Development of FUJI DRI-CHEM v-LIP-P Slide that Has the High

Specificity to Pancreatic Lipase

Kentaro NAKAMURA*, Shigeki KAGEYAMA*, Hideaki TANAKA**,

Kazuya KAWASAKI*, and Kaoru TERASHIMA*

Abstract
The lipase of blood is mainly a deviation enzyme from pancreas, and its activity is a highly evaluated
index for pancreatitis diagnosis in human medical care. However, the sensitivity and specificity for the
pancreatitis of the blood lipase activity is low in the veterinarian medical care for dogs and cats , and making
rapid diagnosis has been difficult to conduct. The Pancreatic Lipase Immunoreactivity (PLI), a blood
diagnosis method, is the most superior in sensitivity and specificity as a pancreatitis marker for the dogs
and cats at present. However, because PLI is not easy and rapid, there is a strong need for the development
of an easy and rapid method of pancreatitis screening. FUJI DRI-CHEM (FDC), a dry chemistry method,
is a system that enables measurement of various enzymic activities in the blood easily and rapidly, which
has widely been used in veterinary care. Now we have developed the FDC v-LIP-P Slide having high
specificity to measurable pancreas lipase. The measurement principle we used is the triglyceride method.
The specificity for the pancreas lipase has also been improved by prescribing colipase, bile salt and sodium
dodecyl benzene sulphonate, which is an anionic detergent. In addition, the reactivity has been improved
by prescribing triolein, an emulsified substrate, to secure the area of the oil-water interface for the reaction
to the pancreatic lipase. It was confirmed that the correlation of FDC v-LIP-P Slide and PLI was excellent.
This shows that this slide is a very effective diagnostic product for the rapid screening of pancreatitis.

1. Introduction activities of multiple different types of lipases would be

evaluated because there are many lipase isozymes other than
While blood lipase as well as amylase and trypsin
the pancreatic lipase including gastric lipase, hepatic lipase
are deviation enzymes from pancreas, blood lipase and
or lipoprotein lipase in blood of these animals.
amylase activities increase significantly in patients with
Hence, Steiner et al. of A&M University of Texas
acute pancreatitis. It is known that lipase activity is more
developed the pancreatic lipase immunoreactivity (PLI)
specific to pancreas and ref lects pancreatic functions
diagnosis method by using an antibody which binds
more specifically than amylase activity does, suggesting
specifically to the pancreatic lipase only and reported that
that increased lipase activity is a highly evaluated index
for diagnosing pancreatitis1). While the mainstream of the method is effective for diagnosing pancreatitis. At
reagents for determining the blood lipase activity is the present, the PLI method is considered to be the most superior
coupled enzyme method, wet chemistry methods include pancreatitis marker in sensitivity and specificity for dogs and
the 1,2-diglyceride substrate method developed by Asahi cats2), 3).
Kasei Corporation and dry chemistry methods include the Nevertheless, the method as only one blood diagnosis
diacetinase method developed by Eastman Kodak Company. method at present for pancreatitis in dogs and cats is not
On the other hand, u nlike hu mans, it has been an easy and rapid procedure with taking a few days to get
demonstrated that the sensitivity and specif icity for measurement results. Because such disadvantages of the
pancreatitis of the blood lipase activity determined by method do not lead to rapid diagnosis, there is a strong
antecedent reagents are low in the veterinary medical care need for developing an easy and rapid method for screening
including diagnosis of pancreatitis in dogs and cats, and it has pancreatitis.
been known that rapid diagnosis of pancreatitis is difficult in Reportedly, the DGGR substrate method, a reagent for
these animals. As the reasons, it is considered that totaled determining the blood lipase activity (a wet chemistry

Original paper (Received December 2, 2010) ** Imaging Materials Production Division

