CN111239311A - Analysis method of acid sugar and acid sugar derivative in infant excrement - Google Patents
Analysis method of acid sugar and acid sugar derivative in infant excrement Download PDFInfo
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- CN111239311A CN111239311A CN201811438772.8A CN201811438772A CN111239311A CN 111239311 A CN111239311 A CN 111239311A CN 201811438772 A CN201811438772 A CN 201811438772A CN 111239311 A CN111239311 A CN 111239311A
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Abstract
The invention relates to an analysis method of acid sugar and acid sugar derivatives in infant excrement. The method is characterized in that: the sample is subjected to water-soluble homogeneous extraction, organic solvent is added for precipitation to remove protein, then the interference of pigment, lactose and other center sugar is removed through an online purification column, and the obtained acid sugar and acid sugar derivatives are subjected to hydrophilic interaction chromatographic separation and liquid phase or mass spectrum detection to obtain the composition and content information of the acid sugar and acid sugar derivatives of the infant excrement. The method has the characteristics of accurate quantification, good reproducibility, stable and controllable operation, high analysis flux and the like, and is suitable for rapid and high-flux analysis of infant excrement.
Description
Technical Field
The invention relates to an analysis method of acid sugar and acid sugar derivatives in infant excrement. Is suitable for the rapid and high-flux analysis of infant excrement.
Technical Field
Oligosaccharides derived from breast milk play a vital role in the growth and development of infants, such as anti-inflammatory, anti-infective, immunomodulating and maintaining intestinal flora balance. Wherein the acid oligosaccharide plays an irreplaceable role in vivo, such as preventing carcinogenesis, promoting normal development of infant brain, and the like.
The precondition for deeply knowing the relationship between the acid sugar in the intestinal tract of the infant and the intestinal flora or other growth and development processes is to analyze the composition and content of the acid sugar, so that the development of a simple, rapid and high-flux quantitative analysis method for the acid sugar is of great significance. However, since the components of the infant excrement are very complex and diverse, the detector is seriously polluted by lipid, protein and pigment components during analysis, the analysis process is seriously interfered by lactose and other neutral sugars in the saccharide components, and the variety of acidic sugars is large, so that the analysis difficulty is increased.
At present, researches on the influence of acidic sugar on infant intestinal flora balance, brain development and other growth and development processes are concerned, but for infant excrement capable of directly measuring the content of the acidic sugar in the infant body, an analysis and quantification method of the acidic sugar is not established yet, mainly because the content of the sugar in the excrement is extremely low, and other complex components including neutral sugar seriously interfere with the detection and quantification of the acidic sugar. Therefore, the invention designs a novel infant excrement purification and analysis method, which uses high-throughput quantitative analysis of infant excrement.
Disclosure of Invention
The invention relates to an analysis method of acid sugar and acid sugar derivatives in infant excrement. The method specifically comprises the following steps: and (3) homogenizing and extracting a sample, precipitating to remove protein, removing interference of pigment, lactose and other center sugar by using an online purification column, and performing hydrophilic interaction chromatographic separation and liquid phase or mass spectrum detection on the obtained acid sugar and acid sugar derivative to obtain the composition and content information of the acid sugar and acid sugar derivative of the infant excrement.
In order to achieve the purpose, the invention adopts the technical scheme that:
the method comprises the steps of carrying out homogeneous extraction and precipitation protein removal on acid sugar and acid sugar derivatives in infant excrement, removing pigments, lactose and neutral sugar through an online system purification column to obtain acid sugar and acid sugar derivative fractions, separating the obtained acid sugar and acid sugar derivative fractions through a hydrophilic interaction chromatography (HILIC) column, detecting components flowing out of the hydrophilic interaction chromatography (HILIC) column through a detection system to obtain structural component analysis of the acid sugar and the acid sugar derivatives in the infant excrement, and obtaining absolute content information of the acid sugar and the acid sugar derivatives in the infant excrement according to a standard curve drawn by each acid sugar and acid sugar derivative under the analysis system.
1. The homogenizing extraction step comprises: adding 0.5-5 times of good solvent such as water into the excrement, vortex shaking for not less than 5 s, ultrasonic treating for not less than 1 min, centrifuging at 500-50000g for 5-500 min, extracting for 1-5 times, and collecting the upper layer of transparent solution to extract saccharide in the excrement
2. The precipitation protein-removing step comprises: taking the clear liquid of the excrement extraction, adding 1-5 times of organic solvent by volume, mixing uniformly, standing for 2-48 hours at 0-10 ℃, centrifuging for not less than 5 minutes at 500-00 g at 0-10 ℃, collecting supernatant, centrifuging for 1-5 times, and evaporating to dryness to obtain a sample. The organic solvent is one or more of methanol, acetonitrile, ethanol and acetone.
