阅读说明：本技术 用于检测原料药中杂质元素的方法 (Method for detecting impurity elements in bulk drugs ) 是由 陈晓强 冯卫 王念彬 黄敏如 何金莲 郝燕娟 黄妍 于 2020-06-19 设计创作，主要内容包括：本发明公开了用于检测原料药中杂质元素的方法,包括步骤a,待测样液制备；步骤b,空白对照样液制备；步骤c,各杂质元素标准曲线绘制；步骤d,ICP-MS测定；步骤e,计算。本发明的检测方法可以同时对镉、铅、砷、汞、钴、钒、镍、钯、铬、锰、铜、钼、锂和锑共14种杂质元素进行检测,检测结果不会受到各元素之间的相互影响与干扰,一次检测即可得到全部杂质元素的含量,不需要对元素分别进行检测,通过改变消解模式和检测模式,提高检测结果的准确度,从而保证本方法的检出限和定量限水平远远高于其他检测方法。在保证检测精度的同时,提高了检测效率。(The invention discloses a method for detecting impurity elements in bulk drugs, which comprises the following steps of a, preparing a sample solution to be detected; step b, preparing a blank reference sample solution; step c, drawing standard curves of all impurity elements; step d, ICP-MS determination; and e, calculating. The detection method can simultaneously detect 14 impurity elements of cadmium, lead, arsenic, mercury, cobalt, vanadium, nickel, palladium, chromium, manganese, copper, molybdenum, lithium and antimony, the detection result is not influenced and interfered by each element, the content of all the impurity elements can be obtained by one-time detection, the elements are not required to be detected respectively, and the accuracy of the detection result is improved by changing a digestion mode and a detection mode, so that the detection limit and the quantitative limit level of the method are far higher than those of other detection methods. The detection precision is guaranteed, and meanwhile the detection efficiency is improved.)

1. The method for detecting impurity elements in the bulk drugs is characterized by comprising the following steps:

step a, preparing a sample solution to be detected:

taking a certain amount of sample, placing the sample in a first digestion tank, adding a gold standard stock solution and a digestion agent, digesting the first digestion tank according to a digestion program, cooling after digestion to obtain a first digestion solution, transferring the first digestion solution to a volumetric flask for constant volume to obtain a sample liquid to be measured;

step b, preparing a blank reference sample solution:

adding a gold standard stock solution and a digesting agent into a second microwave digestion tank, digesting the second digestion tank according to the digestion program, cooling after digestion to obtain a second digestion solution, and transferring the second digestion solution into a volumetric flask for constant volume to obtain a blank reference sample solution;

step c, drawing a standard curve of each impurity element:

testing 26 element working solutions, mercury working solutions and internal standard substance working solutions by using an inductively coupled plasma mass spectrometer, and calculating to obtain a standard curve equation of each impurity element according to a test result;

step d, ICP-MS determination:

testing the sample liquid to be tested and the blank reference sample liquid by using an inductively coupled plasma mass spectrometer, and respectively calculating concentration values of impurity elements in the sample liquid to be tested and the blank reference sample liquid according to a test result and a standard curve equation of each impurity element, wherein the impurity elements comprise lithium elements, and the lithium elements are tested by adopting a standard mode;

step e, calculating:

the content of each impurity element in the sample liquid to be detected is calculated according to the following formula,

in the formula: x is element content, mg/kg;

c1, concentration of each impurity element in the sample solution to be detected, mu g/L;

c0-concentration of each impurity element in blank control sample solution, mu g/L;

v is the volume of the sample with constant volume, mL;

n-dilution multiple of sample liquid to be detected;

m-sample size, g.

