CN105717056A - Method for measuring content of lutein dipalmitate in lutein ester product - Google Patents
Method for measuring content of lutein dipalmitate in lutein ester product Download PDFInfo
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- CN105717056A CN105717056A CN201610189029.8A CN201610189029A CN105717056A CN 105717056 A CN105717056 A CN 105717056A CN 201610189029 A CN201610189029 A CN 201610189029A CN 105717056 A CN105717056 A CN 105717056A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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Abstract
The invention discloses a method for measuring the content of lutein dipalmitate in a lutein ester product. The method comprises steps as follows: the lutein ester product is mixed with ammonia water, trichloromethane and ethanol, a mixture is subjected to ultrasonic processing, and a first mixed solution is obtained; the first mixed solution is subjected to constant volume processing and mixing by the aid of ethanol, and a second mixed solution is obtained; the second mixed solution is diluted with hexyl hydride, and a to-be-measured liquid is obtained; the to-be-measured liquid is detected with a spectrophotometric method, and the content of lutein dipalmitate in the lutein ester product is determined on the basis of a detection result. The method for measuring the content of lutein dipalmitate in the lutein ester product has the advantages of high accuracy, good stability and high repeatability.
Description
Technical field
The present invention relates to field of food.In particular it relates to the method measuring lutein ester product Lutein dipalmitate content.
Background technology
Lutein ester, chemical name is lutein dipalmitate, is a kind of important carotenoid fatty acid ester, character: reddish brown fine particle.Primary structure is to be connected by the long-chain of conjugated double bond containing ten eight carbon atoms by two six-membered carbon rings to form, molecular weight: 1045.71, molecular formula: C72H116O4。
Natural carotenol ester is a kind of important carotenoid fatty acid ester, and main is configured as alltrans, has the trans groups of C=C long-chain, shows gorgeous golden yellow.Both sides do not have active hydroxyl, therefore just determine that lutein ester is a kind of sufficiently stable food color yet.Light, heat and air is highly stable, the applications such as food, beverage, cosmetics can be widely used in, have broad application prospects.Meanwhile, it is after Over emulsfication, it is possible to make water-soluble colorant.Natural carotenol ester will substitute the food grade coloring agent of chemosynthesis, improves the health level of people.
But, the method measuring lutein ester product Lutein dipalmitate content at present still haves much room for improvement.
Summary of the invention
It is contemplated that at least solve one of technical problem of existence in prior art.For this, it is an object of the present invention to provide a kind of method measuring lutein ester product Lutein dipalmitate content.The method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
It should be noted that the present invention is based on the following discovery of inventor and completes:
Adopting in prior art and lutein ester and normal hexane/isopropanol are mixed and sonicated for, but supersound process is after nearly 10 hours, lutein ester still can not be completely dissolved, and then affects the accuracy of result.
In view of this, the present inventor finds through great many of experiments, by changing the solvent dissolving lutein ester, select ammonia, ethanol and chloroform as solvent, alkaline environment is provided for lutein ester, it is thus possible to make lutein ester be completely dissolved, and then can exactly lutein ester be detected.Thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
For this, in a first aspect of the present invention, the present invention proposes a kind of method measuring lutein ester product Lutein dipalmitate content.According to embodiments of the invention, described method includes: mixed with ammonia, chloroform and ethanol by lutein ester product, and the mixture obtained is carried out supersound process, obtains the first mixed liquor；Utilize ethanol that described first mixed liquor is carried out constant volume and mixed, obtain the second mixed liquor；Utilize normal hexane to be diluted by described second mixed liquor, obtain liquid to be measured；And adopt spectrophotography that described liquid to be measured is detected, and determine lutein ester product Lutein dipalmitate content based on testing result.Inventors discovered unexpectedly that, by dissolving in lutein dipalmitate system, addition ammonia is to change the acid-base value of system, for lutein dipalmitate offer alkali condition, it is possible to make lutein dipalmitate be completely dissolved in solvent.Further, changing the organic solvent dissolving lutein dipalmitate, and accelerate when supersound process to dissolve, final lutein dipalmitate can be dissolved completely in solvent, thus ensure that the accuracy of testing result.Thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