3. The stationary phase of the chromatographic column for purifying the acidic sugar and the stationary phase of the HILIC column are polar chromatographic packing, comprise silica gel or polar bonded silica gel packing, and have the following structural formula:
wherein SiO is2Is silica gel, and R is polar group and contains one or more of amino acid, amide, amino group, carboxyl group, glycosyl, zwitter ion and the like.
4. The organic solvent in the eluent is one or more of methanol, acetonitrile, ethanol and acetone.
5. The buffer salt type and its concentration and pH in the mobile phase are as follows:
the selected buffer salt type is one of ammonium formate buffer salt, ammonium acetate buffer salt and ammonium bicarbonate buffer salt, and the concentration of the buffer salt in the mobile phase is 0-200mM, and the pH value is 6-10.
6. The mobile phase gradient was optimized as follows:
and the mobile phase water or the buffer saline solution and the organic solvent are used as eluent, and the mixing ratio is 5/95-95/5.
7. The collection window of the acidic sugar is 0-10BV, and the effluent liquid is collected.
8. The chromatographic operating parameters were optimized as follows: the inner diameter of the chromatographic column is 2.1-10 mm; the flow rate is 0.1-2 BV/min; the column temperature is 15-60 ℃; the detector is a mass spectrometric detector, an electrospray detector or an evaporative light detector.
The invention has the following advantages:
1. high sensitivity and accurate quantification. The method has pertinence in purifying the complex system of excrement and then quantifying the acid sugar, greatly improves the detection limit and the quantification limit, and avoids the pollution and the interference of a large amount of impurities.
2. The flux is high. The purification and analysis process is simple and rapid.
3. The repeatability is good. The chromatographic column stationary phase used in the invention has good stability and is easy to realize automation.
4. The method has wide application range and can be used for analyzing milk and human milk of different mammals.
Drawings
FIG. 1 is a graph showing the online purification and analysis of infant excreta in example 1
FIG. 2 is a graph of sialic acid ion vs. MRM in the quantitative determination of infant excreta in example 3
FIG. 3 is a graph of the 3' SL ion pair MRM in the quantitative determination of infant excrement in example 3
FIG. 4 is a graph of 6' SL ion pair MRM in the quantitative determination of infant excrement in example 3
FIG. 5 is a graph of MRM TIC of an actual sample in the quantitative determination of infant excreta in example 3
Detailed Description
Example 1:
homogeneous extraction process of infant excrement
Adding a certain amount of infant excrement into water with equal mass, carrying out vortex oscillation for 30 seconds, carrying out ultrasonic treatment for 10 minutes, then centrifuging for 5 minutes at 4000g, extracting for 1 time, and taking out the upper transparent solution to finish the extraction of the carbohydrate in the excrement.
Infant excrement precipitation protein-removing process
And taking the sugar extraction clear liquid, adding 2 times of volume of absolute ethyl alcohol solvent, uniformly mixing, standing at 4 ℃ for 12 hours, centrifuging at 4 ℃ for 5 minutes at 4000g, taking out the supernatant, centrifuging for 1 time, and evaporating to dryness to obtain the sugar extraction protein-removed sample.
Infant waste decontamination and analysis process
Directly taking sugar extraction and protein removal liquid samples for analysis. The sample introduction amount is 20 mu L, an SPE purification column uses an amphoteric ion exchange mode stationary phase, the inner diameter of the chromatographic column is 2.1mm, and the flow rate is 1.0 mL/min; the HILIC analysis column adopts amide column with inner diameter of 4.6mm and flow rate of 1.0 mL/min; the column temperature was 30 ℃ and electrospray detection was performed. The mobile phase A is acetonitrile, B is water, and C is 100mM ammonium formate. Acetonitrile is a weak elution solvent, gradient elution is carried out, and elution conditions and valve switching time are as follows:
example 2:
homogeneous extraction and deproteinization process of infant excrement
Adding water with the mass being 2 times of that of the infant excrement into a certain amount of infant excrement, carrying out vortex oscillation for 1 minute, carrying out ultrasonic treatment for 5 minutes, centrifuging 10000g of the infant excrement for 5 minutes, adding 3 times of acetonitrile into the supernatant of the last time, uniformly mixing, standing at 10 ℃ for 2 hours, centrifuging 10000g of the infant excrement at 10 ℃ for 30 minutes, centrifuging for 3 times, and collecting supernatant to obtain the infant excrement protein removal sample.
