CN112268889A - Method for measuring content of vitamin D3 based on Raman spectroscopy - Google Patents
Method for measuring content of vitamin D3 based on Raman spectroscopy Download PDFInfo
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Abstract
The invention belongs to the technical field of functional component analysis, and provides a method for determining the content of vitamin D3 based on Raman spectroscopy, which comprises the following steps: preparing standard solutions with different vitamin D3 concentrations, performing Raman spectrum scanning on the standard solutions, and preparing a linear regression equation according to the vitamin D3 concentration of the standard solutions and the peak area (namely Raman spectrum intensity) of the Raman spectrogram; and directly carrying out Raman spectrum scanning on the sample containing the vitamin D3, and determining the content of the vitamin D3 in the sample according to the linear regression equation and the peak area of the Raman spectrogram of the sample. The method does not need to carry out complicated pretreatment on the sample, has simple operation, is easy to master, saves time and labor, and can quickly and accurately detect the content of the vitamin D3 in the raw material of the vitamin D3 and the preparation product containing the vitamin D3; the method can finish detection only by one Raman spectrometer, greatly reduces the detection cost and is beneficial to popularization and promotion.
Description
Technical Field
The invention relates to the technical field of functional component analysis, in particular to a method for determining the content of vitamin D3 based on Raman spectroscopy.
Background
The vitamin D3 can improve the absorption of calcium and phosphorus by human body. Products such as nutriment prepared from vitamin D3 are suitable for treating rickets in children caused by vitamin D deficiency, and other diseases caused by calcium deficiency in adults, such as osteoporosis.
At present, most of the existing methods for detecting vitamin D3 and vitamin D3 products are high performance liquid chromatography or liquid chromatography-mass spectrometry. The detection method is mainly based on the fourth method of GB5009.82-2016, and adopts a normal phase-reverse phase dual system detection, so that two high performance liquid chromatographs are required. The detection method has the following defects: 1) the method needs to perform pretreatment such as division, crushing, saponification, extraction, washing, concentration, purification, elution, nitrogen blowing constant volume and the like on a sample, and has the disadvantages of complicated steps and long operation time; 2) in the process of purifying a sample by using a normal phase-liquid phase instrument, the outflow time of vitamin D3 in a pipeline is difficult to accurately master, so that the loss of vitamin D3 is easily caused, and the accuracy of a detection result is poor; 3) the detection method has high requirements on a laboratory, two high performance liquid chromatographs and two chromatographic columns are needed, the investment cost and the maintenance cost of the instrument are very high, the detection cost is high, and the popularization is not facilitated.
Disclosure of Invention
The invention aims to provide a method for measuring the content of vitamin D3 based on Raman spectroscopy, which can quickly and accurately detect the content of vitamin D3 in a vitamin D3 raw material and a preparation product containing vitamin D3 and has lower detection cost.
Based on the above, the invention discloses a method for measuring the content of vitamin D3 based on Raman spectroscopy, which comprises the following steps:
step one, preparing standard solutions with different vitamin D3 concentrations;
step two, respectively scanning the standard solutions with different vitamin D3 concentrations by using a Raman spectrometer to obtain Raman spectrograms of the standard solutions, and then determining peak areas (namely Raman spectral intensities) of the Raman spectrograms of the standard solutions;
thirdly, obtaining a linear regression equation according to the concentration of the vitamin D3 in the standard solution and the peak area of the Raman spectrogram;
weighing a quantitative sample containing vitamin D3, scanning the sample by using a Raman spectrometer to obtain a Raman spectrogram of the sample, and determining the peak area of the Raman spectrogram of the sample;
and step five, determining the content of the vitamin D3 in the sample according to the linear regression equation and the peak area of the Raman spectrogram of the sample.
Preferably, in the first step, the concentration of vitamin D3 in the standard solution is 0.3%, 0.5%, 1%, 2%, 3%, respectively.
Further preferably, in the first step, the standard solution is prepared by the following method: weighing vitamin D3 standard substances with different weights, respectively, and dissolving with methanol to obtain standard solutions with vitamin D3 concentrations of 0.3%, 0.5%, 1%, 2%, and 3%.
