CN109655536A - The detection method of content of Sunfat GDC-S and capric acid diglyceride in a kind of medium chain triglyceride - Google Patents
The detection method of content of Sunfat GDC-S and capric acid diglyceride in a kind of medium chain triglyceride Download PDFInfo
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Abstract
The present invention relates to the detection method of content of Sunfat GDC-S and capric acid diglyceride in a kind of medium chain triglyceride, the following steps are included: the preparation of (1) test solution: taking medium chain triglyceride, methanol is added to dissolve and dilute, shakes up, obtains test solution；(2) preparation of reference substance solution: taking Sunfat GDC-S and capric acid diglyceride reference substance, and methanol is added and dissolves and dilutes, obtains mixed reference substance solution；(3) assay: precision measures test solution and mixed reference substance solution injects liquid chromatograph, records chromatogram；By external standard method respectively with the content of Sunfat GDC-S and capric acid diglyceride in calculated by peak area test sample.Measuring method of the invention is practical, and easy to operate, result is accurate, favorable reproducibility, and the quality standard for medium chain triglyceride is worked out and preparation process screening provides data and supports and reference, guarantees science and advance that diglyceride measures.
Description
Technical field
The present invention relates to the detection method of content of Sunfat GDC-S and capric acid diglyceride in a kind of medium chain triglyceride,
Belong to medicinal raw material and field of food detection.
Background technique
Medium chain triglyceride (Medium-chain triglycerides, MCT) also known as midchain oil, are by coconut
The endosperm of Cocos nucifem L. do something in disregard of obstacles part or by oil palm Elaeisguineensis Jacq. dry endosperm extract rouge
The pungent capric acid mixed acid of the fatty acid-that fat oil separates out, the triglyceride mixture with saturated fatty acid made of glycerine esterification.One
As think, typical MCT refer to saturation Trivent OCG or be saturated Triglyceride DDD or be saturated caprylic-capric acid mixing it is sweet
Oily three esters, belong to a kind of modified product of natural grease.It is a kind of safe and reliable product to domestic and international application attestation for many years,
It can be used as emulsifier, lubricant and stabilizer to be widely applied, the original as productions such as drug, food, health care product and cosmetics is auxiliary
Material.The medium chain triglyceride applied in pharmaceutical production at present, be used as in/raw materials for production of long chain fat emulsion injection.
Using medium-chain fatty acid (MCFA) and glycerol as raw material, medical medium chain triglyceride is prepared by chemistry or enzymatic technique
Three esters, wherein diglyceride is one of Main By product.Diglyceride (Diacylglycerol, DG) is a kind of glycerol three
The structured lipid that a fatty acid is replaced by alkyl in ester (Triacylglycerol, TG) is micro in natural plant grease
Ingredient is primarily present 1,3-DG and 1, two kinds of isomers of 2-DG, and content is often less than 5%, be generally recognized as safe food at
Point, while being also a kind of multifunction additive, it is had broad application prospects in industries such as food, medicine, chemical industry.In order to obtain
Meet the middle chain structure ester raw material of medical injection Fat Emulsion requirement, it is necessary to control by-product glycerin diester.
In current patent both at home and abroad, document, there has been no about Sunfat GDC-S in medium chain triglyceride and capric acid glycerol
The report of the absolute quantity measuring method of diester.Currently, most common diglyceride analysis method have thin-layered chromatography (TLC), efficiently
Liquid chromatography (HPLC) and gas chromatography (GC) etc. are several.The shortcomings that the shortcomings that TLC method is that precision is not high, GC method is desirable
High temperature column, the big, poor reproducibility of response fluctuation etc., HPLC method is most suitable for analysis diacylglycerol content.HPLC method is according to detector
A variety of methods such as type generally HPLC-UV, HPLC-ELSD.HPLC-UV is apparent the disadvantage is that UV detector is double to fatty acid
Bond number is very sensitive, therefore the different substance of saturation degree can produce the huge response of difference, in the quantitative analysis for doing grease
Aspect poor feasibility.HPLC-ELSD mass type detection method owned by France, be most suitable for, most general esters analysis method, have point
Debate rate is high, speed is fast, high sensitivity, chromatographic column can Reusability the advantages that.