2. The method for detecting impurity elements in bulk drugs according to claim 1, comprising the following steps:

step a, preparing a sample solution to be detected:

taking 0.2-0.5 g of sample, placing the sample in a first digestion tank, adding 0.1-0.3 ml of gold standard stock solution and 4-13.5 ml of digestion agent, digesting the first digestion tank according to a digestion program, cooling to 70 ℃ after digestion to obtain a first digestion solution, transferring the first digestion solution into a 50ml volumetric flask, cleaning the first digestion tank by using primary water, transferring the first digestion tank into the 50ml volumetric flask, and fixing the volume to obtain a sample solution to be measured;

step b, preparing a blank reference sample solution:

adding 0.1-0.3 ml of gold standard stock solution and 4-13.5 ml of digesting agent into a second microwave digestion tank, digesting the second digestion tank according to the digestion program, cooling to 70 ℃ after digestion to obtain a second digestion solution, transferring the second digestion solution into a 50ml volumetric flask, cleaning the second digestion tank with primary water, transferring the second digestion tank into the 50ml volumetric flask, and carrying out constant volume to obtain a blank reference sample solution;

step c, drawing a standard curve of each impurity element:

testing 26 element working solutions, mercury working solutions and internal standard substance working solutions by using an inductively coupled plasma mass spectrometer, and calculating a standard curve equation of each impurity element according to the test result, wherein the 26 element working solutions comprise 26 element working solutions of 0.5 [ mu ] g/L, 1.0 [ mu ] g/L, 4.0 [ mu ] g/L, 10 [ mu ] g/L, 20 [ mu ] g/L, 50 [ mu ] g/L and 100 [ mu ] g/L, the mercury working solutions comprise mercury working solutions of 0.05 [ mu ] g/L, 0.1 [ mu ] g/L, 0.4 [ mu ] g/L, 1.0 [ mu ] g/L, 1.5 [ mu ] g/L and 2.0 [ mu ] g/L, and the internal standard substance working solution is an internal standard substance working solution of 10 [ mu ] g/L;

step d, ICP-MS determination:

testing the sample liquid to be tested and the blank reference sample liquid by using an inductively coupled plasma mass spectrometer, and calculating to obtain a concentration value of each impurity element in the sample liquid to be tested according to a test result and a standard curve of each impurity element, wherein the impurity element comprises lithium element, and the lithium element is tested by adopting a standard mode;

step e, calculating:

the content of each impurity element in the sample liquid to be detected is calculated according to the following formula,

in the formula: x is element content, mg/kg;

c1, concentration of each impurity element in the sample solution to be detected, mu g/L;

c0-concentration of each impurity element in blank control sample solution, mu g/L;

v is the volume of the sample with constant volume, mL;

n-dilution multiple of sample liquid to be detected;

m-sample size, g.

3. The method for detecting impurity elements in bulk drugs according to claim 1, wherein the impurity elements further comprise some or all of vanadium, cobalt, nickel, copper, arsenic, cadmium, antimony, mercury, lead, palladium, chromium, manganese, and molybdenum.

4. The method for detecting impurity elements in bulk drugs according to claim 3, wherein the test parameters of lithium element are as follows: the radio frequency power is 1550W, the sampling depth is 5.0mm, the carrier gas flow is 1.055L/min, the cooling gas flow is 14.0L/min, the collision gas flow is 0L/min, the atomization temperature is 2.7 ℃, the peristaltic pump speed is 40rpm, and the additional stabilization time is 10 s.

5. The method for detecting impurity elements in bulk drugs according to claim 4, wherein the vanadium, cobalt, nickel, copper, arsenic, cadmium, antimony, mercury, lead, palladium, chromium, manganese and molybdenum elements are tested in a collision reaction tank mode, and the test parameters are as follows: the radio frequency power is 1550W, the sampling depth is 5.0mm, the carrier gas flow is 1.055L/min, the cooling gas flow is 14.0L/min, the collision gas flow is 4.63L/min, the atomization temperature is 2.7 ℃, the pump speed of the peristaltic pump is 40rpm, and the additional stabilization time is 10 s.

6. The method for detecting impurity elements in a bulk drug according to claim 5, wherein the internal standard substance comprises scandium, germanium, indium and rhodium, the scandium element serves as an internal standard substance for vanadium, cobalt, chromium and manganese, the germanium element serves as an internal standard substance for nickel, copper, arsenic and cobalt, the indium element serves as an internal standard substance for cadmium, antimony, mercury and lead, and the rhodium element serves as an internal standard substance for palladium and molybdenum.

