JP4976186B2 - Royal jelly manufacturing method - Google Patents

Royal jelly manufacturing method Download PDF

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JP4976186B2
JP4976186B2 JP2007107112A JP2007107112A JP4976186B2 JP 4976186 B2 JP4976186 B2 JP 4976186B2 JP 2007107112 A JP2007107112 A JP 2007107112A JP 2007107112 A JP2007107112 A JP 2007107112A JP 4976186 B2 JP4976186 B2 JP 4976186B2
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royal jelly
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apicin
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喜久二 山口
透 河野
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Description

本発明は、ローヤルゼリーの製造方法およびその方法によって得られるローヤルゼリーに関する。   The present invention relates to a method for producing a royal jelly and a royal jelly obtained by the method.

ローヤルゼリーはミツバチから採取される食品で、幼虫の育成のために羽化後3〜12日齢の働き蜂が下咽頭腺と大腮腺とから分泌する、乳白色をしたペースト状の物質である。働き蜂となる幼虫は、孵化後3日齢を過ぎるとローヤルゼリーに替わって花粉と蜂蜜を餌として与えられるが、3日齢を過ぎてもローヤルゼリーのみを餌として育てられた幼虫は、一日3000個の卵を産卵する能力を持つ女王蜂へと成長する。働き蜂が産卵から21日で羽化・出房するのに対し、女王蜂は産卵から15日で羽化・出房する。   Royal jelly is a food collected from bees, and is a milky white paste-like substance secreted from the hypopharyngeal gland and large vagina by worker bees 3 to 12 days after emergence for raising larvae. The larvae that serve as worker bees are fed with pollen and honey instead of royal jelly when they pass 3 days after hatching, but 3000 larvae that are fed only with royal jelly after 3 days are fed. Grows into a queen bee with the ability to lay eggs. A worker bee emerges and leaves 21 days after laying eggs, while a queen bee emerges and leaves 15 days after laying eggs.

現在のローヤルゼリーの採乳は、一般的に、故井上丹治氏によって開発された人工王台を用いる方法で行われている。   The current royal jelly milking is generally performed by a method using an artificial king developed by the late Mr. Tanji Inoue.

従来の方法は、3日齢の幼虫を人口王台へ移虫させ、移虫後72時間でローヤルゼリーを採乳するのが、一般的であった。   In the conventional method, a 3-day-old larva was transferred to the population, and royal jelly was collected 72 hours after the transfer.

働き蜂の幼虫への給餌は、3日齢前半までは僅かなローヤルゼリーが与えられるが、3日齢後半時には離乳食にあたら、ハチミツと花粉のミックス食に切り替えられる。ローヤルゼリーが与えられている間は、ローヤルゼリーの全てが吸収され、幼虫の脱糞はないがミックス食に代わると、幼虫は巣房内で脱糞を始め、不純物が混入する。   A small amount of royal jelly is given to the larvae of the worker bees until the first half of the 3rd day, but if they are weaning food at the latter half of the 3rd day, they can be switched to a mixed honey and pollen diet. While the royal jelly is being given, all of the royal jelly is absorbed, and there is no larval defecation, but if it is replaced with a mixed food, the larva begins to defecate in the nest and becomes contaminated.

したがって、育児転換可能日齢の限界は3日齢前半までであって、この間の幼虫であれば、餌をローヤルゼリーに切り替えれば女王蜂に育つのである。それゆえに、3日齢の幼虫が移虫に使用されてきた。   Therefore, the limit of the age at which childcare can be switched is up to the first half of the third day, and if it is a larva during this period, it will grow into a queen bee if the food is switched to royal jelly. Therefore, 3 day old larvae have been used for migrating.

王台は移虫後68〜72時間でほぼ満杯になり、ミツロウで蓋をされるので、移虫後72時間でこれを竹へらなどを用いて回収(採乳)する。   The Odaidai is almost full 68 to 72 hours after the transfer and is covered with beeswax, and is collected (milked) using a bamboo spatula or the like 72 hours after the transfer.

しかし、移虫時の幼虫の大きさや移虫後の採乳時間は、養蜂の手技の便宜で決められてきており、品質管理の観点から検討されたことはなかった。   However, the size of the larvae at the time of migrating and the milking time after the migrating have been determined by the convenience of beekeeping techniques and have not been studied from the viewpoint of quality control.

