ArticlePDF Available

Royal Jelly and Bee Brood: Harvest, Composition, Quality

The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 1
The Royal Jelly Book
For so work the honey-bees
Creatures that by a rule in nature teach
The Act of order to a peopled kingdom.
They have a king and officer of sorts.
Shakespeare, King Henry V
Shakespeare did not know, that the queen was the chief bee officer, royal jelly being the food stuff,
produced by bees for raising her….
Be aware that this online book is only for private use and should not be copied and reprinted as some of the images
are not copyrighted.
I would appreciate your feedback at
Stefan Bogdanov, Muehlethurnen, Switzerland
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 2
Royal Jelly and Bee Brood:
Harvest, Composition, Quality
Stefan Bogdanov
Until the end of the 19
century royal jelly (RJ) was not known as a bee product. RJ is produced by the
hypopharyngeal gland of young worker bees. In 1888 the German von Planta found, that the food of workers,
drones and the queen was different, which was supported also by Haydak
In the sixties and seventies an intensive research by Rembold and coworkers to identify of the key queen substance
was carried out. It became clear, that the main components of the queen and the worker feedings , i.e. proteins,
carbohydrates and lipids are the same, while royal jelly contains more amino acids, nucleotides and vitamines
. The fatty acid pattern of the RJ is also different from the worker jelly
, while the juvenile hormone, present in
RJ plays also a role for the emergence of the queen
Also, it was found that feeding of lower amounts of fructose and glucose produces workers, while feeding of
higher amounts of these sugars produces queens
Another hypothesis incorporating the conflicting results of other researchers, suggested that the “correct” viscosity
of RJ, together with higher consumption is the key factor for queen determination
Thus, today, there is not an accepted unifying hypothesis, which explains how RJ works to produce the bee queen.
A. mellifera queen bees differs from the worker bee in several aspects:
Morphology: Contrary to the workers bees, the queen bee has reproductive organs
Development period: the average queen develops in 15 days, while the worker requires 21 days
Life span: The queen lives for several years, while the worker bees lives several weeks (during summer) to
several months (in winter)
Behaviour: The queen lays several thousand eggs a day, while workers lay only occasionally. Unlike
workers, the queen does not participate in common hive activities.
Mainly the fertility and the longer life span of queen bees created the myth of RJ as a wonder-creating food.
Although RJ has proven biological effects, it is far from having the status of a food creating similar wonders in
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 3
Royal jelly production steps
Images: 1, 2, 4: Y. Kohl; other images: G. Ratia.
1. Introduce drop of royal jelly in each artificial cell (image 1).
2. Take a young larvae (less than three days old) from a comb and place it in the cell (image 1). Introduce frame in
the hive. The bees start raising the queens: Images 2, 3.
3. After 3-4 days the maximum amount of RJ is produced. Take frame with queen cells and uncover the cells. The
queen larvae is in a cell full of RJ. About 0.3 g of RJ is in the cell, ready for harvest: Image 4.
4. The RJ is sucked off with a pipette and filtered (filter mesh-size 0.2 mm). Store clean RJ in a cool dark place:
Images 5 and 6
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 4
It was found that the RJ quality depends on the time of harvesting. Optimum RJ quality is achieved when RJ is
harvested 72 hours after grafting of the larvae
Fresh royal jelly
Normally bees produce small quantities of RJ because they nourish a few queen larvae. In order to produce bigger
RJ amounts beekeepers use bees‘ instinct to produce a new queen, if the old one is away. RJ is produced in
specialised beekeeping units, where up to 500 g per colony can be produced in one season. Two production
methods are used:
Discontinuous method: Take away queen from the colony and introduce a frame with artificial queen cells
containing larvae. This frame will be exchanged every 3 days, while the RJ is harvested. After 3-4 times a
harvest break should be made as the RJ quantity diminishes strongly.
Continuous method: This method allows a permanent production during the whole bee season. A nucleus,
composed of the queen, together with a small part of the colony is placed near the parent colony. After every
second RJ harvest the combs of the nucleus and the queenless mother colony are exchanged, so that the queen
has more room for laying and new nursing bees and larvae can be transferred from the nucleus to the queenless
Presently the continuous method is mostly used for commercial RJ production.
The production steps are the same for both methods.
According to the book “Apiculture in China
the main factors for optimum RJ production are:
Breeding of high RJ producing bee strains
Use of strong colonies
Optimum number of cells per colony. In general about 1 g of royal jelly is produced from 3 to 4 comb
Adjust the cycle royal jelly extraction and use brood of the right age. If royal jelly is extracted every 3 day
the brood should be 1 to 1.5 days old. If the brood is extracted once every other day the brood should be 2
to 2.5 days old
Provide enough feeding and water for the colonies (pollen, nectar). If no enough pollen is available, feed
with pollen substitutes. The production of RJ should not be interrupted unless there is emergency
When RJ is extracted the brood must not be broken when the queen cells are con or the brood is picked out.
After extraction RJ should be filtered with a 100 mesh nylon net and store at –18 C
Details of RJ production are described in recent publications
14, 15, 18, 24, 44-46,
. A system has been developed at Zhejiang University to produce high
yields of royal jelly from a new strain of honey bees by optimising the
different production steps
According to this method the RJ production
methods could be improved by better queen selection, new equipment,
prolongation of the production period, improved manipulations skills,
technology and feeding. Presently, about 150 g per colony over 3 days can
be produced, and 7.7 kg per colony and year
A.m.ligustica and A.m.caucasica are better suited for RJ production than
23, 26
Feeding of the colony during the productions period of the colony should be optimum with sugar syrop or sugar
patties with pollen, but no pollen replacement nutrients should be used
An economic production of royal jelly and its rapid conservation method have been proposed
Freeze-dried royal jelly
Freeze-dried royal jelly is a very hygroscopic powder. It is obtained by evaporating the water
content from the frozen product in vacuum. This is the drying process which best maintains
the original characteristics of the product: it retains the volatile components which would be
removed by evaporation at higher temperatures and does not damage nor denature the
thermolabile components.
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 5
Freeze-drying requires special equipment, ranging from a simple laboratory freeze-drier to large industrial
machines. Though the small laboratory models are normally used for analysis only, small volumes of royal jelly
can be processed adequately with this size of equipment.
