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Picture of bees on honeycomb Beeswax.
Cera Flava and Cera Alba.
Martindales 24th Br.
United States Dispensatory 1926.

Compiled and edited by Ivor Hughes.

Introduction. When a worker bee attends a plant it gathers various substances i.e Nectar, Pollen, Propolis and Wax. It is not questioned that the honey produced by the various species of Apis will differ not only in the seasonal flowers upon which they feed but also in Geo/Climatic factors. We may well understand that the variables involved are of great complexity. Many Pharmacopoeias will carry monographs dealing with tests for beeswax. The tests are based on physical characteristics of the wax being tested e.g.  Description, Solubility, Acid value, Ester value, Melting point and Ratio number.

The test values are not a set figure, for example the melting point for beeswax is usually stated to fall between 62 C and 64 C. Thus the tests  used, make a nod to the variables involved. The point to be made is, that beeswax in the same way as honey, will vary in its chemical and physical  composition. Beeswax is considered to be medicinally inert. This convention of the various Pharmacopoeias is obviously more of a convenience than it is factual. 

T.C Denston, 'A Textbook of Pharmacognosy' 3rd Edition.  states;
" Beeswax is the wax produced by worker bees from nectar and pollen.  These substances are converted in the body of the bee to wax, which then exudes  through wax pores on the ventral surface into 8 small moulds and there hardens  into small scales. When required it is plucked off, made plastic with saliva,  and used for building the honey comb and capping the cells. Beeswax forms about one tenth of the total weight of the honey comb. "

This not, inconsiderable figure representing 3kg for each 30kg of comb.  It must be allowed that in addition to the substances mentioned, there are also further substances collected by the bee, e.g. Volatile Oils, Plant Wax and  Resins. These substances will also vary from each other depending on the source  from which they are collected. To further complicate the matter there is also the problem of chemical spray residues.


Again the British Pharmacopoeia 1958, makes a nod to all of those  variables by usually specifying Cera Alba or bleached wax as the Official substance. Commercially Cera Alba is produced by an acidified solution of chromic acid and then washed in water. Obviously the wax has undergone chemical change. The changes engendered will obviously depend upon the original composition of the wax. The USD 1926 covers the ground well.


MARTINDALES 24th. THE EXTRA PHARMACOPOEIA (Br)

White Beeswax (B.P.). Cera Alba; White Wax (U.S.P.) Cire Blanche; Weisses  Wachs; Cera blanca.

Foreign Pharmacopeias: In all pharmacopeias examined.

Bleached beeswax. A yellowish-white solid with a faint characteristic odour. M.p. 62 to 64 C

Yellow Beeswax (B.P.C.). Cera Flava; Yellow Wax (U.S.P.) Cire Jaune; Gelbes Wachs; Cera amarilla.
Foreign Pharmacopeias: In all pharmacopeias examined

A secretion formed by the hive bee, Apis mellifera and other species of  Apis (Apidae), used by the insect to form the cells of the honeycomb. It is a yellowish-brown solid with an agreeable honey-like odour, brittle when cold,  plastic when warmed. M.p. 62 to 65 C. Insoluble in water; sparingly soluble in alcohol; soluble in chloroform,  warm ether, and fixed and volatile oils.

Uses. Yellow beeswax is used as an ingredient of ointments and enables water to be incorporated to produce water-in-oil emulsions. White beeswax is similarly employed.

Aseptic Surgical Wax (B.P.C.1949). Cera Aseptica Chirurgicalis;  Horsley's Wax; Bone Wax. A sterile mixture of yellow beeswax 7 and olive oil 2, with  phenol 1, all by wt. Cover with a 0.2% w/v solution of mercuric chloride and  protect from light In sterile bottles In a cool place. When required for use the container should be heated to 100 C for 5 minutes and the contents poured into a  0.2% w /v solution of mercuric chloride warmed to 40 C. It is used for  preventing haemorrhage in cranial surgery.

Erlangen Cream (St. Bart's Hosp) White  beeswax 2.5 oz. spermaceti 2 oz. arachis oil 20 fluid oz., water 7 fluid  oz.