7. The method according to claim 6, wherein 10 parts of blank control sample solution are prepared according to the step b, the 10 parts of blank control sample solution are measured according to the steps c to d, 3 times of standard deviation of the measurement result is taken as a detection limit of the method, and 10 times of standard deviation is taken as a quantification limit of the method, wherein:

detection limit: cadmium: 0.001mg/kg, lead: 0.028mg/kg, arsenic: 0.001mg/kg, mercury: 0.004mg/kg, cobalt: 0.001mg/kg, vanadium: 0.001mg/kg, nickel: 0.005mg/kg, palladium: 0.004mg/kg, chromium: 0.013mg/kg, manganese: 0.034mg/kg, copper: 0.006mg/kg, molybdenum: 0.009mg/kg, lithium: 0.01mg/kg, antimony: 0.005 mg/kg;

and (4) quantitative limit: cadmium: 0.01mg/kg, lead: 0.01mg/kg, arsenic: 0.01mg/kg, mercury: 0.01mg/kg, cobalt: 0.01mg/kg, vanadium: 0.01mg/kg, nickel: 0.02mg/kg, palladium: 0.02mg/kg, chromium: 0.05mg/kg, manganese: 0.10mg/kg, copper: 0.10mg/kg, molybdenum: 0.05mg/kg, lithium: 0.05mg/kg, antimony: 0.01 mg/kg.

8. The method for detecting impurity elements in bulk drugs according to claim 7, wherein the digesting agent comprises a nitric acid solution and a hydrogen peroxide solution, and the volume ratio of the nitric acid solution to the hydrogen peroxide solution is 3: 1-4: 1.

9. The method for detecting impurity elements in a bulk drug according to claim 8, wherein the mass fraction of hydrogen peroxide in the hydrogen peroxide solution is 30%, the mass fraction of nitric acid in the nitric acid solution is 65%, and the background value of each impurity element in the nitric acid solution is less than one third of the detection limit.

10. The method for detecting impurity elements in bulk drugs according to claim 1, wherein the digestion procedure is as follows:

(1) heating for 5min to 80 deg.C, and maintaining for 5 min;

(2) heating for 5min to 120 deg.C, and maintaining for 15 min;

(3) heating for 5min to 150 deg.C, and maintaining for 10 min;

(4) the temperature is increased for 5min to 180 deg.C and maintained for 25 min.

Technical Field

The invention relates to the field of detection, in particular to a method for detecting impurity elements in bulk drugs.

Background

The impurity elements refer to substances generated, added or unintentionally introduced in the production or storage process of the bulk drugs. Mainly comes from catalyst residue in drug synthesis, introduction in the drug production and preparation process, introduction of production equipment or a container closed system. These impurity elements do not provide any therapeutic benefit to the patient and may also cause adverse effects or may adversely affect the stability, shelf life of the drug and may cause side effects due to potential toxicity.

In recent years, the limit requirements of various synthetic drugs, animal and plant extracted drugs, nutriments and food on impurity elements are becoming strict at home and abroad, and the export of related industries at home faces huge challenges. The same problem is faced in the synthesis of pharmaceutical products, various impurity elements are often introduced in the process of synthesizing raw medicines of the pharmaceutical products, the control of the impurity elements in the raw medicines by various countries is more and more strict, and the limit requirement of the impurities in the raw medicines after the ICH is added in China is also met internationally. Therefore, it becomes very important to establish an effective detection method for detecting impurity elements in the bulk drugs.

Disclosure of Invention

The invention aims to provide a method for detecting impurity elements in bulk drugs.

