CN101363830B - Method for estimating aptness of royal jelly storage condition - Google Patents
Method for estimating aptness of royal jelly storage condition Download PDFInfo
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- CN101363830B CN101363830B CN2008102234978A CN200810223497A CN101363830B CN 101363830 B CN101363830 B CN 101363830B CN 2008102234978 A CN2008102234978 A CN 2008102234978A CN 200810223497 A CN200810223497 A CN 200810223497A CN 101363830 B CN101363830 B CN 101363830B
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Abstract
The invention provides an evaluation method for the suitability of the storage condition of royal jelly. The suitability of the storage condition of royal jelly is judged by detecting the content of free Gln in the royal jelly and according to the content of free Gln in the royal jelly, and the quality of the royal jelly can be further judged. The storage condition of the royal jelly can be sensitively judged, so as to provide a basis for the manufacturers and the consumers to accurately judge the quality of the royal jelly, and the present royal jelly quality specification can also be completed, in order to effectively monitor the quality of the royal jelly.
Description
Technical field
The present invention relates to a kind of food quality evaluation method, specifically relate to a kind of evaluation method of royal jelly storage requirement adaptability.
Background technology
Royal jelly (Royal jelly) is secreted by nurture honeybee head hypopharyngeal gland (Hypopharyngeal gland) and mandibular gland (Mandibular gland) jointly, has acid, puckery, peppery, sweet and have a little the milky or the flaxen slurry of special aromatic odor.Royal jelly plays an important role in the differentiation of the level type of honeybee, the small grub that the incubating oosperm of being given birth to by queen bee equally becomes, if whole larval phase all by the royal jelly of feeding, only need 16d just can develop into reproductive organs queen bee completely; If only feed royal jelly in first three sky of larval phase, change subsequently and feed honey and pollen, need 21d, and can only develop into the incomplete worker bee of reproductive organs.
The component of royal jelly is very complicated, is rich in protein, carbohydrates and lipid, also comprises the many microcomponents that possibly bring into play important physiological function.Royal jelly contains abundant bioactivator, has such as many-sided nutrition health-care functions such as antifatigue, anti-inflammation, antitumor, hypotensive, growth promotions, has very high using value in fields such as health food, medicine, cosmetics.
Because some active components are responsive to storage temperature and period of storage, royal jelly is had relatively high expectations to storage requirement.If store improper (temperature is too high or period of storage is long), marked change can take place in its physical behavior, chemical constitution and quality thereof, causes its health-care efficacy to reduce even forfeiture.The experiment of bee raising larva shows that storing improperly, royal jelly can not make the bee larva normal development.Long-term clinical verification shows that also the storage requirement that royal jelly experienced is the extremely The key factor of decision royal jelly quality.But owing to lack relevant evaluation method; Quality monitoring department and processing, trading enterprise can't effectively supervise and control in the royal jelly production and the process of circulation so far; The storage requirement and the quality that are difficult to royal jelly is experienced are made evaluation strictly according to the facts, thereby cause on the market royal jelly quality very different.
Summary of the invention
To the problems referred to above, the purpose of this invention is to provide a kind of method that can react storage requirement that royal jelly experiences, evaluation royal jelly quality strictly according to the facts.
Maillard reaction is amine compound in the food (amino acid, peptide and protein) and carbonyls (carbohydrate) abiogenous reaction in food processing and storage.Its course of reaction can make protein and the amino acid in the food lose in a large number.Royal jelly is rich in protein, amino acid and carbohydrates (like fructose, glucose etc.), and it almost is inevitable in the storage process Maillard reaction taking place.In storage process; Some amino acid of free amino acid and constitutive protein matter or polypeptide be prone to participate in reaction such as Mei Lade or react through amino acid whose deaminizating, decarboxylation degrading in the royal jelly, thereby cause that some amino acid contents change in storage process.
Experiment finds that there are linear dependence in royal jelly free amino acid Gln and period of storage and storage temperature, also has linear dependence with the quality of royal jelly.Therefore, the present invention provides a kind of method that can estimate storage requirement that royal jelly experiences, evaluation royal jelly quality strictly according to the facts, exactly.