* Pharmaceutical & Healthcare Research Laboratories Kanagawa Factory
Research & Development Management Headquaters FUJIFILM Corporation
FUJIFILM Corporation Nakanuma, Minamiashigara, Kanagawa 250-0193, Japan
Ushijima, Kaisei-machi, Ashigarakami-gun, Kanagawa
258-8577, Japan
FUJIFILM RESEARCH & DEVELOPMENT (No.56-2011) 5
method) which was developed by Roche Diagnostics and 2.1.2 Assessment of Feasibilities of the DGGR
has becoming common recently, has higher specificity for Substrate Method and the Triglyceride Method
detection of canine pancreatic lipase activity and is more as Dry Chemistry Methods
useful as a method to determine the enzyme activity as The DGGR subst rate assay is a method to use
compared with the 1,2-diglyceride substrate method which 1, 2 - O - d i l a u r y l - r a c - g l y c e r o -3 - g l u t a r i c a c i d - (6’-
had ever been the mainstream of those diagnosis methods4). methylresorufin)-ester as a substrate. The substrate is
It indicates presumably that specific measurement of the decomposed by lipase under the alkaline conditions to form
pancreatic lipase activity can be achieved by using different 1,2-O-dilauryl-rac-glycerol and an unstable intermediate,
substrates, different aids and different conditions of the glutaric acid-(6’-methylresorufin)-ester. Under the alkaline
reaction. conditions, the latter is decomposed spontaneously to form
FUJI DRI-CHM (hereinafter, referred to as FDC) as a glutaric acid and methylresorufin with an absorption peak
dry chemistry method is an easy and rapid assay system to at around 580 nm (Fig. 1). The DGGR substrate assay is a
determine a variety of enzyme activities in blood and has method to measure the rate of increase in absorbance of red
been extensively used now in the veterinary medical care. color of methylresorufin5). Namely, the substrate used for
Hence, we conducted a study to develop an assay method the DGGR substrate method is very unstable, and hence, its
with high specificity for the pancreatic lipase activity in temporal stability was concerned. A substrate DGGR could be
order to address the needs for diagnosing pancreatitis. The dispersed in liquids at slightly acidic pH and was stable for a
present report refers to the results of the study. year in the refrigerator. On the other hand, it was demonstrated
that the substrate was easily decomposed at neutral pH and
2. Points of Development its decomposition rate was accelerated under the slightly
alkaline conditions (pH=8; the optimum pH for lipase) or in the
2.1 Determination of Measurement Principle to
presence of sodium deoxycholate, an activator of lipase. By
be Adopted for Preparation of Slides
NMR and MASS analyzes of decomposition mechanisms of
2.1.1 Selection of Measurement Principle the substrate, it was identified that the ester bond (indicated by
While there are a variety of methods for determining an arrow in Fig. 1) adjacent to the chromophore preferentially
lipase activity, those listed in Table 1 below provide the among 2 ester bonds was preferentially decomposed.
representative colorimetric methods which use visible O O CH3

lights applicable to the FDC analyzer. From various O (CH2)3 O O O

O R
studies conducted hitherto, it has been reported that O R N

the 1,2-diglyceride substrate method 5), 6) as well as the 1,2-O-dilauryl-rac-glycero-3-glutaric acid-(6’-methylresorufin)-ester

＋ OH－
diacetinase method7) have low specificities for the pancreatic Pancreatic lipase
O H O O CH3
lipase. By contrast, the DGGR substrate method has been - (CH2)3
+
O R O O O O
reported to have high specificity4). Additionally, methods O R
N
to use triglyceride, a natural substrate of the pancreatic
Glutaric acid-(6’-methylresorufin)-ester
lipase, have been reported to have higher specificity for the
pancreatic lipase as compared to that to the lipoprotein lipase Spontaneous decomposition ＋ OH－

or the hepatic lipase8). Accordingly, the DGGR substrate -

CH3
O O O O O
method and the triglyceride method were chosen for selection + -
O (CH2)3 O
N
of measurement principle and evaluation was performed for
Methylresorufin Red color，λ=580nm Glutaric acid
preparation of dry chemistry methods.
Fig. 1 Reaction scheme of the DGGR method.

Table 1 The list of the lipase measurement principles.

Maker developed
Name of method Assay method Specificity for pancreatitis Maker adopted the method
the method
Positive reactions may occur Ortho Clinical Diagnostics
Diacetinase method Coupled enzyme
EK also for diseases other than IDEXX Laboratories (exclusive use for
(a dry chemistry method) method pancreatitis7) . animals)
Low specificity for canine Several domestic companies including
1,2-Diglyceride substrate Coupled enzyme pancreatitis, and increased Wako Pure Chemicals Industries
method Asahi Kasei method lipase activity is observed in No.1 share in pharmaceutical market
(a wet chemistry method) heparin-added plasma. for humans and animals

DGGR substrate method Roche Diagnostics, Kyowa Medex Co.