Infant waste decontamination and analysis process
Directly taking a liquid sample for analysis. The sample introduction amount is 100 mu L, an SPE purification column uses an amphoteric ion exchange mode stationary phase, the inner diameter of the chromatographic column is 3.0mm, and the flow rate is 0.4 mL/min; the HILIC analysis column adopts an amino column, the inner diameter of the chromatographic column is 4.6mm, and the flow rate is 1.0 mL/min; the column temperature was 30 ℃ and the detection was carried out by evaporation light. Mobile phase a was methanol, B was water, C was 100mM ammonium formate, pH 4.2. Methanol is a weak elution solvent, gradient elution is carried out under the following conditions and the switching time of a switching valve is as follows:
example 3:
quantitative analysis of sialic acid, 3 'SL, 6' SL in infant faeces
Adding 2 times volume of acetone into a certain amount of infant excrement, carrying out vortex oscillation for 1 minute, carrying out ultrasonic treatment for 10 minutes, adding 4 times volume of acetone into the mixture, uniformly mixing, standing at 4 ℃ for 18 hours, centrifuging at 4 ℃ for 10 minutes at 20000g, centrifuging for 2 times, collecting supernatant to obtain a liquid sample for degreasing and removing protein oligosaccharide, and concentrating to dryness for later use.
Drawing of quantitative standard curve
5ppm sialic acid standard substance (acetonitrile: water 1: 1) is subjected to ion pair optimization in triple quadrupole mass spectrometry, 308.1>170 is selected as a qualitative ion pair, and 308>87 is selected as a quantitative ion pair; sialic acid standards (acetonitrile: water 1: 1) were prepared at concentrations of 0.1. mu.g/mL, 0.2. mu.g/mL, 0.4. mu.g/mL, 1.6. mu.g/mL, 3.2. mu.g/mL, 6.4. mu.g/mL, 12.8. mu.g/mL, and 25.6. mu.g/mL, respectively, and injection was repeated three times for each concentration to obtain an average value and to plot a concentration-response curve as a quantitative curve of sialic acid under the chromatographic conditions.
5ppm 3' SL standard substance (acetonitrile: water 1: 1) is used for ion pair optimization in triple quadrupole mass spectrometry, 632.3>408.2 is selected as a qualitative ion pair, and 308>290.1 is selected as a quantitative ion pair; 3 'SL standard substances (acetonitrile: water 1: 1) with the concentration of 0.1 mu g/mL,0.2 mu g/mL, 0.4 mu g/mL,1.6 mu g/mL,3.2 mu g/mL,6.4 mu g/mL,12.8 mu g/mL and 25.6 mu g/mL are prepared respectively, and the samples are injected for three times to obtain an average value for each concentration so as to draw a concentration-response curve as a quantitative curve of the 3' SL under the chromatographic conditions.
5ppm 6' SL standard substance (acetonitrile: water 1: 1) is subjected to ion pair optimization in triple quadrupole mass spectrometry, 632.3>470.2 is selected as a qualitative ion pair, and 308>290.1 is selected as a quantitative ion pair; 6 'SL standard substances (acetonitrile: water 1: 1) with the concentration of 0.1 mu g/mL,0.2 mu g/mL, 0.4 mu g/mL,1.6 mu g/mL,3.2 mu g/mL,6.4 mu g/mL,12.8 mu g/mL and 25.6 mu g/mL are prepared respectively, and the samples are injected for three times to obtain an average value for each concentration so as to draw a concentration-response curve as a quantitative curve of the 6' SL under the chromatographic conditions.
Infant excrement purification and analysis process
Weighing an excrement extracted protein-removed freeze-dried sample, preparing a solution with the concentration of 100mg/mL by using 50% acetonitrile water, wherein the sample volume is 200 mu L, an SPE purification column uses an amphoteric ion exchange column, the inner diameter of the chromatographic column is 2.1mm, and the flow rate is 0.2 mL/min; the HILIC analysis column adopts amphoteric ion exchange column, the inner diameter of the chromatographic column is 4.6mm, and the flow rate is 1.0 mL/min; the column temperature was 30 ℃ and mass spectrometric detection was carried out. The mobile phase A was ethanol, B was water, C was 50mM ammonium acetate, pH 6.0. Ethanol is a weak elution solvent, gradient elution is carried out, and elution conditions and switching valve switching time are as follows:
Claims (10)
1. an analysis method of acid sugar and acid sugar derivatives in infant excrement is characterized by comprising the following steps: the method comprises the steps of carrying out homogeneous extraction and precipitation protein removal on acid sugar and acid sugar derivatives in infant excrement, removing pigments, lactose and neutral sugar through an online system purification column to obtain acid sugar and acid sugar derivative fractions, separating the obtained acid sugar and acid sugar derivative fractions through a hydrophilic interaction chromatography (HILIC) column, detecting components flowing out of the hydrophilic interaction chromatography (HILIC) column through a detection system to obtain structural component analysis of the acid sugar and the acid sugar derivatives in the infant excrement, and obtaining absolute content information of the acid sugar and the acid sugar derivatives in the infant excrement according to a standard curve drawn by each acid sugar and acid sugar derivative under the analysis system.