Total amount containing diglyceride is no more than 2% in medium chain triglyceride, because of complicated component and content is low, with efficient liquid
Mutually being measured can be by the interference effect at triglycerides principal component peak, and there is also isomers for diglyceride, so
After chromatographic peak is kept completely separate, then carry out Sunfat GDC-S and capric acid diglyceride absolute magnitude measurement difficulty it is very big.
Summary of the invention
It is provided it is an object of the invention to overcome in place of above-mentioned the deficiencies in the prior art pungent in a kind of medium chain triglyceride
The detection method of content of acid triglyceride and capric acid diglyceride, detection method of the invention is practical, easy to operate, result
Accurately, favorable reproducibility, the quality standard for medium chain triglyceride is worked out and preparation process screening provides data and supports and refer to.
To achieve the above object, a kind of the technical solution adopted by the present invention are as follows: Sunfat GDC-S in medium chain triglyceride
With the detection method of content of capric acid diglyceride, comprising the following steps:
(1) preparation of test solution: taking medium chain triglyceride, and methanol is added and dissolves and dilutes, shakes up, obtains test sample
Solution；
(2) preparation of reference substance solution: taking Sunfat GDC-S and capric acid diglyceride reference substance, and methanol dissolution is added simultaneously
Dilution, obtains the mixed reference substance solution of Sunfat GDC-S and capric acid diglyceride；
(3) assay: precision measures test solution and mixed reference substance solution injects liquid chromatograph, records chromatography
Figure；By external standard method respectively with the content of Sunfat GDC-S and capric acid diglyceride in calculated by peak area test sample.
As the preferred embodiment of detection method of the present invention, in the step (1), medium chain triglyceride and methanol
Mass volume ratio be 200mg:10mL.
As the preferred embodiment of detection method of the present invention, in the step (2), precision weighs sad glycerol two
Ester and capric acid diglyceride reference substance are added methanol and dissolve and dilute, and it is respectively 70 that every 1mL methanol, which is made, containing Sunfat GDC-S
μ g, 150 μ g, 210 μ g, 300 μ g, 450 μ g, diglyceride containing capric acid be respectively 30 μ g, 60 μ g, 90 μ g, 120 μ g, 180 μ g it is mixed
Close reference substance solution.
As the preferred embodiment of detection method of the present invention, in the step (3), liquid phase chromatogram condition are as follows:
Chromatographic column: C18 chromatographic column；
Detector: evaporative light scattering detector；
Mobile phase: using volume ratio for 8:2 acetonitrile-water mixed solvent as mobile phase A, it is different for the acetonitrile-of 1:1 with volume ratio
Propanol solvent mixture is that Mobile phase B carries out gradient elution；The condition of the eluent gradient elution are as follows: 0~10min, mobile phase
Middle A (76%), B (24%) in mobile phase；10~30min, A (76% → 30%) in mobile phase, B in mobile phase (24% →
70%)；30~40min, A (30%) in mobile phase, B (70%) in mobile phase；40~50min, A in mobile phase (30% →
76%), B (70% → 24%) in mobile phase；50~60min, A (76%) in mobile phase, B (24%) in mobile phase；
Flow velocity: 1.0mL/min；
Column temperature: 30 DEG C；
It is accurate respectively to measure test solution and 20 μ L of mixed reference substance solution, inject high performance liquid chromatograph, measurement.
As the preferred embodiment of detection method of the present invention, in the step (3), the concrete operations of external standard method
Are as follows: the 20 μ L of mixed reference substance solution that precision measures various concentration injects liquid chromatograph, measures various concentration mixing reference substance
The peak area of solution, using mixed reference substance solution peak area logarithm as ordinate, with corresponding mixed reference substance solution concentration
Logarithm is that abscissa makees standard curve, calculates separately to obtain equation of linear regression；The another accurate 20 μ L of test solution that measures injects liquid
Chromatography measures the peak area of Sunfat GDC-S and capric acid diglyceride in test sample, pungent with regression equation calculation respectively
The content of acid triglyceride and capric acid diglyceride.
