CN105301144A - High-performance liquid chromatography of phosphatidylserine - Google Patents
High-performance liquid chromatography of phosphatidylserine Download PDFInfo
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- CN105301144A CN105301144A CN201510874442.3A CN201510874442A CN105301144A CN 105301144 A CN105301144 A CN 105301144A CN 201510874442 A CN201510874442 A CN 201510874442A CN 105301144 A CN105301144 A CN 105301144A
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Abstract
The invention relates to a high-performance liquid chromatography of phosphatidylserine. The High-performance liquid chromatography comprises the steps of (1) preparing a phosphatidylserine standard solution, and using a chloroform-methanol solution to dissolve and fix the constant volume to be 80 to 400ug/ml; (2) preparing a phosphatidylserine sample solution, using the chloroform-methanol solution to dissolve and fix the volume, and diluting and preparing the sample solution of which the phosphatidylserine content is within the concentration range of standard serial solutions; (3) measuring the standard solution and the sample solution under the condition that the high-performance liquid chromatography cooperates with an evaporative light-scattering detector, performing logarithmetics on the peak area of the standard solution as a y-coordinate, performing logarithmetics on the concentration of the standard solution as an x-coordinate, drawing a double logarithmic standard curve, and finding the concentration of the sample solution from the double logarithmic standard curve according to the peak area. The high-performance liquid chromatography has the following beneficial effects: due to the fact that the high-performance liquid chromatography cooperates with the evaporative light-scattering detector, a simple and effective method is provided for the quality control during a phosphatidylserine production process through the entering of different mobile phase volume ratios.
Description
Technical field
The present invention relates to a kind of HPLC analytical method of phosphatidylserine.
Background technology
Phosphatidylserine has significant curative effect to Improving memory and cognitive ability, senile dementia prevention and cure, depression and alleviation stress; it is a kind of functional health raw material of great exploitation potential for its; the particularly raising aspect of cerebral function, has neuroprotective cell in the brain and improves the effect of brain signal conduction function.The detection method of content of current phosphatidylserine has no report, thus lacks the quality control method to phosphatidylserine.
Therefore, be necessary to propose effective technical scheme, solve the problem.
Summary of the invention
The present invention is directed to above-mentioned the deficiencies in the prior art, a kind of HPLC analytical method of accurate detection phosphatidylserine is provided.
The technical scheme that the present invention solves the problems of the technologies described above is as follows:
A HPLC analytical method for phosphatidylserine, is characterized in that, adopt high performance liquid chromatography to coordinate evaporative light-scattering detector, peak area method measures the content of phosphatidylserine in addition; Specifically comprise the steps:
1) configure phosphatidylserine standard solution, dissolving also constant volume with chloroform-methanol becomes 80-400ug/ml;
2) configure phosphatidylserine sample solution, dissolve and constant volume with chloroform-methanol, dilution is mixed with the sample solution of phosphatidylserine content within the scope of standard items series solution concentration;
3) standard solution, sample solution are measured under high performance liquid chromatography coordinates evaporative light-scattering detector condition, do after logarithm as ordinate with standard solution peak area, concentration of standard solution is horizontal ordinate after doing logarithm, draw double-log typical curve, check in sample solution concentration according to peak area from double-log typical curve.
Preferably, the operating conditions of described high performance liquid chromatography is:
Chromatographic column: the stainless steel column of LiChrosphere100diol, 4um*125mm;
Flow velocity: 1.0mL/min;
Column temperature: 30 DEG C;
Mobile phase: mobile phase A composition volume ratio is: normal hexane: isopropyl alcohol: acetic acid: triethylamine=820:170:10:0.8; Mobile phase B composition volume ratio is: isopropyl alcohol: water: acetic acid: triethylamine=850:140:10:0.8;
Detecting device: evaporative light-scattering detector;
Sample size: 10 μ L.
Beneficial effect: the present invention adopts high performance liquid chromatography to coordinate evaporative light-scattering detector, peak area method measures the content of phosphatidylserine in addition, by entering, for the quality control in phosphatidylserine production run provides a simple effective method of difference flowing phase volume ratio.
Embodiment
In a kind of preferred implementation of the present invention, provide a kind of HPLC analytical method of phosphatidylserine, adopt high performance liquid chromatography to coordinate evaporative light-scattering detector, peak area method measures the content of phosphatidylserine in addition; Specifically comprise the steps:
1) configure phosphatidylserine standard solution, dissolving also constant volume with chloroform-methanol becomes 80-400ug/ml;
2) configure phosphatidylserine sample solution, dissolve and constant volume with chloroform-methanol, dilution is mixed with the sample solution of phosphatidylserine content within the scope of standard items series solution concentration;
3) standard solution, sample solution are measured under high performance liquid chromatography coordinates evaporative light-scattering detector condition, do after logarithm as ordinate with standard solution peak area, concentration of standard solution is horizontal ordinate after doing logarithm, draw double-log typical curve, check in sample solution concentration according to peak area from double-log typical curve.
As one embodiment of the present of invention, the operating conditions of described high performance liquid chromatography is:
Chromatographic column: the stainless steel column of LiChrosphere100diol, 4um*125mm;
Flow velocity: 1.0mL/min;
Column temperature: 30 DEG C;
Mobile phase: mobile phase A composition volume ratio is: normal hexane: isopropyl alcohol: acetic acid: triethylamine=820:170:10:0.8; Mobile phase B composition volume ratio is: isopropyl alcohol: water: acetic acid: triethylamine=850:140:10:0.8;
Detecting device: evaporative light-scattering detector;
Sample size: 10 μ L.