Galen's Wax. Cerat de Galien (Fr. P). White beeswax 130 g. almond oil 535 g.  distilled rose water 330 g. borax 5 g.

Unguentum Aquosum (Chin.P) White beeswax  18g borax 1g. almond oil 61g water to100 g.


The U.S.D. 21st Ed. 1926
The U.S. Monograph also includes the vegetable waxes used for adulterating beeswax.

CER.A ALBA. U.S. Br.
WHITE WAX Cer. Alb.
Yellow Wax, bleached. U.S. White Beeswax is Yellow Beeswax, bleached.  Br.
White Beeswax; Cire Blanche, Fr. Cod. Cera alba, P.G. Weisses Wachs, G.  Cera bianca. It.. Cera blanca, Sp.

CER.A FLA VA. U.S. Br.
YELLOW WAX Cer, Flav. [Beeswax]

Yellow Wax is obtained by melting and purifying the honey-comb of the bee, Apis mellifera,
Linne (Fam Apidae) U.S. Yellow Beeswax is obtained from the honeycomb of  the Hive Bee, Apis mellifera, Linn., and possibly other species of Apis.  Br,

Yellow Beeswax. Cera Citrina. Beeswax. Cire Jaune, Fr. Cod. Cire d'  Abeilies, Fr. Cera Flava, P.G. Gelbes Wachs, G. Cera gialla, Cera vergine, It.  Cera amarilla, Sp.


The name Apis mellifera was given to the honeybee of Linnaeus in 1758,  Systema Naturae 10th edition; subsequently (Ibid. 12th edition,1767), Linnaeus changed the name to mellifica, but according to the established rules of scientific nomenclature the first name must hold, and the revisers of the United States Pharmacopoeia were therefore correct in adopting it. It was at one time doubtful whether the wax which constitutes the walls of the honey-comb is elaborated or merely gathered by the insect. Huber, however, by feeding bees exclusively on honey and water, determined that under these circumstances wax is  produced in the form of scales under the rings of the abdomen. But wax also exists in plants, bearing in this, as in other respects, a close analogy to the  fixed oils.

Besides the official species other bees are used as honey makers. The  extremely vicious, blackish-brown Apis fasciata was kept by the ancient  Egyptians on floating apiaries, which as the season progressed slowly drifted  down the Nile, following the successive opening of the flowers. In Senegal, Apis adansonii, and in Southern Africa, Apis caffra and Apis Scutellata produce honey; while the Apis unicolor of Madagascar has been domesticated in that island and introduced into other parts of the world. In India honey is made in large quantities by Apis dorsata (Apis indica, Apis florea) the largest bees known.

India Wax differs from official wax chiefly in its lower acid value. A wax is also produced in India by the so-called Kota bees,  belonging to the genus Melipona, they are minute stingless insects, which furnish a sticky, dark-colored wax,  resembling in physical and chemical characteristics the propolis of the honey bee rather than the true wax.

Propolis or bee-glue is a brownish resinous substance  sometimes deposited at the base of the hive; apparently it is collected from the resinous exudate of various  trees such as the balsam, poplar and conifers. It has been used in place of the afore mentioned in various preparations. .

1.CERA FLAVA, or Yellow Wax.-This is obtained by  slicing the comb taken from the hive, removing the honey by centrifugal action, and melting the residue in boiling water, which is kept hot for some time in order to allow the impurities to separate and either subside or be dissolved by the water. When the liquid cools the wax concretes, and, having been removed and again melted in boiling water, is strained and poured into  pans or other suitable vessels. The labor saving device is sometimes adopted of  stretching a strainer of cheese cloth upon a hoop and wedging the latter down  into the hot mixture below the level of the water; as this cools, the melted wax  slowly rises through the cloth, and thus a perfectly clean cake of wax is formed  on top on cooling. It is usually brought to market in round flat cakes from  three to six inches in thickness.