According to embodiments of the invention, the volume ratio of described ammonia, chloroform and ethanol is 3:2:5.Thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
According to embodiments of the invention, described supersound process is to carry out in the water-bath of 45 degrees Celsius 30 minutes.Thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
According to embodiments of the invention, the concentration of described ethanol is 95 volume %.Thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
In a second aspect of the present invention, the present invention proposes a kind of method measuring lutein ester product Lutein dipalmitate content.According to embodiments of the invention, described method includes: (1) weighs 0.1g lutein ester in beaker, adds ammonia 3ml, chloroform 2ml, 95% ethanol 5ml, then beaker is placed in 30min in 45 DEG C of ultrasonic water bath pots；(2) mixed liquor obtained for step (1) is proceeded in volumetric flask, be settled to graduation mark with 95% ethanol, mixing；(3) take the obtained solution of 1ml step (2) in volumetric flask, be diluted with normal hexane, mixing, obtain liquid to be measured；(4) with normal hexane, spectrophotometer is returned to zero, the maximum absorbance of described liquid to be measured is measured under 445nm wavelength, and calculate lutein ester product Lutein dipalmitate content based on following equation, computing formula: lutein dipalmitate (%)=(A × B)/(1394 × m), wherein: A absorbance；The extension rate of B sample；M sample quality；1394 specific absorbances.Thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
The additional aspect of the present invention and advantage will part provide in the following description, and part will become apparent from the description below, or is recognized by the practice of the present invention.
Detailed description of the invention
Embodiments of the invention are described below in detail.The embodiments described below is illustrative of, and is only used for explaining the present invention, and is not considered as limiting the invention.
It should be noted that term " first ", " second " are only for descriptive purposes, and it is not intended that indicate or imply relative importance or the implicit quantity indicating indicated technical characteristic.Thus, define " first ", the feature of " second " can express or implicitly include one or more these features.Further, in describing the invention, except as otherwise noted, " multiple " are meant that two or more.
The present invention proposes a kind of method measuring lutein ester product Lutein dipalmitate content.According to embodiments of the invention, the method comprises the following steps, thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
According to embodiments of the invention, the method measuring lutein ester product Lutein dipalmitate content includes:
(1) lutein ester product is mixed with ammonia, chloroform and ethanol, and the mixture obtained is carried out supersound process, obtain the first mixed liquor.
Inventors discovered unexpectedly that, lutein dipalmitate dissolubility in alkaline environment is relatively strong, obtains optimum basic solvent ammonia by great many of experiments optimization.Lutein dipalmitate dissolubility in the solvent containing other alkaline matters is below aqueous ammonia solvent, for instance alkaline sodium hydroxide and weakly alkaline sodium citrate.Further, inventor have found that, lutein dipalmitate dissolubility in the organic solvent containing chloroform is better, additionally, add ethanol in a solvent can be effectively prevented chloroform because of the incidence rate producing phosgene of being heated.Thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
According to embodiments of the invention, the volume ratio of ammonia, chloroform and ethanol is 3:2:5.Inventor have found that, the adding proportion of ammonia, chloroform and ethanol is more significant on testing result impact.Inventor obtains above-mentioned optimum adding proportion through great many of experiments optimization.Thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
According to embodiments of the invention, supersound process is to carry out in the water-bath of 45 degrees Celsius 30 minutes.The purpose of supersound process is to accelerate the rate of dissolution of lutein dipalmitate, and in the water-bath of 45 degrees Celsius, supersound process can make lutein dipalmitate be completely dissolved for 30 minutes, thus ensure that the accuracy of testing result.If bath temperature is too low or ultrasonic time is too short, it is impossible to make lutein dipalmitate be completely dissolved, cause that testing result is on the low side.Thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
According to embodiments of the invention, the concentration of ethanol is 95 volume %.Thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
(2) utilize ethanol that described first mixed liquor is carried out constant volume and mixed, obtain the second mixed liquor.Owing to the volume of the first mixed liquor cannot be determined, so it is carried out constant volume, it is simple to follow-up calculating.
