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Fresenius J Anal Chem (1994) 349:451-453

Fresenius' Journal of

Springer-Verlag 1994

Determination of glycols by HPLC with refractive index detection


L. Nitschke, L. Huber
I Bayerische Landesanstalt ffir Wasserforschung, Kaulbachstrasse 37, D-80539 Mfinchen, Germany

Received: 6 September 1993/Accepted: 1 December 1993

Abstract. A simple and fast HPLC method for the deter- tivated sludge plant was additionally of special interest
mination of glycols is described. It is characterized by a for the control of more common chemical parameters
reversed-phase separation using water as eluent and a such as COD, TOC, ammonia etc. A simple and rapid
refractive index detection. The method was applied to in- HPLC method has been developed for this purpose, re-
vestigate the biodegradation of glycols in a laboratory ac- quiring no special sample pretreatment. The method is
tivated sludge plant and to determine the content of gly- characterized by using a reversed-phase column, water as
cols or alcohols in detergents. The detection limit is eluent, and refractive index detection.
4 rag/1 ethylene glycol or propylene glycol in an aqueous
sample.
Experimental

Chemicals
Introduction All glycols were obtained from Merck, Darmstadt. Sol-
vents for HPLC (water, methanol) were of LiChrosolv
The determination of glycols, especially diethylene glycol
quality and obtained from Merck, too.
(DEG), has become very popular in the eighties when
DEG had been illegally added to wine. In most cases, gas
HPLC procedure
chromatography was used to determine its content in
wine [ 1 - 5 ] . Also thin-layer chromatography [6], 13C- The HPLC equipment consisted of the data system (Kon-
NMR [7] and HPLC (with derivatization to make an UV tron 450) and two pumps (Kontron 420). A refractive in-
or fluorescence detection possible [8]) were applied. Nor- dex detector (Erma ERC-7512) was used for the measure-
mally, glycols are mainly used as intermediate products in ments. The filtered samples were injected by a valve fitted
the chemical industry, as solvents, and antifreezes. Large with a 100 gl loop. A Hypersil ODS column (particle size
amounts of antifreezes containing diethylene glycol and 5 ~m, 250 4.6 ram) was used for the reversed-phase sep-
propylene glycol (PG) are used at airports during the win- aration of the glycols. The mobile phase was water and
ter period for de-icing airplanes and runways. A large the flow rate 2 ml/min. The separation was carried out at
part of the formed mixture of water and glycols is recy- room temperature. The temperature control of the refrac-
cled by distillation at modern airports, especially aque- tive index detector was set to 30C.
ous solutions with a high content of glycols. The rest of
the mixture is treated by the activated sludge process in
waste water treatment plants. Although these glycols are Results and discussion
easily biodegradable by adapted activated sludge, there
are sometimes problems caused by low waste water tem- Reversed-phase chromatograms of an influent and an ef-
peratures in winter and by the load, especially shock- fluent of a laboratory activated sludge plant are shown in
loads. Therefore laboratory activated sludge units were Fig. 1. The influent is an aqueous solution containing a
tested by us to find out the optimal conditions for the prescribed content of nutrients (peptone, beef extract)
biodegradation of glycols, or its restrictions. The deter- and mineral salts (for details see DIN 38412, Teil 26). A
mination of diethylene glycol and propylene glycol in the certain amount of the antifreeze about 88% DEG and
influent and especially in the effluent of a laboratory ac- 270 PG has been added to the solution. The added
amount corresponds to 400 mg/1 DEG and 9 rag/1 PG,
respectively. As can be seen from Fig. 1, PG and DEG can
Correspondence to." L. Nitschke be detected in the influent and effluent of a laboratory
452

