Vanessa Abercrombie1, Gustavo Serrano2, Phil

Analysis of Ethanol and Isopropanol in Alcohol- Stremple2

based hand sanitizers by direct injection GC/FID

191Blue Ravine Rd, Folsom CA 95630, USA, Agilent
Technologies
22850
Centerville Rd, Wilmington, DE 19808, USA
Agilent Technologies

Introduction Results

The most commonly used alcohol-based hand
sanitizers are prepared with isopropyl alcohol (IPA), or
a form of denatured ethanol. The alcohol in both,
ethanol and IPA-based hand sanitizers, is the major
component of the hand sanitizer mixture, and its
concentration in the aqueous solution determines its
effectiveness as disinfecting agents. Other ingredients
in hand sanitizers may include hydrogen peroxide and
glycerin, but most regulatory methods only require
The GC columns used in this study are among the most popular columns used for analysis of alcohols. Both,
the Agilent J&W DB-624 and DB-WAX Ultra Inert, meet column requirements for USP <611>1,2 and ASTM
D3695 methods. In the first set of chromatograms, we see the composition of an IPA-based hand sanitizer
prepared following new guidelines set up by the USFDA. For the first chromatogram, we used a DB-624 UI
connected to a 1-m retention gap. This setup can help with neat injections of aqueous solution by focusing
the water peak prior to reaching the analytical column. Both columns shows good separation and adequate
symmetry for IPA at high concentrations. The complimentary selectivity between both phases is also
pA
noticeable, and the column choice will depend on user preference or the method of choice.
IPA Calibration on J&W DB-624 UI

DB-624 Ultra Inert

50 100%

determination of alcohol content by GC/FID to qualify 45

90%
R² = 0.9974

the efficiency of alcohol-based hand sanitizers. 40

IPA Glycerin
80%

70%

Calculated Conc
60%

Methods for alcohol determination are widely known,

35

50%

and include USP <611> and ASTM D3695. These and 30

40%

other methods generally require a polar or mid-polar 25 30%

GC columns, such as WAX or 624 phases for the better 20

lower glycols impurities
20%

selectivity and response for alcohol compounds. These 15

10%

traditional columns however, have had shortcomings,

0%
20% 30% 40% 50% 60% 70% 80% 90% 100%
10 Expected Conc

particularly with active components in the 0 2 4 6 8 10 12 14 min

alcohol/water matrix. As a consequence, the analysis pA

IPA Calibration on J&W DB-Wax UI

of alcohols suffer from a lack of column inertness,

100%

DB-WAX Ultra Inert

50 R² = 0.9991
90%

which results in tailing peaks and variable, inaccurate 45

IPA 80%

results, in particular, in aqueous samples. 40 Glycerin 70%

Calculated Concentration
35 60%

Here, we will evaluate the use of Agilent J&W Ultra Inert

50%
30

40%

GC columns, DB-624 UI and DB-WAX UI for the analysis 25

30%
lower glycols impurities
of alcohol-based hand sanitizers. These phases offers 20
20%

excellent stability for samples containing high level of 15

10%

water-alcohol mixtures. The unique inertness of this 10 0%

20% 30% 40% 50% 60% 70% 80% 90% 100%

phase delivers sharper, and more symmetric peak

0 2 4 6 8 10 12 14 min Expected Concentration

shapes for alcohols, ensuring better retention time Figure 1. GC/FID chromatograms of an IPA-based hand sanitizer using a DB-624 UI, 30 m x 0.32 mm id x 1.8 µm df (top) and DB-WAX UI, 30 m x 0.25 mm id x
0.25 µm df (bottom). Calibration curves were prepared for IPA in a 30 to 80% v/v range. R2 were 0.997 and 0.999 for DB-624 UI and DB-WAX UI, respectively.
reproducibility for more consistent and accurate
quantifiable results. The chromatograms in Figure 2 are Ethanol-based hand sanitizers obtained using the same set of columns.
Distilleries have begun to make Ethanol based hand sanitizers3. These products, in addition to Ethanol,
contain fusel oils that are by products occurring naturally during the fermentation process. Fusel oils will
often be present and may present themselves as unpleasant odors in the hand sanitizer. The presence of
Experimental these alcohols are apparent with both, the DB-624 UI and DB-WAX UI chromatograms. DB-624 UI has the
pA
selectivity to fully resolve fusel oils if their analysis is required.
Instrumentation 50 EtOH Calibration on J&W DB 624 UI