2. The analytical method of claim 1, wherein: the step of homogeneous extraction of the saccharides and the acid sugar derivatives in the excrement comprises the following steps: adding a good solvent of saccharides with the volume of 0.5-5 times of that of the excrement into the excrement, carrying out vortex oscillation for not less than 5 seconds, carrying out ultrasonic vibration for not less than 1 minute, centrifuging for 5-500 minutes at 50000g, extracting for 1-5 times, and taking out an upper transparent solution to complete the homogeneous extraction of the saccharides and the acidic saccharide derivatives in the excrement.
3. The analytical method of claim 1, wherein: the precipitation protein-removing step comprises the following steps: taking the acid sugar and the acid sugar derivative extraction clear liquid obtained by homogeneous extraction in the excrement, adding 1-5 times of organic solvent by volume, mixing uniformly, standing for 2-48 hours at 0-10 ℃, centrifuging for not less than 5 minutes at 0-10 ℃ and 500-50000g, taking out the supernatant, centrifuging for 1-5 times, and evaporating to dryness to obtain a sample for removing the sugar and the acid sugar derivative protein in the excrement;
the organic solvent is one or more of methanol, ethanol, acetonitrile and acetone.
4. The analytical method of claim 1, wherein: the on-line system comprises a ternary pump, an automatic sample injector, a column incubator, a switching valve (two-position six-way or twelve-way), and a detector.
5. The analytical method of claim 1, wherein: the steps of purifying the acidic sugar and the acidic sugar derivative by the online system purifying column are as follows: taking a fecal sample after biological homogeneous extraction and protein removal by precipitation, dissolving the sample by using 20-90% of organic solvent aqueous solution, taking a hydrophilic material with electrostatic repulsion as a chromatographic column stationary phase, taking a mobile phase of water and organic solvent or buffered salt solution and organic solvent, eluting by using an isocratic or gradient method, and directly feeding acidic sugar and acidic sugar derivative fraction into a serial hydrophilic interaction chromatography (HILIC) column for separation.
6. The analytical method of claim 5, wherein: the method for purifying the acid sugar and the acid sugar derivative in the online system needs to remove lactose and neutral sugar which interfere the detection and pigment and lipid which may pollute the detector, and the optimization operation is as follows:
a) the ratio of the dissolved solution of the excrement sample after biological homogeneous extraction and protein removal by precipitation to the mixed solution of organic solution methanol, ethanol, acetonitrile, acetone and water is 20-90%
b) The purifying column filler is silica gel or silica gel surface bonded polar group filler, and the bonded polar group comprises one or more of amino acid, amide, amino group, carboxyl group, glycosyl, zwitter ion and the like;
c) the organic solvents in the eluent are: one or more of methanol, ethanol, acetonitrile, acetone and isopropyl ketone;
d) the selected buffer salt type is one of ammonium formate buffer salt, ammonium acetate buffer salt and ammonium bicarbonate buffer salt, the concentration of the selected buffer salt in the mobile phase is 0-200mM, and the pH value is 6-10;
e) the ratio of mobile phase water or a buffered saline solution and an organic solvent as an eluent is 5/95-95/5;
f) the inner diameter of the chromatographic column is 2.1-10 mm; the flow rate of the analysis method is 0.1-2 BV/min;
g) the collection window of the acidic sugar and the acidic sugar derivative fraction is 0-10BV, and the effluent liquid is collected.
7. The analytical method of claim 1, wherein: the hydrophilic interaction chromatography (HILIC) column performs the separation as follows: directly collecting the acidic sugar and acidic sugar derivative fraction in HILIC column, separating and analyzing, eluting with water and organic solvent or buffered saline solution and organic solvent under isocratic or gradient amplification, detecting, eluting with water and organic solvent or buffered saline solution and organic solvent under isocratic or gradient method, and detecting.
The hydrophilic interaction chromatographic (HILIC) column is filled with hydrophilic interaction chromatographic material with positive charges on the surface, and comprises one or more of amino acid, acylamino, zwitterion and glycosyl.