Description and Physical Properties. A solid, varying in color from yellow to grayish brown. It has an agreeable, honey-like odor, and a faint, characteristic  taste. When cold, it is somewhat brittle, and when broken presents a dull, granular,  non-crystalline fracture. Yellow Wax is insoluble in water and sparingly soluble  in cold alcohol. Boiling alcohol dissolves the cerotic acid and a portion of the  myricin, which are constituents of Yellow Wax. It is completely soluble in chloroform, ether, and in fixed or volatile oils, partly soluble in cold bezene  or in carbon disulphide, and completely soluble in these liquids at about 30C 

Specific gravity: 0.950 to 0.960 at 25" C. determined as  follows: Melt the Wax at a low temperature, and allow it to fall in separated drops from just above the  surface into alcohol that has been warmed to from 45" to 50" C. Allow the globules to remain  in the alcohol until it has cooled spontaneously to room temperature (20 to 25 C.), then  remove the Wax and keep it at room temperature for twenty-four hours. Prepare a mixture of  four volumes of alcohol and enough distilled water to make ten volumes, and allow it to  stand until free from air bubbles. Moisten the globules of Wax with distilled water by means of a brush, and place them by means of forceps in the alcohol solution just  prepared and contained in a beaker. Then add alcohol, or air-free distilled  water, as required, to the mixture, kept at 25C until the globules of the Wax  float and rest indifferently in the liquid. Finally determine the specific gravity of the alcohol-water mixture. The figure thus obtained represents the  specific gravity of the Wax examined. Yellow Wax melts between 62 and 65"  C.

Boil 1Gm of Yellow Wax for half an hour with 35 cc. of an aqueous solution of sodium hydroxide (1 in 7), the volume being preserved by the occasional addition  of distilled water on cooling, the Wax separates without rendering the liquid opaque, and no  precipitate is produced in the liquid after filtration through glass wool or asbestos on the addition of an excess of hydrochloric acid (fats or fatty acids,  Japan wax, rosin or soap). Warm about 3 Gm. of Yellow Wax, accurately weighed,  in a 200 cc. flask with 25 cc. of neutral dehydrated alcohol until melted, add 1  cc. of phenolphthalein T.S. and while warm titrate with half normal alcoholic  potassium hydroxide until a permanent, faint pink color is produced: the acid  number so obtained is not less than 18 and not more than 24. Now add 25 cc. of half normal alcoholic potassium hydroxide and 50 cc. of alcohol, boil the  mixture for four hours under a reflux condenser, and titrate the excess of the alkali with half-normal hydrochloric acid: the ester number so obtained is not  less than 72 and not more than 77. U.S.

A yellowish-brown solid; somewhat brittle when cold, but becoming plastic by the heat of the hand. Agreeable, honey-like odor. Fracture granular, not crystalline.  Soluble in chloroform and in fixed and volatile oils. Specific gravity 0.958 to  0.970. Melting point 61 to 64 C. Refractive index at 80 C. 1.4380 to 1.4420. When 5 grammes are  boiled for ten minutes with 80 millilitres of an aqueous solution (1 in 10) of sodium  hydroxide, the water lost by evaporation being made up, the resulting solution; cooled and  filtered through asbestos, does not become turbid when acidified with  hydrochloric acid (absence of fats, fatty acids, Japan wax, resin). 5 grammes  heated with 20 millilitres of absolute alcohol and shaken until uniformly  distributed, require for neutralisation at boiling temperature not less than 1.5  and not more than 2.0 millilitres of N/1 alcoholic solution of potassium  hydroxide, solution of phenol phthalein being used as indicator (limits of free  acid). Upon the further addition of 20 millilitres of N /1 alcoholic. solution  of potassium hydroxide, and on well boiling for one and a quarter hours in a  flask to which a reflux condenser is attached, not less than 13.2 and not more  than 13.8 millilitres of N/1 solution of sulphuric acid are required for  neutralisation. When 5 grammes are boiled with a mixture of 15 millilitres of N!1 alcoholic solution of potassium hydroxide N/1 and  15 millilitres of absolute alcohol until completely saponified, the alcohol  evaporated, the residue dissolved in 20 millilitres of glycerin on a water-bath  and 30 millilitres of boiling water added, a clear or translucent solution is  obtained (absence of ceresin, paraffin and other waxes). Not more than 1 per cent. is soluble in boiling water (limit of honey)." Br.

