5794 Lactobionic Acid / Official Monographs
volumetric flask. Dilute with 1% (w/v) hydrochloric acid
tovolume.
Reference solution: Pipet 12.5 ml of the Standard stock
solution into a 100-ml volumetric flask. Dilute with 1%
(w/v) hydrochloric acid to volume.
Acceptance criteria: The Sample solution is clear and not
more intensely colored than the Reference solution.
• 0PncAL ROTATION, Specific Rotation (781 S)
Sample solution: 10 mg/ml of lactobionic Acid. Allow to
stand for 24 h.
Acceptance criteria: +23.0° to +29.0° (anhydrous
substance)
• REDUCING SUCiARs
Sample solution: Dissolve 5.0 g of lactobionic Acid in 25
ml of water with the aid of gentle heat, and cool.
Analysis: To the Sample solution add 20 ml of cupric
citrate TS and a few glass beads. Heat so that boiling
begins after 4 min, and maintain boiling for 3 min. Cool
rapidly, and _add 100 ml of a 2.4% solution of glacial
acetic acid and 20.0 ml of 0.025 M iodine VS. With
continuous shaking, add 25 ml of a mixture of 6 ml of
hydrochloric acid and 94 ml of water. When the
precipitate has dissolved, titrate the excess iodine ;with
0.05 M sodium thiosulfate VS using 1 ml of starch TS,
added toward the end of the titration as an indicator.
Acceptance criteria: NlT 12.8 ml of 0.05 Msodium
thiosulfate VS is required, corresponding to NMT 0.2% of
reducing sugars, as glucose.
• AmCI.ES OF IOTANICAI. OllcilN, .Total Ash (561 ): NMT
0.2%
ADDfflONAL REQUIREMENTS
• PACKACING AND SlORAGE: Preserve in well-closed
containers.
• USP ltERRENCE STANDARDS (11)
USP Lactobionic Acid RS
Anhydrous Ladose
Portions of the monograph text that are national USP text,
and are not part of the harmonized text, are marked with
symbols (' ,) to specify this tact.
~).)~~
1-.-1
OH
DERNfflON
Anhydrous Lactose is O-p-o-galactopyranosyl-(1-+4)-P-D-
glucopyranose (p-lactose), or a mixture of 0-P-D-
galactopyranosyl-(l -+4)-P-D-glucopyranose and O-P-D-
galactopyranosyl-(1-+4)-a-D-glucopyranose (a-lactose).
IDENTIFICATION
• A. INFRARED ABSORP110N (197K)
• •B. THIN-LAYER CHROMATOGRAPHIC IDENTIRCATION TEsr
(201)
Adsorbent: 0.25-mm layer of chromatographie silica gel
Diluent: Methanol and water (3:2)
Standard solution A: 0.5 mg/ml of USP Anhydrous
Lactose RS in Diluent
Standard solution B: Contains 0.5 mg/ml of USP
Dextrose RS, 0.5 mg/ml of USP Anhydrous Lactose RS,
0.5 mg/ml of USP Fructose RS, and 0.5 mg/n,
sucrose RS in Diluent Lof Usp
Sample solution: 0.5 mg/ml of Anhydrous L
Diluent actose in
Application volume: 2 µl
Developing solvent system: Ethylene dichlor'd 1
acetic acid, methanol, and water (10:5:3:i) e, glacial
Spray reag~nt: . 5 mg/ml of thymol in a mixture
and sulfunc ac1d (19: 1) of alcohol
Analysis , .
Samples: Standard solution A, Standard solutio 8
Sample solution n , and
Allow the spots to dry, .and develop the plate in
lined chromatograph1c chamber equilibrated 1~~r-
Developing solvent,system for about-1 h prior t~ ~ the
Allow the chromatogram to develop until the s 1se.
front has moved about three-quarters of the lei ~hnt
the plate. Remove the plate from the chamber _of a
current of warm air, and redevelop the plate i~fr ryhn a
Developing solvent system. Remove the plate frome;h
chamber, mark the solvent front, and dry the plate .e
current of warm air. Spray the plate evenly with
reagent. Heat. the plate at 30° for 0 min.
sp; a
System suitabihty: The test 1s ~ot va_hd unless Standard
solution Bshows four clearly d1s~e~111ble spots,
disregarding any spots at thé ongm.
