percentage of the lipase activity, are 2. Henry, R. J., Sobel, C., and Berkman, S.

and Berkman, S., In view of its economy and apparent

apparently attributable to the pseudo- On the determination of “pancreatitis lipase” unpopularity among other workers it
lipase removal from sera which contain in serum. Clin Chem. 3,77 (1957). seems worthwhile to report here our
both pancreatitis lipase and pseudoli- 3. Verduin, P. A., Punt, J. M. H. M., and experiences.
pase. An example of this is included in Kreutzer, H. H., Studies on the determina- We evaluated the feasibility of using
Table 1. tion of lipase activity. Clin. Chim. Acta 46,11 alcohol USP in this technique by corn-
(1973). paring its ultraviolet absorption profile
On the other hand, the cause for the
occasional slight increases in the assay 4. Kreutzer, H. H., Pennings, A. W., Punt, J. with that of acetonitrile and methanol,
value due to MCE treatment is unclear. M. H. M., and Verduin, P. A., Further studies both of high-performance liquid-chro-
on the determination of lipase activity. Clin.
Random errors were eliminated as a matographic grade. Their ultraviolet
Chim. Acta 60, 273 (1975).
possible cause by the experimental de- absorption (vs. distilled water) was
5. Steinberg, A., Cholesterol-desoxycholic
sign. Another possible cause is reacti- studied in a spectrophotometer with a
acid: A stable antigen for use in a flocculation
vation of reversibly inactivated lipase. test for liver dysfunction. I. Comparison with recorder and digital readout meter
A third possibility is modification of ii- the Hanger cephalin-cholesterol test. J. Lab. (UV-Vis Spectrophotometer, Model
poproteins by MCE so that clearing of Clin. Med. 34, 1049 (1949). 200; Perkin-Elmer Corp., Norwalk,
the substrate is facilitated during the 6. Manual of Laboratory Operations. Lipid Conn.). Their absorbances at six se-
reaction. These increases are slight and Research Clinics Program. Lipid and Lipo- lected wavelengths are summarized in
do not appear to interfere with the di- protein Analysis, DHEW Publication No. Table 1. The acetonitrile showed the
agnostic usefulness of the procedure. (NIH) 75-628, May 1974. best ultraviolet light transparency, but
The odor of MCE is one of its disad- 7. Ononogbu, I. C., and Lewis, B., Lipopro. the alcohol USP appears to be equally as
vantages. Consequently, we tried three tein fractionation by a precipitation method. good as methanol from one source and
other sulfhydryl compounds under A simple quantitative procedure. Clin Chim. to excel that from the other. The greater
similar
conditions: preincubations for 6 Acta 71,397 (1976). absorption of ethanol as compared to
mm at 25 #{176}C
at an inhibitor concentra- acetonitrile did not appear to affect the
J. Lon Pope
tion of 0.1 mol/liter. Reduced gluts- performance of our system. For exam-
thione partly inhibited (about 50-70% Chemistry Laboratory ple, the baseline from a mobile phase of
inhibition) both pancreatitis lipase Saint Joseph Hospital alcohol USP/acidified water (pH = 2.5),
and pseudo-lipase activities. Di- 1835 Franklin St. 20/80 by vol, and reversed-phase column
thiothreitol caused essentially complete Denver, Cob. 80218 (IL-Bondapak C; Waters Associates, Inc.,
inhibition of pseudolipase activity, but Milford, Mass.) is stable, with little
also significantly inhibited pancreatitis noise. We have used this system suc-
lipase. Cysteine caused complete inhi- cessfully to assay salicylates, theophyl-
bition of both pancreatitis lipase and line, and ampicillin in plasma by moni-
pseudolipase activity in all specimens Ethanol As a Mobile-Phase Solvent toring their ultraviolet absorption at
tested. Hence, of those tried, MCE was in High-Performance Liquid- 237, 275, and 212 nm, respectively. For
the best reagent for differentiating Chromatographic Assays the mobile phase containing water, there
pancreatitis lipase and pseudo-lipase. is no need to use absolute ethanol (re-
Table 1 gives representative data il- To the Editor: agent or USP grade) even though it is
lustrating the effectiveness of this MCE also reasonably ultraviolet transpar-
preincubation in removing pseudolipase To date, acetonitrile and methanol are
ent.
activity. The MCE treatment indicated probably the most popular water-mis- In view of the purity and suitability of
above was followed: A 0.2 mol/liter MCE cible organic solvents used in mobile
ethanol to be used as a water-miscible
solution was prepared just before use by phases of high-performance liquid
component in mobile phases and its
dissolving 70 d of MCE in 5 ml of 8.5 chromatography, special grades of them relative cheapness, we recommend that
g/liter NaC1. Equal volumes of serum available from various manufacturers 95% or absolute ethanol should be first
and 0.2 mol/liter MCE were mixed in a being ordinarily used. They are usually tried as a possible solvent in developing
small tube and allowed to stand at room glass-distilled and of very high purity, high-performance liquid-chromato-
temperature for 6 mm. Lipase was then and so they are expensive. In our labo-
graphic assays.
measured in 0.05-mi portions of this ratory the much less expensive alcohol Win L. Chiou
twofold serum dilution according to the USP (95% ethanol) has been extensively
used in the last year in lieu of acetoni- Geoffrey W. Peng
directions given by the Perkin-Elmer Roger L. Nation
Corp. for serum analysis with the Amy- trile or methanol for assays of many
lase-Lipase Analyzer. If necessary, as drugs in plasma, such as theophylline, Clinical Pharmacokinetic Laboratory
with high pancreatitis lipase activity, we chloramphenicol, salicylates, acetami- Department of Pharmacy
further diluted the serum in 0.2 mol/liter nophen, ampicillin, and sulfisoxazole. University of Illinois Medical Center
MCE in order to obtain meter readings The results have been very satisfactory. Chicago, Ill. 60612
between 1 and 3 kU/liter on the instru-
ment. Amylase was measured with the Table 1. Absorbances (A) of Acetonltrile, Methanol, and Ethanol
Amylase-Lipase Analyzer according to Compared
the directions given by the Perkin- A at 350 nm 325 nm 300 nm 254 nm 225 nm 210 nm
Elmer Corp. Amylase and the Tietz- Acetonitrile8 0 -0.002 -0.002 -0.00 1 0.006 0.039
Fiereck (1) lipase assay values are in-
cluded as evidence for or against pan-
Methanol5 0 0 0.002 0.020 0.190 0.550
creatic involvement. The normal ranges Methanol b 0 0 0.005 0.025 0.304 0.706
applying here are: nephelometric lipase, Ethanol, 95% 0 0 0.005 0.020 0.199 0.554
0-1.5 kU/liter; Tietz lipase, 0-1.0 kU/ Ethanol, absolute” 0 0 0.011 0.035 0.322 0.636
liter; and nephelometric amylase, 5GlaIstlIIed; Btwdick and Jackson Laboratory, Muskegon, Mich.
150-2000 U/liter. HPLC-grade; FIsher Scientific Co., Fair Lawn, N. J.
References C Alcohol, USP; Medical Center General Stores, UniversIty of Illinois at the Medical Center, Chicago, ill.

1. Tietz, N. W., and Fiereck, E. A., A specific 5Alcohol, dehy&ated; USP, Medical Center General Stores, University of Illinois at the Medical Center.
method for serum lipase determination. Clin. Chicago, III.
Chim. Acta 13,352 (1966).

CLINICAL CHEMISTRY, Vol. 23, No. 12, 1977 2355