For drying, the royal jelly is first diluted with some clean water. This leads to a more regular and complete loss of
water, particularly if large quantities are freeze dried in one batch. No such preparation is necessary if royal jelly is
dried directly in the sales vial. During the final drying phase, in order to achieve more complete removal of residual
water, the substrate can be warmed very slightly, but never above 35
After freeze-drying, the royal jelly becomes extremely hygroscopic and must be protected from the humidity of the
environment by storage in an air-tight container. Larger processors handle freeze-dried royal jelly only in
controlled atmospheres, i.e. air conditioned rooms with very low humidity. Depending on the final use of the dried
royal jelly, a carrier base or stabilizer can be added at this point to reduce the hygroscopicity of the dried product,
e.g. cyclodextrin
Freeze-dried royal jelly marketed directly to the consumer is usually presented in separate vials one or more for a
liquid solvent and others containing the dry phase. This is the best solution for conservation without chemical
preservatives. The liquid phase can be pasteurized and packed aseptically, without damaging the heat sensitive
royal jelly. Freeze dried RJ has the same biological properties as fresh one
Freshness has been attributed a great importance for RJ quality. Royal jelly can be spoiled easily if not properly
stored. Immediately after harvest it should be placed in dark vessel and stored 0 - 5°C. Stored under these
conditions its quality remains OK for half an year. Deterioration of royal jelly can be prevented by storing RJ in
Argon after harvesting
. After longer storage it will turn rancid. Frozen royal jelly can be lyophilised as it can be
transported more easily in the dry state. If frozen, it can be stored for 2-3 years without loosing of its quality.
Chauvin states that the physical properties of RJ change after 20 hours after harvest, if left at ambient temperature
. That means that RJ should be stored in the cold immediately after harvest. According to Chauvin RJ the
biological properties of RJ in what regards its capability to induce hyperglycaemia, remain intact only for 1 month,
if stored at about 4°C. On the other hand Krylov tested whole RJ, stored for one year at 5 °C and found out that its
antimyocard activity, measure was not different, in comparison to fresh RJ
. Recently it was also shown that only
storage of RJ in frozen state prevents decomposition of biologically active RJ proteins
On the other hand, storage experiments of fresh RJ and FTIR measurements of protein degradation showed that
after 21 months of storage at -20°C the protein begins to decompose. When RJ is stored at 4 °C RJ should be
stored for a maximum of 7 weeks
Experiments have shown that the enzyme glucose oxidase enzyme contained in RJ is influenced both by storage
temperature and time
2, 6
. At 4°C there was small reduction of enzyme activity, while at 20°C it decreases
significantly after one month and degrades completely after one year
. At 37 and 50°C this decrease is faster
. The
determination of glucose oxidase is analytically very simple and thus within the capabilities of all laboratories. This
method could be used to evaluate the product’s freshness; however, further investigation must first be conducted
into the natural variability of this component in the fresh product.
Recently it was proposed that furosine content can be used as a marker for RJ freshness
. The initial content of this
compound is very low in fresh royal jelly. Specifically, the content rose to as high as 500 mg/100g of protein after
18 months’ storage at room temperature and 50 mg/100g at 4°C. Samples taken from store shelves showed values
ranging from 40 to 100 mg/100g protein. The value of furosine, a product of Maillard’s reaction, proved very low
(from 0 to 10 mg/100g of protein) in freshly produced RJ samples but increases over time and in relation to
temperature. A limit of 50 mg furosine / 100g protein could be used for fresh RJ. A specific RJ protein,
decomposing upon storage can also be used as a freshness marker
. A cheap and fast method based on a
chromogenic reaction of RJ and HCl has been proposed
Improvement of storability
From the above findings it is clear that RJ is an unstable product. Freeze drying is the most important technological
method in order to improve the storability of RJ. However, there is a loss of volatile substances, as reported by
Vahonina, 1995 in
. Stabilisation can be achieved by mixing 1 to 2 % of RJ into honey, where all enzymatic
activity is stopped.
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 6
As reported in
the Russian Braines found out in 1968, that RJ can be bound to a mixture of lactose and glucose,
which improves its durability. In Russia RJ is often offered in such lactose-glucose pills under the name of Apilac.
The method of Braines was improved as follows: Six part of frozen RJ are added to one part of dried glucose-
lactose (1:1), then the mixture containing 50 mg/kg L-ascorbic acid as an antixodant is dried until 4 % humidity.
This product is stable at 4 to 8
C for 2 years
9, 10
Royal jelly is a viscous jelly substance. It is partially soluble in water with a density of 1.1 g/mL. Its colour is
whitish to yellow, the yellow colour increasing upon storage. Its odour is sour and pungent, the taste being sour and
sweet. The sensory characteristic is an important quality criterion. Old royal jelly, which has not been properly
stored tends to be darker and a rancid taste can develop. For optimum quality it should be stored in frozen state.
The viscosity varies according to water content and age - it slowly becomes more viscous when stored at room
temperature or in a refrigerator at 50
. The increased
viscosity appears to be related to an increase in water
insoluble nitrogenous compounds, together with a reduction in soluble nitrogen and free amino acids
. These
changes are apparently due to continued enzymatic activities and interaction between the lipid and protein
There is no international standard for royal jelly, while some countries like Brazil, Bulgaria, Japan, Switzerland and
Uruguay have established national ones. A working group of the International Honey Commission is working on
the elaboration of an international standard. A first work in view of establishment of a standard has been
Composition of royal jelly
Fresh lyophilized
Water % 60 - 70
< 5
Lipids % 3 - 8 8 – 19
10-hydroxy-2-decenoic acid % > 1,4 > 3,5
Protein % 9 – 18 27 – 41
Fructose + glucose+ sucrose % 7 – 18
Fructose % 3 – 13
Glucose % 4 – 8
Sucrose % 0,5 – 2,0
Ash % 0,8 – 3,0 2 – 5
pH 3,4 – 4,5 3,4 – 4,5
Acidity (ml 0.1N NaOH/g) 3,0 – 6,0
The water content, with 60-70 % is the main component of royal jelly. The dry substance is composed of
carbohydrates, proteins, amino acids and fat. Smaller quantities of minerals and vitamins are also present (see
Proteins and peptides
With 17 to 45 % of the RJ dry weight they are the main substance class of RJ
, and also the principal nitrogenous
compounds, accounting for about 97-98 % of them
. About 60 % of them are water-soluble
Free amino acids represent only 0.6 - 1.5% , the majority of which belong to the L series. The most representative
are proline and lysine
7, 65
. Upon storage at 4°C for 10 months no significant changes of amino acids were
encountered, while after room temperature storage proline and lysine content increased
. This is due probably to
proteolytic enzyme activity.
A number of enzymes have been characterised: glucose oxidase
2, 62
, invertase
2, 73
, acid and alkaline phosphatase,
alpha and beta esterase, leucine aminopeptiadase,valin aminopeptiadase lipase, phosphoamidase,
and superoxide
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 7
The lipids with 3 to 19 % of the RJ dry weight
7, 38
, are second in importance after the proteins. 80 to 90 % of the
lipid fraction consists of free fatty acids, the rest being neutral lipids, sterols, hydrocarbons
31, 36, 38, 40, 41
Most of the organic acids are free with rather unusual structure rarely encountered in nature, mono- and dihydroxy
acids and dicarboxylic acids with 8 and 10 carbon atoms
37, 38
. The identification of this fraction – in particular as
regards the pattern and quantitative analysis of free organic acids – is believed to represent the criteria of choice for
defining the genuineness of RJ
6, 11
. The main acid 10-hydroxy-2-decenoic (HDA) is an unsaturated acid, which is
determined for the evaluation of RJ genuinely (see Quality parameters and Standard)
HDA and also the other fatty acids of RJ have antibacterial properties
53, 67
, thus contributing to the relatively low
content of bacteria in this product.