Roche Diagnostics Resorufin substrate High specificity for human and
(a wet chemistry method) method canine pancreatitis4) . No.2 share in pharmaceutical market
for humans
Triglyceride method Specificity of triglyceride for the
Coupled enzyme
(a wet chemistry method/a Asahi Kasei pancreatic lipase is higher than
method
dry chemistry method) that for hepatic lipase or LPL8) .

6 Development of FUJI DRI-CHEM v-LIP-P Slide that Has the High Specificity to Pancreatic Lipase
 In wet chemistry methods, a substrate unstable against
some pH would be stored before using in solutions under
Substrate emulsion
emalsion
the pH conditions where it is stable. Upon analysis by
an autoanalyzer, it would be mixed with another reaction
reagent at pH 8 which is the optimal pH for lipase to enable
Bile salt
a measurement of lipase activity. By contrast, in case of
the FDC slide, all reagents necessary for measurement of colipase
N-terminal domein
lipase activity have to be packed onto the slide as described C-terminal domein
in section 2.3. Accordingly, it has to contain buffering Pancreatic lipase
agents and alkaline components necessary for preparing “Lid” domein
an environment at pH 8 in addition to reaction reagents. It Fig. 2 Schematic view of the reaction of the pancreas lipase.
would be very difficult to store DGGR as a substrate in the
slide under such conditions.
Since a combined formulation of bile salt and colipase has
In contrast, in the case of the FDC slide prepared by using
been used also for the DGGR substrate method with high
triglyceride as a substrate, it was demonstrated that there
specificity for the pancreatic lipase, formulation of the dry
was no change in its reactivity in a 2- or 3 week accelerated
chemistry method was designed based on such a formulation.
stability test at 45°C even at pH 8, suggesting its extremely
higher stability as compared with DGGR. 2.2.2 Study on Formulations for the Dry Chemistry
Method with High Specificity for the Pancreatic
B ased on the above results, it was determined that
Lipase
development of a dry chemistry method would be conducted
for the triglyceride method. Study on formulations was performed with the use of
triolein as a reaction substrate and furthermore, by using
2.2 Study on Improvement of Reactivity and colipase and bile salt. While sodium deoxycholate was
Specificity of the Pancreatic Llipase used mainly as bile salt, a combined formulation of sodium
2.2.1 Reaction Mechanism of the Pancreatic Lipase5) deoxycholate and water soluble sodium taurodeoxycholate
was used because water solubility of the former was not
One of the enzymatic properties of the pancreatic lipase is
so high. In addition, various reagents were formulated
that the enzyme requires colipase, a protein with a molecular
to achieve the following reaction scheme because the
weight of 11kDa, for its efficient enzymatic activity.
measurement principle of this assay was the coupled enzyme
Colipase is secreted from pancreas as procolipase which is
method (Fig. 3).
then transformed to active colipase by the action of trypsin
in the intestinal lumen for binding to lipase. The pancreatic Lipase, Colipase
lipase is also characterized by another property distinctly Triolein + 2H2O 2-Monoglyceride + 2 Oleic acid

different from those of other esterases, that is, it exerts its MGLP
2-Monoglyceride + H2O Glycerol + Oleic acid
high enzymatic activity only after an interface is formed
between fatty acid esters and the water layer. Accordingly, GK, MgCl2
Glycerol + ATP L-α-glycerophosphate + ADP
the lipase activity would be influenced by the presence of
such substances as bile salt which affect on the emulsion GPO
L-α-glycerophosphate + O2 H2O2 + Dihydroxyacetone phosphate
formation9), 10).
The unique properties of the pancreatic lipase are POD
Diarylimidazole leuco dye + H2O2 Blue color dye + 2H2O
considered attributable to its characteristic structure referred
to as “a lid” domain which covers the active center of this Fig. 3 Reaction scheme of the FDC v-LIP-P Slide.
enzyme. It is considered that, by ordinary, the “lid” is closed
and its enzymatic activity is only modest when it stands In canine samples, was demonstrated a correlation of the
alone. However, its conformation changes in the presence of FDC slide prepared according to the above formulation and
colipase and bile salt and its active center would be exposed the DGGR substrate method.
as shown in Fig. 2. Then, the hydrophobic domain of the While these 2 methods correlated relatively well with
active center binds to substrates stably to exert enzymatic each other [correlation coefficient (r) = 0.930], there were
activity 11), 12). Under the physiological conditions, the separations of some data at the lower lipase concentration
pancreatic lipase is considered to form such a complex with range among those provided by the FDC slide and especially
colipase, bile salt and lipids for digesting fats. those obtained by the DGGR substrate method [Liquitech
Lipase Color II (Roche Diagnostics), hereinafter, referred to