Sunfat GDC-S has 1,3- Sunfat GDC-S, 1,2- Sunfat GDC-S, two isomers, so 1,
3- Sunfat GDC-S, 1,2- Sunfat GDC-S content calculate when, while using Sunfat GDC-S as reference substance, it is pungent
Acid triglyceride content is sum of the two.Similarly, capric acid diglyceride has 1,3- capric acid diglyceride, 1,2- capric acid diglyceride
Two isomers, so when the content of 1,3- capric acid diglyceride, 1,2- capric acid diglyceride calculates, while using capric acid
For diglyceride as reference substance, capric acid diacylglycerol content is sum of the two.
As the preferred embodiment of detection method of the present invention, the specification of the C18 chromatographic column be 250mm ×
4.6mm, 5 μm.
As the preferred embodiment of detection method of the present invention, the drift tube temperature of the detector is 72 DEG C, is carried
Gas velocity is 2.0mL/min, and yield value 4, mode is shunt mode.
As the preferred embodiment of detection method of the present invention, the content of the Sunfat GDC-S is 1,3- octanoic acid
The sum of diglyceride, content of 1,2- Sunfat GDC-S are all made of Sunfat GDC-S conduct pair in mixed reference substance solution
According to；The content of the capric acid diglyceride is the sum of 1,3- capric acid diglyceride, the content of 1,2- capric acid diglyceride, is all made of
Capric acid diglyceride is as control in mixed reference substance solution.
Compared with the existing technology, the invention has the benefit that
(1) present invention establishes a kind of simplicity, accurately and reliably measuring method, using liquid chromatogram isolation technics, evaporation
Light scattering detector detection, the content of triglyceride component in medium chain triglyceride is calculated using liquid phase external standard method, and test sample is straight
It connects and is dissolved with methanol, is not required to by hydrolysis, oxidizing ferment, derivatization, esterification, i.e., using liquid phase measurement and to calculate its pungent
The content of acid triglyceride and capric acid triglyceride component；
(2) the triglyceride component chromatographic peak peak shape of liquid chromatographic system good separating effect of the invention, test sample is good,
Separating degree is excellent between each chromatographic peak, reaches baseline separation, therefore the detection method can accurately and effectively measure medium chain triglyceride sample
The content of triglyceride component in product；
(3) measuring method of the invention is practical, and it is medium chain triglyceride that easy to operate, result is accurate, favorable reproducibility
Quality standard work out and preparation process screen provide data support and reference, guarantee diglyceride measurement science and it is advanced
Property.
Detailed description of the invention
Fig. 1 is Sunfat GDC-S reference substance solution HPLC map of the present invention.
Fig. 2 is capric acid diglyceride reference substance solution HPLC map of the present invention.
Fig. 3 is the separated component 1,3- Sunfat GDC-S HPLC map of medium chain triglyceride of the present invention.
Fig. 4 is the separated component 1,2- Sunfat GDC-S HPLC map of medium chain triglyceride of the present invention.
Fig. 5 is the separated component 1,3- capric acid diglyceride HPLC map of medium chain triglyceride of the present invention.
Fig. 6 is the separated component 1,2- capric acid diglyceride HPLC map of medium chain triglyceride of the present invention.
Fig. 7 is medium chain triglyceride test sample HPLC map of the present invention.
Fig. 8 is medium chain triglyceride test sample HPLC map (0~20min enlarged drawing) of the present invention.
Fig. 9 be medium chain triglyceride repeatability of the present invention measurement mixed reference substance solution HPLC map (0~20min is put
Big figure).
Figure 10 is that medium chain triglyceride repeatability of the present invention measures test sample HPLC map (0~20min enlarged drawing).
Figure 11 is that 0 hour property of medium chain triglyceride stability of the present invention measures the (0~20min amplification of test sample HPLC map
Figure).
Figure 12 is that 24 hours property of medium chain triglyceride stability of the present invention measure the (0~20min amplification of test sample HPLC map
Figure).
Specific embodiment
Purposes, technical schemes and advantages in order to better illustrate the present invention, below in conjunction with the drawings and specific embodiments pair
The present invention is described further.
