: including dissolving surfactant sodium lauryl sulfate or polysorbate 80 in water, wherein the concentration range of sodium lauryl sulfate is 0.35%~0.45%, and the concentration range of polysorbate 80 is 0.35%~0.45%. is 0.8%~1.0%;

(2) Preparation of the sample to be tested: including taking the megestrol acetate oral suspension to be tested and fully shaking, and degassing under reduced pressure for 30-60 minutes to obtain the sample to be tested;

(3) Determination of dissociation amount: add the sample to be tested into the dissolving medium through the automatic sample adding device, in which a syringe is used to measure 5~20ml of the sample to be tested, and a silicone hose is used to connect the syringe and the sampling needle, and the head of the sampling needle is placed At 0.5cm~1.5cm below the liquid level of the dissolving medium, the inner diameter of the injection needle is about 1.5mm, adjust the injection speed of the syringe pump so that the injection time is controlled at 8~15s, and the speed of the stirring blade of the dissolving device is set to 30 rpm/ 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, and 60 minutes, respectively, and measure the corresponding dissociation amount according to high performance liquid chromatography or ultraviolet spectrophotometry.

According to some specific embodiments of the present invention, wherein, step (1) further comprises the step of adjusting the pH value of the dissolving medium by using a buffer salt selected from the group consisting of hydrochloric acid, phosphate, acetate, glacial acetic acid, sodium hydroxide and One or more of the potassium hydroxide is mixed, and the amount of the buffer salt is to adjust the pH value of the prepared dissolving medium to 1.0-7.0.

It can be understood that in step (1), when preparing the dissolving medium, you can add Buffer salt is added or not.

According to some specific embodiments of the present invention, wherein, step (1) comprises dissolving the surfactant in water, and then adding an appropriate buffer salt to prepare a dissolving vehicle.

According to some specific embodiments of the present invention, wherein, step (1) comprises dissolving the surfactant in water to prepare a dissolving medium, and dissolving the surfactant in water and further adding a buffer salt to prepare a dissolving medium, so as to be respectively prepared into The added amount of buffer salt is respectively 0, and the pH value of the prepared dissolving medium is adjusted to 1.2, 4.5 and 6.8 dissolving medium, and the buffer salt is selected from hydrochloric acid, phosphate, acetate, glacial acetic acid, sodium hydroxide and A mixture of one or more of potassium hydroxide.

It can be understood that the dissolving medium in step (1) can include eight kinds:

According to some specific embodiments of the present invention, there are four kinds of dissolving media prepared by sodium lauryl sulfate, which are: 0.4% sodium lauryl sulfate aqueous solution (without adding buffer salt); sodium lauryl sulfate The mass concentration is 0.4%, and the pH of the prepared dissolving medium is adjusted to 1.2 by adding buffer salt; the mass concentration of sodium lauryl sulfate is 0.4%, and the pH of the prepared dissolving medium is adjusted to 4.5 by adding buffer salt; The mass concentration of sodium alkyl sulfate is 0.4%, and buffer salt is added to adjust the pH of the prepared dissolving medium to 6.8;

According to some specific embodiments of the present invention, there are four kinds of dissolving media prepared by polysorbate 80, respectively: 0.9% polysorbate 80 aqueous solution (without adding buffer salt); polysorbate 80 mass concentration of 0.9%, The pH of the prepared dissolving medium was adjusted to 1.2 by adding buffer salt; the mass concentration of polysorbate 80 was 0.9%, and the pH of the prepared dissolving medium was adjusted to 4.5 by adding buffer salt; the mass concentration of polysorbate 80 was 0.9%, The pH of the prepared dissolving medium was adjusted to 6.8 by adding buffer salt.

According to some specific embodiments of the present invention, wherein, step (1) does not add buffer The preparation process of the dissolving medium liquid of rush salt and sodium lauryl sulfate with a mass concentration of 0.4% (or a polysorbate 80 mass concentration of 0.9%) is as follows: take 4.0 g of sodium lauryl sulfate (or 9.0 g of polysorbate). Sorbitan 80) is placed in a measuring flask, add 700~800ml of water to dissolve, add water to dilute to 1000ml, mix well, and degas.