Wherein, the use of mobile phase is important concerning the HPLC analytical method of phosphatidylserine of the present invention.Concrete, mobile phase A composition volume ratio is: normal hexane: isopropyl alcohol: acetic acid: triethylamine=820:170:10:0.8; Mobile phase B composition volume ratio is: isopropyl alcohol: water: acetic acid: triethylamine=850:140:10:0.8; Working time is different, and adopt different eluent gradients, namely the volume ratio of mobile phase A and Mobile phase B is different according to the difference of time, is divided into six working times, specifically in table 1.
Table 1 mobile phase and gradient
| Step | Working time/min | Mobile phase A/% | Mobile phase B/% |
| 1 | 0 | 100 | 0 |
| 2 | 20 | 50 | 50 |
| 3 | 25 | 0 | 100 |
| 4 | 29 | 0 | 100 |
| 5 | 30 | 100 | 0 |
| 6 | 35 | 100 | 0 |
The content detection result of phosphatidylserine, substitutes into the percentage composition of phosphatidylserine in following publicity calculation sample:
X＝CV/W(1-F)*10
-6*100％
Wherein,
The percentage composition of phosphatidylserine in X-sample;
The concentration of phosphatidylserine in C-sample solution, ug/ml;
The cumulative volume of V-sample diluting liquid, ml;
W-sample volume, g;
Moisture in F-sample; This moisture detecting method is according to the mensuration of moisture in GB50093-2010 food.
The present invention can summarize with other the concrete form without prejudice to spirit of the present invention or principal character.Therefore, no matter from which point, above-mentioned embodiment of the present invention all can only be thought explanation of the present invention and can not limit the present invention, claims indicate scope of the present invention, and scope of the present invention is not pointed out in above-mentioned explanation, therefore, any change in the implication suitable with claims of the present invention and scope, all should think to be included in the scope of claims of the present invention.
Claims (2)
1. a HPLC analytical method for phosphatidylserine, is characterized in that, adopt high performance liquid chromatography to coordinate evaporative light-scattering detector, peak area method measures the content of phosphatidylserine in addition; Specifically comprise the steps:
1) configure phosphatidylserine standard solution, dissolving also constant volume with chloroform-methanol becomes 80-400ug/ml;
2) configure phosphatidylserine sample solution, dissolve and constant volume with chloroform-methanol, dilution is mixed with the sample solution of phosphatidylserine content within the scope of standard items series solution concentration;
3) standard solution, sample solution are measured under high performance liquid chromatography coordinates evaporative light-scattering detector condition, do after logarithm as ordinate with standard solution peak area, concentration of standard solution is horizontal ordinate after doing logarithm, draw double-log typical curve, check in sample solution concentration according to peak area from double-log typical curve.
2. HPLC analytical method as claimed in claim 1, it is characterized in that, the operating conditions of described high performance liquid chromatography is:
Chromatographic column: the stainless steel column of LiChrosphere100diol, 4um*125mm;
Flow velocity: 1.0mL/min;
Column temperature: 30 DEG C;
Mobile phase: mobile phase A composition volume ratio is: normal hexane: isopropyl alcohol: acetic acid: triethylamine=820:170:10:0.8; Mobile phase B composition volume ratio is: isopropyl alcohol: water: acetic acid: triethylamine=850:140:10:0.8;
Detecting device: evaporative light-scattering detector;
Sample size: 10 μ L.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107688073A (en) * | 2017-11-02 | 2018-02-13 | 威海百合生物技术股份有限公司 | A kind of detection method of phosphatidylserine content |
| CN107727755A (en) * | 2017-09-06 | 2018-02-23 | 南通励成生物工程有限公司 | A kind of detection method of phosphatidylserine |
| CN108931595A (en) * | 2018-06-20 | 2018-12-04 | 广东省测试分析研究所（中国广州分析测试中心） | The measuring method of phosphatidylserine content in a kind of gelatin gel candy |
| CN111721848A (en) * | 2019-03-21 | 2020-09-29 | 仙乐健康科技股份有限公司 | Method for measuring content of phosphatidylserine |
| CN111855844A (en) * | 2020-07-02 | 2020-10-30 | 武汉迈特维尔生物科技有限公司 | Method for analyzing phosphatidylserine |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107727755A (en) * | 2017-09-06 | 2018-02-23 | 南通励成生物工程有限公司 | A kind of detection method of phosphatidylserine |
| CN107688073A (en) * | 2017-11-02 | 2018-02-13 | 威海百合生物技术股份有限公司 | A kind of detection method of phosphatidylserine content |
| CN107688073B (en) * | 2017-11-02 | 2020-12-04 | 威海百合生物技术股份有限公司 | Method for detecting content of phosphatidylserine |
| CN108931595A (en) * | 2018-06-20 | 2018-12-04 | 广东省测试分析研究所（中国广州分析测试中心） | The measuring method of phosphatidylserine content in a kind of gelatin gel candy |
| CN108931595B (en) * | 2018-06-20 | 2021-05-11 | 广东省测试分析研究所（中国广州分析测试中心） | Method for determining content of phosphatidylserine in gelatin type gel candy |
| CN111721848A (en) * | 2019-03-21 | 2020-09-29 | 仙乐健康科技股份有限公司 | Method for measuring content of phosphatidylserine |
| CN111855844A (en) * | 2020-07-02 | 2020-10-30 | 武汉迈特维尔生物科技有限公司 | Method for analyzing phosphatidylserine |
| CN111855844B (en) * | 2020-07-02 | 2021-04-30 | 武汉迈特维尔生物科技有限公司 | Method for analyzing phosphatidylserine |
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Application publication date: 20160203 |