Determination of Impurities in Citalopram Hydrobromide by HPLC

(1)

in 1000 13i of ic~cil<r; 38 volumes of rtterknt2oi and 7 vglumes of acetoizi[~r,-il~, with the pH acijustzd to 6.5

-

with ortiiopilosphori ocld, - fioiv rare: 0.8 ml per niiniite, . - spectropl~otomctcr sct at 239 i!ml

- irjection volume: 20 PI.

Inject reference solution (b). The test is not valid unless the resolution between impurit)i £3 and citalopram is not less than 3.0.

Injeci reference solution jaj. Tile test is not valid u n h s the c~lumn eficiency is not less r'nan 5iX0 tifieoretical plates, the tailing factor is nor more than 1.5 and the relative standard deviation for replicate injections is not more than 5.0 per cent.

. . .

. .

^"

[:.lfer/?ai s&z:l&& sali!!ioc. ,\ 0.025 per cent wl\: soltit:.oi; 61

d i n ~ e f / ? ~ ~ i - ( ! - t i ~ ~ ~ j ~ i - j, j - ~ ~ p l ~ ~ i ; ~ ~ ~ u l l ~ ~ l ) ~ ~ ~ z i ~ z e j ~ ~ d r a c h ~ u ~ . i ~ ' ~ fiS(iiiaiopi-~n; Ltzpi!i-ic: CRSj iii? the solvent mixtu:?.

Chromatographic system

- a stainless sicel ci.i;l~rtiit 25 cfil s 3.6 mm, ^wirh octadecylsilane bonded to porous silica (5 pm), - column temperature: 4S":

- mobile ?has" : 9.077 per cent wi'v soiution of dodecyiiri~ner,~j:Ic~in~~:ot~i~ii~z bromide in rile solven!

~llL~.iure,

- 8 0 i ~ rate: I ml per minute, - spectrophotometcr set at 254 nin, - injectionvolume: 10 pi.

Inject reference solution (a) and the test solution. [n the chromatogram obtained .with the test solution, the area of any secondary peak isnot more than 2.5 times the area of the peak

. in the chromatogram obtained with the reference solution (a) (0.25 per pnt) and the sum of the areas of all the secondary peaks is not more than 8 times the area of the peak in the chromatogram obtained with the reference solution (a)

.

(0.8 per cent).

Uniformity of content (For tablets coflfait?in,o 10 mg or less).

Conlply with the test stated under Tablets.

Determine by liquid chromatography (2.4.14).

Use the chromatogaphic system described in the Assay using the following test solution.

inject the reference solution. The relative retention time for citalopram impurity C is about 1.36 and the resolution behveen citalopram and citalopram impurity C is not less than'l.5. The test is not valid unless ths column efficiency is not less than 2000 thearetical plates and the relative standard deviation for replicate injections is not more than 1 :5 per cent.

Inject the referince solution and the test solution.

Calculate the content of C10H21FN20 in the tablets.

Storage. Store protected from moisture, at a temperature not exceeding 30'.

Labelling. The label states the strength in terms of the equii:alent amount of citalopram.

i

Test solution. Powder one tablet, disperse in 10 ml of a

-

0.142 per cent wlv solution of anhydrozis dibasic sodizrtn

plzosphate, add 40 ml of methanol and mix with rhe aid of

CiticO[ine Sodium

ultrasound for 5 minutes. Add sufficient volume of the internal standard solution and dilute stepwise, if necessaq with the solvent mixture to obtain a solution containing 0.0 1 per cent

W/V ofcitaloprarn and 0.0025 per cent wlv of internal standard

solution and filter.

N5

Calculate the content of Cz,HzrFNzO in the tabiet. - 0

A ^N

Other tests. Comply with the tests stated under Tablets.

Assay. Determine by liquid chromatography (2.4.14).

+

^H3C.^cH3

I

/\/ O-f-O-lm-fJ

f(

0 - 0 - Solvenf mixturee

80

volumes of mefh~nol and 20 volumes of a -

Na -

0.142 per cent wlv solution of anhydrous dibasic sodium H3C

~d

^'OH

- phosphate.

~ e s t sol&on. Weigh and powder 20 tablets. Weigh accurately C14H@4Na0,,P2 Mol. Wt. 5 10.3

(2)

'mr~hyI ester of cy1idin?-5'-inonaphospha1e: 5-CLIP estcr.

A.d?e.izmUne by infrared absorption specfr~p!?oiornzv (2.4.6). :. L > ~ ~ L I ~ L ; - - .,.