FUJIFILM RESEARCH & DEVELOPMENT (No.56-2011) 7

as the DGGR substrate method] (Fig. 4a). lipase was assessed by developing a method to prepare
Separations of those data observed in the lower lipase emulsions of toriolein in advance. For preparing triolein
concentration range were considered attributable to emulsions, sodium dodecyl benzene sulfonate was used as a
insufficient formation of an interface between fatty acid detergent, while gelatin and polyvinyl pyrrolidone were used
esters and the water layer where lipase exerts its activity. as protective colloids. Emulsification was performed with a
Accordingly, effects of addition of a detergent were assessed high pressure emulsifier/disperser to prepare emulsions with
in order to provide a better oil-water interface as the a mean particle diameter of about 200 nm.
reaction field for lipase. A point to consider for selecting a Reactivities of the FDC slides containing the dissolved
detergent is to find out a detergent that does not affect the type of triolein and the emulsified type of triolein were
reactivity of lipases other than the pancreatic lipase. On assessed for the control sample of the pancreatic lipase
the contrary, anionic detergents including sodium dodecyl with high activity (800 U/L) and that with low activity (40
sulfate are known to inhibit the activity of lipases other than U/L) to compare the reaction rates provided by using these
the pancreatic lipase 13), 14), 15). In results of assessments different states of substrate. In result, it was demonstrated
conducted based on these reports, sodium dodecyl benzene that reactivity for the pancreatic lipase of the FDC slide
sulfonate among anionic detergents was found to elevate the containing the emulsified type of substrate is approximate
activity of the pancreatic lipase without changing the activity 30% higher than that containing the dissolved type of
of lipases other than the pancreatic lipase. In the assessment substrate (Table 2).
of a correlation between above 2 methods performed with the Based on the above results, the method was selected to
same canine sample as that used in the aforementioned test, apply and dry the emulsions of toriolein prepared in advance
improvement was observed in the lower lipase concentration for preparing the FDC slide (a dry chemistry method).
range with very good correlation coefficient [(r)=0.977] being
Table 2 Reactivity comparison by difference of state of substrate
observed (Fig. 4b). (triolein) in FDC v-LIP-P Slide.
ａ. Colipase×1、SDBS×0　（ｎ=46） ｂ. Colipase×1、SDBS×1　(n=46) Dissolved Emulsified
1800 1800 type type
y = 1.0985x - 6.935
R2 = 0.8658
1500 1500 Difference in reaction rate ( ODr/min) 0.07 0.091
r = 0 .9 30
FDC v-LIP-P (U/L)
FDC v-LIP-P (U/L)

1200 1200 Percent difference in reaction rate

of emulsified type relative to that of 100% 130%
900
900 dissolved type
600 y = 1.3313x - 9.2031
600
R2 = 0.9551

300 300 r = 0.97 7

0
0 300 600 900 1200 1500 1800
0
0 300 600 900 1200 1500 1800
2.3 Structure of the FDC Slide
DGGR substrate method (H7180,U/L) DGGR substrate method (H7180,U/L)
The structure of the FDC v-LIP-P slide is shown in Fig. 5.
Fig. 4 Canine sample correlation of FDC v-LIP-P Slide and
DGGR method (a. without sodium dodecylbenzene
【Spreading layer】
sulphonate (SDBS), b. with SDBS). Triolein emulsion
(Triorein, Sodium Dodecylbenzene Sulphonate,
Gelatine, Polyvinylpyrrolidone)
Colipase
2.2.3 Improvement of Reactivity of the Pancreatic Bile acid
Sodium Dodecylbenzene Sulphonate
Lipase [Emulsification of Substrate (Toriolein)] Monoglyceride lipase
Calcium chloride
Formation of an interface between fatty acid ester and
the water layer is important for expression of lipase activity. 【Reagent layer】
Polymer
However, it was anticipated that the reaction rate of lipase Glycerol kinase, Magnesium chloride
Adenosine triphosphate disodium salt
would be decreased in the case of dry chemistry methods, Glycerol-3-phosphate oxidase
Peroxidase
because it would be difficult to secure the effective area Diarylimidazole leuco dye