Embodiment 1
The specificity of detection method is tested
A kind of reality of the detection method of content of Sunfat GDC-S and capric acid diglyceride in medium chain triglyceride of the present invention
Apply example, comprising the following steps:
(1) preparation of test solution: taking 250mg medium chain triglyceride, and 10mL methanol is added and dissolves and dilutes, shakes up,
Obtain medium chain triglyceride sample solution；The 1,3- Sunfat GDC-S for taking medium chain triglyceride structural identification to separate respectively
(MTG-DG-1), 1,2- Sunfat GDC-S (MTG-DG-2), 1,3- capric acid diglyceride (MTG-DG-5), 1,2- capric acid glycerol
Diester (MTG-DG-6) 10mg is added 10mL methanol and dissolves and dilute, shakes up, obtain diglyceride test solution；
(2) preparation of reference substance solution: take Sunfat GDC-S and capric acid diglyceride be respectively 10mg, 10mg, it is molten
In 1mL methanol, Sunfat GDC-S reference substance solution and capric acid diglyceride reference substance solution is made；
(3) it measures:
Liquid phase chromatogram condition are as follows:
Chromatographic column: C18 chromatographic column (250mm × 4.6mm, 5 μm)；
Detector: (drift tube temperature is 72 DEG C, flow rate of carrier gas 2.0mL/min to evaporative light scattering detector, and yield value is
4, mode is shunt mode)；
Mobile phase: using volume ratio for 8:2 acetonitrile-water mixed solvent as mobile phase A, it is different for the acetonitrile-of 1:1 with volume ratio
Propanol solvent mixture is that Mobile phase B carries out gradient elution；The condition of the eluent gradient elution are as follows: 0~10min, mobile phase
Middle A (76%), B (24%) in mobile phase；10~30min, A (76% → 30%) in mobile phase, B in mobile phase (24% →
70%)；30~40min, A (30%) in mobile phase, B (70%) in mobile phase；40~50min, A in mobile phase (30% →
76%), B (70% → 24%) in mobile phase；50~60min, A (76%) in mobile phase, B (24%) in mobile phase；
Flow velocity: 1.0mL/min；
Column temperature: 30 DEG C；
Precision measures the Sunfat GDC-S reference substance solution and each 20 μ of capric acid diglyceride reference substance solution of various concentration
L injects liquid chromatograph, records chromatogram.
Sunfat GDC-S reference substance solution, capric acid diglyceride reference substance solution are measured, medium chain triglyceride structure is true
It is sweet to demonstrate,prove 1,3- Sunfat GDC-S (MTG-DG-1), 1,2- Sunfat GDC-S (MTG-DG-2), the 1,3- capric acid separated
1,3- octanoic acid glycerol two in oily diester (MTG-DG-5), 1,2- capric acid diglyceride (MTG-DG-6) and test solution
Ester, 1,2- Sunfat GDC-S, 1,3- capric acid diglyceride, 1,2- capric acid diglyceride, the results are shown in Table 1.Sad glycerol
Diester reference substance solution HPLC map, capric acid diglyceride reference substance solution HPLC map, medium chain triglyceride separated component
Separated component 1,2- Sunfat GDC-S HPLC map, the middle chain of 1,3- Sunfat GDC-S HPLC map, medium chain triglyceride
The separated component 1,2- capric acid glycerol of the separated component 1,3- capric acid diglyceride HPLC map of triglycerides, medium chain triglyceride
Diester HPLC map, medium chain triglyceride test sample HPLC map are respectively as shown in Fig. 1~7, medium chain triglyceride test sample
0~20min enlarged drawing of HPLC map is as shown in Figure 8.