Dieterich examined hundreds of specimens annually, and found the wax to  vary in sp. gr.
from 0.963 to 0.966;' He also determined the specific gravities of the following substances some of which are used to adulterate wax.

white wax, 0.973; cera japonica, Japan wax, 0.975; ceresin, half white,  0.920; ozokerite, crude, 0.952; rosin, common, 1.108; cacao butter, 0.980 to 0.981; pure  rosin, 1.045; beef suet, 0.952 to 0.953; yellow wax, 0.963 to 0.964, ceresin,  white, 0.918; ceresin, yellow, 0.922 . spermaceti, 0.960; French rosin, 1.104 to  1.105 ; paraffin, med. hard, 0.913 to 0.914; stearin, A No.1, 0.971 to 0.972;  mutton suet, 0.961.

2. CERA ALBA,
Bleached Yellow Wax, or White Wax. The color of yellow wax is discharged by exposing it, with an extended surface, to the combined  influence of air, light and moisture. The process of bleaching is carried on to a considerable  extent in this country. The wax, previously melted, is made to fall in streams  upon a revolving cylinder, kept constantly wet, upon which it concretes, forming  thin ribbon-like layers. These, having been removed, are spread upon linen cloths stretched on frames and exposed to the air and light; care being taken to water and occasionally turn them. In a few days they are partially bleached;  but, to deprive the wax completely of color, it is necessary to repeat the whole process once, if not oftener. When sufficiently white, it is melted and cast  into small circular cakes. Yellow wax may also be decolorized by treatment with animal charcoal or with potassium permanganate, potassium dichromate and  sulphuric acid, and hydrogen peroxide. White wax sometimes contains one or more  free fatty acids, consequent probably upon the employment of alkalies in  bleaching it, which render it an unfit ingredient in the unctuous preparations of certain salts. It  may be deprived of these acids by means of alcohol.

WHITE WAX is less unctuous to the touch than the yellow,  soft and ductile at 35 C. and fusible at 65 C. retaining its fluidity at a lower  temperature.

Description and Physical Properties. A yellowish-white solid, somewhat translucent in thin layers. It has a faint, characteristic odor, is free from rancidity,  and is nearly tasteless. The acid value is not less than 17 and not more than  23, and the ester value not less than 72 and not more than 79, as determined by  the methods under Cera Flava. In other respects White Wax has the  characteristics of, and responds to, the tests for identity and purity under  Cera Flava. U.S. Hard, nearly white, translucent cakes or masses. 5 grammes  heated with a mixture of 10 millilitres of alcohol (90 per cent.) and 10  millilitres of absolute alcohol) and shaken until evenly distributed, require  for neutralisation at boiling temperature not less than 1.5 and not more than  2.2 millilitres of N/1alcoholic solution of potassium hydroxide, solution of  phenolphthalein being used as indicator (limits of free acid). In other respects  White Beeswax responds to the tests described under Cera Flava. By a great heat  it is partly volatilised and partly decomposed; and, when flame is applied to  its vapor, it takes fire and burns with a clear bright light. It is insoluble in  water, and in cold alcohol or ether, but is slightly soluble in boiling alcohol  and ether, which deposit it in a great measure upon cooling. The volatile and  fixed oils dissolve it with facility, rosin readily unites with it by fusion,  and soaps are formed by the action of sodium and potassium hydroxides. It is not  affected by the acids at ordinary temperatures, but is converted into a black  mass when boiled with concentrated sulphuric acid.

Composition. The chief constituent of beeswax is myricin  (myricyl palmitate); melting point 64C. It also contains cerotic acid (formerly called cerin),  C26H52O2, and melissic acid, C30H60O2. There are also present about 6 per cent. of hydrocarbons  of the paraffin series, including heptacosane, C27H56, and hentracontane, C31H64. Myricyl  alcohol, C30H61OH, is liberated when myricyl palmitate is saponified, as  glycerol is liberated during the saponification of ordinary fats; when fused  with potash lime it is converted into potassium melissate, KC30H59O2. Hot  alcohol extracts the cerotic and melissic acids ,from beeswax. Saponification,  of course, decomposes the myricyl palmitate.