ローヤルゼリーの化学成分については、働き蜂の生理的状態、ローヤルゼリーの採取条件や保存状態、分析方法等によりかなりの変動が認められるが、生ローヤルゼリーの平均的な成分組成(重量%)は、水分が約66%、タンパク質が約13%、糖質が約14.5%、脂質が約4.5%であり、その他の成分(ビタミンB1、ビタミンB2、ビタミンB6、ニコチン酸、葉酸、パントテン酸、ビオチン、イノシトール、アセチルコリン、γ−アミノ酪酸など)が約2%である。   The chemical composition of royal jelly varies considerably depending on the physiological state of the worker bee, the collection and storage conditions of the royal jelly, the analysis method, etc., but the average composition of raw royal jelly (% by weight) 66%, protein about 13%, carbohydrates about 14.5%, lipids about 4.5%, other ingredients (vitamin B1, vitamin B2, vitamin B6, nicotinic acid, folic acid, pantothenic acid, biotin , Inositol, acetylcholine, γ-aminobutyric acid, etc.) is about 2%.

ローヤルゼリーには、ラットなどの実験動物を使用した研究から、抗腫瘍作用、体重増加及び成長促進作用、血清コレステロール及び脂質低下作用、血流量増加及び血圧効果作用、創傷治癒促進作用、性ホルモン様作用、性中枢老化防止作用、放射線照射動物の延命効果、抗菌作用、酵素作用などの薬理効果が報告されている。また、ヒトに対する臨床的効果として、高血圧症などの慢性疾患の改善、放射線や化学療法剤の副作用軽減作用、発育促進作用、不定愁訴症候群や自律神経失調症の改善作用などが報告されている。また、ローヤルゼリーを化粧品に利用し美肌法などの美容効果の目的でも使用されている。   Royal jelly has been studied in rats and other experimental animals from anti-tumor effects, weight gain and growth promoting effects, serum cholesterol and lipid lowering effects, blood flow increases and blood pressure effect effects, wound healing promotion effects, and sex hormone-like effects. In addition, pharmacological effects such as sexual central aging preventive action, life-prolonging effect of irradiated animals, antibacterial action, and enzyme action have been reported. As clinical effects on humans, improvement of chronic diseases such as hypertension, reduction of side effects of radiation and chemotherapeutic agents, growth promotion, improvement of indefinite complaint syndrome and autonomic ataxia have been reported. In addition, royal jelly is used for cosmetics, and it is also used for the purpose of cosmetic effects such as skin beautification.

ローヤルゼリーの生理活性成分としては、10−ヒドロキシデセン酸が知られている。従来の採乳方法によるローヤルゼリーは、およそ1.7〜2.2%程度の10−ヒドロキシデセン酸を含有する。   As a physiologically active ingredient of royal jelly, 10-hydroxydecenoic acid is known. The royal jelly by the conventional milking method contains about 1.7 to 2.2% of 10-hydroxydecenoic acid.

従来は、10−ヒドロキシデセン酸の含量がローヤルゼリーの品質の指標として用いられてきた。しかし、粉末ローヤルゼリーの製造の際に網に残った10−ヒドロキシデセン酸の結晶体を微粒子状の粉末とし、大量生産された劣悪低活性のローヤルゼリー原乳(10−ヒドロキシデセン酸1.4%以下)に、人工的に作られた10−ヒドロキシデセン酸粉末を後から添加することによって10−ヒドロキシデセン酸含有量を2.0%以上にした、紛い物が横行していた。したがって、10−ヒドロキシデセン酸含量に替わる科学的に信用のおける新たなローヤルゼリーの品質指標が求められていた。   Conventionally, the content of 10-hydroxydecenoic acid has been used as an indicator of the quality of royal jelly. However, the 10-hydroxydecenoic acid crystalline substance remaining in the net during the production of the powder royal jelly is made into a fine powder, and the poorly active royal jelly raw milk mass-produced (10-hydroxydecenoic acid 1.4% or less) ), 10-hydroxydecenoic acid powder made artificially was added later, and the content of 10-hydroxydecenoic acid was increased to 2.0% or more. Therefore, a new scientifically reliable quality indicator of royal jelly that replaces the 10-hydroxydecenoic acid content has been sought.

