EUROPEAN PHARMACOPOEIA 11.

1 Lactose

04/2023:1061 Acidity or alkalinity. Dissolve 6.0 g by heating in 25 mL

of carbon dioxide-free water R, cool and add 0.3 mL of
phenolphthalein solution R1. The solution is colourless. Not
more than 0.4 mL of 0.1 M sodium hydroxide is required to
change the colour of the indicator to pink or red.
LACTOSE (1) Specific optical rotation (2.2.7): + 54.4 to + 55.9 (anhydrous
substance).
Lactosum Dissolve 10.0 g in 80 mL of water R with heating at 50 °C.
Allow to cool and add 0.2 mL of dilute ammonia R1. Allow to
stand for 30 min and dilute to 100.0 mL with water R.
Absorbance : proteins and light-absorbing impurities
C12H22O11 Mr 342.3 (2.2.25).
[63-42-3]
Test solution (a). Solution S.
DEFINITION Test solution (b). Dilute 1.0 mL of test solution (a) to 10.0 mL
O-β-D-Galactopyranosyl-(1→4)-β-D-glucopyranose or mixture with water R.
of O-β-D-galactopyranosyl-(1→4)-α-D-glucopyranose and Spectral range : 400 nm for test solution (a) and 210-300 nm
O-β-D-galactopyranosyl-(1→4)-β-D-glucopyranose. for test solution (b).
♦CHARACTERS Results :
Appearance : white or almost white, crystalline powder. – at 400 nm : maximum 0.04 for test solution (a) ;
Solubility : freely soluble in water, practically insoluble in – from 210 nm to 220 nm : maximum 0.25 for test
ethanol (96 per cent).♦ solution (b) ;
– from 270 nm to 300 nm : maximum 0.07 for test
IDENTIFICATION solution (b).
First identification : A, ◊D. Water (2.5.12): maximum 1.0 per cent, determined on 1.00 g,
Second identification : B, C, D.◊ using a mixture of 1 volume of formamide R and 2 volumes of
A. Infrared absorption spectrophotometry (2.2.24). methanol R as solvent.
Comparison : anhydrous lactose CRS. Sulfated ash (2.4.14): maximum 0.1 per cent, determined on
◊B. Thin-layer chromatography (2.2.27). 1.0 g.
Solvent mixture : water R, methanol R (40:60 V/V). Microbial contamination
Test solution. Dissolve 10 mg of the substance to be TAMC : acceptance criterion 102 CFU/g (2.6.12).
examined in the solvent mixture and dilute to 20 mL with Absence of Escherichia coli (2.6.13).
the solvent mixture.
Reference solution. Dissolve 10 mg of anhydrous lactose CRS ◊FUNCTIONALITY-RELATED CHARACTERISTICS
in the solvent mixture and dilute to 20 mL with the solvent This section provides information on characteristics that are
mixture. recognised as being relevant control parameters for one or
Plate : TLC silica gel plate R. more functions of the substance when used as an excipient
(see chapter 5.15). Some of the characteristics described in
Mobile phase : water R, methanol R, glacial acetic acid R, the Functionality-related characteristics section may also be
methylene chloride R (10:15:25:50 V/V/V/V); measure the present in the mandatory part of the monograph since they
volumes accurately, as a slight excess of water produces also represent mandatory quality criteria. In such cases, a
cloudiness. cross-reference to the tests described in the mandatory part is
Application : 2 μL ; thoroughly dry the points of application. included in the Functionality-related characteristics section.
Development A : over 3/4 of the plate. Control of the characteristics can contribute to the quality
Drying A : in a current of warm air. of a medicinal product by improving the consistency of the
manufacturing process and the performance of the medicinal
Development B : immediately, over 3/4 of the plate, after
product during use. Where control methods are cited, they are
renewing the mobile phase.
recognised as being suitable for the purpose, but other methods
Drying B : in a current of warm air. can also be used. Wherever results for a particular characteristic
Detection : spray with a solution of 0.5 g of thymol R in a are reported, the control method must be indicated.
mixture of 5 mL of sulfuric acid R and 95 mL of ethanol The following characteristics may be relevant for lactose used
(96 per cent) R ; heat at 130 °C for 10 min. as filler/diluent in solid dosage forms (compressed and powder).
Results : the principal spot in the chromatogram obtained Particle-size distribution (2.9.31 or 2.9.38).
with the test solution is similar in position, colour and size
to the principal spot in the chromatogram obtained with Bulk and tapped density (2.9.34). Determine the bulk density
the reference solution. and the tapped density. Calculate the Hausner ratio.
C. Dissolve 0.25 g in 5 mL of water R. Add 5 mL of ammonia R α-Lactose and β-lactose. Gas chromatography (2.2.28).
and heat in a water-bath at 80 °C for 10 min. A red colour Silylation reagent : dimethyl sulfoxide R, N-trimethylsilylimida-
develops. zole R, pyridine R (19.5:22:58.5 V/V/V).
D. Water (see Tests).◊ Test solution. Introduce 10 mg of the substance to be examined
into a vial with a screw cap and add 4 mL of the silylation
TESTS reagent. Sonicate for 20 min at room temperature, allow to
Solution S. Dissolve 1.0 g in boiling water R, allow to cool and cool and transfer 400 μL to an injection vial. Add 1 mL of
dilute to 10.0 mL with water R. pyridine R, close the vial and mix well.
Appearance of solution. Solution S is clear (2.2.1) and not Reference solution. Prepare a mixture of α-lactose
more intensely coloured than reference solution BY7 (2.2.2, monohydrate R and β-lactose R to obtain an anomeric ratio
Method II). of about 1:1 based on the labelled anomeric contents of the
(1) This monograph has undergone pharmacopoeial harmonisation. See chapter 5.8. Pharmacopoeial harmonisation.