Schwalb (Ann; Chem., ccxxiv, 225) found melissic acid and some  unsaturated acids of the oleic series; also higher solid hydrocarbons of the saturated series, such as C27H56 and C31H64.

Kebler states that the hydrocarbons in beeswax amount to from 12.72 to  13.78 per cent.
(A.J.P. 1893,587.) T. Marie believes that cerotic acid, as ordinarily obtained, is a mixture of two distinct acids. He gives C30H60O2 as the formula for melissic acid and C25H50O2 for pure cerotic acid.

Adulterations
. Wax has been variously adulterated. Heavy  substances sink to the bottom of the vessel when the wax is melted. Starch meal, and other insoluble substances remain behind when the Wax is dissolved in oil of turpentine or petroleum  benzin. Water, which, is said to be sometimes fraudulently incorporated with it, by agitation  when partially melted, is driven off by heat. Fatty substances render lime water turbid when agitated with it and allowed to stand. The best test for fats, fatty acids, or rosin is the  pharmacopeia test of boiling with sodium hydroxide solution and testing the filtrate obtained  after cooling. For the detection of rosin, cold alcohol is sufficient. It dissolves the  rosin, and yields it on evaporation, attended with a very small portion of pure  Wax, which yields 2.4 per cent. to cold alcohol. F. Jean states that a few drops  of sulphuric acid added to the melted wax adulterated with rosin, cause a red color, or, if only 1 per cent. be present, a green tint.
(A. J. P., 1881,p. 307.)

To detect Japan wax, Dullo boils together for a minute 10 Gm. of  wax, 120 Gm. of water, and 1 Gm. of sodium hydroxide; if Japan wax be present, a  soap will immediately form, which will slowly solidify on cooling. Beeswax does  not saponify under these circumstances. Ceresin, a principle obtained from  ozokerite (see Part II), is also employed as an adulterant, and is manufactured  largely for that purpose in Vienna. It is only native paraffin, and of course  answers to the tests for that substance. There are other less precise methods of detecting adulterations. Thus, spermaceti and lard  render wax softer and less cohesive, of a smoother and less granular fracture, and of a  different odor when heated. The melting point and specific gravity are lowered  by tallow, suet, lard, and especially by paraffin. Legrip's cereometer is based  upon the altered specific gravity of wax when adulterated. Anyone may apply this  principle by making such a mixture of alcohol and water that pure wax will  neither sink nor rise in it, but remain wherever placed. Adulterated wax would  either float or sink in this liquid. The most valuable information, however, is obtained by the determination of the chemical constants-viz., acid value, ester  value, and saponification value. In yellow wax the iodine value is also of use;  in white wax the bleaching process has altered the bodies which absorb the  iodine; (See also Chem. Ztg., 1893, 918; Proc. A. Ph. A., 1894, 957 ; D. C.,  1898,33; Am. Drug., 1898,97, and 1904, 103,105.) According to G. Brehner (Ztsch. Off Ch., 1912, No.5), who has  investigated many samples of beeswax of normal and abnormal character, beeswax may be  divided into three groups, according to source, as follows:

(1) Ordinary beeswax in which the ratio of acid number to ester number is  1:3.6 to 1 : 4.
(2) Beeswax of African origin in which the ratio is 1 : 3.
(3) Beeswax of Indian origin in which the ratio is 1 : 12. A Leijs  (J.P.C. 1912, No.12) describes a method-based on the relative solubility in  fuming hydrochloric acid, which, he claims, enables him with certainty to separate paraffin,  ceresin, and carnauba wax readily from beeswax. Under the name of White Wax No.2 or White Wax for technical use, there has 'been found in commerce a product put  up in exact imitation of white beeswax, and consisting of over 90 per cent. of  paraffin.     

 

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