(3) utilize normal hexane to be diluted by described second mixed liquor, obtain liquid to be measured.Owing to the second mixed liquor Lutein dipalmitate content is higher, if directly it being detected, upper limit of detection will be exceeded, so needing it is diluted, in order to subsequent detection.
(4) adopt spectrophotography that liquid to be measured is detected, and determine lutein ester product Lutein dipalmitate content based on testing result.According to embodiments of the invention, under 445nm wavelength, measure the maximum absorbance of liquid to be measured, and determine lutein ester product Lutein dipalmitate content according to maximum absorbance value.Thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
It addition, the present invention proposes the another kind of method measuring lutein ester product Lutein dipalmitate content.According to embodiments of the invention, the method includes: (1) weighs 0.1g lutein ester in beaker, adds ammonia 3ml, chloroform 2ml, 95% ethanol 5ml, then beaker is placed in 30min in 45 DEG C of ultrasonic water bath pots；(2) mixed liquor obtained for step (1) is proceeded in volumetric flask, be settled to graduation mark with 95% ethanol, mixing；(3) take the obtained solution of 1ml step (2) in volumetric flask, be diluted with normal hexane, mixing, obtain liquid to be measured；(4) with normal hexane, spectrophotometer is returned to zero, the maximum absorbance of liquid to be measured is measured under 445nm wavelength, and calculate lutein ester product Lutein dipalmitate content based on following equation, computing formula: lutein dipalmitate (%)=(A × B)/(1394 × m), wherein: A absorbance；The extension rate of B sample；M sample quality；1394 specific absorbances.Thus, the method accuracy measuring lutein ester product Lutein dipalmitate content according to embodiments of the present invention is strong, good stability and repeatability high.
Below in conjunction with embodiment, the solution of the present invention is made an explanation.It will be understood to those of skill in the art that the following examples are merely to illustrate the present invention, and should not be taken as limiting the scope of the invention.Unreceipted concrete technology or condition in embodiment, technology or condition described by the document in this area or carry out according to product description.Agents useful for same or the unreceipted production firm person of instrument, be can pass through city available from conventional products.
Embodiment 1
In this embodiment, lutein ester product Lutein dipalmitate content is measured in following manner:
1, reagent:
Ammonia (analytical pure), chloroform (analytical pure), ethanol (95%) (analytical pure)
2, instrument and equipment:
Ultraviolet-uisible spectrophotometer；
Quartz colorimetric utensil (10mm)；
Volumetric flask (100ml)；
Beaker (100ml)；
Analytical balance；
Ultrasonic water bath pot (temperature-adjustable).
3, analytical procedure
(1) 0.1g lutein ester is accurately weighed in 100ml beaker, add ammonia 3ml, chloroform 2ml, ethanol (95%) 5ml, then beaker is placed in 30min in 45 DEG C of ultrasonic water bath pots (with surface plate, beaker being built reduction solvent to volatilize).
(2) mixed liquor obtained for step (1) is proceeded in 100ml volumetric flask, be settled to graduation mark with ethanol (95%), mixing.
(3) take the obtained solution of 1ml step (2) in 100ml volumetric flask, add normal hexane by diluted sample to scale, mixing, obtain liquid to be measured.
(4) with normal hexane, spectrophotometer is returned to zero, under 445nm wavelength, measure the maximum absorbance of liquid to be measured, and calculate lutein ester product Lutein dipalmitate content based on following equation.
Computing formula: lutein dipalmitate (%)=(A × B)/(1394 × m)
Wherein: A=absorbance；The extension rate of B=sample；M=sample quality；1394=specific absorbance.
Comparative example 1
Measure lutein ester product Lutein dipalmitate content according to the method for embodiment 1, be distinctive in that,
A, " ammonia 3ml, chloroform 2ml, ethanol (95%) 5ml " is replaced with " 50mL normal hexane/isopropanol (volume ratio is 95:5) solution ".
B, sonication treatment time are 10 hours.
In detection process, the solution Lutein ester that the step (1) of embodiment 1 is obtained is completely dissolved, and in the step of comparative example 1 (1), still it can be seen that particulate material after ultrasonic 10 hours, and then found out by table 1, the obtained lutein dipalmitate content of comparative example 1 is relatively low, illustrates that it is not completely dissolved, causes that testing result is on the low side.
