CN109752471A - A kind of detection method of calcitriol - Google Patents
A kind of detection method of calcitriol Download PDFInfo
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- CN109752471A CN109752471A CN201910017625.1A CN201910017625A CN109752471A CN 109752471 A CN109752471 A CN 109752471A CN 201910017625 A CN201910017625 A CN 201910017625A CN 109752471 A CN109752471 A CN 109752471A
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- calcitriol
- mobile phase
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- methanol
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Abstract
The present invention relates to a kind of detection methods of calcitriol, belong to Pharmaceutical Analysis field.The detection method of the calcitriol of the application detects calcitriol by HPLC method, and the HPLC method carries out gradient elution: chromatographic column: C18 column using following condition；Mobile phase A: methanol；Mobile phase B: methanol and acetonitrile；Sample volume: 5-10 μ L；Flow velocity: 1.5-2.0mL/min；Column temperature: 30-40 DEG C；Detection wavelength: 255nm.Screening is optimized to the condition of HPLC method in the present invention, it is determined that optimal mobile phase and condition of gradient elution, the range of linearity are 0.25-5 μ g/ml, in a few days day to day precision RSD value is respectively less than 2%, and precision is high, reproducible, average recovery rate is 99.73%, RSD value less than 2%, and the rate of recovery is high.
Description
Technical field
The present invention relates to Pharmaceutical Analysis fields, and in particular to a kind of detection method of calcitriol.
Background technique
Calcitriol (Calcitriol, 1), chemical name 9,10- open loop cholesteric -5Z, 7E, 10 (19)-triolefin -1 α, 3
β, 25- triol.Calcitriol is one of most important metabolic activity product of vitamine D3 in human body, can promote suction of the enteron aisle to calcium
It receives, and adjusts the calcification of sclerotin.Calcitriol is produced by Roche Holding Ag, Switzerland earliest, is listed within 1978, trade name
" Rocaltrol ", for treating low postmenopausal osteoporosis, chronic kidney, postoperative hypothyroidism, idiopathic first
Gland hypofunction, pseudohypothyroidism, vitamin D dependent ricket, hypophosphatemic vitamin D resistance type Gou by shape
Crooked disease etc..Calcitriol has reached many decades for osteoporosis in Japan, New Zealand, Australia, Italy, British Clinical.
Since calcitriol is endogenous material, curative for effect, safety and stability, therefore be the choice drug of osteoporosis.
The detection method of calcitriol is less in the prior art, and generally existing accuracy and repeated low problem.
Summary of the invention
The object of the present invention is to provide a kind of calcitriol detection methods precise and stable, repeatability is high.
The application solve the technical problem technical solution be a kind of calcitriol detection method, by HPLC method to bone
Change triol to be detected, the HPLC method carries out gradient elution using following condition:
Chromatographic column: C18 column；
Mobile phase A: methanol；
Mobile phase B: methanol and acetonitrile；
Sample volume: 5-10 μ L；
Flow velocity: 1.5-2.0mL/min；
Column temperature: 30-40 DEG C；
Detection wavelength: 255nm.
Preferably, the volume ratio of methanol and acetonitrile is 35-50:50-65 in the Mobile phase B.
It is further preferred that the volume ratio of methanol and acetonitrile is 40:60 in the Mobile phase B.
Wherein, the condition of the gradient elution are as follows:
| Time (min) | Mobile phase A (V%) | Mobile phase B (V%) |
| 0 | 70 | 30 |
| 3 | 70 | 30 |
| 10 | 60 | 40 |
| 20 | 52 | 48 |
| 30 | 40 | 60 |
| 30.1 | 0 | 100 |
| 40 | 40 | 60 |
| 50 | 60 | 40 |
。
Preferably, the sample volume is 10 μ L.
Preferably, the column temperature is 38 DEG C.