General Notices (1) apply to all monographs and other texts 4713
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Lactose monohydrate EUROPEAN PHARMACOPOEIA 11.1

α-lactose monohydrate and the β-lactose. Introduce 10 mg of ♦CHARACTERS

the mixture into a vial with a screw cap and add 4 mL of the Appearance : white or almost white, crystalline powder.
silylation reagent. Sonicate for 20 min at room temperature, Solubility : freely soluble in water, practically insoluble in
allow to cool, and transfer 400 μL to an injection vial. Add ethanol (96 per cent).♦
1 mL of pyridine R, close the vial and mix well.
Precolumn : IDENTIFICATION
– material : intermediate-polarity deactivated fused silica ; First identification : A, ◊D.
– size : l = 2 m, Ø = 0.53 mm. Second identification : B, C, D.◊
Column : A. Infrared absorption spectrophotometry (2.2.24).
– material : fused silica ; Comparison : lactose monohydrate CRS.
– size : l = 15 m, Ø = 0.25 mm ; ◊B. Thin-layer chromatography (2.2.27).
– stationary phase : phenyl(5)methyl(95)polysiloxane R (film Solvent mixture : water R, methanol R (40:60 V/V).
thickness 0.25 μm). Test solution. Dissolve 10 mg of the substance to be
Carrier gas : helium for chromatography R. examined in the solvent mixture and dilute to 20 mL with
Flow rate : 2.8 mL/min. the solvent mixture.
Temperature : Reference solution. Dissolve 10 mg of lactose
monohydrate CRS in the solvent mixture and dilute to
Time Temperature
20 mL with the solvent mixture.
(min) (°C)
Plate : TLC silica gel plate R.
Column 0-1 80
Mobile phase : water R, methanol R, glacial acetic acid R,
1-3 80 → 150 methylene chloride R (10:15:25:50 V/V/V/V); measure the
3 - 15.5 150 → 300
volumes accurately, as a slight excess of water produces
cloudiness.
15.5 - 17.5 300 Application : 2 μL ; thoroughly dry the points of application.
Injection port 275 or use cold on-column Development A : over 3/4 of the plate.
injection Drying A : in a current of warm air.
Detector 325 Development B : immediately, over 3/4 of the plate, after
Detection : flame ionisation. renewing the mobile phase.
Injection : 0.5 μL, splitless or by cold on-column injection. Drying B : in a current of warm air.
Relative retention with reference to β-lactose (retention Detection : spray with a solution of 0.5 g of thymol R in a
time = about 12 min) : α-lactose = about 0.9. mixture of 5 mL of sulfuric acid R and 95 mL of ethanol
(96 per cent) R ; heat at 130 °C for 10 min.
System suitability : reference solution :
Results : the principal spot in the chromatogram obtained
– resolution : minimum 3.0 between the peaks due to α-lactose with the test solution is similar in position, colour and size
and β-lactose. to the principal spot in the chromatogram obtained with
Calculate the percentage content of α-lactose using the the reference solution.
following expression : C. Dissolve 0.25 g in 5 mL of water R. Add 5 mL of ammonia R
100Sa and heat in a water-bath at 80 °C for 10 min. A red colour
Sa + Sb develops.
D. Water (see Tests).◊
Calculate the percentage content of β-lactose using the
following expression : TESTS
100Sb Solution S. Dissolve 1.0 g in boiling water R, allow to cool and
Sa + Sb dilute to 10.0 mL with water R.
Appearance of solution. Solution S is clear (2.2.1) ◊and not
Sa = area of the peak due to α-lactose ; more intensely coloured than reference solution BY7 (2.2.2,
Sb = area of the peak due to β-lactose. Method II)◊.
Acidity or alkalinity. Dissolve 6.0 g by heating in 25 mL
Loss on drying (2.2.32). Determine on 1.000 g by drying in of carbon dioxide-free water R, cool and add 0.3 mL of
an oven at 80 °C for 2 h.◊ phenolphthalein solution R1. The solution is colourless. Not
more than 0.4 mL of 0.1 M sodium hydroxide is required to
04/2023:0187 change the colour of the indicator to pink or red.
Specific optical rotation (2.2.7): + 54.4 to + 55.9 (anhydrous
substance).
Dissolve 10.0 g in 80 mL of water R with heating at 50 °C.
Allow to cool and add 0.2 mL of dilute ammonia R1. Allow to
LACTOSE MONOHYDRATE(2) stand for 30 min and dilute to 100.0 mL with water R.
Absorbance : proteins and light-absorbing impurities
Lactosum monohydricum (2.2.25).
Test solution (a). Solution S.
Test solution (b). Dilute 1.0 mL of test solution (a) to 10.0 mL
C12H22O11,H2O Mr 360.3 with water R.
DEFINITION Spectral range : 400 nm for test solution (a) and 210-300 nm
O-β-D-Galactopyranosyl-(1→4)-α-D-glucopyranose for test solution (b).
monohydrate. Results :
(2) This monograph has undergone pharmacopoeial harmonisation. See chapter 5.8. Pharmacopoeial harmonisation.

4714 See the information section on general monographs (cover pages)

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