of the oil-water interface due to aggregation and union 【Transparent support】

PET film
of emulsified substrate as well as loss of water as a
dispersion medium caused by application and drying of the Fig. 5 Structure of FDC v-LIP-P Slide.
emulsified substrate on the slide, even when a homogeneous
emulsification was achieved in the presence of a detergent The FDC v-LIP-P slide is composed of 2 layers including
and protective colloids. In the previous assessment of the a reagent layer and a spreading layer set on a transparent
prepared slides, toriolein dissolved in such an organic solvent support film made of polyethylene terephthalate. While the
as ethanol was applied and dried on the slide, whereas bile reagent layer consists of coupled enzymes which react with
salt and sodium dodecyl benzene sulfonate, required for glycerin generated in the spreading layer and color forming
formation of the reaction field, were added to the slide dyes, the spreading layer contains toriolein emulsion which
separately. On the contrary, reactivity of the pancreatic is a substrate of the pancreatic lipase, colipase and bile acid

8 Development of FUJI DRI-CHEM v-LIP-P Slide that Has the High Specificity to Pancreatic Lipase
necessary for expression of the pancreatic lipase activity, by using the diacetinase method or the 1,2-diglyceride method
sodium dodecyl benzene sulfonate as a reaction aid and a showing low correlations with the PLI method were higher
coupled enzyme. When a test sample was applied on the FDC in many cases than the data obtained by the PLI method,
v-LIP-P Slide, it spreads uniformly in the spread layer to react especially in the case of test samples with low PLI value.
with the substrate (triolein). 2-Monoglyceride generated by Hence, it was suggested that lipase isozymes other than the
the action of lipase is decomposed by the monoglyceride lipase pancreatic lipase may be measured by these 2 methods.
to form glycerol and fatty acid. Glycerol is converted to L-α- It is presumed that both of the DGGR substrate method
glycerophosphoric acid by the action of the glycerol kinase in and the FDC v-LIP-P slide show high specificity for the
the presence of ATP and Mg2+. L-α-glycerophosphoric acid pancreatic lipase from the results that very good correlations
is oxidized by the action of the glycerol-3-phosphate oxidase were observed among these 2 methods and the PLI method by
to generate hydrogen peroxide. By the action of peroxidase, using canine test samples. Hence, a correlation was assessed
hydrogen peroxide oxidizes diaryl imidazole type of leuco- between the DGGR substrate method and the FDC v-LIP-P
pigment to generate a blue pigment. The rate of formation Slide by using canine and feline test samples (Fig. 7a).
of this blue pigment is determined by measuring the changes By using canine test samples, a very good correlation
in the reflected optical density at 650 nm from 3.0 min to 5.0 was observed between the DGGR substrate method and the
min which, then, would be converted to lipase activity by a FDC v-LIP-P slide with a correlation coefficient (r) = 0.993.
calibration curve built in a dedicated instrument. Although the quantities of test samples with intermediate
and high lipase content were limited in the case of feline
3. Confirmation of the Effectiveness of samples, a good correlation was observed between the
FDC v-LIP-P Slide DGGR substrate method and the FDC v-LIP-P slide with a
correlation coefficient (r) = 0.983 (Fig. 7b).
3.1 Confirmation of Specificity for the
Pancreatic Lipase ａ. FDC v-LIP-P slide (a dry chemistry method) ｂ. Diacetinase method
(a dry chemistry method, IDEXX Laboratories)
(N=40) （N=40）
800 6000
In order to validate the effectiveness of FDC v-LIP-P
5000
slide developed this time, we confirmed a correlation of data
FDC v-LIP-P slide　（U/L)

600

IDEXX LIP slide　（U/L)

