CN103487455A - Novel method for quick quantitative determination of hydroxy components in olive oil - Google Patents

Abstract

The invention discloses a novel method for quick quantitative determination of hydroxy components in olive oil and belongs to the field of edible fat and oil detection. Based on the method of the 19 FNMR technological invention, the method, disclosed by the invention, can detect the content of a hydroxy compound in olive oil in a simple, quick and quantitative manner and can be widely applied to detection of content of the hydroxy compound in other edible fat and oil. A fluorine derivation reagent- fluoro benzoyl chloride has substitution reaction with the hydroxy compound to enable the hydroxy compound to be subjected to fluorine derivatization. A Fourier superconductivity nuclear magnetic resonance spectrometer is adopted to draw a 19 FNMR atlas for the hydroxy compound subjected to fluorine derivatization. Manual integration is performed on the atlas, and the content of the hydroxy compound can be accurately calculated. According to the novel method disclosed by the invention, the result is accurate and reliable, the method is simple and practical, and the operation is simple and easy. The novel method can provide valuable reference for quality analysis of olive oil and can also be applied to detection of content of the hydroxy compound in other edible fat and oil.

Description

A kind of method of hydroxy-containing compounds in novel Quantitative detection olive oil

Technical field

The invention belongs to edible oil and fat detection technique field, specifically detect the method for hydroxy-containing compounds in olive oil.

Background technology

Olive oil is that in edible oil and fat commonly used, nutritive value is more a kind of than more rich.Because olive oil contains abundant unsaturated fatty acid and microcomponent, make it there is very high edibility and health-care efficacy.Be described as " liquid golden ", " Mediterranean sweet dew " in west.Hydroxy-containing compounds is the important component part of olive oil microcomponent, comprises the materials such as monoglyceride, diglyceride, straight-chain fatty alcohol class, sterols (cupreol, Δ-5-avenasterol, cholesterol), phenols (tocopherol, tyrosol, hydroxytyrosol, lignin etc.).

The fatty acid overwhelming majority in olive oil is all that the form with glycerate exists, wherein sweet three esters account for more than 90%, the content of monoglyceride is lower than 1%, DAG is mainly with 1, the form of 3-DAG exists, but due to the differences such as the increase along with the storage time, refining, the place of production, 1 in olive oil, the 3-DAG can turn to 1,2-DAG by isomery.Research shows that 1,3-DAG and the ratio of total DAG can be used as the important evidence of olive oil classification.

Clinical research shows, the plant sterol in diet can suppress small intestine to the absorption of cholesterol and reduce the level of Blood Cholesterol.Modern study shows that plant sterol also has anti-inflammatory, antibiotic, antiulcer, anti-oxidant and antitumor activity.What the free content of phytosterol in virgin oil was subject to storage time and condition of storage affects the meeting esterification, and the free plant sterol in refined olive oil also esterification can occur in addition.Therefore the content that detects the free plant sterol in olive oil has important references to the quality assessment of olive oil and is worth, and is also to detect the adulterated important indicator of olive oil.

Aldehydes matter in olive oil (

, hydroxytyrosol, olive bitter principle etc.) be natural antioxidant, olive oil is without adding other synthetic antioxidants again, and aldehydes matter is also the main matter of giving the olive oil particular tastes.Epidemiological study shows that aldehydes matter can be good at pre-anti-cancer and coronary heart disease, also has antidotal effect.Because the content of phenolic compounds in olive oil is low, complex structure, the detection method of traditional method is difficult to detect fast and accurately content.Aldehydes matter in refined olive oil descends owing to through high temperature, deodorization etc., processing content.

Acidity is for measuring the index of the shared ratio of free fatty acid in each hectogram grease, free fatty acid is again free fatty acid, is the fatty ingredient of " not behaving oneself " in grease, and acidity is too high easily causes oxidation, thereby cause spoiled by rancid oil or fat, edible not.The standard demonstration of International Olive Oil Council and European Union, the oleic acid content of Extra Virgin is lower than 0.8, and virgin oil is lower than 2.0, and refined olive oil is due to process chemical deacidification process, and acidity is lower than 0.3.The acidity of detection olive oil and other greases can judge the ratio index of the free fatty acid of grease, easily judges whether to become sour.Also can produce aldehyde compound in the Oxidation of Fat and Oils process, the content that detects aldehyde compound can judge the degree of oxidation of grease, virgin oil does not pass through high temperature, high pressure, so aldehyde compound content is all lower, the aldehyde group content of refined olive oil is more much higher than virgin oil, and this is one of important indicator of judgement olive oil grade.