According to some specific embodiments of the present invention, wherein, in step (1), the pH value of the dissolving medium is 1.2, and the sodium lauryl sulfate is the dissolving medium with a mass concentration of 0.4% (or a polysorbate 80 mass concentration of 0.9%). The preparation process is as follows: Weigh 4.0g sodium lauryl sulfate (or 9.0g polysorbate 80) into a measuring bottle, add 700~800ml water to dissolve, add 7.65ml hydrochloric acid, add water to dilute to 1000ml, mix well, Degassing, and that's it.

According to some specific embodiments of the present invention, wherein, in step (1), the pH value of the dissolving medium is 4.5, and the sodium lauryl sulfate is the dissolving medium with a mass concentration of 0.4% (or a polysorbate 80 mass concentration of 0.9%). The preparation process is as follows: Weigh 2.99g of sodium acetate and place it in a measuring flask, add 14ml of 2mol/L acetic acid solution, and dilute to 400ml with water; weigh 4.0g of sodium lauryl sulfate (or 9.0g of polysorbate 80) and set it In another volumetric flask, add 500ml of water to dissolve, mix the solutions in the two volumetric flasks, dilute to 1000ml with water, mix well, and degas.

According to some specific embodiments of the present invention, wherein, in step (1), the pH value of the dissolving medium is 6.8, and the mass concentration of sodium lauryl sulfate is 0.4% (or the mass concentration of polysorbate 80 is 0.9%). The preparation process is as follows: weigh 6.0 g of sodium dihydrogen phosphate and place it in a measuring flask, add 112 ml of 0.2 mol/L sodium hydroxide solution, and dilute to 400 ml with water; weigh 4.0 g of sodium dodecyl sulfate (or 9.0 g of poly sorbate 80) in another measuring bottle, add 500 ml of water to dissolve, mix the solutions in the two measuring bottles, dilute to 1000 ml with water, mix well, and degas.

According to some specific embodiments of the present invention, wherein, step (3) further comprises drawing a dissolution curve with the measured dissolution amount to evaluate the in vitro dissolution characteristics of the sample to be tested.

According to some specific embodiments of the present invention, using the dissociation curve drawn in step (3), the similarity factor f2 is used to evaluate the degree of similarity between different formulations and process preparations and the original preparation.

According to some specific embodiments of the present invention, the in vitro dissolution curve of the sample to be tested is used to evaluate the influence of product formulation or process changes on the quality of the megestrol acetate oral suspension.

According to some specific embodiments of the present invention, the in vitro dissolution curve of the sample to be tested is used to predict the difference in the in vivo bioequivalence of the product.

According to some specific embodiments of the present invention, wherein, the method further includes the step of preparing a reference substance solution, and in step (3), the reference substance and the sample to be tested are respectively added to the dissolving medium through the automatic sample adding device, and the solution is according to the dissolving device. The stirring paddle method was used to measure the samples respectively, and the samples were taken at different sampling time points to measure the dissociation amount at different time points.

According to some specific embodiments of the present invention, wherein, the method further comprises:

(1) preparation of dissolving medium: comprising the step of dissolving surfactant in water to prepare dissolving medium, wherein the surfactant comprises sodium dodecyl sulfate (SDS) or polysorbate 80 (tween 80), When the surfactant is sodium lauryl sulfate, the mass concentration range of sodium lauryl sulfate is 0.35% to 0.45%, and when the surfactant is polysorbate 80, the mass concentration range of polysorbate 80 is 0.8%~1.0%;

(2) prepare the reference substance solution;

(3) Preparation of the sample to be tested: including taking the megestrol acetate oral suspension to be tested and shaking well, and degassing under reduced pressure to obtain the sample to be tested;

(4) Determination of dissolution amount: including adding the reference substance and the sample to be tested through the automatic sample adding device respectively In the dissolving medium, the measurement was carried out according to the stirring paddle method of the dissociation device, and the samples were taken at different sampling time points to measure the dissociated amount at different time points.

To sum up, the present invention provides a discriminative method for measuring the dissolution curve of megestrol acetate suspension. The assay method of the present invention has the following advantages:

(1) in the present invention, dissolving medium liquid is set to 0.4% sodium lauryl sulfate solution or 0.9% polysorbate 80 solution, and rotating speed is set to 30 rev/min, shows by a large number of experiments: adopt this dissolving condition to measure The dissolution curve is more discriminating than the USP method, and can appropriately distinguish the quality of the generic drug from the original drug.