ローヤルゼリーに含まれるタンパク質のうち、約75%が水溶性であり、残りの約25%が水不溶性タンパク質である。現在までに性質や構造が明らかにされたタンパク質には、抗菌作用を持つロイヤリシン(分子量5,500)、培養細胞に対して細胞増殖促進作用を有する糖タンパク質であるアピシン(分子量350,000)及び分子量55,000の糖タンパク質(55k−RJGP)、分子量57,000の糖タンパク質であるロイヤラクチンがある。しかし、いずれのタンパク質が品質管理に有用であるかは知られていなかった。   Of the protein contained in royal jelly, about 75% is water-soluble and the remaining about 25% is water-insoluble protein. Proteins whose properties and structures have been elucidated so far include royaricin (molecular weight 5,500) having antibacterial action, apicin (molecular weight 350,000), a glycoprotein having cell growth promoting action on cultured cells, and There is a glycoprotein (55k-RJGP) with a molecular weight of 55,000 and royalactin which is a glycoprotein with a molecular weight of 57,000. However, it was not known which protein was useful for quality control.

したがって、ローヤルゼリーの品質管理の適切な指標の開発及びその指標に基づく、最適な移虫時の幼虫の大きさや移虫後の採乳時間の決定が待たれていた。   Therefore, the development of an appropriate index for quality control of royal jelly and the determination of the optimum larvae size at the time of migrating and milking time after migrating based on the index have been awaited.

本発明は、ローヤルゼリーの品質管理の適切な指標、及び最適な移虫時の幼虫の大きさや移虫後の採乳時間を提供する。   The present invention provides an appropriate index for quality control of royal jelly, and the optimum larvae size at the time of migrating and milking time after migrating.

本発明の発明者らは、上記課題を解決するために鋭意努力した結果、ローヤルゼリー品質指標(Index of Quality of Royal Jelly:IQR)やアピシン含有量及び/又はアピシン含有量/可溶性タンパク総量比がローヤルゼリーの品質管理の適切な指標であることを見出し、そして、最適な移虫時の幼虫の大きさが48時間齢であり、最適な移虫後の採乳時間が移虫から40、52又は68時間後であることを見出し、本発明を完成させた。この製造工程は、山口喜久二式自然養蜂と呼ばれる農法の最も重要な工程である。   As a result of diligent efforts to solve the above-mentioned problems, the inventors of the present invention have a royal jelly having a royal jelly quality index (Index of Quality of Royal Jelly: IQR), apicin content and / or apicin content / total soluble protein ratio. And the optimal larvae size at the time of migrating is 48 hours, and the optimum milking time after migrating is 40, 52 or 68 It was found that it was time later and the present invention was completed. This manufacturing process is the most important process of the farming method called Yamaguchi Kikuji natural beekeeping.

すなわち、本発明は、下記:
1.王台に孵化後25〜30時間齢の幼虫を移虫する工程を含む、ローヤルゼリーの製造方法、
2.さらに、移虫から40、52又は68時間後に、王台に分泌されたローヤルゼリーを採乳する工程を含む、上記1に記載の製造方法、
3.ローヤルゼリーの品質指標が1.5以上である、上記1又は2に記載の製造方法で製造されたローヤルゼリー、
4.アピシン含有量が1500mAU以上であり、及び/又はアピシン含有量/可溶性タンパク総量比が150以上である、上記1又は2に記載の製造方法で製造されたローヤルゼリー。
に関する。
That is, the present invention provides the following:
1. A method for producing royal jelly, comprising a step of transferring 25-30 hour-old larvae after hatching on the royal palace,
2. The method according to 1 above, further comprising the step of milking the royal jelly secreted by the royal family 40, 52, or 68 hours after the transfer.
3. The royal jelly manufactured by the manufacturing method according to the above 1 or 2, wherein the quality index of the royal jelly is 1.5 or more,
4). The royal jelly manufactured with the manufacturing method of said 1 or 2 whose apicin content is 1500 mAU or more and / or whose apicin content / soluble protein total amount ratio is 150 or more.
About.

以上のように、本発明に示すローヤルゼリー製造方法は、高品質なローヤルゼリーの製造に有用であり、この方法を用いて製造されたローヤルゼリーからは高い生理活性が得られる。   As described above, the method for producing a royal jelly shown in the present invention is useful for producing a high-quality royal jelly, and a high physiological activity can be obtained from the royal jelly produced using this method.