• The analyses were performed using an Agilent 7890 DB-624 Ultra Inert
100%

Glycerin
EtOH R² = 0.9955

GC equipped with a flame ionization detector (FID).

45
90%

• Sample introduction was done using an Agilent 40

Calculated Concentration

80%

7693A automatic liquid sampler with 5 µL syringe.35 Fusel oils

70%

GC Conditions 30
60%

25
Method 1 50%

20
40%

Column Agilent J&W DB-624 Ultra Inert, 30 m x 15 40% 50% 60% 70%
Expected Concentration
80% 90% 100%

0.32 mm x 1.8 µm (p/n 123-1334UI) 10

Deactivated fused silica, 1 m x 0.32 mm 0 2 4 6 8 10 12 14 min

(p/n 160-2325-1) pA

Carrier Helium, constant flow, 6 mL/min (73 cm/s) 50

EtOH Calibration on J&W DB- Wax UI

DB-WAX Ultra Inert

100%

Oven 40 °C (5 min), 20 °C/min to 225 °C (2.5 min).

45
EtOH Glycerin
R² = 0.9945

Runtime: 16.75 min

90%
40
Calculated Concentration

35 80%

Inlet Split mode, 140 °C, split ratio 40:1 30

Fusel oils 70%

Inlet liner Ultra Inert, flow pressure drop with glass

25 60%

wool (p/n 5190-2295) 20

50%

Injection 1 µL 15
40%
40% 50% 60% 70% 80% 90% 100%
Expected Concentration
10

0 2 4 6 8 10 12 14 min

Method 2 Figure 2. GC/FID chromatograms of an Ethanol hand sanitizer obtained from a distillery. The chromatograms were obtained using a DB-624 UI, 30 m x 0.32 mm id
x 1.8 µm df (top) and DB-WAX UI, 30 m x 0.25 mm id x 0.25 µm df (bottom). Calibration curves were prepared for EtOH in a 50 to 90% v/v range. R2 were 0.995 and
0.994 for DB-624 UI and DB-WAX UI, respectively.
Column Agilent J&W DB-WAX Ultra Inert,
30 m x 0.25 mm x 0.25 µm (p/n 122-7032UI)
Conclusions References
Carrier Helium, constant flow, 6 mL/min (80 cm/s)
This study shows the positive impact the Agilent J&W 1.Isopropyl Alcohol, Monograph US
Oven 40 °C (5 min), 20 °C/min to 225 °C (2.5 min). DB-WAX UI and DB-624 UI columns have on the Pharmacopeia, 2020
Runtime: 16.75 min quantification of alcohols in alcohol-based hand 2.USP <611> Alcohol Determination,
sanitizers. Retention times and peak shapes for the Monograph US Pharmacopeia, 2020
Inlet Split mode, 140 °C, split ratio 40:1 alcohols were consistent over the course of several
direct sample injections, even in the presence of water 3.Temporary policy for preparation of certain
Inlet liner Ultra Inert, flow pressure drop with glass being injected as a solvent. Calibration curves up to 90% alcohol-based hand sanitizers products
wool (p/n 5190-2295) of the alcohol compound with an R2 > 0.99 were during the public health emergency, FDA
Injection 1 µL prepared. If analysis of fused oils is required, DB-624 UI
gives a better resolution for these compounds, however, © Agilent Technologies, Inc 2020
both columns, show superior inertness and stability for Printed in USA, April 15, 2020
the analysis of alcohol-based hand sanitizers. 5994-1923EN
DE.4822453704