Detect at present hydroxyl material in olive oil and be mainly vapor-phase chromatography, ¹h NMR method, ¹³c NMR method.Due to ¹h NMR has overlap peak, ¹³the C NMR relaxation time is long insensitive, like this spectrum analysis and quantitatively all have very large challenge.Gas chromatographic detection needs first by the free sterol esterification, owing in olive oil, itself, just containing some esterification plant sterol, so the free plant sterol of detection will first detect total plant sterol, could further detecting the plant sterol that dissociates.This procedure very complicated and taking time and effort, require all higher to the operation of experiment and the control of condition.

The present invention is based on ¹⁹the method of F NMR technological invention can Simple fast quantitatively detects the content of hydroxy-containing compounds in olive oil.

Summary of the invention

The present invention is based on ¹⁹the method kit of F NMR technological invention can Simple fast quantitatively detects the content of hydroxy-containing compounds in olive oil, can extensively apply to the detection of other edible oil and fat.

The technical solution adopted in the present invention is: fluorine derivation reagent 4-fluorobenzoyl chloride and hydroxy-containing compounds generation substitution reaction make hydroxy-containing compounds fluorine derivatization.Use German BRUKER AVANCE/AV 500 MHz Fourier NMR spectrometer with superconducting magnet that the hydroxy-containing compounds of fluorine derivatization is done ¹⁹f NMR collection of illustrative plates.

Use nuclear magnetic spectrum process software MestReNova V6.1 to carry out manual integration to the hydroxy-containing compounds image, calculate hydroxy-containing compounds content in sample.

A kind of method of hydroxy-containing compounds in novel Quantitative detection olive oil, it is characterized in that thering is following process and step:

A. the selection of the fluorine derivatization reagent of hydroxyl compound: described fluorine derivation reagent comprises 4-fluorobenzoyl chloride, 2,4,6-trifluorobenzoyl chloride, 3,5-difluoro benzoyl chloride.

B. the preparation of storing solution: described storing solution is the mixed liquor that deuterochloroform and pyridine volume ratio are 1.5:1 or 1.6:1, the storage reagent dosage is 10.0ml, store reagent and contain 0.1mg phenyl-hexafluoride 20mg tert-butyl group Pyrogentisinic Acid, be placed on waterproof in molecular sieve and store.Phenyl-hexafluoride, tert-butyl group Pyrogentisinic Acid are interior mark.

C. the reaction treatment of olive oil sample: described olive oil sample injected volume 100mg to 150mg, be placed in the 5mm nuclear magnetic tube and add the above-mentioned storage reagent of 0.5ml to 0.6ml and 30 μ L fluorine derivation reagent, reaction mixture is placed under room temperature and reacted 30min, after complete reaction is complete, of nuclear magnetic resonance spectrometer, make immediately ¹⁹f NMR collection of illustrative plates.

D. utilize nuclear magnetic resonance spectrometer to do ¹⁹f NMR collection of illustrative plates: utilize nuclear magnetic resonance spectrometer to do ¹⁹f NMR collection of illustrative plates obtains in BRUKER AVANCE/AV 500 MHz Fourier NMR spectrometer with superconducting magnet operations; Probe temperature is 26 ± 1 ° of C; Typical spectrum parameter is as follows: 90 pulse width 19.3 μ s, sweep length 100 kHz, relaxation delay 1s, memory size 64 K.Each spectrum accumulated samples 64 times.All collection of illustrative plates, before carrying out Fourier transform, enlarge live width 0.3Hz tuning drift.

E. use nuclear magnetic resonance map process software MestReNova V6.1 to carry out manual integration to the hydroxy-containing compounds image, calculate the content of hydroxy-containing compounds in sample: computing formula is:

i _dG with i _i be respectively the integration of diglyceride and 4-fluorobenzoyl chloride, m _dG for the molecular weight of diglyceride, m is the actual use amount of olive oil in experiment, a for interior target mM amount.

The invention has the beneficial effects as follows: can simply quantitatively detect fast the content of hydroxy-containing compounds in olive oil.Result accurately and reliably, the method simple possible, simple to operate easy to learn, the quality analysis that can be olive oil provides of great value reference, also can be used for detecting the content of the hydroxy-containing compounds of other edible oils.

Figure of description

Fig. 1, utilization ¹⁹f NMR detects the collection of illustrative plates of glycerine.

figure2, use ¹⁹f NMR detects the collection of illustrative plates of hydroxytyrosol.

figure3, use ¹⁹f NMR detects the collection of illustrative plates of palmityl alcohol.

figure4, use ¹⁹f NMR detects

collection of illustrative plates.

figure5. use ¹⁹f NMR detects 1,2-diolein, 1, the collection of illustrative plates of 3-diolein.

figurehydroxytyrosol,

, palmityl alcohol etc. ¹⁹f NMR collection of illustrative plates

Embodiment

below with reference to the embodiment form, foregoing of the present invention is described further.