According to one aspect of the present invention, there is provided a method for detecting impurity elements in a bulk drug, comprising the following:

step a, preparing a sample solution to be detected:

taking a certain amount of sample, placing the sample in a first digestion tank, adding a gold standard stock solution and a digestion agent, digesting the first digestion tank according to a digestion program, cooling after digestion to obtain a first digestion solution, transferring the first digestion solution to a volumetric flask for constant volume to obtain a sample liquid to be measured;

step b, preparing a blank reference solution:

adding a gold standard stock solution and a digesting agent into a second microwave digestion tank, digesting the second digestion tank according to the digestion program, cooling after digestion to obtain a second digestion solution, and transferring the second digestion solution into a volumetric flask for constant volume to obtain a blank reference sample solution;

step c, drawing a standard curve of each impurity element:

testing 26 element working solutions, mercury working solutions and internal standard substance working solutions by using an inductively coupled plasma mass spectrometer, and calculating to obtain a standard curve equation of each impurity element according to a test result;

step d, ICP-MS determination:

testing the sample liquid to be tested and the blank reference sample liquid by using an inductively coupled plasma mass spectrometer, and respectively calculating concentration values of impurity elements in the sample liquid to be tested and the blank reference sample liquid according to a test result and a standard curve equation of each impurity element, wherein the impurity elements comprise lithium elements, and the lithium elements are tested by adopting a standard mode;

step e, calculating:

the content of each impurity element in the sample liquid to be detected is calculated according to the following formula,

in the formula: x is element content, mg/kg;

c1, concentration of each impurity element in the sample solution to be detected, mu g/L;

c0-concentration of each impurity element in blank control sample solution, mu g/L;

v is the volume of the sample with constant volume, mL;

n-dilution multiple of sample liquid to be detected;

m-sample size, g.

In some embodiments, a method for detecting impurity elements in a drug substance, comprises the following:

step a, preparing a sample solution to be detected:

taking 0.2-0.5 g of sample, placing the sample in a first digestion tank, adding 0.1-0.3 ml of gold standard stock solution and 4-13.5 ml of digestion agent, digesting the first digestion tank according to a digestion program, cooling to 70 ℃ after digestion to obtain a first digestion solution, transferring the first digestion solution into a 50ml volumetric flask, cleaning the first digestion tank by using primary water, transferring the first digestion tank into the 50ml volumetric flask, and fixing the volume to obtain a sample solution to be measured;

step b, preparing a blank reference solution:

adding 0.1-0.3 ml of gold standard stock solution and 4-13.5 ml of digesting agent into a second microwave digestion tank, digesting the second digestion tank according to the digestion program, cooling to 70 ℃ after digestion to obtain a second digestion solution, transferring the second digestion solution into a 50ml volumetric flask, cleaning the second digestion tank with primary water, transferring the second digestion tank into the 50ml volumetric flask, and carrying out constant volume to obtain a blank reference sample solution;

step c, drawing a standard curve of each impurity element:

testing 26 element working solutions, mercury working solutions and internal standard substance working solutions by using an inductively coupled plasma mass spectrometer, and calculating a standard curve equation of each impurity element according to the test result, wherein the 26 element working solutions comprise 26 element working solutions of 0.5 [ mu ] g/L, 1.0 [ mu ] g/L, 4.0 [ mu ] g/L, 10 [ mu ] g/L, 20 [ mu ] g/L, 50 [ mu ] g/L and 100 [ mu ] g/L, the mercury working solutions comprise mercury working solutions of 0.05 [ mu ] g/L, 0.1 [ mu ] g/L, 0.4 [ mu ] g/L, 1.0 [ mu ] g/L, 1.5 [ mu ] g/L and 2.0 [ mu ] g/L, and the internal standard substance working solution is an internal standard substance working solution of 10 [ mu ] g/L;

step d, ICP-MS determination:

testing the sample liquid to be tested and the blank reference sample liquid by using an inductively coupled plasma mass spectrometer, and calculating to obtain a concentration value of each impurity element in the sample liquid to be tested according to a test result and a standard curve of each impurity element, wherein the impurity element comprises lithium element, and the lithium element is tested by adopting a standard mode;

step e, calculating:

the content of each impurity element in the sample liquid to be detected is calculated according to the following formula,

in the formula: x is element content, mg/kg;

c1, concentration of each impurity element in the sample solution to be detected, mu g/L;

c0-concentration of each impurity element in blank control sample solution, mu g/L;

v is the volume of the sample with constant volume, mL;

n-dilution multiple of sample liquid to be detected;

m-sample size, g.