(2) In the present invention, it is stipulated that the preparation method of the sample to be tested is as follows: shake vigorously, fully mix the suspension, and degas under reduced pressure for 30-60 minutes. If the suspension is placed for a long time, sedimentation and stratification will occur. If the mixing time is short, the state and content of the samples in different positions in the bottle cannot be guaranteed to be consistent; after vigorous shaking and mixing, a large number of tiny air bubbles will appear in the suspension. For the determination of the dissolution curve, the amount of dissolution at the same sampling time point will increase, and the degree of discrimination of the dissolution curve will become worse. A large number of experiments show that the use of the preparation method of the sample to be tested in the present invention can improve the discriminating power of the dissolution curve determination method while ensuring the homogeneity of the test sample.

(3) In the present invention, an automatic sample adding device is adopted for the sample adding method for the determination of the dissolution amount, which eliminates the interference of the difference of the sample adding speed between different personnel and different times, and significantly improves the precision and accuracy of the dissolution curve method.

(4) In the present invention, the sample adding position, the sample adding speed, and the diameter of the needle used for sample adding are specified to ensure the reproducibility of the dissolution curve.

(5) The method for determining the dissolution curve of the suspension provided by the method of the present invention has the characteristics of science, durability, high precision, good reproducibility, etc. Consistency evaluation work, the consistency of drug batch quality, and the consistency of drug quality before and after process changes provide reliable guarantees.

1 is a schematic diagram of the dissolution curve of the original preparation provided in the embodiment of the present invention 1 with USP method 1, and a schematic diagram of the dissolution curve of the original preparation of the dissolution medium in the process of SDS concentration screening and dissolution device rotational speed screening;

Fig. 2 is the dissociation curve schematic diagram of the original preparation agent and self preparation agent that provided in the embodiment of the present invention 1 adopting different processing methods to measure;

Fig. 3 is the dissociation curve schematic diagram of the original preparation agent provided in the embodiment of the present invention 1 using different sample addition times to measure;

Fig. 4 is the dissociation curve schematic diagram of the original preparation agent provided in the embodiment of the present invention 1 adopting different sample adding positions to measure the dissociation curve;

5 is a schematic diagram of the dissolution curve of the original preparation provided in Example 1 of the present invention using a needle with different apertures to measure;

6 is a schematic diagram of the dissolution curves of the original preparation provided in Example 1 of the present invention using automatic sample addition and manual sample addition;

Fig. 7 adopts the method for the determination of the dissolution curve of megestrol acetate oral suspension with a discriminative power provided in Example 2, the measured original preparation and the megestrol acetate prepared by 3 different prescriptions and processes Dissolution curves of oral suspensions;

Fig. 8 adopts a kind of method for measuring the dissolution curve of megestrol acetate oral suspension with discriminative power provided in Example 2, and measures the dissolution curve diagrams of the original trituration agent in 4 kinds of different pH dissolution media;

Fig. 9 is the automatic sample adding device adopted in the present invention.

The implementation process and beneficial effects of the present invention are described in detail below through specific examples, which are intended to help readers better understand the essence and characteristics of the present invention, and are not intended to limit the scope of implementation of the present case.

Example 1

In the prior art, in the United States Pharmacopoeia (USP<42>), the dissolution method for measuring megestrol acetate oral suspension is recorded, and USP method 1 is used to measure the dissolution curve of the original preparation: with 0.5% dodecane Sodium sulphate (SDS) aqueous solution is the dissolving medium, add 900 ml into the dissociation cup, set the speed of the dissociation device to 25 rpm, and when the temperature of the dissolving medium reaches 37±5 °C, use a syringe to suck 4 ml of mixed acetic acid The megestrol oral suspension was injected into the dissolving cup on the surface of the dissolving medium, and 5 ml of the solution was taken at 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, and 60 minutes, respectively. Filtration with microporous membrane, taking the subsequent filtrate as the test solution, and measuring the dissociation amount by high performance liquid chromatography. The chromatographic conditions are as follows:

Detector: UV detector;

Chromatographic column: take ODS-3, 4.6mm×150mm, 5μm as the chromatographic column;

Detection wavelength: 292nm;

Column temperature: 35℃;

Mobile phase: take acetonitrile-water (65:35) as mobile phase;

Flow rate: 1.2ml/min.