Preferably, in step three, the linear regression equation takes the vitamin D3 concentration of the standard solution as the ordinate and the peak area of the Raman spectrum as the abscissa.
Further preferably, in step three, the linear regression equation is: y is 0.018 x-0.013.
Preferably, in the fourth step, when the sample containing vitamin D3 is powder, the sampling weight of the powder is 20-30 mg.
Preferably, in the fourth step, when the sample containing vitamin D3 is a soft capsule, the sampling weight of the content of the soft capsule is 240 mg-260 mg.
Preferably, in the second step and the fourth step, the detection parameters of the raman spectrometer are as follows: the laser source is 785nm, the laser power is 1000mW, the exposure time is 1 second, and the exposure times are 25 times; the Raman spectrometer is 150-2700 cm-1Scanning within the displacement range.
Preferably, in the second step and the fourth step, the vitamin D3 is 1555cm in length when the Raman spectrometer is used for detection-1Has characteristic peaks.
Compared with the prior art, the invention at least comprises the following beneficial effects:
the invention relates to a method for measuring vitamin D3 content based on Raman spectroscopy, which comprises the following steps: preparing standard solutions with different vitamin D3 concentrations, performing Raman spectrum scanning on the standard solutions, and preparing a linear regression equation according to the vitamin D3 concentration of the standard solutions and the peak area of the Raman spectrogram; and directly carrying out Raman spectrum scanning on the sample containing the vitamin D3, and determining the content of the vitamin D3 in the sample according to the linear regression equation and the peak area of the Raman spectrogram of the sample. Compared with the existing detection method, such as high performance liquid chromatography, the detection method provided by the invention does not need to carry out pre-treatment such as division, crushing, saponification, extraction, washing, concentration, purification, elution and nitrogen blowing to fix volume on a sample, but directly carries out Raman spectrum scanning on the sample after weighing a certain amount of the sample containing vitamin D3, is very simple to operate, easy to master, time-saving and labor-saving, and can be used for quickly and accurately detecting the content of vitamin D3 in a vitamin D3 raw material and a preparation product containing vitamin D3; in addition, the existing detection method needs two high performance liquid chromatographs and two stainless steel chromatographic columns, and the detection cost is very high, but the detection method can complete the detection only by one Raman spectrometer, so the detection cost is greatly reduced, and the popularization and the promotion are facilitated, so the detection method has wide market application prospect.
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FIG. 1 shows standard solutions of different vitamin D3 concentrations at 1555cm-1A nearby raman spectrum; wherein a, b, c, D and e respectively represent the concentration of vitamin D3 in standard solution of 0.3%, 0.5%, 1%, 2% and 3% at 1555cm-1Nearby raman spectra.
Fig. 2 is a linear regression equation of vitamin D3 concentration of the standard solution and raman spectrum intensity of the standard solution.
FIG. 3 is a Raman spectrum of vitamin D3 soft gelatin capsule and vitamin D3 standard; wherein, 1 and 2 respectively represent Raman spectra of vitamin D3 soft capsule and vitamin D3 standard substance.
Detailed Description
The present invention will be described in further detail with reference to specific embodiments in order to make the above objects, features and advantages more apparent and understandable.
The method for measuring the content of vitamin D3 based on raman spectroscopy in this embodiment specifically includes the following steps:
firstly, preparation of samples and instruments:
vitamin D3 standard: cholecalciferol (C)27H44O, CAS number: 511-28-4), purity>98 percent; or a standard substance that is certified by a country and awarded a standard substance certificate.
Vitamin D3 soft capsules: (20200515, Nysberg Guangxi (Guangdong) Biotech GmbH).
The instrument comprises the following steps: raman spectroscopy (suzhou excel precision instruments science and technology ltd).
Secondly, the determination steps are as follows:
s1 preparation of standard solutions with different vitamin D3 concentrations: respectively weighing vitamin D3 standard substances with different weights, and respectively adding methanol to dissolve the weighed vitamin D3 standard substances to prepare standard solutions with vitamin D3 concentrations of 0.3%, 0.5%, 1%, 2% and 3%.