The separating resulting of 1 medium chain triglyceride of table
As shown in Table 1, Sunfat GDC-S, capric acid diglyceride reference substance and middle chain are measured under chromatographic condition of the invention
Sunfat GDC-S, the capric acid triglyceride component appearance time separated in triglycerides are corresponding, and (octanoic acid containing 1,3- is sweet
Oily diester, 1,2- Sunfat GDC-S, 1,3- capric acid diglyceride, 1,2- capric acid diglyceride), with medium chain triglyceride for examination
Sunfat GDC-S, capric acid triglyceride component appearance time in product is corresponding.1,3- Sunfat GDC-S, 1,2- octanoic acid are sweet
Oily diester is isomer, so using simultaneously when the content of 1,3- Sunfat GDC-S, 1,2- Sunfat GDC-S calculates
For Sunfat GDC-S as reference substance, Sunfat GDC-S content is sum of the two；Similarly, 1,3- capric acid diglyceride, 1,2-
Capric acid diglyceride is isomer, so when the content of 1,3- capric acid diglyceride, 1,2- capric acid diglyceride calculates, together
For Shi Caiyong capric acid diglyceride as reference substance, capric acid diacylglycerol content is sum of the two.As can be seen from the results respectively at swarming point
From degree, theoretical cam curve meet the requirements, illustration method specificity is good, it is possible to using external standard method calculate Sunfat GDC-S,
The content of capric acid diglyceride.
Embodiment 2
The linear property of detection method is tested
A kind of reality of the detection method of content of Sunfat GDC-S and capric acid diglyceride in medium chain triglyceride of the present invention
Apply example, comprising the following steps:
(1) preparation of test solution: taking 200mg medium chain triglyceride, and 10mL methanol is added and dissolves and dilutes, shakes up,
Obtain test solution；
(2) preparation of reference substance solution: precision weighs Sunfat GDC-S and capric acid diglyceride reference substance, and methanol is added
It dissolves and dilutes, it is respectively 70 μ g, 150 μ g, 210 μ g, 300 μ g, 450 μ g that every 1mL methanol, which is made, containing Sunfat GDC-S, contains the last of the ten Heavenly stems
Acid triglyceride is respectively the mixed reference substance solution of 30 μ g, 60 μ g, 90 μ g, 120 μ g, 180 μ g；
(3) it measures:
Liquid phase chromatogram condition are as follows:
Chromatographic column: C18 chromatographic column (250mm × 4.6mm, 5 μm)；
Detector: (drift tube temperature is 72 DEG C, flow rate of carrier gas 2.0mL/min to evaporative light scattering detector, and yield value is
4, mode is shunt mode)；
Mobile phase: using volume ratio for 8:2 acetonitrile-water mixed solvent as mobile phase A, it is different for the acetonitrile-of 1:1 with volume ratio
Propanol solvent mixture is that Mobile phase B carries out gradient elution；The condition of the eluent gradient elution are as follows: 0~10min, mobile phase
Middle A (76%), B (24%) in mobile phase；10~30min, A (76% → 30%) in mobile phase, B in mobile phase (24% →
70%)；30~40min, A (30%) in mobile phase, B (70%) in mobile phase；40~50min, A in mobile phase (30% →
76%), B (70% → 24%) in mobile phase；50~60min, A (76%) in mobile phase, B (24%) in mobile phase；
Flow velocity: 1.0mL/min；
Column temperature: 30 DEG C；
Each 20 μ L of mixed reference substance solution that precision measures various concentration injects liquid chromatograph, records chromatogram, measures
The peak area of various concentration mixed reference substance solution, using mixed reference substance solution peak area logarithm as ordinate, with corresponding
Mixed reference substance solution log concentration is that abscissa makees standard curve, calculates separately to obtain equation of linear regression；It the results are shown in Table 2, from
As a result it is found that the calibration curve equation of Sunfat GDC-S: y=2.0467x+5.7424, correlation coefficient r2=0.9984, show
This method is in good linear in 0.07285mg/ml~0.4371mg/ml concentration range；With capric acid diglyceride peak area pair
Numerical value is ordinate, makees standard curve using corresponding concentration of standard solution logarithm as abscissa, the results are shown in Table 2, as can be known from the results,
The calibration curve equation of capric acid diglyceride: y=1.3592x+5.5447, correlation coefficient r2=0.9990, show that this method exists
It is in good linear in 0.03025mg/ml~0.1815mg/ml concentration range.