The other fatty acids are all saturated mono- and dihydroxy-, mono- and dicarboxylic acids have not been
quantified exactly, but can be roughly estimated to be around 0.5 to to 1 g/ 100 g
These are third in importance, composed of mainly fructose, glucose and sucrose
39, 42, 66
, with some traces of
maltose, trehalose, melibiose, ribose and erlose also being found
39, 42
The ash content (minerals) represents 0.8 to 3 % of RJ fresh matter.
The major elements are K, P, S, Na, Ca, Al, Mg, Zn, Fe, Cu and Mn but there are trace amounts (0.01-1 mg/100 g)
of Ni, Cr, Sn, W, Sb,Ti and Bi. The sodium content of RJ varies between 11 and 14 mg/ 100 g.
The concentrations of vitamins in RJ are distributed over a broad spectrum; vitamins showing fairly uniform values
are riboflavin, thiamine, niacin and folic acid. Likewise present but with greater variations are pyridoxine, biotin,
pantothenic acid and inositol.
Only traces of vitamin C are present, while the fat soluble vitamins like vitamine A,D, E and K are absent
Other minor components
Numerous minor compounds, belonging to diverse chemical categories, have been identified in royal jelly. Among
these are two heterocyclic substances, biopterine and neopterine at 25 and 5 µg/g of fresh weight respectively
These compounds are found in the food of worker bee larvae too, but at about one tenth of these concentration.
Other substances identified include several nucleotides as free bases (adenosine, uridine, guanosine, iridin and
cytidine) the phosphates AMP, ADP, and ATP
, acetylcholine (1 mg/g dry weight,
and gluconic acid (1.4 % of
fresh weight,
. Benzoic acid (8-15 mg/kg) has also been found
. Small amounts of malic, lactic and citric acid
have also been found
. RJ has antioxidant activity
for which polyphenols are responsible
There is no international standard. Some countries as Brazil, Bulgaria, Japan, Switzerland and Uruguay have
national standards
Sensory requirements
Sensory test Requirements
Colour White to yellow, yellow colour increases with storage
Odour Sour, pungent
Taste Sour, sweet
Consistency A viscous jelly
Visual purity Pollen, very few wax and larvae particles
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 8
Quality requirements for royal jelly and standards
Parameter Requirement
Fresh RJ Lyophylized RJ Reference, analysis
General quality
Humidity Max. 70 g/100g Max. 5 g/100g
10-HDA (HPLC) > 1,9 g / 100 g > 3,5 g / 100 g
12, 28, 32
Furosine (HPLC,GC-MS) Max 50 mg / 100 g* Max 50 mg / 100 g*
50, 54, 77
Purity, Authenticity Corresponds to requirements, see
5, 61
A possible adulterant of royal jelly is bee brood. Bee brood has a similar composition as RJ (see bee brood section)
as to the type of components, but their concentration is different, allowing the proof of bee brood adulterated RJ.
The authenticity of royal jelly is a major quality issue. There are methods for the determination of the authenticity
of RJ
61, 78, 79
Irradiation of RJ which can deteriorate RJ quality can be traced by Electron Spin Resonance spectral
Royal jelly can be contaminated by antibiotics by improper beekeeping practices
. Best quality of royal jelly can be
achieved in certified organic beekeeping.
Fresh, lyophilised
Indicate protein-, carbohydrate and fat content, 10 g RJ correspond to 30 calories
Intake (See Report on health claims, intake)
One full tea spoon of fresh royal jelly is approx. 10 g, determine dosage of RJ on spatula
Fresh RJ: adults: 100 – 250 mg per day; children: half dose
Warning: It is recommended that people who are susceptible to allergies or asthma should avoid intake of royal
Shelf life
Fresh royal jelly:
6 months, if stored in the refrigerator (3 to 5 ºC)
2 years if stored in the freezer (< - 18 ºC)
Lyophylised royal jelly
One year if stored in the refrigerator (3 to 5 ºC)
At least 2 years if stored in the freezer (< - 18 ºC)
Fresh or lyophylised royal jelly in honey
Two years at room temperature, if honey-RJ total humidity is less than 18 %.
Royal Jelly is product, which is very well known in East Asia, while it is much less known in other parts of the
world. There are no official statistics on RJ trade. Some figures are given by Crane. In 1984 the annual production
was about 700 t. In the same year Japan produced 46 tons of RJ. According to these figures China and Taiwan
account for approximately 60 % and 20 % of the world production, the rest is produced in Korea, Japan, Eastern
Europe, Italy, France
In America Mexico is the largest RJ producer
. In Europe RJ is produced mainly in Eastern Europe, Italy and
France. According to a recent article published in 2009 about 3000 tons of RJ are produced annually in China
Thus, today about 4-5000 tons annually are produced world-wide.
In the sixties the whole sale price of RJ was 180 to 400 $ per kg
. In 1993 the wholesale price of Chinese royal
jelly was 50-80 $ per kg, in 2010 it is around 20 to 40 $, according to offers in Internet. Thus, there has been an
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 9
enormous price decline. Efforts are necessary to insure RJ quality and achieve higher prices which are by all means
deserved by such a valuable product.
Bee brood (BB) which is the least recognised bee product in terms of use
for human nutrition. Drone brood (DB), instead of bee brood should be
harvested in order to keep bee populations stable.
In earlier cultures this product was probably of second greatest importance
after honey. Bee brood could therefore serve as a direct food source once the
beekeeper has no more need for extra bees or brood, or when undesired
colonies have to be removed. Honeybee brood of all ages is eagerly
consumed by honey hunters in Africa and Asia and is generally considered a
delicious treat. The brood is said to form a considerable part of their diet
(Hill et al., 1984 and Bailey, 1989; as cited in
). In China and Japan, drone
larvae are canned for export or, after being covered in chocolate. DB is eaten
most often together with the combs or the pupae can be picked out and pickled or boiled. Indeed, bee brood is
particularly rich in protein.
In 1960 it was estimated that 132 tons of bee brood is destroyed before winter just in the Canadian provinces of
Alberta, Manitoba and Sasketchewan. They wanted to develop a market for this food and found out that the most
accepted form was deep fat frying of the brood. When brood was prepared by either shallow frying in butter or deep-
fat frying in vegetable cooking fat and tested by a panel of Canadians, "Most reactions were favourable and some were
eulogistic; initial prejudice proved easier to overcome than we had expected. When the tasters were asked to compare
the material to some more familiar food, those most commonly mentioned were walnuts, pork crackling, sunflower
seeds, and rice crispies. In a later, larger taste test, deep-fat fried, butter fried, and baked preparations were highly rated
while smoked, pickled, and brandied were much less preferred
In many Asian and African countries fresh DB is considered a delicacy. A bee brood products named “bakuti”, is
produced in Nepal, described in 1990 by Burgett: Sections of comb are placed in a woven, fabric bag and hand
squeezed over an open container that collects the liquid phase. This is then heated and gently stirred until it
becomes the consistency of soft scrambled eggs. The odor and flavor of bakuti, Dr. Burgett describes as "nut like."