Examination of the linear relationship of S2: adjusting instrument parameters of the Raman spectrometer: the laser source is 785nm, the laser power is 1000mW, the exposure time is 1 second, and the exposure times are 25 times; taking the standard solutions with different vitamin D3 concentrations in the step S1, respectively placing the standard solutions into sample tanks of a Raman spectrometer, and respectively adopting the Raman spectrometer with adjusted parameters to be 150-2700 cm-1Scanning within a displacement range, sampling each standard solution with the vitamin D3 concentration for 25 times, and then taking an average value; wherein, referring to figure 1, in the Raman spectrogram, the concentration of vitamin D3 in the scanned standard sample is increased continuously, and the standard sample is 1555cm-1The intensity of the characteristic peak is continuously enhanced, and the concentration of the vitamin D3 in the standard sample and the intensity of the characteristic peak of the Raman spectrogram present an obvious linear rule; thus, the standard solution is usedThe concentration of vitamin D3 (i.e., vitamin D3 content) in the solution was plotted on the ordinate and the raman spectrum intensity of the standard solution was plotted on the abscissa, to obtain a linear regression equation, y, of 0.018x-0.013, see fig. 2.
Raman spectrum of vitamin D3 in S3 sample: taking the vitamin D3 soft capsule as a sample, respectively taking 20-30 mg of a vitamin D3 standard substance and 250 +/-10 mg of the content of the vitamin D3 soft capsule, respectively carrying out Raman spectrum scanning on the weighed content of the vitamin D3 standard substance and the vitamin D3 soft capsule according to the scanning parameters of the step S2 to obtain a Raman spectrum diagram, and referring to a diagram 3, the content of the vitamin D3 standard substance and the vitamin D3 soft capsule is 1555cm in the Raman spectrum diagram-1This is so that vitamin D3 in the contents of the vitamin D3 soft capsule can be detected qualitatively.
S4 calculation of results: and (3) obtaining the peak area of the Raman spectrogram of the vitamin D3 soft capsule to obtain the value of x in the linear regression equation, substituting the value into the linear regression equation y of 0.018x-0.013, and calculating to obtain the concentration of the vitamin D3 in the vitamin D3 soft capsule, namely the content (y).
And (3) performance testing:
the determination method of this example and the current national standard method (i.e. the fourth method of GB 5009.82-2016) were used to determine the vitamin D3 content in three batches of vitamin D3 soft capsules, and the results are shown in table 1 below:
TABLE 1
As can be seen from Table 1, the measurement method of this example has no significant difference from the measurement results of the existing national standard method, and therefore, the measurement method of this example can be used to replace the existing national standard method to detect the content of vitamin D3 in the sample; the determination method of the embodiment can quickly and accurately detect the content of the vitamin D3 in preparation products such as a raw material of the vitamin D3, powder containing the vitamin D3, soft capsules containing the vitamin D3 and the like, and greatly reduces the detection cost.
While preferred embodiments of the present invention have been described, additional variations and modifications of these embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the embodiments of the invention.
The technical solutions provided by the present invention are described in detail above, and the principle and the implementation of the present invention are explained in this document by applying specific examples, and the descriptions of the above examples are only used to help understanding the method and the core idea of the present invention; meanwhile, for a person skilled in the art, according to the idea of the present invention, there may be variations in the specific embodiments and the application scope, and in summary, the content of the present specification should not be construed as a limitation to the present invention.
Claims (9)
1. A method for measuring the content of vitamin D3 based on Raman spectroscopy is characterized by comprising the following steps:
step one, preparing standard solutions with different vitamin D3 concentrations;
scanning the standard solutions with different vitamin D3 concentrations by using a Raman spectrometer to obtain Raman spectrograms of the standard solutions, and determining peak areas of the Raman spectrograms of the standard solutions;
thirdly, obtaining a linear regression equation according to the concentration of the vitamin D3 in the standard solution and the peak area of the Raman spectrogram;
weighing a quantitative sample containing vitamin D3, scanning the sample by using a Raman spectrometer to obtain a Raman spectrogram of the sample, and determining the peak area of the Raman spectrogram of the sample;
and step five, determining the content of the vitamin D3 in the sample according to the linear regression equation and the peak area of the Raman spectrogram of the sample.
2. The method for measuring the content of vitamin D3 based on Raman spectroscopy according to claim 1, wherein in the first step, the concentration of vitamin D3 in the standard solution is 0.3%, 0.5%, 1%, 2%, 3%, respectively.