8. The analytical method of claim 7, wherein: optimizing chromatographic column operating parameters including flow rate, column temperature and detector, and valve switching time, the specific operations are as follows:
a) the flow rate is 0.1-2 BV/min;
b) the inner diameter of the chromatographic column is 2.1-10mm
c) The column temperature is 15-60 ℃;
d) the detector is mass spectrum, electrospray or evaporative light;
e) the valve switching time is 0-10 BV.
9. The analytical method of claim 1, wherein: the methods for obtaining the composition and structure information of the acidic sugar and the acidic sugar derivative include mass spectrometry detection, electrospray detector detection and evaporative light detector detection.
10. The analytical method of claim 1, wherein: the detection modes for obtaining the standard curve of the acidic sugar and the acidic sugar derivative under the analysis system comprise a mass spectrum detector, an electrospray detector and an evaporative light detector.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008309699A (en) * | 2007-06-15 | 2008-12-25 | Hitachi High-Technologies Corp | Two-dimensional liquid chromatograph of ion exchange and normal phase column |
| CN101421012A (en) * | 2004-04-23 | 2009-04-29 | 吉恩康国际股份有限公司 | Method of separating components in a sample using silane-treated silica filter media |
| US20100003699A1 (en) * | 2008-01-18 | 2010-01-07 | Glykos Finland Ltd. | Tissue carbohydrate compositions and analysis thereof |
| CN105431733A (en) * | 2013-07-11 | 2016-03-23 | 台湾神隆股份有限公司 | Analytical method for detecting sulfated oligosaccharides |
| CN105755073A (en) * | 2014-12-15 | 2016-07-13 | 中国科学院大连化学物理研究所 | High-efficiency separation preparation method for preparing N-linked polysaccharide from ovalbumin, and polysaccharide |
| CN106866749A (en) * | 2015-12-13 | 2017-06-20 | 中国科学院大连化学物理研究所 | A kind of preparation method of breast milk oligosaccharides |
| CN107621399A (en) * | 2016-07-14 | 2018-01-23 | 北京三元食品股份有限公司 | A kind of method of oligosaccharide in detection breast milk |
| CN108699018A (en) * | 2015-11-25 | 2018-10-23 | 德玛医药（Bc）有限公司 | The analysis of the preparation of double dehydrated galactitols and its derivative or the like and resolving method |
-
2018
- 2018-11-28 CN CN201811438772.8A patent/CN111239311A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101421012A (en) * | 2004-04-23 | 2009-04-29 | 吉恩康国际股份有限公司 | Method of separating components in a sample using silane-treated silica filter media |
| JP2008309699A (en) * | 2007-06-15 | 2008-12-25 | Hitachi High-Technologies Corp | Two-dimensional liquid chromatograph of ion exchange and normal phase column |
| US20100003699A1 (en) * | 2008-01-18 | 2010-01-07 | Glykos Finland Ltd. | Tissue carbohydrate compositions and analysis thereof |
| CN105431733A (en) * | 2013-07-11 | 2016-03-23 | 台湾神隆股份有限公司 | Analytical method for detecting sulfated oligosaccharides |
| CN105755073A (en) * | 2014-12-15 | 2016-07-13 | 中国科学院大连化学物理研究所 | High-efficiency separation preparation method for preparing N-linked polysaccharide from ovalbumin, and polysaccharide |
| CN108699018A (en) * | 2015-11-25 | 2018-10-23 | 德玛医药（Bc）有限公司 | The analysis of the preparation of double dehydrated galactitols and its derivative or the like and resolving method |
| CN106866749A (en) * | 2015-12-13 | 2017-06-20 | 中国科学院大连化学物理研究所 | A kind of preparation method of breast milk oligosaccharides |
| CN107621399A (en) * | 2016-07-14 | 2018-01-23 | 北京三元食品股份有限公司 | A kind of method of oligosaccharide in detection breast milk |
Non-Patent Citations (3)
| Title |
|---|
| JINGYU YAN ET AL: "Profiling of Sialylated Oligosaccharides in Mammalian Milk Using Online Solid Phase Extraction-Hydrophilic Interaction Chromatography Coupled with Negative-Ion Electrospray Mass Spectrometry", 《ANAL. CHEM.》 * |
| MARIA LORNA A. DE LEOZ ET AL: "A quantitative and comprehensive method to analyze human milk oligosaccharide structures in the urine and feces of infants", 《ANAL BIOANAL CHEM》 * |
| 刘峰 等: "HILIC-MS/MS法测定人尿中内源性糖类化合物浓度", 《药物分析杂志》 * |
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