Acceptance criteria: The principal spot from the Sompk
solution corresponds in appearance and R, value to that
from Standard solution A.,
OTHER COMPONENTS
• •coNnNT OF ALPHA AND BETA ANOMERS
Silylation reagent: Dimethyl sulfoxide, pyridine, and
trimethylsilylimidazole (19 .5: 58.5: 22)
Standard solution: Prepare a mixture of alpha-lactose
monohydrate and beta-lactose having an anomeric ratio
of about 1:1 based on the labeled anomeric contents of
the alpha-lactose monohydrate and the beta-lactose.
lntroduce 10 mg of this mixture into a vial with a screw
cap. Add 4 ml of Silylation reagent. Sonicate for 20 min at
room temperature. Transfer 400 µl to an injection vial.
Add 1 ml of pyridine. Close the vial, and mix well.
Sample solution: lntroduce 1O mg of Anhydrous Lactose
into a vial with a screw cap. Add 4 ml of Silylation
reagent. Sonicate for 20 min at room temperature. ..
Transfer 400 µl to an injection vial. Add 1 ml of pyndme.
Close the vial, and mix·well.
Chromatographie system
(See Chromatography (621 ), System Suitability.)
Mode: GC
Detector: Flame ionization
Columns
Precolumn:1 0.53-mm x 2-m intermediate polarity
deactivated fused silica . . . film
Analytical: 2 0.25-mm x 15-m G27 on fused sihca,
thickness 0.25 µm
Temperatures
Detector: 325° . •ection
Injection port: 275° or use cold on-column inJ
Column: See Table 1.
1
2
Restek Guard column is suitable.
Varian CP-Sil 8 CB is suitable.
 Official Monographs I Lactose 5 795

Table 1
Instrumental conditions
remperature Final
Hold Time at Fi-
nal
Mode: Vis
1~1v•
1 Ramp Temperature T emperature 1nalytlcal wavelength: 400 nm
~ra'""' (°/min) (·) (min) Acceptance crlterla: NMT 0.04 for the absorbance .
fifl' (')
80 divided by the path length in centimeters; and the cl~nty
so of the Sample solution is the same as that of water or its
35 150 opalescence is not more pronounced than that of the
so Reference suspension, and it is not more colored than the
12 300 2
150 Reference solution.
• Loss ON DRYINC. (7 31 )
. gas· Helium Analysis: Dry a sample at 80° for 2 h.
z
c,r11er . s ml/min Acceptance criteria: NMT 0.5%
flO rate.
w . O5 L
•on volume: . • µ • WATER DEIUIMINATION, Method I (921)
1o!~on type: Sphtless or by cold on-column injection Sample solution: Anhydrous Lactose in a mixture of
io,..-- sultability methanol and formamide (2:1)
systernle· standard solution Acceptance criteria: NMT 1 .0%
~f billty requirements
s;esolution: NLT 3.0 between the peaks due to alpha-
• MICROBIAL ENUMERATION TESTS (61) and TESTS FOR
SPf.CIRm MKROORGANISMS (62): The total aerobic
lactose and beta-lactose , microbial count is NMT 102 du/g and •the total
combined molds and yeasts count is NMT 50 cfu/g, . lt
All~~~e: Sample solution meets the requirements of the test for absence of
[NOTE-The rel_ative retention time with reference to Escherichia coti.
tieta-lactose 1s about 0.9 for.alpha-lactose (retention • PROTBN AND LIC.HT-ABSORBINC. IMPURfflES
time = about 12 min).] (See Ultraviolet-Visible Speetroscopy (857).)
Calculate the percentage content of alpha-lactose: Sample solution: 1% solution (w/v)
Instrumental conditions
Result = S J(S a+ Sb) x 100 Mode: UV
Wavelength range: 210-300 nm
5, = area of the peak due to alpha-lactose Acceptance criterla: NMT 0.25 for the absorbance
Sb = area of the peak due to beta-lactose divided by the path length in centimeters at 210-220 nm;
NMT 0.07 for the absorbance divided by the path length
Calculate the percentage content of beta-lactose: in centimeters at 270-300 nm
• ACIDrrY OR AlKAUNrrY
Result = S J(S a+ Sb) x 100 Sample solution: Dissolve 6 g by heating in 25 ml of
carbon dioxide-free water, cool, and add 0.3 ml of
S, = area of the peak dùe to alpha-lactose phenolphthalein TS.