The results show that the dissociation result using USP method 1 is too fast (see Figure 1), and the dissociation amount has reached 85% in 15 minutes, and the dissociation method is indistinguishable.

In order to explore a more suitable method to make the dissolution curve of megestrol acetate oral suspension achieve better discriminating power, in further research experiments, the changes of SDS concentration, rotation speed, sample processing methods, The effect of sample addition method on the dissolution curve of megestrol acetate oral suspension.

A large number of experiments have shown that when the concentration of SDS in the dissolution medium is 0.4% and the rotation speed is 30 r/min, the measured dissolution curve is better than the USP method, and can appropriately distinguish the quality of generic drugs and original drugs. It can scientifically distinguish the difference between generic drug prescription and process, as shown in Figure 1.

If the suspension is placed for a long time, sedimentation and stratification will occur. For example, if the mixing time is short, the state and content of the samples in different positions in the bottle cannot be guaranteed to be consistent, thus affecting the precision of the dissolution curve method. A large number of tiny bubbles will appear inside. At this time, when the dissolution curve is measured, the amount of dissolution at the same sampling time point will increase, and the degree of discrimination of the dissolution curve will become worse. Experiments show that after the suspension is vigorously shaken and mixed, and then degassed under reduced pressure for 30-60 minutes, the dissolution amount of the same sampling point within 30 minutes becomes smaller, and the dissolution curve is better. See Table 1, Figure 2 ;

[Table 1] Differences in the amount of dissociation between different sample treatments

A large number of experiments have shown that when the suspension is added, the parameters such as the speed of sample addition, the sample addition position, and the diameter of the needle of the sample injection needle will all affect the dissolution curve. Add the sample needle at 0.5cm-1.5cm below the liquid surface of the dissolving medium, and complete the sample addition in 10-15 seconds.

In the present invention, the dissociated sample adopts the laboratory syringe pump automatic sample adding device shown in FIG. 9, which eliminates the interference of different personnel and the difference of sample adding speed at different times, and can ensure that the sample is always added at a uniform speed, and the first and second points can be controlled. The RSDs were all lower than 5%, which significantly improved the precision and accuracy of the dissolution curve method (as shown in Table 2).

[Table 2] Dissolution curves and precision results of different sample addition methods

Example 2

The present embodiment provides a method for measuring the dissolution curve of megestrol acetate oral suspension, comprising the following steps:

Step (1) preparation of dissolving medium:

The dissolving medium is 0.4% sodium lauryl sulfate (or 0.9% polysorbate 80) aqueous solution, and its preparation process is: take 4.0g sodium lauryl sulfate (or 9.0g polysorbate 80) and place it in the amount In the bottle, add 700ml of water to dissolve, add water to dilute to 1000ml, mix well and degas.

The dissolving medium is pH 1.2, containing 0.4% sodium lauryl sulfate (or 0.9% polysorbate 80) aqueous solution, and its preparation process is: weigh 4.0g sodium lauryl sulfate (or 9.0g polysorbate 80) ) in a measuring flask, add 700~800ml of water to dissolve, add 7.65ml of hydrochloric acid, add water to dilute to 1000ml, mix well, and degas.

The dissolving medium is pH 4.5, containing 0.4% sodium lauryl sulfate (or 0.9% polysorbate 80) aqueous solution, and its preparation process is: weigh 2.99g of sodium acetate and place it in a measuring bottle, add 2mol/L acetic acid solution 14ml, add water to dilute to 400ml; take 4.0g sodium lauryl sulfate (or 9.0g polysorbate 80) and place it in another measuring bottle, add 500ml water to dissolve, and mix the solutions in the two measuring bottles , diluted with water to 1000ml, mixed, and degassed.