To make it more acceptable to the U.S. palate, he mixes an equal volume of Philadelphia brand cream cheese and
serves the preparation on crackers. In Zimbabwe the Shona use three kinds of hive, recognized as mukuyo
(honeycombs), the machinda (bee pupae), and the pfuma (royal jelly). "Only the mukuyo honey is taken home, that
from the machinda hive is either eaten on the spot or thrown away and that from the pfuma eaten there and then."
A cake-like mass made from honey boiled with millet, and called chihungwe, is eaten as a delicacy or may be taken
to other villages and sold or bartered for grain
BB is regularly sold alongside honey in markets in many parts of Asia
Different aspects of harvesting bee brood as food were investigated. To insure uniformity of larval age at harvest time,
brood rearing was concentrated in certain frames by confining queens in frames having queen excluder walls. Every
fourth day the comb filled with eggs was removed from the cage and replaced by an empty brood comb. Brood was
allowed to develop until most of the larvae were capped (9-11 days). Cells can be uncapped with a thin serrated knife,
and larvae are extracted easily and efficiently by spraying the comb with one or more jets of water. Larvae are
removed from both sides of the comb and allowed to fall onto a cloth filter such as cheesecloth. After the water is
shaken from the cells, the dark empty brood combs can be returned to the queens. The queens prefer them and they
encourage maximum egg production. The author states that it is possible to harvest at least one pound of larvae per
week from each producing queen
Harvesting of DB, which has to be taken away from bee colonies in the frame of alternative Varroa control is a good
occasion for harvesting.
Apilarnil is a Romanian product, based on drone bee larvae and its food. Its composition is similar to the one of
royal jelly
but outside Romania there is no published scientific data on this product.
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 10
The composition of Bulgarian bee brood and royal jelly after
is given in the table below:
Mean±SD and ranges (n=7)
Royal Jelly Drone brood
Water content, %
63.39±1.75 (61.00-65.20) 70.97±0.72 (70.30-72.30
Protein, %
16.73±1.29 (14.65-18.33) 9.35±0.63 (8.12-10.00)
Fructose, %
4.88±0.37 (4.24-5.35) 0.11±0.11 (0.00-0.34
Glucose, %
3.46±0.58 (2.70-4.15) 6.74±0.65 (5.92-7.88)
Sucrose, % 1.53±0.55 (0.59-2.05) 0.05±0.07 (0.00-0.18)
Total sugars, % 9.86±0.93 (8.47-10.80) 6.92±0.70 (6.22-8.22)
pH 3.95±0.09 (3.80-4.02) 6.49±0.14 (6.23-6.63)
Total acidity, ml 0.1 N NaOH 4.07±0.30 (3.68-4.42) 0.88±0.15 (0.74-1.10)
Electrical conductivity, uS/cm 205.14±8.73 (194.00-219.00) 161.43±10.67 (144.00-178.00)
There are no quality requirements for bee brood, as it is generally not sold on the market.
Storage and shelf life
This topic was studied in detail by Burimistrova
. The conclusions of the studies are:
The maximum storage of fresh drone brood (FDB) at room temperature is one hour, then it has to be cooled
or frozen
The maximum storage at 4 to 8
C is 12 to 24 hours
At -20
C it can be stored until 3 months
For better storage six parts FDB it is added to one part of dried glucose-lactose (1:1) the mixture containing 50
mg/kg L-ascorbic acid as an antioxidant is dried until 4 to % humidity. This product is stable at 4 to 8
C for 2
1. ASENCOT, M; LENSKY, Y (1988) The effect of soluble sugars in stored royal jelly on the differentiation of female
honeybee (Apis mellifera L.) larvae to queens. Insect Biochemistry 18 (2): 127-133.
2. BAGGIO, A; DAINESE, N (1998) La qualita della gelatina reale nella conservazione. Industrie Alimentari 37 (375):
3. BAGGIO, A; DAINESE, N (1998) Royal jelly quality during storage. Industrie Alimentari 37 (375): 1290.
4. BALKANSKA, R; KARADJOVA, I; IGNATOVA, M (2014) Comparative analyses of chemical composition of
royal jelly and drone brood. Bulgarian Chemical Communications 46 (2): 412-416.
5. BOGDANOV, S (2006) Contaminants of bee products. Apidologie 38 (1): 1-18.
(2002) Valutazione di produzioni apistiche: gelatina reale e cera, In Sabatini, A G; Bolchi Serrini, G; Frilli,
F; Porrini, C (eds) Il ruolo della ricerca in apicoltura, Litosei; Bologna; pp 321-329.
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 11
7. BOSELLI, E; CABONI, M F; SABATINI, A G; MARCAZZAN, G L; LERCKER, G (2003) Determination and
changes of free amino acids in royal jelly during storage. Apidologie 34 (2): 129-137.
8. BURGETT, M (1990) Bakuti - A Nepalese culinary preparation of giant honey bee brood. The Food Insect Newsletter
3 (2): 1-2.
9. BURIMISTROVA, L; AGAFONOV, A; BUDNIKOVA, N; HARITONOVA, M (2008) Methods for the stabilisation
of biologically active components royal jelly (Russian), Apitherapy today, Ribnoe, 13.Oct.2008: pp 175-
10. BURIMISTROVA, L (1999) Physico-chemical and biological appreciation of drone brood. PhD Ryazan Medical
University, Russia; 159pp.
11. CABONI, M F; SABATINI, A G; LERCKER, G (2004) La gelatina reale: origine, proprietà e composizione/Royal
jelly:origin, properties and composition. APOidea 1: 72-79.
12. CAPARICA-SANTOS, C; MARCUCCI, M C (2007) Quantitative determination of trans-10-Hydroxy-2-Decenoic
Acid (10-HDA) in Brazilian royal jelly and commercial products containing royal jelly. Journal of
Apicultural Research 46 (3): 149-153.
13. CHAUVIN, R (1987) La ruche et l'homme. Calmann-Lévy, France
14. CHEN, C T; CHEN, P L (1999) Effect of fructose, sucrose and queen age on the royal jelly production of honeybee,
Apis mellifera L. Plant Protection Bulletin (Taipei) 41 (1): 59-66.
15. CHEN, S L; SU, S K; LIN, X Z (2002) An introduction to high-yielding royal jelly production methods in China. Bee
World 83 (2): 69-77.
16. CHEN, Y (1993) Apiculture in China. Aricultural Publishing House Beijing
17. CRANE, E (1990) Bees and beekeeping: Science, practice and world resources. Cornell University Press Ithaca, New
18. FERT, G (1999) The production of royal jelly. Bull.Techn.Apicole 26 (3): 109-120.
19. GARY, N E (1961) Mass production of honeybee larvae. Gleanings in Bee Culture 89 (9): 550-552.
20. HAYDAK, M H (1943) Larval Food and development of castes in the honey-bee. Journal of Economic Entomology
36 (5): 778-792.
