CN102095817A - Method for measuring trigonelline hydrochloride in trigonella tincture - Google Patents
Method for measuring trigonelline hydrochloride in trigonella tincture Download PDFInfo
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- CN102095817A CN102095817A CN 201110002937 CN201110002937A CN102095817A CN 102095817 A CN102095817 A CN 102095817A CN 201110002937 CN201110002937 CN 201110002937 CN 201110002937 A CN201110002937 A CN 201110002937A CN 102095817 A CN102095817 A CN 102095817A
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Abstract
The invention provides a method for measuring trigonelline hydrochloride in trigonella tincture, and is characterized in that: a sample passes through a filter membrane of which the thickness is 0.45 microns, and is directly fed and detected by using a NH2 column as an analysis column, and water and acetonitrile as mobile phases by a high performance liquid chromatography, wherein a detector is a diode array detector. The method for measuring the trigonelline hydrochloride in the trigonella tincture specifically comprises the following steps of: (1) drawing a standard working curve: preparing standard solution at at least five concentrations of a trigonelline hydrochloride standard substance, detecting the content of the trigonelline hydrochloride standard substance by using a high performance liquid chromatograph, drawing the standard working curve and calculating to obtain a regression equation of the standard working curve; and (2) preparing a sample: directly injecting the sample after the trigonella tincture passes through the filter membrane of which the thickness is 0.45 microns, respectively substituting the obtained peak areas into the regression equation of the standard working curve, and calculating to obtain the content of the trigonelline hydrochloride in the sample. The method has the advantages that: the pre-treatment step is simple, and the repetitiveness and the recovery rate are high, so the method is applicable to analysis of a large batch of samples; and the blank of analysis and detection of the trigonelline hydrochloride in the trigonella tincture is filled.
Description
Technical field
The present invention relates to alkaloidal detection method, is a kind of method that detects trigonelline in the trigonella tincture.
Background technology
Fenugreek (Trigonellafoenum-graecum L.) has another name called Trigonella foenum-graecum, fragrant clover, Lu Bazi, vanilla, snow Sha, originates in West Asia, north African, and in China, fenugreek is a kind of traditional conventional Chinese medicine.Get fenugreek seed through baking, pulverize the back and get tincture with certain density alcohol extract.Other has tincture and the medicinal extract that utilizes the fenugreek grass to produce, and its fragrance is different from seed extract.Be usually used in the food flavor, can prepare the essence of cocoa, coffee, nut, maple maple and caramel note.In cigarette smoke, can suppress cigarette pungent pungency, cover assorted gas, correct to inhale flavor, increase hay sample baking fragrance.Odor characteristic in the flue gas is inhaled flavor and is characterized as increase body note, nutty, sweet, maple maple perfume (or spice) for increasing body note, this perfume (or spice) of maple maple.
Trigonelline is the main alkaloid of fenugreek medicinal material, has hypoglycemic and antitumor action.At present, the detection of trigonelline mainly contains liquid phase chromatography.Detect the content of trigonelline in the trigonella tincture, the inherent quality of controlling trigonella tincture is had certain reference value, yet the trigonelline in the trigonella tincture detects and do not appear in the newspapers as yet.
Summary of the invention
Purpose of the present invention just is being based on above-mentioned condition and the assay method of trigonelline in a kind of trigonella tincture of providing, has filled up the blank of analyzing and testing trigonelline in the trigonella tincture.
The objective of the invention is to be achieved through the following technical solutions: the assay method of trigonelline is as follows in the trigonella tincture: sample is crossed direct injected behind the 0.45 μ m filter membrane, with NH
2Post is an analytical column, and water and acetonitrile are moving phase, adopts high performance liquid chromatography to detect, and wherein detecting device is a diode array detector.