(D
O)
03 C
C DEG 0
0 C1
g1 (.0

[_

--A

B
] I = I ]
0 3 6 5 iO
time [mini time [min]
Fig. 1. Chromatograms of (A) influent (400 mg/1 diethylene glycol and Fig. 3. Chromatogram of a mixture of (/) 100mg/1 glycerol, (2)
9 mg/1 propylene glycol are added to the nutrient solution), (B) effluent 150 mg/1 propylene glycol, (3) 200 rag/1 diethylene glycol, (4) 400 mg/1
of a laboratory activated sludge plant. H P L C conditions: column: propan-2-ol, (5) 250 mg/1 triethylene glycol in water. For HPLC condi-
Hypersil ODS (2504.6 mm I.D., particle size 5 gm); injection: 100 gl; tions see Fig. 1
eluent: water; flow rate: 2 ml/min; detection: refractive index

biodegradation) from the stationary phase. The frequen-


activated sludge plant with the described HPLC method. cy of the cleaning procedure depends on the composition
There are no interferences from other substances. The of the injected samples. In our case it is necessary to rinse
broad peak at the beginning of the chromatogram has the column with methanol for I h (flow rate 2 ml/min)
been ascribed to the content of inorganic ions in the after 10 to 15 chromatograms have been recorded.
aqueous solutions. There ions very weakly interact with The calibration plot of propylene glycol is shown in
the column material, i.e. they are transported more or less Fig. 2. The correlation coefficient and the 3a detection
with the solvent front. On the other hand, organic sub- limit were calculated to 0.9996 and 4 mg/1, respectively. A
stances added as nutrients are adsorbed on the C~8 col- similar calibration was obtained from DEG. The repro-
umn material. By using water as eluent these substances ducibility was determined with an aqueous solution con-
have a long retention time and pass the column very slow- taining 100mg/1 PG. A relative standard deviation of
ly. For this reason, glycols can be detected in the influent 0.6% was obtained. The concentration of DEG was de-
and effluent of laboratory activated sludge plants with- termined to be 7.3 mg/1 in the effluent as shown in Fig. 1.
out interferences from other substances. After recording This means that the biodegradation of DEG in the labo-
several chromatograms, the column has to be rinsed with ratory activated sludge plant amounts to over 98%. The
methanol or another appropriate solvent to elute the ad-
sorbed organic substances (widely unknown products of

(D PG
0
peak area [mV*min] C
6 0
CI
(,9

C.

propan-2-ol

[ I [
O 3 6 9
0 50 100 150 200 250 300 350 tme [mini
concentration [mg/I]
Fig. 4. Chromatogram of a detergent containing 570 propylene glycol
Fig. 2. Calibration plot of propylene glycol in water and propan-2-ol each. For HPLC conditions see Fig. 1
453

concentration of PG in the same sample is below the de- mine glycols and alcohols with a limited length of the
tection limit. It should be pointed out that the H P L C alkyl chain in the presence of other organic substances
method described here is also applicable to the determi- such as nutrients of a laboratory activated sludge plant or
nation of other glycols or alcohols without interferences surfactants.
by other organic substances. An important requirement,
however, is a limited length of the alkyl chain of the gly-
cols and alcohols (maximum 4 up to 6 - C H 2 - groups),
depending also on the number of polar groups in the
molecules. A chromatogram of a sample containing 4 dif- References
ferent glycols and one alcohol is shown in Fig. 3. The
peaks are well resolved. 1. Holzer H (1985) Ern/~hrung 9:568-569
2. Kaiser RE, Rieder RI (1985) J High Res Chromatogr 8:863-865
Based on the German detergents law, our institute and 3. Werkhoff P, Bretschneider W (1986) Z Lebensm Unters Forsch
certain other authorities have to check the declared for- 182:298 - 302
mulations of detergents. The above H P L C method is sat- 4. Lehmann G, Ganz J (1985) Z Lebensm Unters Forsch 181:362
isfactorily applicable to determine glycols and alcohols in 5. Caccamo F, Di Corcia A, Samperi R (1986) J Chromatogr 354:
the presence of surfactants. A chromatogram of a deter- 478-481
gent containing propylene glycol and propan-2-ol is 6. Klaus R, Fischer W (1987) Chromatographia 23:137-140
7. Rapp S, Spraul M, Humpfer E (1986) Z Lebensm Unters Forsch
shown in Fig. 4. 182:19
In summary, it may be emphasized that the described 8. Bayliss MA, Homer RB, Shepard MJ (1988) J Chromatogr
H P L C method is a fast and simple procedure to deter- 445:403- 408