21. HENSCHLER, D (1956) [Identification of choline esters in biological material, especially acetylcholine in royal jelly
of bee]. Hoppe-Seyler's Zeitschrift für physiologische Chemie 305 (1): 34-41.
22. HOCKING, B; MATSUMURA, F (1960) Bee brood as food. Bee World 41: 113-120.
23. JÉANNE, F (2002) La gelée royale. Technique de production. Bulletin Téchnique Apicole 29 (2): 87-90.
24. JÉANNE, F (2002) Royal jelly. Method of production. Bull.Techn.Apicole 29 (2): 87-90.
25. JENTER, K (2002) New and economic production of royal jelly and its rapid conservation using a revised method.
Bienenpflege (5): 177-179.
26. JENTER, K (2002) New and economic production of royal jelly and its rapid conservation using a revised method
2004. Bienenpflege (5): 177-179.
27. KAMAKURA, M; FUKUDA, T; FUKUSHIMA, M; YONEKURA, M (2001) Storage-dependent degradation of 57-
kDa protein in royal jelly: a possible marker for freshness. Bioscience, Biotechnology and Biochemistry 65
(2): 277-284.
28. KIM, J; LEE, J (2010) Quantitative Analysis of Trans-10-Hydroxy-2-Decenoic Acid in Royal Jelly Products
Purchased in Usa by High Performance Liquid Chromatography. Journal of Apicultural Science 54 (1): 77-
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 12
29. KIM, J K; SON, J H; OH, H S (1989) Analysis of organic acids in honey and royal jelly. Korean Journal of
Apiculture 4 (2): 105-111.
30. KIM, J-G; SON, J-H (1996) The quantity of superoxide dismutase (SOD) in fresh royal jelly. Korean Journal of
Apiculture 11 (1): 8-12.
31. KODAI, T; UMEBAYASHI, K; NAKATANI, T; ISHIYAMA, K; NODA, N (2007) Compositions of royal jelly II.
Organic acid glycosides and sterols of the royal jelly of honeybees (Apis mellifera). Chemical &
Pharmaceutical Bulletin 55 (10): 1528-1531.
32. KOSHIO, S; ALMEIDA-MURADIAN, L B (2003) HPLC application for 10-HDA determination in pure royal jelly
and honey with royal jelly. Quimica Nova 26 (5): 670-673.
33. KRELL, R (1996) Value-added products from beekeeping. FAO Food and Agriculture Organization of the United
Nations Roma; 409 pp
34. KRYLOV, V; SOKOLSKII C. (2000) Royal jelly (in Russian). Agroprompoligrafist Krasnodar; 214 pp
35. LEE, A; YEH, M; WEN, H; CHERN, J; LIN, J; HWANG, W (1999) The application of capillary electrophoresis on
the characterization of protein in royal jelly. Journal of Food and Drug Analysis 7 (1): 73-82.
36. LERCKER, G; CABONI, M F; VECCHI, M A; SABATINI, A G; NANETTI, A (1992) Characterizaton of the main
constituents of royal jelly. Apicoltura (8): 27-37.
37. LERCKER, G; CABONI, M F; VECCHI, M A; SABATINI, A G; NANETTI, A (1992) Characterizaton of the main
constituents of royal jelly
410. Apicoltura (8): 27-37.
38. LERCKER, G; CABONI, M F; VECCHI, M A; SABATINI, A G; NANETTI, A (1993) Caratterizzazione dei
principali costituenti della gelatina reale. Apicoltura 8: 27-37.
della frazione glucidica della gelatina reale e della gelatina delle api operaie in relazione all'eta larvale.
Apicoltura 1: 123-139.
40. LERCKER, G; CAPELLA, P; CONTE, L S; RUINI, F (1981) Components of royal jelly: I. Identification of the
organic acids. Lipids 16 (12): 912-919.
41. LERCKER, G; CAPELLA, P; GIORDANI, G (1982) Components of royal jelly II: The lipid fractions hydrocarbons
and sterols. Journal of Apicultural Research 21 (3): 178-184.
determination of royal jelly by high resolution gas chromatography (HRGC). Food Chemistry 19: 255-264.
43. LERCKER, G; VECCHI, M A; PIANA, L; NANETTI, A; SABATINI, A G (1984) Composition de la fraction
lipidique de la gelée de larves d'abeilles reines et ouvrières (Apis mellifera ligustica Spinola) en fonction de
l'age des larves. Apidologie 15 (3): 303-314.
44. LI, J (2000) Technology for royal jelly production. American Bee Journal 140 (6): 469-472.
45. LI, J (2001) Technologie der Produktion von Weiselfuttersaft. Deutsches Bienen Journal 9 (2): 55-57.
46. LI, J; CHEN, S; ZHONG, B; SU, S (2003) Optimizing royal jelly production. American Bee Journal 143 (3): 221-
47. LI, J K; WANG, T; PENG, W J (2007) Comparative analysis of the effects of different storage conditions on major
royal jelly proteins. Journal of Apicultural Research 46 (2): 73-80.
48. LIHONG, C (2009) Advances in propolis research and propolis industry in China. J.Royal Inst Thailand 1: 136-151.
(2014) Fast analysis of polyphenols in royal jelly products using automated TurboFlow (TM)-liquid
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 13
chromatography-Orbitrap high resolution mass spectrometry. Journal of Chromatography B-Analytical
Technologies in the Biomedical and Life Sciences 973: 17-28.
50. MARCONI, E; CABONI, M F; MESSIA, M C; PANFILI, G (2002) Furosine: a suitable marker for assessing the
freshness of royal jelly. Journal of agricultural and food chemistry 50 (10): 2825-2829.
52. MATSUKA, M (1993) Content of benzoic acid in royal jelly and propolis. Honeybee Science 14 (2): 79-80.
53. MELLIOU, E; CHINOU, I (2005) Chemistry and bioactivity of royal jelly from Greece. J.Agricultural & Food
Chemistry 53: 8987-8992.
54. MESSIA, M C; CABONI, M F; MARCONI, E (2005) Storage stability assessment of freeze-dried royal jelly by
furosine determination. Journal of agricultural and food chemistry 53 (11): 4440-4443.
55. NAGAI, T (2001) [Properties and functions of gluconic acid and its salts]. Honeybee Science 22 (4): 171-174.
(2015) The Lyophilization Process Maintains the Chemical and Biological Characteristics of Royal Jelly.
(2014) Comparison between local and commercial royal jelly - use of antioxidant activity and 10-hydroxy-2-
decenoic acid as quality parameter. Journal of Apicultural Research 53 (1): 116-123.
58. REMBOLD, H (1987) Die Aufklärung der Kastenentstehung im Bienenstaat, In von Detfurth, H (ed.) Ein Panorama
der Naturwissenschaften, Boehringer Mannheim, GmbH.Mannheim; pp 167-231.