2 linear relationship test result of table
Embodiment 3
The repetitive test of detection method
A kind of reality of the detection method of content of Sunfat GDC-S and capric acid diglyceride in medium chain triglyceride of the present invention
Apply example, comprising the following steps:
(1) preparation of test solution: taking 6 parts of same lot number medium chain triglyceride 200mg, and the dissolution of 10mL methanol is added simultaneously
Dilution, shakes up, obtains test solution；
(2) preparation of reference substance solution: precision weighs Sunfat GDC-S and capric acid diglyceride reference substance, and methanol is added
It dissolves and dilutes, it is respectively 70 μ g, 150 μ g, 210 μ g, 300 μ g, 450 μ g that every 1mL methanol, which is made, containing Sunfat GDC-S, contains the last of the ten Heavenly stems
Acid triglyceride is respectively the mixed reference substance solution of 30 μ g, 60 μ g, 90 μ g, 120 μ g, 180 μ g；
(3) it measures:
Liquid phase chromatogram condition are as follows:
Chromatographic column: C18 chromatographic column (250mm × 4.6mm, 5 μm)；
Detector: (drift tube temperature is 72 DEG C, flow rate of carrier gas 2.0mL/min to evaporative light scattering detector, and yield value is
4, mode is shunt mode)；
Mobile phase: using volume ratio for 8:2 acetonitrile-water mixed solvent as mobile phase A, it is different for the acetonitrile-of 1:1 with volume ratio
Propanol solvent mixture is that Mobile phase B carries out gradient elution；The condition of the eluent gradient elution are as follows: 0~10min, mobile phase
Middle A (76%), B (24%) in mobile phase；10~30min, A (76% → 30%) in mobile phase, B in mobile phase (24% →
70%)；30~40min, A (30%) in mobile phase, B (70%) in mobile phase；40~50min, A in mobile phase (30% →
76%), B (70% → 24%) in mobile phase；50~60min, A (76%) in mobile phase, B (24%) in mobile phase；
Flow velocity: 1.0mL/min；
Column temperature: 30 DEG C；
Each 20 μ L of mixed reference substance solution that precision measures various concentration injects liquid chromatograph, records chromatogram, measures
The peak area of various concentration mixed reference substance solution, using mixed reference substance solution peak area logarithm as ordinate, with corresponding
Mixed reference substance solution log concentration is that abscissa makees standard curve, calculates separately to obtain equation of linear regression；Another accurate measure supplies
20 μ L of test sample solution injects liquid chromatograph, measures the peak area of Sunfat GDC-S and capric acid diglyceride in test sample, point
Not with the content of 6 batches of Sunfat GDC-Ss and capric acid diglyceride of regression equation calculation.
Medium chain triglyceride repeatability measures 0~20min enlarged drawing such as Fig. 9 institute of the HPLC map of mixed reference substance solution
Show, 0~20min enlarged drawing of test sample HPLC map is as shown in Figure 10.
The results are shown in Table 3 for the content repeatability of Sunfat GDC-S, and the content repeatability result of capric acid diglyceride is such as
Shown in table 4.By table 3 and table 4 it is found that Sunfat GDC-S and capric acid diglyceride in this method measurement median chain triglyceride oil
Content is reproducible.
The content repeatability result of 3 Sunfat GDC-S of table
The content repeatability result of 4 capric acid diglyceride of table
Embodiment 4
The stability test of detection method
A kind of reality of the detection method of content of Sunfat GDC-S and capric acid diglyceride in medium chain triglyceride of the present invention
Apply example, comprising the following steps:
(1) preparation of test solution: taking 1 part of medium chain triglyceride 200mg, and 10mL methanol is added and dissolves and dilutes, shakes
It is even, obtain test solution；
(2) it measures:
Liquid phase chromatogram condition are as follows:
Chromatographic column: C18 chromatographic column (250mm × 4.6mm, 5 μm)；
Detector: (drift tube temperature is 72 DEG C, flow rate of carrier gas 2.0mL/min to evaporative light scattering detector, and yield value is
4, mode is shunt mode)；
Mobile phase: using volume ratio for 8:2 acetonitrile-water mixed solvent as mobile phase A, it is different for the acetonitrile-of 1:1 with volume ratio
Propanol solvent mixture is that Mobile phase B carries out gradient elution；The condition of the eluent gradient elution are as follows: 0~10min, mobile phase
Middle A (76%), B (24%) in mobile phase；10~30min, A (76% → 30%) in mobile phase, B in mobile phase (24% →
70%)；30~40min, A (30%) in mobile phase, B (70%) in mobile phase；40~50min, A in mobile phase (30% →
76%), B (70% → 24%) in mobile phase；50~60min, A (76%) in mobile phase, B (24%) in mobile phase；
Flow velocity: 1.0mL/min；
Column temperature: 30 DEG C；
Precision is measured with a test solution, respectively at 0 hour, 2 hours, 4 hours, 8,12 hours, and 24 small time-divisions
The inaccurate 20 μ L that draw inject liquid chromatograph, record chromatogram, measure peak area of each component, and it is sad in 24 hours to calculate separately
The RSD of diglyceride peak and capric acid diglyceride area is respectively 2.5% and 2.9%, the results are shown in Table 5, illustrates sample solution
Interior for 24 hours relatively stable.0~20min enlarged drawing of 0 hour property measurement test sample HPLC map of medium chain triglyceride stability is such as
Shown in Figure 11,0~20min enlarged drawing such as Figure 12 institute of 24 hours property measurement test sample HPLC maps of medium chain triglyceride stability
Show.