本発明において、孵化後25〜30時間齢の幼虫とは、孵化した後、25〜30時間を経過した幼虫である。従来2日齢と呼ばれていた25時間齢〜48時間齢では、得られるローヤルゼリーの品質に顕著な違いがあることが判ったため、移虫用の働き蜂の幼虫を時間齢で規定したものである。このときの幼虫の大きさは1mm以内である。   In the present invention, larvae 25 to 30 hours old after hatching are larvae that have passed 25 to 30 hours after hatching. Since it was found that there was a marked difference in the quality of the royal jelly obtained at 25 to 48 hours, which was conventionally called 2 days old, the bee larvae for migrating insects were defined by time age. . The size of the larvae at this time is within 1 mm.

本発明の製造方法では、移虫から40、52又は68時間後に、王台に分泌されたローヤルゼリーを採乳する。好ましくは、移虫から40時間後に、王台に分泌されたローヤルゼリーを採乳する。   In the production method of the present invention, royal jelly secreted by the royal family is collected 40, 52 or 68 hours after the transfer. Preferably, the royal jelly secreted by the royal family is collected 40 hours after the transfer.

従来は、養蜂作業の開始時刻は朝露の無くなる朝9時頃であることから、開始時刻から計算すると移虫後40時間目は夜の11時となってしまい、養蜂作業を行うのが不可能な時刻であることから、一般的養蜂家は昼過ぎの、働き蜂が巣箱から飛び立ち、巣内に内勤蜂のみとなっている頃を見計らって移虫作業を行っていた。すなわち、午後1〜3時頃に移虫作業を行い、移虫後48時間又は72時間後の午後1〜3時頃に採乳を行っていたのである。これに対して、本発明の方法では、夜中の12時頃に移虫を行って、40時間後の午後4時に採乳を行うことになる。   Conventionally, the start time of beekeeping work is around 9:00 in the morning when there is no morning dew, so calculating from the start time means that the 40th hour after migrating is 11:00 at night, making it impossible to perform beekeeping work. Because it was a long time, beekeepers were carrying out migrating work in the afternoon, in anticipation of worker bees flying out of the hive and only house bees in the nest. That is, migrating work was carried out around 1 to 3 pm, and milking was carried out around 1 to 3 pm 48 hours or 72 hours after migrating. In contrast, in the method of the present invention, migrating is performed around 12:00 in the middle of the night, and milking is performed at 4 pm after 40 hours.

本発明においてローヤルゼリー品質指標とは、ローヤルゼリー中のアピシン含有量(mAU)とアルブミン含有量(mAU)の比、すなわち、
アピシン含有量/アルブミン含有量
である。
In the present invention, the royal jelly quality index is the ratio of apicin content (mAU) to albumin content (mAU) in the royal jelly,
Apicin content / albumin content.

また、本発明においてローヤルゼリー品質指標とは、
ローヤルゼリー中のアピシン含有量(mAU)、及び/又はローヤルゼリー中のアピシン含有量(mAU)と可溶性タンパク総量(mg/ml)との比、すなわち、アピシン含有量/可溶性タンパク総量
である。
In the present invention, the royal jelly quality index is
The apicin content in royal jelly (mAU) and / or the ratio of apicin content in royal jelly (mAU) to total soluble protein (mg / ml), ie, apicin content / total soluble protein.

アピシンは、ヒトや動物の培養細胞の増殖を促進し、細胞死を防ぐ作用を有することが知られている(ローヤルゼリー、美容食品の開発と展望、猪居武監修、シーエムシー出版、2004年、221−233)。   Apisin is known to promote the growth of cultured cells of humans and animals and to prevent cell death (Royal Jelly, Development and Prospect of Beauty Foods, Supervised by Takeshi Aoi, CM Publishing, 2004, 221-233).

ローヤルゼリー品質指標は、0.5以上であることが好ましく、より好ましくは、1.0以上であり、さらに好ましくは、1.5以上である。したがって、ローヤルゼリー品質指標0.5未満は、等外であり、0.5以上〜1未満が三級であり、1以上〜1.5未満が二級であり、1.5以上が一級であるというように評価することができる。   The royal jelly quality index is preferably 0.5 or more, more preferably 1.0 or more, and further preferably 1.5 or more. Therefore, the royal jelly quality index of less than 0.5 is not equal, 0.5 to less than 1 is tertiary, 1 to less than 1.5 is secondary, and 1.5 or more is primary. It can be evaluated as follows.