59. REMBOLD, H; DIETZ, A (1965) Biologically active substances in royal jelly. Vitamines and Hormones 23: 359-383.
60. REMBOLD, H; LACKNER, B (1978) Vergleichende Analyse von Weiselfuttersäften. Mitteilungen der Deutschen
Gesellschaft für allgemeine und angewandte Entomologie 1 (2/3/4): 299-301.
Quality and standardisation of royal jelly. JAAS 1: 1-6.
62. SANO, O; KUNIKATA, T; KOHNO, K; IWAKI, K; IKEDA, M; KURIMOTO, M (2004) Characterization of royal
jelly proteins in both Africanized and European honeybees (Apis mellifera) by two-dimensional gel
electrophoresis. Journal of agricultural and food chemistry 52 (1): 15-20.
63. SASAKI, M; TSURUTA, T; ASADA, S (1987) Role of physical property of royal jelly in queen differentiation of
honeybee, In Eder, J; Rembold, H (eds) Chemistry and biology of social insects, German Federal Republic,
Verlag J. Papemy; Munich, Germany; pp 306-307.
64. SCHMIDT, J O; BUCHMANN, S L (1992) Other products of the hive. in: The Hive and the Honey Bee (Graham,
J.M., Editor) Dadant & Sons, Hamilton, IL. unknown: 927-988.
65. SERRA BONVEHI, J (1990) Studies on the proteins and free amino acids of royal jelly. Anal.Bromatol. 42 (2): 353-
66. SERRA BONVEHI, J (1992) Sugars, acidity and pH of royal jelly. Anal.Bromatol. 44 (1): 65-69.
67. SERRA BONVEHI, J; ESCOLA JORDA, R (1991) Study of the microbiological quality and bacteriostatic activity of
queen food (royal jelly): effect of organic acids. Deutsche Lebensmittel-Rundschau 87 (8): 256-529.
68. SESTA, G; LUSCO, L (2008) Refractometric determination of water content in royal jelly. Apidologie 39 (2): 225-
69. STANGACIU, S; HARTENSTEIN, E (2004) Sanft Heilen mit Bienen - Produkten. Haug Verlag
The Royal Jelly Book, Chapter 1
Bee Product Science, April 2016 14
70. STOCKER, A; SCHRAMEL, P; KETTRUP, A; BENGSCH, E (2005) Trace and mineral elements in royal jelly and
homeostatic effects. Journal of Trace Elements in Medicine and Biology 19 (2-3): 183-189.
71. TAKENAKA, T; YATSUNAMI, K; ECHIGO, T (1986) Changes in quality of royal jelly during storage. Nippon
Shokuhin Kogyo Gakkaishi 33 (1): 1-7.
of royal jelly protein degradation during storage using Fourier-transform infrared (FTIR) spectroscopy.
Journal of Apicultural Research 51 (2): 185-192.
73. THRASYVOULOU, A T (1982) Biochemical and biological aspects of honey bee (Apis mellifera L.) larval food.
The Pennsylvania State University. The Graduate School. Department of Entomology Pennsylvania, USA;
pp 1-208.
74. TSENG, C; YU, Z; LI, C (1994) Preparation of royal jelly powders and property characterization of the products
during storage. Journal of the Chinese Agricultural Chemical Society 32 (1): 113-124.
QUARANTOTTO, G (1993) Sali minerali nel nutrimento larvale di api regine e operaie (Apis mellifera
ligustica Spinola). Apicoltura 8: 39-54.
76. WOO, K S; LEE, H S; YOON, S Y K J (1998) [Comparative study of royal jelly production in single storey and
multiple storey hives]. Korean Journal of Apiculture 13 (2): 101-104.
77. WYTRYCHOWSKI, M; PAISSE, J O; CASABIANCA, H; DANIELE, G (2014) Assessment of royal jelly freshness
by HILIC LC-MS determination of furosine. Industrial Crops and Products 62: 313-317.
BRION, B (2013) Physicochemical characterisation of French royal jelly: Comparison with commercial
royal jellies and royal jellies produced through artificial bee-feeding. Journal of Food Composition and
Analysis 29 (2): 126-133.
79. WYTRYCHOWSKI, M; DANIELE, G; CASABIANCA, H (2012) Combination of sugar analysis and stable isotope
ratio mass spectrometry to detect the use of artificial sugars in royal jelly production. Analytical and
Bioanalytical Chemistry 403 (5): 1451-1456.
80. YAMAOKI, R; KIMURA, S; OHTA, M (2014) Electron spin resonance spectral analysis of irradiated royal jelly.
Food Chemistry 143: 479-483.
81. ZHENG, H Q; HU, F L; DIETEMANN, V (2010) Changes in composition of royal jelly harvested at different times:
consequences for quality standards. Apidologie DOI: 10.1051/apido/2010033
82. ZHENG, H Q; WEI, W T; WU, L M; HU, F L; DIETEMANN, V (2012) Fast Determination of Royal Jelly Freshness
by a Chromogenic Reaction. Journal of Food Science 77 (6): S247-S252.
... The chemical composition of fresh drone brood is similar to that of royal jelly. Drone brood homogenate is characterized by a higher water content but its protein and carbohydrate content is lower than that in royal jelly [32,33]. A comparison of the physicochemical and chemical properties of fresh drone homogenate, lyophilizate (apilarnil) and fresh royal jelly is presented in Table 1. ...
... Lowering the level of alanine transaminase, alkaline phosphatase and bilirubin in the blood serum Rats [53] DNA obtained from drone brood protects liver tissue against the hepatotoxic effects of acetylsalicylic acid, buserelin and carbon tetrachloride Rats [54] Fetal shielding properties Protect the fetus against the harmful effects of acetylsalicylic acid Rats [55] Healing effect in nervous and mental diseases Improves the mental state of patients with depressive neurosis, fatigue, anorexia, feelings of helplessness Humans [43] Positive effect on the symptoms of neurastemia (fatigue, weariness, dizziness) Humans [56] Improvement in memory, reduction of psychomotor lability, return of sphincter control Humans [43] Improvement of neurovegetative and sexual functions in elderly people Humans [32] ...
... According to Burmistrova [68], the storability of fresh bee brood can be improved, by binding fresh drone brood to a glucose/lactose adsorbent with the additive of L-ascorbic acid as an antioxidant (50 mg/kg). If the mixture is dried until 4% humidity, product is stable at 4 to 8 • C for 2-3 years [32,68]. Due to the different production technologies (extraction, lyophilization, thermal drying, adsorption, stabilization, mixing), the preparations based on the drone brood have a non-permanent qualitative and quantitative composition, have different severity of nutritional and therapeutic properties, characterized by a different shelf life [16]. ...