The content repeatability result of 5 capric acid diglyceride of table
The above embodiments are merely illustrative of the technical solutions of the present invention rather than limiting the scope of the invention, although ginseng
The present invention is explained in detail according to preferred embodiment, those skilled in the art should understand that, it can be to of the invention
Technical solution is modified or replaced equivalently, without departing from the spirit and scope of technical solution of the present invention.
Claims (8)
1. the detection method of content of Sunfat GDC-S and capric acid diglyceride in a kind of medium chain triglyceride, which is characterized in that
The following steps are included:
(1) preparation of test solution: taking medium chain triglyceride, and methanol is added and dissolves and dilutes, shakes up, it is molten to obtain test sample
Liquid；
(2) preparation of reference substance solution: taking Sunfat GDC-S and capric acid diglyceride reference substance, and the dissolution of addition methanol is simultaneously dilute
It releases, obtains the mixed reference substance solution of Sunfat GDC-S and capric acid diglyceride；
(3) assay: precision measures test solution and mixed reference substance solution injects liquid chromatograph, records chromatogram；
By external standard method respectively with the content of Sunfat GDC-S and capric acid diglyceride in calculated by peak area test sample.
2. detection method as described in claim 1, which is characterized in that in the step (1), medium chain triglyceride and methanol
Mass volume ratio is 200mg:10mL.
3. detection method as described in claim 1, which is characterized in that in the step (2), precision weighs Sunfat GDC-S
With capric acid diglyceride reference substance, methanol is added and dissolves and dilutes, it is respectively 70 μ that every 1mL methanol, which is made, containing Sunfat GDC-S
G, 150 μ g, 210 μ g, 300 μ g, 450 μ g, diglyceride containing capric acid be respectively 30 μ g, 60 μ g, 90 μ g, 120 μ g, 180 μ g it is mixed
Close reference substance solution.
4. detection method as described in claim 1, which is characterized in that in the step (3), liquid phase chromatogram condition are as follows:
Chromatographic column: C18 chromatographic column；
Detector: evaporative light scattering detector；
Mobile phase: using volume ratio for 8:2 acetonitrile-water mixed solvent as mobile phase A, be acetonitrile-isopropanol of 1:1 with volume ratio
Mixed solvent is that Mobile phase B carries out gradient elution；The condition of eluent gradient elution are as follows: 0~10min, A in mobile phase
(76%), B (24%) in mobile phase；10~30min, A (76% → 30%) in mobile phase, B (24% → 70%) in mobile phase；
30~40min, A (30%) in mobile phase, B (70%) in mobile phase；40~50min, A (30% → 76%) in mobile phase, stream
B (70% → 24%) in dynamic phase；50~60min, A (76%) in mobile phase, B (24%) in mobile phase；
Flow velocity: 1.0mL/min；
Column temperature: 30 DEG C；
It is accurate respectively to measure test solution and 20 μ L of mixed reference substance solution, inject high performance liquid chromatograph, measurement.