また、ローヤルゼリー品質指標は、アピシン含有量500mAU以上かつアピシン含有量/可溶性タンパク総量比75以上であることが好ましく、より好ましくは、1000mAU以上かつ100以上であり、さらに好ましくは、1500mAU以上かつ150以上である。したがって、アピシン含有量500mAU以上かつアピシン含有量/可溶性タンパク総量比75未満は、等外であり、アピシン含有量500mAU以上かつアピシン含有量/可溶性タンパク総量比75以上〜100未満が三級であり、アピシン含有量1000mAU以上かつアピシン含有量/可溶性タンパク総量比100以上〜150未満が二級であり、アピシン含有量1500mAU以上かつアピシン含有量/可溶性タンパク総量比150以上が一級であるというように評価することができる。   The royal jelly quality index is preferably an apicin content of 500 mAU or more and an apicin content / total soluble protein ratio of 75 or more, more preferably 1000 mAU or more and 100 or more, and further preferably 1500 mAU or more and 150 or more. It is. Therefore, an apicin content of 500 mAU or more and an apicin content / soluble protein total ratio of less than 75 are exceptional, an apicin content of 500 mAU or more and an apicin content / soluble protein total ratio of 75 to 100 are tertiary, Evaluation is made such that the apicin content is 1000 mAU or more and the apicin content / total soluble protein ratio is 100 to less than 150, and the apicin content is 1500 mAU or more, and the apicin content / soluble protein total ratio is 150 or more. be able to.

本発明を下記の実施例により詳細に説明するが、本発明はこれに限定されるわけではない。   The present invention will be described in detail by the following examples, but the present invention is not limited thereto.

ローヤルゼリー品質指標を以下のように評価した。
実験材料および実験方法
1.人工王台を利用した自然養蜂を単一の菜の花畑を利用して実験養蜂を行った。使用した蜂は西洋蜂である。人工王台に西洋蜂の幼虫(48時間齢または72時間齢)を移虫し、移虫後24時間から72時間まで4時間ごとに人工王台に蓄積されたローヤルゼリーを採取し、可溶性タンパクを抽出し、ゲルろ過、陰イオン交換HPLCによって分画・精製し、SDS−PAGE、2次元電気泳動パターンを分析した。さらにMALDI−TOF/TOF MSによって質量分析を行った。
2.HPLCを使用して、山口喜久二養蜂によって採乳された原乳ローヤルゼリーおよび山口喜久二養蜂ではない方法(非山口喜久二養蜂)によって採乳された市販の原乳ローヤルゼリー(アスケン株式会社、株式会社ウェルネスジャパン、ミナミヘルシーフーズ株式会社、株式会社サクラ印はちみつ、森川健康堂株式会社、株式会社山田養蜂場、株式会社神州及び松本等の12社の製品)の可溶性タンパクを比較検討した。
The royal jelly quality index was evaluated as follows.
Experimental materials and experimental methods A natural beekeeping using an artificial royal king was conducted using a single rape field. The bees used are western bees. Western bee larvae (48 hours or 72 hours old) were transferred to the artificial kingdom, and the royal jelly accumulated in the artificial kingdom was collected every 4 hours from 24 to 72 hours after transferring, and soluble protein was collected. Extraction, fractionation and purification by gel filtration and anion exchange HPLC were performed, and SDS-PAGE and two-dimensional electrophoresis patterns were analyzed. Furthermore, mass spectrometry was performed by MALDI-TOF / TOF MS.
2. Raw milk royal jelly collected by Kikuji Yamaguchi beekeeping using HPLC, and commercially available raw milk jelly collected by a method that is not beekeeping Yamaguchi Kikuji (non-Yamaguchi Kikuji beekeeping) (Askene Co., Ltd. The soluble proteins of Wellness Japan, Minami Healthy Foods Co., Ltd., Sakura Ink Honey Co., Ltd., Morikawa Kendo Co., Ltd., Yamada Bee Farm Co., Ltd., Shinshu Co., Ltd., Matsumoto Co., Ltd.) were compared.

実験結果
1.人工王台から経時的に採取したローヤルゼリーの可溶性タンパクはゲルろ過によって普遍的に5つのピークが検出された。特にピーク2は、Major Royal Jelly Protein 1 (アピシン)単独のピークであると同定された。
Experimental results The royal jelly soluble protein collected over time from the artificial kingdom was universally detected with five peaks by gel filtration. In particular, peak 2 was identified as the peak of Major Royal Jelly Protein 1 (Apicin) alone.