(1). store the preparation of reagent: preparation deuterochloroform and pyridine volume ratio are that 1.5:1 stores reagent 10.0ml, store reagent and contain 0.1mg phenyl-hexafluoride 20mg tert-butyl group Pyrogentisinic Acid, are placed on waterproof in molecular sieve and store.Phenyl-hexafluoride, tert-butyl group Pyrogentisinic Acid are interior mark.

(2). the preparation of nuclear-magnetism sample: accurately weigh olive oil 150mg and be placed in the 5mm nuclear magnetic tube and add the above-mentioned storage reagent of 0.5ml and 30 μ L4-fluorobenzoyl chlorides, reaction mixture is placed under room temperature and reacted 30min, after complete reaction is complete, use immediately nuclear magnetic resonance spectrometer to do ¹⁹f NMR collection of illustrative plates.

(3). magnetic resonance detection: ¹⁹f NMR collection of illustrative plates obtains in BRUKER AVANCE/AV 500 MHz Fourier NMR spectrometer with superconducting magnet operations.Probe temperature is 25 ℃.In this research, typical spectrum parameter is as follows: 19.3 μ s of 90 pulse widths, sweep length 100 kHz, relaxation delay 1s, memory size 64 K.Each spectrum accumulated samples 64 times.All collection of illustrative plates, before carrying out Fourier transform, enlarge live width 0.3Hz tuning drift.

The model compound fluorine derivative ¹⁹under the chemical shift of F NMR (ppm) is shown in table 1.

The fluorine derivative of table 1. model compound ¹⁹f nmr chemical displacement (ppm)

Compound 19The displacement of F nmr chemical

1,2-diolein 107.94

1,3-diolein 107.84

Glycerine 107.89(alpha-position hydroxyl)

107.71(β-position hydroxyl)

Hydroxytyrosol 107.28(phenolic hydroxyl group)

108.54(alcoholic extract hydroxyl group)

107.62

Palmityl alcohol 108.95

Stearyl alcohol 108.95

Free fatty acid 103.90

Aldehyde radical 105.13

The detection of relevant olive oil sample

By Shanghai City Entry-Exit Inspection and Quarantine Bureau, provided from 37 olive oil samples of five countries such as Spain, Italy, Greece, Tunisia, Turkey, 12 samples of Spain wherein, 13 samples of Italy, 5 samples of Greece, 4 samples of Turkey, 3, Tunisia sample.Sample 1 is Extra Virgin to sample 24, and sample 25 is mixed olive oil to sample 37.

The result statistics that detection computations obtains exists table 2in.

Table 2. ¹⁹the content of the diglyceride that the olive oil of F NMR detection different regions, different brackets obtains.

。

Claims (1)

1. the method for hydroxy-containing compounds in a novel Quantitative detection olive oil, it is characterized in that thering is following process and step:

A. the selection of the fluorine derivatization reagent of hydroxyl compound: described fluorine derivation reagent comprises 4-fluorobenzoyl chloride, 2,4,6-trifluorobenzoyl chloride, 3,5-difluoro benzoyl chloride.

B. the preparation of storing solution: described storing solution is the mixed liquor that deuterochloroform and pyridine volume ratio are 1.5:1 or 1.6:1, the storage reagent dosage is 10.0ml, store reagent and contain 0.1mg phenyl-hexafluoride 20mg tert-butyl group Pyrogentisinic Acid, be placed on waterproof in molecular sieve and store.Phenyl-hexafluoride, tert-butyl group Pyrogentisinic Acid are interior mark.

C. the reaction treatment of olive oil sample: described olive oil sample injected volume 100mg to 150mg, be placed in the 5mm nuclear magnetic tube and add the above-mentioned storage reagent of 0.5ml to 0.6ml and 30 μ L fluorine derivation reagent, reaction mixture is placed under room temperature and reacted 30min, after complete reaction is complete, of nuclear magnetic resonance spectrometer, make immediately ¹⁹f NMR collection of illustrative plates.

D. utilize nuclear magnetic resonance spectrometer to do ¹⁹f NMR collection of illustrative plates: utilize nuclear magnetic resonance spectrometer to do ¹⁹f NMR collection of illustrative plates obtains in BRUKER AVANCE/AV 500 MHz Fourier NMR spectrometer with superconducting magnet operations; Probe temperature is 26 ± 1 ° of C; Typical spectrum parameter is as follows: 90 pulse width 19.3 μ s, sweep length 100 kHz, relaxation delay 1s, memory size 64 K.Each spectrum accumulated samples 64 times.All collection of illustrative plates, before carrying out Fourier transform, enlarge live width 0.3Hz tuning drift.

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