5. detection method as described in claim 1, which is characterized in that in the step (3), the concrete operations of external standard method are as follows:
The 20 μ L of mixed reference substance solution that precision measures various concentration injects liquid chromatograph, measures various concentration mixed reference substance solution
Peak area, using mixed reference substance solution peak area logarithm as ordinate, with corresponding mixed reference substance solution log concentration
Make standard curve for abscissa, calculates separately to obtain equation of linear regression；The another accurate 20 μ L of test solution that measures injects liquid phase color
Spectrometer measures the peak area of Sunfat GDC-S and capric acid diglyceride in test sample, sweet with regression equation calculation octanoic acid respectively
The content of oily diester and capric acid diglyceride.
6. detection method as claimed in claim 4, which is characterized in that the specification of the C18 chromatographic column is 250mm × 4.6mm,
5μm。
7. detection method as claimed in claim 4, which is characterized in that the drift tube temperature of the detector is 72 DEG C, carrier gas
Flow velocity is 2.0mL/min, and yield value 4, mode is shunt mode.
8. detection method as claimed in claim 5, which is characterized in that the content of the Sunfat GDC-S is that 1,3- octanoic acid is sweet
The sum of oily diester, content of 1,2- Sunfat GDC-S are all made of Sunfat GDC-S in mixed reference substance solution and are used as control；
The content of the capric acid diglyceride is the sum of 1,3- capric acid diglyceride, the content of 1,2- capric acid diglyceride, is all made of mixed
It closes capric acid diglyceride in reference substance solution and is used as control.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000018944A1 (en) * | 1998-09-25 | 2000-04-06 | Conlinco, Inc. | Triacylglycerols of enriched cla content |
| CN101708156A (en) * | 2009-12-10 | 2010-05-19 | 中国人民解放军第二军医大学 | Camptothecin medicament injection solution and injection and preparation method thereof |
| CN102539586A (en) * | 2010-12-14 | 2012-07-04 | 上海良友（集团）有限公司 | Method for isolating and detecting oxidized triglyceride (ox-TG) of edible vegetable oil and application of the method |
| CN106018611A (en) * | 2016-06-21 | 2016-10-12 | 广州白云山汉方现代药业有限公司 | Testing method for detecting content of medium chain triglyceride with gas chromatography internal standard method |
| CN107334063A (en) * | 2017-07-11 | 2017-11-10 | 桂林医学院 | A kind of oatmeal containing medium chain fatty acid ester and preparation method thereof |
-
2018
- 2018-11-21 CN CN201811390546.7A patent/CN109655536B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000018944A1 (en) * | 1998-09-25 | 2000-04-06 | Conlinco, Inc. | Triacylglycerols of enriched cla content |
| CN101708156A (en) * | 2009-12-10 | 2010-05-19 | 中国人民解放军第二军医大学 | Camptothecin medicament injection solution and injection and preparation method thereof |
| CN102539586A (en) * | 2010-12-14 | 2012-07-04 | 上海良友（集团）有限公司 | Method for isolating and detecting oxidized triglyceride (ox-TG) of edible vegetable oil and application of the method |
| CN106018611A (en) * | 2016-06-21 | 2016-10-12 | 广州白云山汉方现代药业有限公司 | Testing method for detecting content of medium chain triglyceride with gas chromatography internal standard method |
| CN107334063A (en) * | 2017-07-11 | 2017-11-10 | 桂林医学院 | A kind of oatmeal containing medium chain fatty acid ester and preparation method thereof |
Non-Patent Citations (5)
| Title |
|---|
| INDRASENA, WM 等: "Qualitative and quantitative analysis of lipid classes in fish oils by thin-layer chromatography with an Iatroscan flame ionization detector (TLC-FID) and liquid chromatography with an evaporative light scattering detector (LC-ELSD)", 《JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES》 * |
| 樊巧英 等: "甘油二酯异构体高效液相色谱分析法的建立", 《中国油脂》 * |
| 籍淑贤 等: "食用植物油中甘油三酯色谱分析方法研究进展", 《分析测试学报》 * |
| 舒庆 等: "《生物柴油科学与技术》", 31 December 2012, 冶金工业出版社 * |
| 赵欣欣 等: "甘油二酯的制备及其高效液相色谱的检测技术", 《食品工业》 * |
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