2.ピーク2(アピシン)はHPLCによってその量をmAUとして定量的に示すことが可能であるが、人工王台の中で成長する幼虫がアピシンを特異的に消費していることが明らかとなった。消費すなわちアピシン減少パターンは時間的に変動があるが、その時間的推移は一定であり、24時間から40時間までの間に一回目の大きな減少、次に40時間から52時間までの間に2回目の大きな減少、次に52時間から68時間までの3回目の大きな減少、最後に68時間から72時間までの大きな減少が認められた。従って、王台中のローヤルゼリーのアピシンル量は移虫後24時間、40時間、52時間、68時間目に豊富に存在する一方、それ以外の時間帯では幼虫によって急速に消費されていることが明らかとなった。   2. Peak 2 (Apicin) can be quantitatively shown by HPLC as mAU, but it was revealed that the larvae growing in the artificial king are specifically consuming apicin. The consumption or apicin reduction pattern varies over time, but its time course is constant, the first large decrease between 24 and 40 hours, then 2 between 40 and 52 hours. There was a large decrease for the first time, followed by a third large decrease from 52 to 68 hours, and finally a large decrease from 68 to 72 hours. Therefore, it is clear that royal jelly in the royal palace is abundant at 24 hours, 40 hours, 52 hours, and 68 hours after transfer, but is rapidly consumed by larvae at other times. became.

3.72時間齢を移虫した王台中のローヤルゼリーに含まれるアピシン量を48時間齢を移虫した場合と比較した結果、アピシン含有量は48時間齢に比べて半分以下で有意に低値であった。   As a result of comparing the amount of apicin contained in royal jelly in the royal kingdom that was transferred at 72 hours, compared with the case where 48 hours were transferred, the amount of apicin was less than half that at 48 hours and was significantly lower. there were.

4 製品化されたローヤルゼリーの原乳ローヤルゼリー中のアピシン含有量を山口喜久二養蜂と非山口喜久二養蜂において比較した結果、山口喜久二養蜂による原乳(5ロット)では平均1828mAU(1543〜2006mAU)であるのに対し、非山口喜久二養蜂における原乳(12ロット)では平均428mAU(27〜1225mAU)と有意に低値であった。また、ローヤルゼリー品質保証の一つであるアピシン含有量/可溶性タンパク総量比でも比較した結果、山口喜久二養蜂では平均158(134〜173)であったが、非山口喜久二養蜂では平均40(2.5〜114)と有意に低値であった。
また、IQRについても、山口喜久二養蜂では1.511であったが、非山口喜久二養蜂では0.124〜1.203と有意に低値であった。
4 As a result of comparing the content of apicin in the raw milk jelly of commercialized royal jelly in Yamaguchi Kikuji and non-Yamaguchi Kikuji beekeeping, average 1828 mAU (1543-2006 mAU) in raw milk (5 lots) by Yamaguchi Kikuji beekeeping On the other hand, in raw milk (12 lots) in non-Yamaguchi Kikuji beekeeping, the average was 428 mAU (27-1225 mAU), which was significantly lower. In addition, as a result of comparison in the ratio of apicine content / total amount of soluble protein, which is one of the quality assurance of royal jelly, the average was 158 (134-173) for Kikuji Yamaguchi bee but 40 (2 .5 to 114).
In addition, IQR was 1.511 for Yamaguchi Kikuji beekeeping, but non-Yamaguchi Kikuji beekeeping was significantly low at 0.124 to 1.203.

本発明のローヤルゼリーの製造方法は、高品質のローヤルゼリーの製造のために有用である。   The method for producing a royal jelly of the present invention is useful for producing a high-quality royal jelly.

Claims (1)

王台に孵化後48時間齢の幼虫を移虫する工程、及び
移虫から40、52又は68時間後に、王台に分泌されたローヤルゼリーを採乳する工程
を含む、アピシンを含むローヤルゼリーの製造方法。
Transferring larvae 48 hours after hatching to the royal palace ; and
A method for producing royal jelly containing apicin, comprising the step of milking royal jelly secreted by the royal family 40, 52 or 68 hours after the transfer.
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