Full-text available
Drone brood homogenate is a little-known bee product used in folk medicine to treat various health problems. It is a very nutritious milky substance with high content of nutrients: proteins, lipids, fatty acids, carbohydrates, vitamins (A, B, E and D), and minerals. Moreover, when collected on early stage of larvae development, it is, most of all, rich source of sex hormone (testosterone, progesterone and estradiol). Some beekeepers consider drone brood as a waste product, although in some countries they use it to fight Varroa. Meanwhile, in many scientific reports a curative effect of bee drone homogenate in treating urgent global health problems have been confirmed, including ovarian dysfunction in women and male infertility, thyroid and immunity disorders, as well as malnutrition in children. A few dietary supplements based on drone brood are available online. Many patents relating to drone brood-based dietary supplements have been filed in Russia, but their prevalence in EU countries is still limited. Further research is needed to fully recognize the pharmacological activity and increase the use of drone brood.
... When it comes to lipids content, these macromolecules represent 7-18% of RJ bioactive compounds [40] and 3-19% of RJ dry matter [41]. The lipid content encompasses more ...
... When it comes to lipids content, these macromolecules represent 7-18% of RJ bioactive compounds [40] and 3-19% of RJ dry matter [41]. The lipid content encompasses more than 80% of fatty acids, phospholipids (0.4-0.8%), phenols (4-10%), steroids (3-4%) and waxes (5-6%). ...
Full-text available
Royal jelly (RJ) is one of the most valued natural products and is known for its health-promoting properties. Due to its therapeutic effects, it has been used in medicine since antiquity. Nowadays, several studies indicate that RJ acts as a powerful antimicrobial agent. Indeed, researchers shed light on its antioxidant and anticancer activity. RJ’s biological properties are related to its bioactive compounds, such as proteins, peptides, phenolic, and fatty acids. The aim of this review is to highlight recent findings on RJ’s main bioactive compounds correlated with its health-promoting properties. The available literature suggests that these bioactive compounds can be used as an alternative approach in order to enhance human health. Moreover, throughout this paper, we underline the prominent antibacterial effect of RJ against several target bacterial strains. In addition, we briefly discuss other therapeutic activities, such as antioxidative and anticancer effects, of this outstanding natural product.
... When it comes to lipids content, these macromolecules represent 7-18% of RJ bioactive compounds [40] and 3-19% of RJ dry matter [41]. The lipid content encompasses more ...
... When it comes to lipids content, these macromolecules represent 7-18% of RJ bioactive compounds [40] and 3-19% of RJ dry matter [41]. The lipid content encompasses more than 80% of fatty acids, phospholipids (0.4-0.8%), phenols (4-10%), steroids (3-4%) and waxes (5-6%). ...
Full-text available
TTürkiye farklı iklim ve coğrafi şartlara sahip olduğu için çok çeşitli bal arı ırklarının merkezi konumundadır. Türkiye’de yaygın olarak Anadolu arısı (Apis mellifera anatolica M.), Kafkasya arısı (Apis mellifera caucasia G.), Suriye Arısı (Apis mellifera syrica), İran arısı (Apis mellifera meda Skorikov, 1929) ırkları bulunmaktadır. Arıcılığın geçmişi Taş Devri ve eski medeniyet çağlarına kadar dayanmaktadır. İspanya'da yapılan kazılarda ortaya çıkarılan M.Ö. 7000 yıllarına ait arı fosil ve resimleri eski çağ insanlarının doğal yaşamlarında arılardan yararlandığını kanıtlamaktadır. Arının kültüre alındığı ilk yer Mısır olup, arıcılığın ise ilk defa 7000 yıl önce Orta Anadolu’da başladığı bilinmektedir. Literatürde Mısırlılarda arıcılığın daha da geliştiği, 4.000 yıl öncesinde balın malların alım satımında takas malzemesi olduğu ve vergi ödemede kullanıldığı belirtilmektedir. Günümüzde arıcılık sektörü oldukça gelişmiş ve bal dışında diğer arı ürünlerinin önemi de araştırmacılar tarafından keşfedilmiştir. Bal dışındaki diğer arı ürünleri arasında propolis, arı sütü, polen, arı ekmeği (Perga), arı zehiri ve balmumu yer almaktadır. Bu çalışmada arı ürünlerinin tanıtılması, beslenme ve sağlık açısından önemlerinin vurgulanması amaçlanmaktadır.
Türkiye farklı iklim ve coğrafi şartlara sahip olduğu için çok çeşitli bal arı ırklarının merkezi konumundadır. Türkiye’de yaygın olarak Anadolu arısı (Apis mellifera anatolica M.), Kafkasya arısı (Apis mellifera caucasia G.), Suriye Arısı (Apis mellifera syrica), İran arısı (Apis mellifera meda Skorikov, 1929) ırkları bulunmaktadır. Arıcılığın geçmişi Taş Devri ve eski medeniyet çağlarına kadar dayanmaktadır. İspanya'da yapılan kazılarda ortaya çıkarılan M.Ö. 7000 yıllarına ait arı fosil ve resimleri eski çağ insanlarının doğal yaşamlarında arılardan yararlandığını kanıtlamaktadır. Arının kültüre alındığı ilk yer Mısır olup, arıcılığın ise ilk defa 7000 yıl önce Orta Anadolu’da başladığı bilinmektedir. Literatürde Mısırlılarda arıcılığın daha da geliştiği, 4.000 yıl öncesinde balın malların alım satımında takas malzemesi olduğu ve vergi ödemede kullanıldığı belirtilmektedir. Günümüzde arıcılık sektörü oldukça gelişmiş ve bal dışında diğer arı ürünlerinin önemi de araştırmacılar tarafından keşfedilmiştir. Bal dışındaki diğer arı ürünleri arasında propolis, arı sütü, polen, arı ekmeği (Perga), arı zehiri ve balmumu yer almaktadır. Bu çalışmada arı ürünlerinin tanıtılması, beslenme ve sağlık açısından önemlerinin vurgulanması amaçlanmaktadır.
Drone brood is a little-known bee product which is frequently considered as a male equivalent of royal jelly and is sometimes used as its adulterant. The aim of the study was to compare the chemical composition and biological activity of both bee products originated from the same apiaries (n = 3) limiting the influence of genetic and environmental factors. Moreover, for drone brood study covered testing three stages of larval development (days 7, 11, and 14). The comparison included mineral composition (ICP-OES method), protein content and protein profile (SDS-PAGE), testosterone and estradiol content (ELISA tests). HPTLC method was used to analyze of sugar, amino acids, and polyphenolic profile of drone brood and royal jelly. Moreover, their antioxidant and enzymatic properties were compared. A lot of similarities between drone brood and royal jelly were found in terms of chemical components. However, drone brood was more abundant in iron and manganese, reducing sugars and some amino acids, especially proline, tyrosine, and leucine. It contained more testosterone (especially on the 14th day) and estradiol (on the 7th day). The greatest differences in the enzymatic activities and polyphenolic profile were found. Diastase and α-glucosidase activity were found as specific enzymes of the drone brood. Similarly, ferulic and ellagic acids were characteristic for brood and were not present in royal jelly. The study showed a lot of similar features for both tested bee products, however, some specific markers which can serve to differentiate drone brood and royal jelly were found.