Sb = area of the peak due to beta-lactose Acceptance criteria: The solution is colorless, and NMT
0.4 ml of O. 1 N sodium hydroxide is required to produce
a pink or red color.
• OPTICAL ROTATION, Specific Rotation (781 S)
INIITIES Sample solution: Dissolve 10 g by heating in 80 ml of
•IEslluE ON k.NmON (281 ): NMT 0.1% water to 50°. Allow to cool, and add 0 .2 ml of 6 N
ammonium hydroxide. Allow to stand for 30 min, and
IPECIICTEm dilute with water to 100 ml.
'CWrrt AND COI.OR OF SOLUllON Acceptance criteria: +54.4° to +55.9°, calculated on the
Hydrazine sulfate solution: Dissolve 1.0 g of hydrazine anhydrous basis, at 20°
suijate in water, and dilute to 100.0 ml. Allow to stand
for~h. ADDfflONAL REQUIREMENTS
Hexamethylenetetramine solution: ln a 100-ml ground- • •PACllAC.INC. AND STORAC.l: Preserve in tight containers.
tass stoppered flask dissolve 2.5 g of • I.ABlUNC.: Where the labeling indicates the relative
quantities of alpha- and beta-lactose, determine
P,examethylenetetramine in 25 .0 ml of water.
~rnary opalescent suspension: To the compliance using Content of Alpha and Beta Anomers.
ramethylenetetramine solution in the flask add 25.0 ml Where the labeling states the particle size distribution it
also indicates the d ,o, d so, and d 90 values and the ra~ge
t
th e Hydrazine sulfate solution. Mix and allow to stand
24 h. This suspension is stable for 2 months, provided for each.
~s stored in a glass container free from surface defects. • USP REFERE.NCl STANDARDS (11)
we~I su~pension must not adhere to the glass and must be USP Dextrose RS
Stan mixed before use. USP Fructose RS
da rd opalescence: Dilute 15.0 ml of the Primary USP Anhydrous Lactose RS
:lesc~nt suspension to 1000.0 ml with water. This USP Sucrose RS
to t~sion is freshly prepared and may be stored for up
Rtleren . d
oPo/e ce suspension: To 5 .0 ml of the Standa~
Use scence add 95 .0 ml of water. Mix and shake before
~~en · es
llllot ce solution: To 6.0 ml of ferric chlonde . , 2 ·5
Cs ad cobaltous chloride es, and 1.0 ml of cupnc sulfate
Slnlpl d hydrochloric acid (1 o g/l HCI) to make 1OOO ml.
tocil.e SOlutlon: 1 g in 10 ml of boiling water. Allow to
 5796 Lactose/ Official Monographs

10 , du/g, and it 171e~ts the requirements of the te

Lactose Monohydrate
Portions of the monograph text that are national U5P text,
and are not part of the harmonized text, are marked with
symbols (' ,) to specify this tact.
DEFINmON
Lactose Monohydrate is the monohydrate of O-P-D-
galactopyranosyl-(1 ....,4)-a-D-glucopyranose. [NOTE-Lactose
Monohydrate may be modified as to ils physical
characteristics. lt may contain varying proportions of
amorphous lactose.]
IDENTIFICATION
• A. INFRARED ABSORPTION (197K)
• '8. THIN-LAYER (HROMATOCRAPHIC IDENTIFICATION TEsT
(201 )
Diluent: Methanol and water (3:2)
Standard solution A: 0.5 mg/ml of USP Lactose
Monohydrate RS in Diluent
Standard solution B: 0.5 mg/ml each of USP Dextrose
RS, USP Lactose Monohydrate RS, USP Fructose RS, and
USP Sucrose RS in Diluent
Sample solution: 0.5 mg/ml of Lactôse Monohydrate in
Diluent .
Adsorbent: 0.25-mm layer of chromatographie silica gel
Application volume: 2 µL
Developing solvent system: Ethylene dichloride, glacial
acetic acid, methanol, and water (10:5:3:2)
Spray reagent: 5 mg/ml of thymol in a mixture of alcohol
and sulfuric acid (19:1)
Analysis
Samples: Standard solution A, Standard solution 8, 11nd
Sample solution
Allow the spots to dry, and develop the plate in a paper-
lined chromatographie chamber equilibrated.with .