The dissolving medium is pH 6.8, containing 0.4% sodium lauryl sulfate (or 0.9% polysorbate 80) aqueous solution, and its preparation process is as follows: weigh 6.0g of sodium dihydrogen phosphate and place it in a measuring bottle, add 0.2mol/ L sodium hydroxide solution 112ml, add water to dilute to 400ml; weigh 4.0g of sodium lauryl sulfate (or 9.0g of polysorbate 80) and place it in another measuring bottle, add 500ml of water to dissolve, mix the two Mix the solution in the bottle, dilute it with water to 1000ml, mix well, degas, and that's it.

When measuring the dissolution curve of megestrol acetate oral suspension, due to the difference in pH value, it is necessary to determine the dissolution curve of megestrol acetate oral suspension in each pH dissolution vehicle.

Step (2) reference substance solution preparation:

Accurately weigh about 18mg of megestrol acetate reference substance in a 100ml measuring bottle, add about 2.5ml methanol to dissolve, dilute to the scale with the dissolving medium in step 1, and make about 180 μg acetic acid in every 1ml. Megestrol solution, shake well, as the reference solution.

Step (3) Preparation of sample to be tested:

The megestrol acetate suspension to be tested is shaken vigorously to make the suspension fully mixed, and then degassed under reduced pressure for 30-60 minutes to obtain the sample to be tested.

Step (4) preparation of test solution:

Take the dissolving medium in step (1), according to the second method of dissociation measurement method (stirring paddle method), and set the rotating speed to 30 rev/min. Use a disposable syringe to suck up about 20ml of the freshly mixed suspension (sample solution) in step (3), put it on the laboratory dual-channel syringe pump (shown in Figure 9), and use a silicone connecting hose to connect the syringe and add the sample Needle (needle with an inner diameter of 1.5mm), and make the silicone connection hose to the sampling needle to be filled with the sample solution (no air bubbles). The sample time is 10 seconds. When the temperature of the dissolution cup reaches 37 ± 5 °C, the sample is automatically added, and the time is measured, and 5 ml of the solution is taken at 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, and 60 minutes, respectively. The chaoate was filtered through a 0.45 μm microporous membrane, and the subsequent filtrate was taken as the test solution.

Step (5) Dissolution determination method:

The dissociation amount was determined by high performance liquid chromatography or ultraviolet spectrophotometry.

The chromatographic conditions for high performance liquid chromatography are:

Detector: UV detector;

Chromatographic column: take ODS-3, 4.6mm×150mm, 5μm as the chromatographic column;

Detection wavelength: 292nm;

Column temperature: 35℃;

Mobile phase: take acetonitrile-water (65:35) as mobile phase;

Flow rate: 1.2ml/min.

The detection conditions of UV spectrophotometry are: detection wavelength 292nm.

Step (6) Determination of Dissolution Curve

Make need testing solution according to step (3) and step (4), precisely measure reference substance solution and need testing solution 10 μ l, inject liquid chromatograph, and record chromatogram, press external standard method with peak area The cumulative dissolution of megestrol acetate at 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes and 60 minutes of the original preparation was calculated respectively. Repeat the above steps twice, calculate the average cumulative dissociation amount of the original trituration agent at 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, and 60 minutes, and draw the original trituration agent in step (1) different pH dissolution media Dissolution curves in liquid.

Step (7) Evaluation:

Using the similarity factor method, the similarity factor f2 was calculated using the average cumulative dissolution data of the original preparation and the imitation preparation at 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes and 60 minutes. It is used to compare the similarity between the dissolution curve of the generic preparation and the original preparation; when the similarity factor f2 is used to judge the similarity, when the similarity factor f2 is greater than or equal to 50, the dissolution curves of the generic preparation and the original preparation are similar, and the similarity factor f2 When it is less than 50, the dissolution curves of the generic preparation and the original preparation are not similar.

In the further optimization plan, it is possible to preferentially select the dissolution data of the generic preparation and the original preparation at four time points of 10, 15, 20, and 30 minutes, and calculate the value of the similarity factor f2, which can not only achieve the same effect, but also simplify the The calculation process is convenient to use the value of the similarity factor f2 to compare the similarity of the dissolution curve of the generic preparation with the dissolution curve of the original preparation.