In some embodiments, the impurity elements further include some or all of vanadium, cobalt, nickel, copper, arsenic, cadmium, antimony, mercury, lead, palladium, chromium, manganese, and molybdenum. The method can detect 14 impurity elements of lithium, vanadium, cobalt, nickel, copper, arsenic, cadmium, antimony, mercury, lead, palladium, chromium, manganese and molybdenum in the bulk drugs.

In some embodiments, the test parameters for lithium are: the radio frequency power is 1550W, the sampling depth is 5.0mm, the carrier gas flow is 1.055L/min, the cooling gas flow is 14.0L/min, the collision gas flow is 0L/min, the atomization temperature is 2.7 ℃, the peristaltic pump speed is 40rpm, and the additional stabilization time is 10 s.

In some embodiments, vanadium, cobalt, nickel, copper, arsenic, cadmium, antimony, mercury, lead, palladium, chromium, manganese, and molybdenum are tested using the collision cell model with test parameters: the radio frequency power is 1550W, the sampling depth is 5.0mm, the carrier gas flow is 1.055L/min, the cooling gas flow is 14.0L/min, the collision gas flow is 4.63L/min, the atomization temperature is 2.7 ℃, the pump speed of the peristaltic pump is 40rpm, and the additional stabilization time is 10 s.

In some embodiments, the internal standard includes scandium, germanium, indium, and rhodium, scandium as the internal standard for vanadium, cobalt, chromium, and manganese, germanium as the internal standard for nickel, copper, arsenic, and cobalt, indium as the internal standard for cadmium, antimony, mercury, and lead, and rhodium as the internal standard for palladium and molybdenum. Different internal standard substances are selected for different impurity elements, so that the error in the detection process can be further reduced, and the detection accuracy and precision are improved.

In some embodiments, 10 blank control samples are prepared according to step b, and the 10 blank control samples are measured according to steps c-d, with 3 times the standard deviation of the measurement as the detection limit and 10 times the standard deviation as the quantitation limit of the method, wherein:

detection limit: cadmium: 0.001mg/kg, lead: 0.028mg/kg, arsenic: 0.001mg/kg, mercury: 0.004mg/kg, cobalt: 0.001mg/kg, vanadium: 0.001mg/kg, nickel: 0.005mg/kg, palladium: 0.004mg/kg, chromium: 0.013mg/kg, manganese: 0.034mg/kg, copper: 0.006mg/kg, molybdenum: 0.009mg/kg, lithium: 0.01mg/kg, antimony: 0.005 mg/kg;

and (4) quantitative limit: cadmium: 0.01mg/kg, lead: 0.01mg/kg, arsenic: 0.01mg/kg, mercury: 0.01mg/kg, cobalt: 0.01mg/kg, vanadium: 0.01mg/kg, nickel: 0.02mg/kg, palladium: 0.02mg/kg, chromium: 0.05mg/kg, manganese: 0.10mg/kg, copper: 0.10mg/kg, molybdenum: 0.05mg/kg, lithium: 0.05mg/kg, antimony: 0.01 mg/kg.

In some embodiments, the digesting agent comprises a nitric acid solution and a hydrogen peroxide solution, and the volume ratio of the nitric acid solution to the hydrogen peroxide solution is 3: 1-4: 1. The nitric acid solution and the hydrogen peroxide solution can play a good role in digestion, the volume ratio of the nitric acid solution to the hydrogen peroxide solution is 3: 1-4: 1, the bulk drugs can be completely digested, all impurity elements in the bulk drugs are digested, and subsequent content detection is facilitated.

In some embodiments, the mass fraction of hydrogen peroxide in the hydrogen peroxide solution is 30%, the mass fraction of nitric acid in the nitric acid solution is 65%, and the background value of each impurity element in the nitric acid solution is less than one third of the detection limit. The 30% hydrogen peroxide solution has strong oxidation effect and anti-bumping effect, and can completely digest the raw material medicine and ensure the safety of the digestion process. The background value of each impurity element in the nitric acid paste is lower than one third of the detection limit of the method, so that the better detection precision can be ensured, the influence of the background value in the nitric acid solution on the detection result is reduced, and the problem of overhigh detection limit caused by overhigh background value in the nitric acid solution is avoided.