Developing solvent system for about l h prior to us~.
Allow the chromatogram to develop until the solven_t
front has moved about three-quarters of the length of
the plate. Remove the plate from the chamber, dry in a
current of warm air, and redevelop the plate in fresh
Developing solvent system. Remove the plate from the
chamber, mark the solvent front, and dry the plate in a
current of warm air. Spray the plate evenly with Spray
reagent. Heat the plate at 130° for 10 min.
System suitability: The test is not valid unless the
chromatogram of Standard solution Bshows four clearly
discernible spots, disregarding any spots at the origin: ·
Acceptance criteria: The principal spot from the Sample
solution corresponds in appearance and R, value to that
from Standard solution A.•
IMPURITIES
• RfSIDUE ON IGNmON (281 )
Analysis: Asample ignited at a temperature of 600 ± 50°
Acceptance criteria: NMT 0.1%
SPECIFIC TESTS
• (LAIIJTY AND (OLOR OF 5oumoN
Sample solution: 1 g in 1Oml of boiling water
absence of Escherich1a col,., st for
• OPTICAL ROTATION,_Specific Rotation (781 S)
Sample solution: Dissolve 10 g by heating in 80
water to 50°. Allow_to cool, and add 0.2 ml of 6~Lof
ammonium hydrox1de. Allow to stand for 30 rnin
dilute with water to 100 ml. , and
Acceptance criteria: +54:4° to +5\9°, calculated
0
anhydrous basis, determmed at 20 n the
• ACIDnY OR AI.J<ALl~nY .
Sample solution: Dissolve 6 g by heatmg in 25 rnL
carbon dioxide-free water, cool, and add 0.3 rnL 0 1 f°
phenolphthalein !S. . .
Acceptance critena: _The solut10~ 1s col?rless, and NM
0.4 ml of 0.1 N sodium hydrox1de VS 1s required to T
produce a pink or red color.
• 'LOSS ON DRYING (731 )
Analysis: Dry~ sa~ple at 80° for 2 h.
Acceptance cntena
Monohydrate: NMT 0.5%
Monohydrate, modified: NMT 1.0%,
• WATER DETERMINATION, Method I (921 )
sample solution: Prepare a preparation containing
Lactose Monohydrate in a mixture of methanol and
formamide (2:1).
Acceptance criteria: 4.5%-5.5%
• PRomN AND "IJGHT-ABSORBING IMPUllfflES
(See Ultraviolet-Visible Spectroscopy (857).)
Sample solution: 1% (~/v) .
Analysis: Measure the hght absorption of the Sample
solution in the range of 210-300 nm.
Acceptance criteria: •The absorbance divided by the path
length, in cm, is NMT 0.25 in the range of 210-220 nm
and is NMT 0.07 in the range of 270-300 nm.
ADDmONAL REQUIREMENTS
• 'PACKAGING AND STORAGE: Preserve in tight containers.
• l.ABEUNG: Where the labeling states the particle size
distribution, it also indicates the d, 0, d5o, and d90 values
and the range for each. For modified Lactose
Monohydrate, also label it to indicate the method of
modification.
• USP REFEIIENCE STANDARDS (11 )
USP Dextrose RS
USP Fructose RS
USP Lactose Monohydrate RS
USP Sucrose RS
Lanolln, Anhydrous
-see Lano/in General Monographs
Lanolln, Modlfled-see Modified Lano/in General
Monographs

_ ________
Analysis: The Sample solution is clear and nearly colorless.
Determine the absorbance of this solution at a
"'
..c:
a.
...01Ill
wavelength of 400 nm.
Acceptance criteria: The absorbance divided by the path
length, in cm, is NMT 0.04.
• • MICROBIAL ENUMERATION TEsTs (61 ) and TEsTs FOR
___
Lanolln Alcohols
__;_ _;_

0 SPEORED MICROORc.ANISMS (62): The total aerobic (8027-33-6].

C: microbial count does not exceed 1 x 102 du/g, the total DEFINmON
0 combined molds and yeasts count does not exceed 5 x Lanolin Alcohols is a mixture of aliphatic alcohols,
2 triterpenoid alcohols, and sterols, obtained by the