Full-text available
Royal jelly is not only a natural health substance for human beings, but also a lucrative hive-product for beekeepers. Achieving a higher royal jelly yield is an important pursuit for beekeepers engaged in commercial royal jelly production. Since breeding a higher production royal jelly bee in the 1980s in China, royal jelly production has had a burgeoning growth. In this article, a special queen excluder and factory production method are introduced that will enable collection of up to 10 kilos of royal jelly per colony.
We report the analysis of Brazilian commercial bee products using high performance liquid chromatography, and a faster method, to detect trans-10-Hydroxy-2-Decenoic Acid or 10-Hydroxy-2-E-decenoic acid (10-HDA) in royal jelly. Determination was carried out using aqueous phosphoric acid 0.1% (pH 2.5)/methanol (50/50) as the mobile phase, and a LiChrospher® RP-18 column with detection at 215 nm. A good linearity was shown between the concentration of 10-HDA and peak area in the concentration range of 5.0–50.0 mg/L (r 0.99, n = 5). The within-day and between-day relative standard deviation (RSD) were1 % and 3 %, respectively.Presentamos el análisis de productos apícolas comerciales brasileños mediante cromatografía líquida de alta ejecución, junto con otro método más rápido, para detectar los ácidos trans-10-hidroxi-2-decenoico o 10-hidroxi-2-E-decenoico (10-HDA) en la jalea real. La determinación se llevó a cabo usando ácido acuoso fosfórico al 0,1% (pH2,5)/metanol (50/50) como fase móvil, y columnas LiChrospher® RP-18 con detección a 215 nm. Una correcta linealidad se observó entre la concentración de 10-HDA y el área del pico en el rango de concentración de 5,0–50,0 mg/L (r 0.99, n = 5). La desviación estándar relativa (DER) durante el día y entre los días de análisis fue del 1% y 3% respectivamente.
The changes in quality of royal jelly stored at 50°C for 2h 30′, 5h, 7h 30′ and a: 37°C for 7 and 15 days were investigated. Browning, pH, acidity, enzimatic activity (diastase, invertase, glucose-oxidase) as quality indexes were analized. During the ageing the colour of royal jelly changed: the browning reaction increased proportionally to both temperature and storage time. pH and acidity values didn't change. Glucose-oxidase, the only enzyme detected in royal jelly, was subjected to an exponential decrease: the same value of glucose-oxidase in the sample stored at 37°C for 7 days was reached after 7 hours at 50°C.
Quantitative analysis of trans-10-hydroxy-2-decenoic acid (10-HDA) in pure royal jelly creams and dietary supplements of royal jelly available in the United States was carried out via reversed phase high performance liquid chromatography (HPLC). The target compound, 10-HDA, in samples and an internal standard, methyl 4-hydroxybenzoate (MHB), were separated using a Zorbax Eclipse XDB-C18 column (150 × 4.6 mm) with a mobile phase composed of methanol and an aqueous solution of phosphoric acid at 25°C. The flow rate was 1.0 mL/min and the UV detection was performed at 215 nm. The average recovery rate of 10-HDA was 97.4 - 100.4% with the relative standard deviation (RSD) of 2.4 - 3.4% over the concentrations ranging from 10 to 80 μg/mL. The limit of detection (LOD) and limit of quantification (LOQ) were found to be about 0.05 and 0.25 μg/mL, respectively. Our results show that the concentration of 10-HDA lies between 1.85 and 2.18% for pure royal jelly creams and between 0.43 and 6.28% for royal jelly supplements.
The objective of this study was to characterize the protein fractions in royal jelly made from Apis mellifera ligustica from middle and southern areas of Taiwan. The total nitrogen content of fresh royal jelly was 2.46%, and the total amino acid nitrogen was 2.34%, suggesting that the nitrogen compound in royal jelly was mostly derived from protein . The nitrogen content of free amino acid in royal jelly was 0.11%, and the amino type nitrogen was 0.20%, indicating that the protein in royal jelly existed mainly in the form of large moleculars. To characterize the protein, royal jelly was dissolved in 0.1 M phosphate buffer(pH 7.0), followed by centrifugation, ammonia sulfate precipitation and dialysis to separate the protein into water soluble and water insoluble fractions. Water soluble fraction accounts for more than 60 % of the total protein in royal jelly, and was further investigated by DEAE-Sephacel, SDS-PAGE and capillary electrophoresis. By DEAE-Sephacel, two fraction peaks (F1 and F2) were identified and collected. By SDS-PAGE, F1 fraction was further separated into two bands, and the molecular weight was determined to be 50 KDa and 44 KDa, whereas F2 fraction was shown to have only one band with molecular weight of 55 KDa. By capillary zone electrophoresis, four poorly-separated peaks were observed in F1 fraction, and two well-separated peaks in F2 fraction. By capillary gel electrophoresis, two peaks were identified in F1 fraction, of which the molecular weight was estimated to be 59 KDa and 73 KDa. By contrast, only one peak was identified in F2 fraction, of which the molecular weight was estimated to be 118 KDa. However, by capillary isoelectric focusing, 6 peaks were identified in F1 fraction with pI of 6.9, 6.7, 6.3, 5.9, 5.7 and 5.5, respectively. Of that, pI of 4.8 and 4.7 were identified in F2 fraction.
An experiment was carried out to analyse changes in the protein components of royal jelly (RJ) under different storage conditions, based on two dimensional electrophoresis (2-DE). The proteins identified were compared to those proteins already identified in the proteome complement of the RJ. The results showed that the total detected protein spots were 75, 45, 63 and 69, with molecular weight in the range of 7.64-72.33 kDa, isoelectric point 4.95-8.70, in the 2-DE image of RJ protein components stored at -20°C for 80 days, 4°C for 80 days, room temperature for 30 days, room temperature for 80 days, respectively. The spot of major royal jelly protein, apalbumin I, was saturated in all images in this experiment, indicating that temperature has no significant effects on it. The spots number and the quantity of apalbumin 2 and apalbumin 3 did not increase or decrease following the temperature trend, suggesting they are also sensitive to temperature. However, spots of apalbumin 4 and glucose oxidase were observed only in the image of -20°C for 80 days, and spots of apalbumin 5 were detected in the images of -20°C and 4°C for 80 days, indicating they are the proteins most sensitive to storage temperature and thus may be potential freshness markers for RJ, and that the best way to maintain quality of RJ is under freezing conditions.
Quantitative differences were found in the chemical composition of royal jelly samples collected in spring and summer by the same producer. The main difference was in the free fatty acids, which showed a marked increase in 10-hydroxydecanoic acid in summer. The sterol and hydrocarbon fractions were also investigated. The most significant sterols were identified by gas chromatography—mass spectrometry. The hydrocarbon components, identified by their retention times, were a homologous series of straight-chain compounds, from C16 to C33, with even and odd numbers of carbon atoms. Considerable amounts of some hydrocarbons were found. Of the sterols identified, the most abundant was 24-methylene cholesterol; stigmasterol, β-sitosterol, Δ5-avenasterol, Δ7-avenasterol and cholesterol were also present.