In some embodiments, the digestion procedure is as follows:

(1) heating for 5min to 80 deg.C, and maintaining for 5 min;

(2) heating for 5min to 120 deg.C, and maintaining for 15 min;

(3) heating for 5min to 150 deg.C, and maintaining for 10 min;

(4) the temperature is increased for 5min to 180 deg.C and maintained for 25 min. Under the digestion program, the bulk drugs can be completely digested, so that impurity elements in the bulk drugs are completely dissolved in the digestion solution, the digestion time is shortened, and the subsequent detection is facilitated.

The invention has the beneficial effects that: the detection method can quickly and accurately detect the impurity elements in the raw material medicines, reduce the influence of the raw material medicines on the detection result, reduce the impurity element loss caused by digestion by changing the digestion mode, reduce the background value of the digestion agent, and further ensure the accuracy of the detection result; the atomic weight of the lithium element is 7, and the detection amount of the lithium element is relatively low relative to the actual content in the conventional detection mode, even the lithium element cannot be detected, so that the lithium element is not accurately detected. According to the invention, the standard mode is adopted to test the lithium element, so that the lithium element in the sample liquid can be accurately detected, and the accuracy of the method is ensured. And a new detection mode is adopted, so that all impurity elements are detected in the most suitable mode, the loss of the impurity elements caused by the detection mode is avoided, and the accuracy of a detection result is improved, so that the detection limit and the quantitative limit level of the method are far higher than those of other detection methods. The detection method can simultaneously detect 14 impurity elements of cadmium, lead, arsenic, mercury, cobalt, vanadium, nickel, palladium, chromium, manganese, copper, molybdenum, lithium and antimony, the detection result is not influenced and interfered by each element, the content of all the impurity elements can be obtained by one-time detection, the elements are not required to be detected respectively, and the detection efficiency is improved while the detection precision is ensured.

Drawings

Fig. 1 is a linear graph of a standard curve of a part of impurity elements obtained by detecting 12 impurity elements in a raw material fosfomycin trometamol according to a method for detecting impurity elements in a raw material drug in an embodiment of the present invention.

Fig. 2 is a linear graph of a standard curve of a part of impurity elements obtained by detecting 12 impurity elements in a raw material fosfomycin trometamol according to the method for detecting impurity elements in a raw material drug of the present invention.

Fig. 3 is a linear graph of a standard curve of part of impurity elements obtained by detecting 10 impurity elements in cefuroxime sodium as a raw material drug by using the method for detecting impurity elements in a raw material drug according to an embodiment of the present invention.

Fig. 4 is a linear graph of a standard curve of part of impurity elements obtained by detecting 10 impurity elements in cefuroxime sodium as a raw material drug by using the method for detecting impurity elements in a raw material drug according to an embodiment of the present invention.

Fig. 5 is a linear graph of a standard curve of part of impurity elements obtained by detecting 10 impurity elements in cefuroxime sodium as a raw material drug according to the method for detecting impurity elements in a raw material drug of the present invention.

Fig. 6 is a linear graph of a standard curve of a part of impurity elements obtained by detecting 10 impurity elements in the raw material cefepime hydrochloride/L-arginine according to the method for detecting impurity elements in the raw material drug of the present invention.

Fig. 7 is a linear graph of a standard curve of a part of impurity elements obtained by detecting 10 impurity elements in the raw material cefepime hydrochloride/L-arginine according to the method for detecting impurity elements in the raw material drug of the present invention.

Fig. 8 is a linear graph of a standard curve of a part of impurity elements obtained by detecting 10 impurity elements in the raw material cefepime hydrochloride/L-arginine according to the method for detecting impurity elements in the raw material drug of the present invention.

Detailed Description

The present invention will be described in further detail with reference to the accompanying drawings and examples.