The impact on testing result of table 1 different solvents
Embodiment 2
Liquid stability analysis to be measured
After being placed 1 minute, 3 minutes, 6 minutes, 9 minutes by liquid to be measured obtained for the step (3) of embodiment 1 respectively, carrying out the detection of step (4), result is as shown in table 2.It can be seen that lutein dipalmitate had good stability in 9 minutes, thus ensure that the accuracy of testing result.
Table 2 stability analysis
Embodiment 3
Repeatability is analyzed
Repeating to detect according to the method for embodiment 1 by same sample, result is as shown in table 3.It can be seen that maximum extreme difference is 0.41, further relate to its repeatability better.
Table 3 repeatability is analyzed
Comparative example 2
In this comparative example, measuring lutein ester product Lutein dipalmitate content according to the method for embodiment 1, be distinctive in that, the addition of ammonia, chloroform and ethanol is as shown in table 4.It can be seen that ammonia according to embodiments of the present invention, chloroform and ethanol addition can fully dissolve lutein dipalmitate, thus measuring its content exactly.
The impact on testing result of the addition of table 4 ammonia, chloroform and ethanol
In the description of this specification, specific features, structure, material or feature that the description of reference term " embodiment ", " some embodiments ", " example ", " concrete example " or " some examples " etc. means in conjunction with this embodiment or example describe are contained at least one embodiment or the example of the present invention.In this manual, the schematic representation of above-mentioned term is necessarily directed to identical embodiment or example.And, the specific features of description, structure, material or feature can combine in one or more embodiments in office or example in an appropriate manner.Additionally, when not conflicting, the feature of the different embodiments described in this specification or example and different embodiment or example can be carried out combining and combining by those skilled in the art.
Although above it has been shown and described that embodiments of the invention, it is understandable that, above-described embodiment is illustrative of, it is impossible to be interpreted as limitation of the present invention, and above-described embodiment can be changed, revises, replace and modification by those of ordinary skill in the art within the scope of the invention.
Claims (5)
1. the method measuring lutein ester product Lutein dipalmitate content, it is characterised in that including:
(1) weigh 0.1g lutein ester in beaker, add ammonia 3ml, chloroform 2ml, 95% ethanol 5ml, then beaker is placed in 30min in 45 DEG C of ultrasonic water bath pots；
(2) mixed liquor obtained for step (1) is proceeded in volumetric flask, be settled to graduation mark with 95% ethanol, mixing；
(3) take the obtained solution of 1ml step (2) in volumetric flask, be diluted with normal hexane, mixing, obtain liquid to be measured；
(4) with normal hexane, spectrophotometer is returned to zero, under 445nm wavelength, measures the maximum absorbance of described liquid to be measured, and calculate lutein ester product Lutein dipalmitate content based on following equation,
Computing formula: lutein dipalmitate (%)=(A × B)/(1394 × m),
Wherein: A absorbance；The extension rate of B sample；M sample quality；1394 specific absorbances.
2. the method measuring lutein ester product Lutein dipalmitate content, it is characterised in that including:
Lutein ester product is mixed with ammonia, chloroform and ethanol, and the mixture obtained is carried out supersound process, obtain the first mixed liquor；
Utilize ethanol that described first mixed liquor is carried out constant volume and mixed, obtain the second mixed liquor；
Utilize normal hexane to be diluted by described second mixed liquor, obtain liquid to be measured；And
Adopt spectrophotography that described liquid to be measured is detected, and determine lutein ester product Lutein dipalmitate content based on testing result.
3. method according to claim 2, it is characterised in that the volume ratio of described ammonia, chloroform and ethanol is 3:2:5.
4. method according to claim 2, it is characterised in that described supersound process is to carry out in the water-bath of 45 degrees Celsius 30 minutes.
5. method according to claim 2, it is characterised in that the concentration of described ethanol is 95 volume %.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113588577A (en) * | 2021-07-30 | 2021-11-02 | 河北东之星生物科技股份有限公司 | Method for detecting content of lutein in chrysanthemum granules |
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Application publication date: 20160629 |