Beneficial effects of the present invention:
The detection method of calcitriol of the invention detects calcitriol using HPLC method.The present invention is to HPLC method
Condition screening is optimized, it is determined that optimal mobile phase and condition of gradient elution, the range of linearity be 0.25-5 μ g/ml,
In a few days day to day precision RSD value is respectively less than 2%, and precision is high, and reproducible, average recovery rate 99.73%, RSD value is less than
2%, the rate of recovery is high.
Detailed description of the invention
Fig. 1 is calcitriol detection method standard curve (n=3).
Specific embodiment
Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.In the following examples, the experimental methods for specific conditions are not specified, usually according to conventional strip
Part or according to the normal condition proposed by manufacturer.Unless otherwise stated, otherwise all percentage, ratio, ratio or number is pressed
Poidometer.
Unless otherwise defined, all professional and scientific terms as used herein and meaning familiar to those skilled in the art
Justice is identical.In addition, any method similar to or equal to what is recorded and material can be applied to the method for the present invention.Wen Zhong
The preferred implement methods and materials are for illustrative purposes only.
The preparation of 1 reference substance solution of embodiment
Calcitriol reference substance 5mg is accurately weighed into 100mL volumetric flask, adds mobile phase A to be diluted to scale, obtains stock solution
a；The accurate stock solution a 1mL that measures adds mobile phase A to be diluted to scale, obtains stock solution b (in every 1mL solution into 10mL volumetric flask
Calcitriol is about 5 μ g).
The preparation of 2 test solution of embodiment
10 calcitriol soft capsules (0.25 μ g/) is taken, soft capsule is cut as in 50mL volumetric flask, flowing is added
Sufficiently dissolution content of phase A ultrasound (calcitriol is about 0.5 μ g in every 1mL solution).
The foundation of 3 calcitriol detection method of embodiment
(1) standard curve and the range of linearity
Accurate stock solution b 0.5,1.0,2.0,4.0,6.0, the 10.0mL of measuring uses mobile phase into 10mL volumetric flask respectively
Dilution, shakes up constant volume, and acquired solution Calcitriol concentrations are respectively 0.25,0.5,1,2,3,5 μ g/ml.The above solution is distinguished
10 μ L of sample introduction carries out HPLC detection in 255nm, repeats 3 batches.It is mapped with peak area A to concentration C, carries out linear regression, obtain y=
254681x-21.212, R2=0.9999, standard curve are shown in Fig. 1.
According to result above it is found that the detection method of calcitriol of the invention is linearly closed within the scope of 0.25-5 μ g/ml
System is good.
(2) day to day precision in day
Accurate stock solution b 0.5,4.0, the 10.0mL of measuring with flowing phase dilution, constant volume, shakes into 10mL volumetric flask respectively
Even, acquired solution Calcitriol concentrations are respectively 0.25 μ g/ml (low concentration), 0.5 μ g/ml (middle concentration), 5 μ g/ml (highly concentrated
Degree), by the above solution distinguish 10 μ L of sample introduction, in 255nm carry out HPLC detection, respectively at 0,2,4,6,8h and 0,1,2,3,4d into
Row HPLC measurement, calculates RSD value, and experimental result is shown in Table 1.
1 calcitriol detection method of table in a few days day to day precision RSD (n=3)
According to result above it is found that calcitriol detection method of the invention in a few days day to day precision RSD value is respectively less than
2%, precision is high.
(3) repeated
By the calcitriol test sample of 2 μ g/ml with HPLC analysis method METHOD FOR CONTINUOUS DETERMINATION 9 times, RSD value is 0.87%, is less than
2%, calcitriol detection method of the invention is reproducible.
(4) rate of recovery
The accurate stock solution b 0.5mL and 4.0mL that measures with flowing phase dilution, constant volume, shakes into 10mL volumetric flask respectively
Even, acquired solution Calcitriol concentrations are respectively 0.25 μ g/ml and 0.5 μ g/ml, as reference substance solution, 10 μ L of sample introduction, in
255nm carries out HPLC detection.