Assay method specifically may further comprise the steps:
(1) standard working curve is formulated: 1.848 * 10
-3Prepare the standard solution of at least five concentration of trigonelline standard items in the-2.31mg/mL scope with methyl alcohol, utilize high performance liquid chromatograph respectively the content of the standard items in the standard solution of above-mentioned all concentration to be detected again, and according to the peak area of standard items and corresponding concentration drawing standard working curve and calculate its regression equation;
(2) specimen preparation: trigonella tincture is crossed direct injected behind the 0.45 μ m filter membrane, utilize high performance liquid chromatograph to adopt the condition identical respectively sample solution to be detected again with step (1), and bring into respectively according to the peak area that obtains in the regression equation of standard working curve, calculate the content in the sample.
The efficient liquid phase chromatographic analysis condition is as follows:
Chromatographic column: Agilent, NH
2Analytical column (150 mm * 4.6 mm, 5 μ m);
Moving phase: A is a water, and B is an acetonitrile;
Condition of gradient elution: 90%B begins, and 75%B behind the 8min, 50%B behind the 18min, 30%B behind the 23min, 23min are with 90%B balance pillar 5min, and analysis time is 28min altogether;
Flow velocity 1mL/min;
Sample size 5 μ L;
30 ℃ of column temperatures;
Detect wavelength: 265nm.
Why the present invention selects for use with NH
2Post is an analytical column, is that polarity is very big because trigonelline is an inner salt, keeps minimumly on the C18 post, when analyzing trigonelline, be difficult for separating with impurity, so, adopt the NH of polar stationary phase
2Post can strengthen the retention time of trigonelline on post.
And when selecting moving phase for use, select the mixed system of first alcohol and water at first for use, find that the trigonelline chromatographic peak produces hangover, and when adopting acetonitrile to be moving phase, trigonelline chromatographic peak profile symmetry.Final acetonitrile and the water of adopting utilizes NH as moving phase
2Post separates trigonelline.
This method advantage is: pre-treatment is simple, and the repeatability and the recovery are fine, are applicable to the analysis of batch samples.Filled up the blank of analyzing and testing trigonelline in the trigonella tincture.
Description of drawings
Fig. 1 is a trigonelline standard specimen chromatogram.
Fig. 2 is trigonella tincture sample chromatogram figure.
Among the figure: 1 is trigonelline.
Embodiment
Below by embodiment the present invention is further specified:
Chromatographic condition: Agilent 1200 liquid chromatographies, configuration DAD detecting device detects wavelength 265nm.Chromatographic column: Agilent, NH
2Analytical column (150 mm * 4.6 mm, 5 μ m).Moving phase: water (A), acetonitrile (B); Gradient elution, 90%B begins, 75%B behind the 8min, 50%B behind the 18min, 30%B behind the 23min, 23min is with 90%B balance pillar 5min.Analysis time is 28min altogether.Flow velocity: 1.0mL/min.30 ℃ of column temperatures.Sample size 5 μ L.
1) formulation of standard working curve:
Trigonelline is prepared typical curve with methyl alcohol, and mother liquor is the 2.31mg/mL methanol solution.Typical curve is 5,5,5 and 2 times of mother liquor stepwise dilutions, and typical curve has 5 check points.The standard curve range 1.848 * 10 of trigonelline
-3-2.31mg/mL, standard equation are y=6995.5x+26.585, and R2=1, y are chromatographic peak area, and x is trigonelline concentration (mg/mL).
2) sample detection:
Trigonella tincture is crossed direct injected behind the 0.45 μ m filter membrane.
Fig. 1 is the standard specimen chromatogram of trigonelline.Fig. 2 is faenum graecum tincture sample chromatogram figure.
Measure trigonelline content in the trigonella tincture parallel 5 times according to the described method of experimental section, relative standard deviation (RSD) sees Table 1.Adopt the recovery of standard addition of three different interpolation levels to see Table 2.Snr computation quantitative limit according to 10 times.Trigonelline quantitatively is limited to 1.232 * 10 in the trigonella tincture
-4Mg/mL.
Adopt the method for trigonelline content in the HPLC-DAD mensuration trigonella tincture simple after the direct filtration, the repeatability and the recovery are good, detect sensitivity, are suitable for the detection of trigonelline content in the trigonella tincture sample in enormous quantities.