.,

^.-^,^.---z~lc,ri:.p!i.~jplio~i'rplidlidai-: ^. 5-CMP rnorpholidzic.

Compare the spectrum with that obtained ^ii:iihcii:icoli~re

sadiumRSor \%rith the nferencesoectmmofcilica[ine jodium. Inject reference solution ^(a).The rest is not valid ~in!ess the 0. It gives the reactions of sodium salt (2.3.1). column efficiency is not less than 6000 rixoretical plates and

the tailing factor is not more than 1.5.

-

^Tests

!

Inject reference solution (b). The test is not valid ~lnless rhe ^.

I

pH (2.4.24i. 6.5 to 7.5. deternlineci on 20 percertr ~t.!\!s~iiiiion resoilition betu.~een the ciric~line impuriky

B

and principal peak

in V;Rlei'. is not less thzn 7.C.

I

I ~ ~substances. ~j ~ ~ ~by liquid chromatography inject reference solution jaj and fhe test soiution. The area ~ d~ ~ ~ ~ i ~ ~ of d

-

(24.14). any secondary peak is not more than 0.4 times the area of t11e

principal peak in the chromatogram obtained with reference Test solri~ion. Dissolve 100 mg of the substance under solution (a) (0.2 per cent) and the sum ofthe areas of all the examination in wuter-and dilute to 50.0 ml with rc:aalef: secondary peaks is not more than 2 times the area of principal _{- .} Rejbence .sol~rriot7 ^fa).A 0.001 per cent w!'v solution of peak in the chromatograin obtained wit6 reference solution

ciiicoline sodilrm RS in ~aret: (a) (1.0 percent).

~ g e k c e rolurioiz (b). A soltlrion contaming 0.2 per cent LVIV of citicoiine soh'izm7 RSand 0.002 per cent wlv of citicoiirze inzpnri& B RS in watel:

- a stainless steel column 25 cm s 4.6 mm, packed with

-' octadecylsila~le bonded to porous silica (5 pin), - coluinn temperarure: 30";

- sample temperature: 10°,

-- mobile phase: A. a mixture of I \-olumeofmetAa~ioland 99 volumzj of0.2 per cent vlv solution offormic mid, adjusred to pH 7.5 with rr-ieihj.i~imj~ze,

' B. meil7ano1, - flow rate: 0.5 mi per minute,

- a gradient programme using the conditions given below

Iron (2.3.14j. ?gcomplies with the limit test for iron (10 ppm):

Chlorides(2.3.12). 0.5 gcomplies with the limit test for chlorides

.

^.

(500ppm). " ^. .

Water (2.3.13). Not morz than 5.0 per cent,'defem~inzd on 0.5 g.

Heavy metals (2.3.13). 2 g complies with the limit test for heat:

metals. Method B (10 ppm).

Assay. Dctcrminc by liquid chromatography (2.4.14).

&st solution.

iss solve

50 mg 0.f the substance under,

examination in water anddilute to 100.0 mi with ~t~oltnler. a Reference solufion. A 0.05 per cent \v/v solution of cificofiile

socfiunr RS in water.

- spectrophorometer set at 276 nm, Chromatographic system !

-

injection volume: 1.0

PI.

- a stainless steel coiumn 25 cm s 1.6 mm, packed with

:

Tm~e ~ o b i l e phase A Mobile phase B octadecylsilane bonded to porous silica (5 pm),

1

(in mim.) (per cent vlv) ^{. ..}(per cent vlv) ^-column temperature: 30°, - sample temperature: lo0,

I 0 100 0 - mobile phase: a mixture o f 1 volume ofn~ethanol and 99

20 100 - 0 volumes of 0.2 per cent yIv solution of foi-mic acid,

40 75 25

. .

- adjusted to pH 7.5 with triethyIatnitre,

45 75 25 - flow rate: 0.5 rnl per minute,

--- ^... .

47 100 0 - spectrophotomcfr set at 276 nm,

ldcs!i~etliyl cyiidinz-5'-dipliosphocholine sodiu

Q

(3)

. .