The present invention provides a kind of method of estimating aptness of royal jelly storage condition, may further comprise the steps:
1) measures free Gln content in the royal jelly;
2) judge the storage requirement that royal jelly experiences according to the free Gln content of royal jelly.
The method of estimating aptness of royal jelly storage condition of the present invention, judge period of storage and the temperature that royal jelly experiences through free Gln content:
When free Gln content during, be equivalent to royal jelly and under 25 ℃ of conditions, store and surpassed 1 month less than 1.08 ± 0.10mg/g.Here; The upper and lower bound of under 25 ℃ of conditions, storing the free Gln content when having surpassed 1 month with royal jelly is as two threshold values; Estimate the freshness of royal jelly; Promptly when free Gln content is less than or equal to 0.98mg/g, be equivalent to royal jelly and under 25 ℃ of conditions, store and surpassed 1 month, be stale royal jelly; Free Gln content is fresh royal jelly more than or equal to the royal jelly of 1.18mg/g; Free Gln content be in 0.98 and 1.18mg/g between royal jelly between wherein, be more stale royal jelly.
Amino acid content is measured can adopt amino-acid analyzer or liquid chromatograph.
According to long-term scientific research and practical experience, royal jelly is long preservation at high temperature, it is generally acknowledged that royal jelly can not surpass 1 month 25 ℃ of preservations, can not be above 3 months 4 ℃ of following preservations.The period of storage of royal jelly has surpassed this cycle and just has been considered to experience unsuitable storage, and it is stale that royal jelly also just is considered to.Therefore, we can select for use royal jelly 25 ℃ of threshold values of preserving 1 month free Gln content as estimating aptness of royal jelly storage condition and freshness.Through various article test, find that the free Gln content of royal jelly when preserving 1 month for 25 ℃ is 1.08 ± 0.10mg/g.Therefore, select the upper limit 1.18mg/g of the free Gln content of royal jelly when preserving 1 month for 25 ℃ and lower limit 0.98mg/g to store two threshold values of adaptability and freshness as estimating royal jelly.
The method of estimating aptness of royal jelly storage condition of the present invention has following advantage:
1) storage requirement that can react royal jelly strictly according to the facts and experienced, thermograde and time gradient all can show difference on free Gln content height, thereby confirm the quality of royal jelly, and whether judge whether to experience unsuitable storage and storage requirement qualified etc.;
2) highly sensitive, analytical approach is simple.
Description of drawings
Fig. 1 is the UPLC chromatogram of 26 seed amino acid standard models.
Fig. 2 is royal jelly free amino acid Gln changes of contents figure in 25 ℃ of storage processes.
Fig. 3 is a free amino acid chromatogram of under different temperatures, storing 6 months royal jelly, is respectively the royal jelly sample that is stored in 25 ℃, 4 ℃ and-18 ℃ from top to bottom.
Embodiment
Following examples are used to explain the present invention, but are not used for limiting scope of the present invention.
With 26 commercial seed amino acid standard models: histidine (His), serine (Ser), arginine (Arg), glycocoll (Gly), asparatate (Asp); Glutamic acid (Glu), threonine (Thr), alanine (Ala), oxylysine (Hylys), proline (Pro); Halfcystine (Cys), lysine (Lys), tyrosine (Tyr), methionine (Met), valine (Val); Isoleucine (Ile), leucine (Leu), phenylalanine (Phe), taurine (Tau), GABA (GABA); Aminoisobutyric acid (AABA), ornithine (Om), glutamine (Gln), asparagine (Asn), hydroxyproline (Hypro) and tryptophane (Trp) (sigma company) are dissolved in the pure water; Final concentration is 5pmoles/ μ L (wherein Cys is 2.5pmoles/ μ L), measures with the Ultra Performance Liquid Chromatography appearance, obtains the standard liquid chromatography, sees Fig. 1.