3. The method for measuring the content of vitamin D3 based on Raman spectroscopy according to claim 2, wherein in the first step, the standard solution is prepared by: weighing vitamin D3 standard substances with different weights, respectively, and dissolving with methanol to obtain standard solutions with vitamin D3 concentrations of 0.3%, 0.5%, 1%, 2%, and 3%.
4. The method for determining the content of vitamin D3 by Raman spectroscopy according to claim 1, wherein in step three, the linear regression equation has the vitamin D3 concentration of the standard solution as the ordinate, and the peak area of the Raman spectrum as the abscissa.
5. The method for measuring the content of vitamin D3 based on Raman spectroscopy according to claim 4, wherein in step three, the linear regression equation is as follows: y is 0.018 x-0.013.
6. The method for measuring the content of vitamin D3 based on Raman spectroscopy according to claim 1, wherein in step four, when the sample containing vitamin D3 is powder, the sampling weight of the powder is 20-30 mg.
7. The method for measuring vitamin D3 content based on Raman spectroscopy of claim 1, wherein in step four, when the sample containing vitamin D3 is soft capsule, the sampling weight of the content of the soft capsule is 240 mg-260 mg.
8. The method for measuring the content of vitamin D3 based on Raman spectroscopy as claimed in claim 1, wherein in step two and step four, the detection parameters of the Raman spectrometer are as follows: the laser source is 785nm, the laser power is 1000mW, the exposure time is 1 second, and the exposure times are 25 times; the pull rodThe Raman spectrometer is 150-2700 cm-1Scanning within the displacement range.
9. The method for determining vitamin D3 content by Raman spectroscopy as claimed in claim 1, wherein in step two and step four, vitamin D3 is 1555cm-1Has characteristic peaks.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102253130A (en) * | 2010-12-24 | 2011-11-23 | 天津天狮生物发展有限公司 | Method for measuring vitamins A, D and E in compound vitamin |
| CN106033064A (en) * | 2015-03-10 | 2016-10-19 | 上海信谊药厂有限公司 | A cholecalciterol cholesterol measuring method |
| CN106323942A (en) * | 2016-09-08 | 2017-01-11 | 福建师范大学 | Fast detecting method of grease peroxidation value |
| CN106383106A (en) * | 2016-09-08 | 2017-02-08 | 福建师范大学 | Quick detection method of trace antioxidant in grease |
| CN108802000A (en) * | 2018-03-16 | 2018-11-13 | 上海交通大学 | A kind of lossless quick cholecalciferol-cholesterol content quantitative method based on the full spectrum analysis of Raman |
-
2020
- 2020-11-13 CN CN202011267572.8A patent/CN112268889A/en active Pending
- 2020-11-13 CN CN202310046160.9A patent/CN115901727A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102253130A (en) * | 2010-12-24 | 2011-11-23 | 天津天狮生物发展有限公司 | Method for measuring vitamins A, D and E in compound vitamin |
| CN106033064A (en) * | 2015-03-10 | 2016-10-19 | 上海信谊药厂有限公司 | A cholecalciterol cholesterol measuring method |
| CN106323942A (en) * | 2016-09-08 | 2017-01-11 | 福建师范大学 | Fast detecting method of grease peroxidation value |
| CN106383106A (en) * | 2016-09-08 | 2017-02-08 | 福建师范大学 | Quick detection method of trace antioxidant in grease |
| CN108802000A (en) * | 2018-03-16 | 2018-11-13 | 上海交通大学 | A kind of lossless quick cholecalciferol-cholesterol content quantitative method based on the full spectrum analysis of Raman |
Non-Patent Citations (4)
| Title |
|---|
| 国家药典委员会: "《中华人民共和国药典四部》", 31 May 2020, 中国医药科技出版社 * |
| 戴健等: "胆维丁原料中共晶与游离态维生素D_3和胆固醇含量的定量分析", 《上海医药》 * |
| 肖新月 等: "《化学药品对照品图谱集—红外、拉曼、紫外光谱》", 31 August 2014 * |
| 赖源发 等: "维生素C片的拉曼光谱法测定", 《中国医药工业杂志》 * |
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