Citicdine f~jection

1il.jeci rei'crence S * O ~ Z ~ I D ~ I (aj and the test soiuiion. in thc chrorna:ogram obtained wit\\ the tes: sdlution, h e area of Citicoiine Sodium iiji?iectior! ^{I . .}- peak due io Y-c!.iidyIic acid ~nuitiplyin~ by correction factor Citicoline Injection isa sterile soiution of Ciiico!ine Sodium in of 0.7 is not nore than- 1 ^{. j}tines ifie area of the principal peak Water for Injectians. ^iilthe chromatogam obtained with reference solution (aj (1.5 per centj. The area of any other secondary peak is not more Citicoline Injection toniains not less than 90.0 per cent and than 0. the area ofthe plincipal peak in thechromatomm not more than i 10.0 per cent of the stated alnountof ciLk0lhel obtainf.j i,,:ith rcferenc. (a) (0.5 per ;en[) and the C14H~s14~\1401 1Pl

. r

usual strength 259 ins per ^ill!.

ldentifica tion

. . ~

ot the arzas or' ail il~e secondaq peaks olhei ~lrat! (lie f '- iytidyji; ~ c i s peak is noi more [hall twice the ace; of iht

principai peak in the chroni?iogram obtained with reference solution (2) (2.0 per cent).

In the Assay, the principal peak in the chrdmatogram obtailled Other tests. Coinply with the tests stated under Parenteral with the test solution corresponds to the principal peak In tile Preparations (Iqections).

chromatogram obtained ivith the reference solution. Bacterial endotosins (2.2.3 j. Not Inore than 0.175 Endotoxill Unit per rng of Citicoline.

Tests

Assay. Detenine by liquid ckronlatogiaphy (2.4.14). - . .

~H~(2.4.21). 6.3 to 8.0.

:LOTE--L)etela7ine ~vnter- content of'citicolir7e sodiuin RS Related Substances, Deternine by liquid chr-otnarogaphy before l,xe and calcltlale [he pntenc-,

+2.4.14). ^.. ^-

NOTE-Deter111me water cot?[ent ofciticoline so~fiwn RS before zne and calculate rlze i~orenc?:

Test solution. Dilutea volume of injection containing 150 mg of citicolinc to 50.0 ml with the mobile phase. Dilute 5.0 ml of this solution to 50.0 ml with the mobile phase and filter.

Refpazce sollrlion (a). Dissolve a quantity,of citicoline sodium RSin the mobile phase to obtain a solution containing 0.0005 percent wlv of citicoline.

Reference solution (b). Asolution containing 0.00025 per cent W/V each of citicoline sodium RS and.5 '-c~tidvlic acid RS in the mobile phase.

Tatsolrrtion. Mix the content of 10 containers. Dilute a volume ofthe injection containing 250 mg ofciticoline with the mobile phase and dilute to 50.0 ml with the mobile phase. Dilute 1.0 ml ofthis solution to 100.0 mi with the mobile phase and filter.

Reference solu~iot~. Dissolve a qua11 ti ty of cificolirze sodirrnl RS in the lnobile phase to obtain a solution containi~~g 0.005 per cent w!v of citicoline.

Use chromatographic system as described under Related substances.

Equ~librate the column with mobile phase for at least 90 minutes.

inject the reference solution. The test is not valid unless the tailing factor is not more than 2.0 and the relative standard deviation for replicate injections is not more than 2.0 per cent.

---a stainless steel column 25 crn x 4.6 mln. packed with

Inject the reference solutio~l and ^thetest solution.

octadecylsilane bonded to porous silica ( 5 pm). ^.

- mobileohase:amixture of9 j wlumes of buffer jo[ution Calculate the contellt of Ci.IHJW.4IP: in the injection.

prepared by dissolving 1.697 g of tetrabu~lammoniuin Storage. Store protected from light, ar a temperature not hydrogen sulphafe in 1000 ml of 11-ater. add I! ml of ^'exceeding2j0.

frkfh~famine and adjusted pH 6.0 with difufeacefic ~ ~TIle label states b ((I ~ )e ~ illterms of the i ~ ~ acid a d ⁵volumes of nzethanol, equivalent amount ofciticolix~e; (2) the preparation is intended

-. - - flow rate: 0.8 ml per minute, for intramuscular and intravenous injection only.

-

spectrophotometer set at 770 nm,

t. t ' S b LLcb.,-.'by-v. .,

--.'=~T*~-TY- . ~ ~ ~ ~ . ~ ~ x : r ; . . ; : r ~ ~ ~ ~ ~ ~ ~ e d - ~ p ! ~ ~ . ~ , ~ .

.

^'.' TsE/ets are manufictured

@!octa,-er.r, ,.i.,hiixr coa~phying ^withthe ^.

@the m n ~ g r ~ i p i z ^cii-enot intercI~angeable,

.

$yE&\\.c: ^.%..Je:.

solution (aj and the resolution b z ~ c e n the obtained i ~ i t h reference solution (b).