The royal jelly of newly gathering (camellia royal jelly, Zhejiang) is divided into 5 parts, stores down at 25 ℃, respectively on the same day of gathering, gather and measured free aminoacid content in back 1 month, 3 months, 6 months and 10 months.
Take by weighing the 1.0g royal jelly in the 50mL plastic centrifuge tube, be accurate to 0.01g.Add the 25mL90% ethanolic solution, vortex vibration 2min, behind the ultrasonic Extraction 3min, the centrifugal 10min of 5000rpm gets supernatant.In sediment, add the 25mL90% ethanolic solution again, repeat above step twice, merge supernatant.Utilize Rotary Evaporators with the liquid evaporate to dryness at 40 ℃, residue dissolves with ultrapure water, and is settled to 50mL, is analyzing liquid sample.(Millipore, MA USA) filter the back and measure with the Ultra Performance Liquid Chromatography appearance with 0.22 μ m filter membrane.
The result shows, the same day of gathering, gathers back 1 month, 3 months, 6 months and the content of 10 months royal jelly free amino acid Gln is respectively 1.34,1.08,0.64,0.06,0.02mg/g, downtrending in gradient (Fig. 2).Explain that the free Gln content of royal jelly can well react the influence that the period of storage of royal jelly causes.
The royal jelly of newly gathering (rape royal jelly, Henan) being divided into 3 parts, storing down at-18 ℃, 4 ℃ and 25 ℃ respectively, is 6 months mensuration free amino acids in storage period respectively.
Take by weighing the 1.0g royal jelly in the 50mL plastic centrifuge tube, be accurate to 0.01g.Add the 25mL90% ethanolic solution, vortex vibration 2min, behind the ultrasonic Extraction 3min, the centrifugal 10min of 5000rpm gets supernatant.In sediment, add the 25mL90% ethanolic solution again, repeat above step twice, merge supernatant.Utilize Rotary Evaporators with the liquid evaporate to dryness at 40 ℃, residue dissolves with ultrapure water, and is settled to 50mL, is analyzing liquid sample.(Millipore, MA USA) filter the back and measure (Fig. 3) with the Ultra Performance Liquid Chromatography appearance with 0.22 μ m filter membrane.
The result shows; When be 6 months storage period; The content that is stored in the royal jelly free amino acid Gln of-18 ℃, 4 ℃ and 25 ℃ is respectively 1.25,1.10 and 0.06mg/g; Downtrending is in gradient explained that free amino acid Gln content can react well to store the ground temperature conditions to influence that royal jelly caused.
Gather respectively 50 parts in fresh camellia royal jelly (Zhejiang), twigs of the chaste tree royal jelly (Beijing), acacia royal jelly (Shandong), rape royal jelly (Qinghai) sample respectively, totally 200 samples were stored 1 month down at 25 ℃.Take by weighing the 1.0g royal jelly then in the 50mL plastic centrifuge tube, be accurate to 0.01g.Add the 25mL90% ethanolic solution, vortex vibration 2min, behind the ultrasonic Extraction 3min, the centrifugal 10min of 5000rpm gets supernatant.In sediment, add the 25mL90% ethanolic solution again, repeat above step twice, merge supernatant.Utilize Rotary Evaporators with the liquid evaporate to dryness at 40 ℃, residue dissolves with ultrapure water, and is settled to 50mL, is analyzing liquid sample.(Millipore, MA USA) filter the back and measure with the Ultra Performance Liquid Chromatography appearance with 0.22 μ m filter membrane.
The result shows; The free Gln content of the camellia royal jelly (Zhejiang) when storing 1 month down for 25 ℃, twigs of the chaste tree royal jelly (Beijing), acacia royal jelly (Shandong), rape royal jelly (Qinghai) is respectively 1.05 ± 0.10mg/g, 1.09 ± 0.12mg/g, 1.10 ± 0.05mg/g, 1.08 ± 0.11mg/g; Explanation is when storing 1 month down for 25 ℃; The free Gln content of the royal jelly of separate sources remains between 0.98～1.18mg/g basically, keeps stable, and average content is 1.08 ± 0.10mg/g.