The accurate test solution 5mL and 10.0mL that measure with flowing phase dilution, constant volume, shakes into 10mL volumetric flask respectively
Even, acquired solution Calcitriol concentrations are respectively 0.25 μ g/ml (low concentration), 0.5 μ g/ml (middle concentration), 5 μ g/ml (highly concentrated
Degree), as reference substance solution, 10 μ L of sample introduction carries out HPLC detection in 255nm.The results are shown in Table 2.
2 calcitriol detection method rate of recovery experimental result (n=3) of table
According to result above it is found that calcitriol detection method average recovery rate of the invention is that 99.73%, RSD value is small
In 2%, the rate of recovery is high.
The foregoing is merely illustrative of the preferred embodiments of the present invention, the substantial technological content model being not intended to limit the invention
It encloses, substantial technological content of the invention is broadly defined in the scope of the claims of application, any technology that other people complete
Entity or method also or a kind of equivalent change, will if identical with defined in the scope of the claims of application
It is considered as being covered by among the scope of the claims.
Claims (6)
1. a kind of detection method of calcitriol, which is characterized in that detected by HPLC method to calcitriol, the HPLC
Method carries out gradient elution using following condition:
Chromatographic column: C18 column；
Mobile phase A: methanol；
Mobile phase B: methanol and acetonitrile；
Sample volume: 5-10 μ L；
Flow velocity: 1.5-2.0mL/min；
Column temperature: 30-40 DEG C；
Detection wavelength: 255nm.
2. the detection method of calcitriol according to claim 1, wherein the body of methanol and acetonitrile in the Mobile phase B
Product is than being 35-50:50-65.
3. the detection method of calcitriol according to claim 2, wherein the body of methanol and acetonitrile in the Mobile phase B
Product is than being 40:60.
4. the detection method of calcitriol according to claim 1-3, wherein the condition of the gradient elution
Are as follows:
。
5. the detection method of calcitriol according to claim 1-4, wherein the sample volume is 10 μ L.
6. the detection method of calcitriol according to claim 1-4, wherein the column temperature is 38 DEG C.
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| CN201910017625.1A CN109752471A (en) | 2019-01-07 | 2019-01-07 | A kind of detection method of calcitriol |
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ID=66405265
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103142554A (en) * | 2013-03-21 | 2013-06-12 | 青岛正大海尔制药有限公司 | Calcitriol controlled-release capsule and preparation method thereof |
| CN107782810A (en) * | 2016-08-26 | 2018-03-09 | 人福普克药业（武汉）有限公司 | The method for detecting calcitriol content in calcitriol soft capsule |
| CN107782808A (en) * | 2016-08-26 | 2018-03-09 | 人福普克药业（武汉）有限公司 | Determine the method and system of calcitriol soft capsule dissolution rate |
-
2019
- 2019-01-07 CN CN201910017625.1A patent/CN109752471A/en not_active Withdrawn
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103142554A (en) * | 2013-03-21 | 2013-06-12 | 青岛正大海尔制药有限公司 | Calcitriol controlled-release capsule and preparation method thereof |
| CN107782810A (en) * | 2016-08-26 | 2018-03-09 | 人福普克药业（武汉）有限公司 | The method for detecting calcitriol content in calcitriol soft capsule |
| CN107782808A (en) * | 2016-08-26 | 2018-03-09 | 人福普克药业（武汉）有限公司 | Determine the method and system of calcitriol soft capsule dissolution rate |
Non-Patent Citations (4)
| Title |
|---|
| JESUSA JOYCE N. CIRUNA等: "Separation from Related Compounds and Assay of Calcipotriol by High-Performance Liquid Chromatography", 《JOURNAL OF CHROMATOGRAPHIC SCIENCE》 * |
| 李季等: "骨化三醇软胶囊的稳定性研究", 《中国药业》 * |
| 郝晶晶等: "RP-HPLC 测定骨化三醇胶丸中的骨化三醇及反式骨化三醇", 《中国药学杂志》 * |
| 陈阳生等: "HPLC测定骨化三醇胶丸的含量", 《药物分析杂志》 * |
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