Example 3

According to a kind of discriminative megestrol acetate suspension dissolution curve determination method provided in the embodiment 2, the megestrol acetate oral suspension of the original preparation, different prescriptions, different processes and the final prescription technology was dissolved respectively curves were determined, where,

In step (1), the dissolving medium used is in 0.4% aqueous sodium dodecyl sulfate solution;

In step (5), specifically adopt high performance liquid chromatography to carry out dissociation measurement;

Through the measurement, the dissolution results of the original preparation, different formulations, different process formulations and final formulations were obtained, as shown in Table 3 and Figure 7.

[Table 3] The method of the present invention measures the comparison of the dissolution results of different preparations

It can be seen from the measurement results that the dissolution curve method provided by the present invention has good discriminating power for megestrol acetate oral suspensions of different prescriptions and processes, and can effectively distinguish the differences brought by different batches of prescriptions and processes.

Example 4

According to a kind of discriminative megestrol acetate suspension dissolution curve determination method provided in Example 2, firstly, the original preparation was subjected to different pH dissolution curve measurement, wherein,

In step (1), the dissolving media used are respectively: 0.4% sodium dodecyl sulfate aqueous solution described in Example 2, pH 1.2, containing 0.4% sodium dodecyl sulfate solution, pH 4.5, containing 0.4% Sodium dodecyl sulfate solution, pH 6.8, containing 0.4% sodium dodecyl sulfate solution;

In step (5), specifically adopt high performance liquid chromatography to carry out dissociation measurement;

Through the measurement, the dissolution curves of the original preparation agent in the four dissolving media were obtained. As shown in Figure 8, it can be seen that the dissolution curves of the original preparation agent in the four dissolution media are very close, and the method provided by the present invention can be used. The dissolution curve of the preparation is used as a reference to effectively evaluate the similarity and quality consistency of the generic preparation.

In a further example, according to a discriminative method for measuring the dissolution curve of megestrol acetate suspension provided in Example 2, the final formulation in Example 3 was subjected to the dissolution curves of different pH dissolution mediators. The similarity between the original preparation and the final formulation was evaluated by the similarity factor f2. The results are shown in Table 4.

[Table 4] Similarity evaluation between the original preparation and the self-developed preparation

Among them, the original preparation is the commercially available megestrol acetate oral suspension is Megace®, specification: 40mg/ml;

The prescription and proportion of the self-prepared preparation are the same as the original preparation. The preparation process includes: mixing megestrol acetate, polyethylene glycol 1450, polysorbate 80, xanthan gum, sucrose, sodium benzoate, citric acid (monohydrate) ), sodium citrate (dihydrate), lemon essence, etc. are mixed with water and homogenized.

The present invention further replaces the sodium lauryl sulfate of Example 2-4 with polysorbate 80, and achieves substantially the same effect as Example 2-4.

To sum up, the dissolution curve method of the present invention not only has good discriminating power, but also can be used to evaluate the influence of product prescription or process change on the quality of megestrol acetate oral suspension, and to predict the difference in the in vivo bioequivalence of the product. , which can provide in vitro basis for quality control and similarity assessment of generic preparations.

Claims (16)