Claims (3)
1. the assay method of trigonelline in the trigonella tincture is characterized in that: sample is crossed direct injected behind the 0.45 μ m filter membrane, with NH
2Post is an analytical column, and water and acetonitrile are moving phase, adopts high performance liquid chromatography to detect, and wherein detecting device is a diode array detector.
2. the assay method of trigonelline in the trigonella tincture according to claim 1, it is characterized in that: this assay method specifically may further comprise the steps:
(1) standard working curve is formulated: 1.848 * 10
-3Prepare the standard solution of at least five concentration of trigonelline standard items in the-2.31mg/mL scope with methyl alcohol, utilize high performance liquid chromatograph respectively the content of the standard items in the standard solution of above-mentioned all concentration to be detected again, and according to the peak area of standard items and corresponding concentration drawing standard working curve and calculate its regression equation;
(2) specimen preparation: trigonella tincture is crossed direct injected behind the 0.45 μ m filter membrane, utilize high performance liquid chromatograph to adopt the condition identical respectively sample solution to be detected again with step (1), and bring into respectively according to the peak area that obtains in the regression equation of standard working curve, calculate the content in the sample.
3. the assay method of trigonelline in the trigonella tincture according to claim 1 and 2, it is characterized in that: the efficient liquid phase chromatographic analysis condition is as follows:
Chromatographic column: Agilent, NH
2Analytical column (150 mm * 4.6 mm, 5 μ m);
Moving phase: A is a water, and B is an acetonitrile;
Condition of gradient elution: 90%B begins, and 75%B behind the 8min, 50%B behind the 18min, 30%B behind the 23min, 23min are with 90%B balance pillar 5min, and analysis time is 28min altogether;
Flow velocity 1mL/min; Sample size 5 μ L; 30 ℃ of column temperatures; Detect wavelength: 265nm.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 201110002937 CN102095817A (en) | 2011-01-08 | 2011-01-08 | Method for measuring trigonelline hydrochloride in trigonella tincture |
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| CN 201110002937 CN102095817A (en) | 2011-01-08 | 2011-01-08 | Method for measuring trigonelline hydrochloride in trigonella tincture |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107319134A (en) * | 2017-06-14 | 2017-11-07 | 江苏隆达生物科技有限公司 | A kind of feed addictive and preparation method for improving animal reproduction performance and resistance against diseases |
| CN107632092A (en) * | 2017-09-14 | 2018-01-26 | 上海上药第生化药业有限公司 | The separation method and its application of carnitine and trigonelline |
| CN114384183A (en) * | 2022-01-18 | 2022-04-22 | 秦皇岛海关技术中心 | Trigonelline detection method and application thereof in honey identification |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101089166A (en) * | 2006-06-12 | 2007-12-19 | 罗海章 | Natural enzymolysis process of trigonella seed |
-
2011
- 2011-01-08 CN CN 201110002937 patent/CN102095817A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101089166A (en) * | 2006-06-12 | 2007-12-19 | 罗海章 | Natural enzymolysis process of trigonella seed |
Non-Patent Citations (1)
| Title |
|---|
| 《中国中药杂志》 20020331 赵怀清 等 《高效液相色谱法测定胡芦巴中胡芦巴碱的含量》 第27卷, 第3期 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107319134A (en) * | 2017-06-14 | 2017-11-07 | 江苏隆达生物科技有限公司 | A kind of feed addictive and preparation method for improving animal reproduction performance and resistance against diseases |
| CN107319134B (en) * | 2017-06-14 | 2018-10-26 | 江苏隆达生物科技有限公司 | A kind of feed addictive and preparation method improving animal reproduction performance and disease resistance |
| CN107632092A (en) * | 2017-09-14 | 2018-01-26 | 上海上药第生化药业有限公司 | The separation method and its application of carnitine and trigonelline |
| CN114384183A (en) * | 2022-01-18 | 2022-04-22 | 秦皇岛海关技术中心 | Trigonelline detection method and application thereof in honey identification |
| CN114384183B (en) * | 2022-01-18 | 2024-02-27 | 秦皇岛海关技术中心 | Method for detecting trigonelline and application of trigonelline in honey identification |
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Application publication date: 20110615 |