Embodiment 4
Gather 5 parts of fresh camellia royal jelly (Zhejiang), twigs of the chaste tree royal jelly (Beijing), acacia royal jelly (Shandong), rape royal jelly (Qinghai) sample respectively, stored 3 months down at 4 ℃.Take by weighing the 1.0g royal jelly then in the 50mL plastic centrifuge tube, be accurate to 0.01g.Add the 25mL90% ethanolic solution, vortex vibration 2min, behind the ultrasonic Extraction 3min, the centrifugal 10min of 5000rpm gets supernatant.In sediment, add the 25mL90% ethanolic solution again, repeat above step twice, merge supernatant.Utilize Rotary Evaporators with the liquid evaporate to dryness at 40 ℃, residue dissolves with ultrapure water, and is settled to 50mL, is analyzing liquid sample.(Millipore, MA USA) filter the back and measure with the Ultra Performance Liquid Chromatography appearance with 0.22 μ m filter membrane.
The result shows; The free Gln content of the camellia royal jelly (Zhejiang) when storing 3 months down for 4 ℃, twigs of the chaste tree royal jelly (Beijing), acacia royal jelly (Shandong), rape royal jelly (Qinghai) is respectively 1.02 ± 0.09mg/g, 1.08 ± 0.11mg/g, 1.07 ± 0.11mg/g, 1.03 ± 0.13mg/g; Explanation is when storing 3 months down for 4 ℃; The free Gln content of the royal jelly of separate sources remains between 1.00～1.15mg/g basically, keeps stable, and average content is 1.04 ± 0.09mg/g.
Experimental example
Take by weighing commercially available royal jelly sample: A (Shandong, lot number 20070815) respectively, B (Beijing, lot number 20080409), 5 parts in C (Hubei, lot number 20070106) and D (Zhejiang, lot number 20071013) sample were stored 1 month down at 25 ℃.Accurately take by weighing the 1.0g royal jelly then in the 50mL plastic centrifuge tube, be accurate to 0.01g.Add the 25mL90% ethanolic solution, vortex vibration 2min, behind the ultrasonic Extraction 3min, the centrifugal 10min of 5000rpm gets supernatant.In sediment, add the 25mL90% ethanolic solution again, repeat above step twice, merge supernatant.Utilize Rotary Evaporators with the liquid evaporate to dryness at 40 ℃, residue dissolves with ultrapure water, and is settled to 50mL, is analyzing liquid sample.(Millipore, MA USA) filter the back and measure with the Ultra Performance Liquid Chromatography appearance with 0.22 μ m filter membrane.
The result shows that the free Gln content of four kinds of commercially available royal jelly samples is respectively A, 1.26mg/g; B, 1.37mg/g; C, 0.96mg/g; D, 0.65mg/g; Thereby judge that A and B quality grade are fresh, C and D quality grade are stale.Wherein, the product batch number of sample D is 20071013, possibly under uncomfortable condition, store a period of time but still stale reason is the preceding royal jelly of enterprise production processing.Test findings is consistent with the result of organoleptic examination.
Claims (2)
1. the method for an estimating aptness of royal jelly storage condition is characterized in that, may further comprise the steps:
1) measures free Gln content in the royal jelly;
2) judge period of storage and the temperature that royal jelly experiences through free Gln content:
When free Gln content is less than or equal to 0.98mg/g, is equivalent to royal jelly and under 25 ℃ of conditions, stores and surpassed 1 month, be stale royal jelly; Free Gln content is fresh royal jelly more than or equal to the royal jelly of 1.18mg/g; Free Gln content be in 0.98 and 1.18mg/g between royal jelly between wherein, be more stale royal jelly.
2. the method for estimating aptness of royal jelly storage condition as claimed in claim 1 is characterized in that, amino acid content is measured and adopted amino-acid analyzer or liquid chromatograph.
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| Emanuele BOSELLI et al..Determination and changes of free amino acids in royal jelly during storage.《Apidologie》.2003,(第34期),129-137. * |
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