A discriminative method for determining the dissolution curve of a megestrol acetate suspension, wherein the method comprises the following steps: (1) preparation of a dissolving medium: comprising dissolving a surfactant in water to prepare a dissolving medium Step, wherein surfactant comprises sodium lauryl sulfate or polysorbate 80, when surfactant is sodium lauryl sulfate, the mass concentration range of sodium lauryl sulfate is 0.35%～0.45%, when When the surfactant is polysorbate 80, the mass concentration range of polysorbate 80 is 0.8% to 1.0%; (2) preparation of the sample to be tested: including taking the megestrol acetate oral suspension to be tested and fully shaking it up , the sample to be tested is obtained by degassing under reduced pressure; (3) Determination of dissociation amount: including adding the sample to be tested into the dissolving medium through the automatic sample adding device, wherein the sample adding position is located 0.5~1.5 below the liquid level of the dissolving medium. cm, according to the stirring paddle method of the dissociation device, sampling at different sampling time points, and measuring the dissociation amount at different time points. The method according to item 1 of the scope of application, wherein step (2) comprises vigorously shaking the megestrol acetate oral suspension to be tested, so that the suspension is sufficiently shaken, and then decompressed and decompressed. gas for 30~60 minutes, and the vacuum degree is maintained at -50~-80KPa to obtain the sample to be tested. The method according to item 1 of the scope of the application, wherein step (3) comprises using an automatic sample adding device to add the sample to be tested into the dissolving medium, wherein the automatic sample adding device is at least composed of a syringe, a syringe pump, a sample adding needle and a Connect the hoses. The method according to item 1 of the claimed scope, wherein the sample addition amount in step (3) is in the range of The circumference is 0.5ml~10ml. The method according to item 3 of the scope of the application, wherein the sample adding position in step (3) is located 1 cm below the liquid surface of the dissolving medium; the inner diameter of the needle tip of the sample adding needle is 0.5 mm to 1.54 mm. The method according to claim 1, wherein the sampling time points in step (3) include 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, and 60 minutes. The method according to item 1 of the claimed scope, wherein the method for determining the dissociation amount in step (3) includes high performance liquid chromatography or ultraviolet spectrophotometry. The method according to any one of items 1 to 7 of the scope of the application, wherein the method comprises the following steps: (1) preparation of a dissolving medium: comprising mixing the surfactant sodium dodecyl sulfate or poly Sorbitan 80 is soluble in water, wherein the concentration range of sodium lauryl sulfate is 0.35%~0.45%, and the concentration range of polysorbate 80 is 0.8%~1.0%; (2) Preparation of samples to be tested: including taking the acetic acid to be tested The megestrol oral suspension is fully shaken, and the sample to be tested is prepared by degassing under reduced pressure for 30 to 60 minutes; (3) Determination of dissociation amount: the sample to be tested is added to the dissolving medium through an automatic sample adding device, wherein Use a syringe to measure 5~20ml of the sample to be tested, connect the syringe and the injection needle with a silicone rubber hose, and place the head of the injection needle at 0.5cm~1.5cm below the liquid surface of the dissolving medium. The inner diameter of the injection needle is about 1.5mm. , adjust the injection speed of the syringe pump so that the injection time is controlled at 10~15 seconds, and the speed of the stirring blade of the dissolution device is set to 30 rpm, take samples at 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, and 60 minutes, respectively, and measure the corresponding dissolution amount according to high performance liquid chromatography or ultraviolet spectrophotometry. The method according to any one of items 1 to 7 of the claimed scope, wherein step (1) further comprises the step of adjusting the pH value of the dissolving medium by using a buffer salt, the buffer salt is selected from hydrochloric acid, phosphoric acid Mixing of one or more of salt, acetate, glacial acetic acid, sodium hydroxide and potassium hydroxide, the amount of the buffer salt is to adjust the pH value of the prepared dissolving medium to 1.0~7.0. The method of claim 9, wherein step (1) comprises dissolving the surfactant in water, and then adding an appropriate buffer salt to prepare a dissolving medium. The method according to any one of items 1 to 7 of the claimed scope, wherein step (1) comprises dissolving the surfactant in water to prepare a dissolving medium, and dissolving the surfactant in water without adding Or further add a buffer salt to prepare a dissolving medium, to prepare a dissolving medium with a buffer salt addition amount of 0 respectively, and adjusting the pH value of the prepared dissolving medium to 1.2, 4.5 and 6.8, the buffer salt is selected from the group consisting of: A mixture of one or more of hydrochloric acid, phosphate, acetate, glacial acetic acid, sodium hydroxide and potassium hydroxide. The method according to item 1 of the claimed scope, wherein step (3) further comprises drawing the measured dissociation amount into a dissociation curve to evaluate the in vitro dissociation characteristics of the sample to be tested. A method as described in claim 12, wherein the evaluation method adopts a similarity factor f2 is performed. The method according to item 12 of the claimed scope, wherein the in vitro dissolution curve of the sample to be tested is used to evaluate the influence of product formulation or process changes on the quality of the megestrol acetate oral suspension. The method of claim 12, wherein the in vitro dissolution curve of the sample to be tested is used to predict the difference in the in vivo bioequivalence of the product. The method according to item 1 of the scope of the application, wherein the method further comprises the step of preparing a reference substance solution, and in step (3), the reference substance and the sample to be tested are respectively added to the dissolving medium through an automatic sample adding device , according to the stirring paddle method of the dissociation device, respectively, take samples at different sampling time points, and measure the dissociation amount at different time points.

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