4000
generated by the FDC v-LIP-P slide and those generated by the
400 3000
PLI method (Spec cPL: IDEXX Laboratories, Inc., referred to
2000
as the PLI method), which is said to be only one effective blood 200 r=0 .931
1000 r = 0.31 4
diagnosis method of pancreatitis in dogs and cats. In addition,
0 0
correlations of data were confirmed among those generated by 0 200 400 600 800 0 200 400 600 800
PLI method (an immunoreactivity diagnosis method) （μg/L) PLI method (an immunoreactivity diagnosis method) （μg/L)
the FDC v-LIP-P slide, the diacetinase method (VetTest Slide DGGR substrate method 1,2-Diglyceride substrate method
ｃ. (a wet chemistry method, Roche Diagnostics) ｄ. (a wet chemistry method, Wako Pure Chemical Industries)
LIPA: IDEXX Laboratories, Inc., referred to as the diacetinase 800
（N=40)
1400
（N=24)

method) which is a conventional blood lipase assay method 1200

600
as a dry chemistry method, the 1,2-glyceride method (Lipase 1000
Roche LIP（U/L)

Wako LIP（U/L)

800
Color Auto Test Wako kit: Wako Pure Chemical Industries, 400
600
referred to as the 1,2-diglyceride method) and the DGGR 400
200 r = 0.972 ｒ = 0.40 1
substrate method as wet assay methods (Fig. 6). 200

A good correlation was observed between the FDC v-LIP-P 0 0

0 200 400 600 800 0 200 400 600 800
Slide and the PLI method with a correlation coefficient (r) = PLI method (an immunoreactivity diagnosis method) （μg/L) PLI method (an immunoreactivity diagnosis method) （μg/L)

0.931. On the other hand, a low correlation was found between Fig. 6 Canine sample correlation of PLI and various measurement
the diacetinase method, as another dry chemistry method and of the lipase activity (a. FDC v-LIP-P Slide (dry chemistry),
b. Diacetinase method (dry chemistry), c. DGGR method
the PLI method, with a correlation coefficient (r) = 0.314.
(wet chemistry), d.1,2-Diglyceride method (wet chemistry)).
With respect to wet chemistry methods, a good correlation
was observed between the DGGR substrate method and the ａ. Canine test sample, correlation among multi-samples ｂ. Feline test sample, correlation among multi-samples
(n=61) (n=30)
PLI method with a correlation coefficient (r) = 0.972, and it 1200
y = 0.986 x + 3.041
800
2
y = 1.044 x + 5.577
R = 0.986 700
was confirmed that the pancreatic lipase could be measured 1000 2
R = 0.966

r=0.993 600 r=0.983

FDC v-LIP-P slide (U/L)

FDC v-LIP-P slide (U/L)

specifically by this method as reported in articles. On the 800

500

other hand, it was confirmed that the correlation coefficient (r) 600 400

= 0.401 when a correlation between the 1,2-diglyceride method 400

300

200
and the PLI method was assessed. 200
100

Based on these results, it is supposed that the FDC v-LIP-P 0 0

0 200 400 600 800 1000 1200 0 100 200 300 400 500 600 700 800
Slide showing a good correlation with the PLI method is a DGGR substrate method (U/L)　H-7180 DGGR substrate method (U/L)　H-7180

method with high specificity for the pancreatic lipase similarly Fig. 7 Correlation of FDC v-LIP-P Slide and DGGR method
as the DGGR substrate method. By contrast, data provided (a. canine samples, b. feline samples).

FUJIFILM RESEARCH & DEVELOPMENT (No.56-2011) 9

 As for feline test sample, correlations have not been the pancreatic lipase. 1,2-O-dilauryl-rac-glycero-3-glutaric
confirmed between the PLI method and the FDC v-LIP-P acid-(6’-methylresorufin)-ester, a substrate to be used for
Slide or the DGGR substrate method. Moreover, further the DGGR method, is very degradable and unstable at pH 8,
studies would be needed on specificity of these methods i.e. optimum pH for the pancreatic lipase, whereas toriolein,
for the pancreatic lipase, since sufficient quantities of test a substrate of the triglyceride method, is stable at pH 8.0.
samples with intermediate and high lipase content were In the case of the FDC slide, it has to contain all reagents
not subjected to the assessment. It was supposed, however, necessary for measurement of lipase activity. Since triolein,
that isozymes other than the pancreatic lipase would not be a substrate of the triglyceride method, was stable at pH 8, i.e.
measured by these methods because a high correlation was under its storage condition, it was considered applicable as
observed between these 2 methods. a substrate of a dry chemistry method and the triglyceride
method was selected as the measurement principle.
3.2 Confirmation of Simultaneous Reproducibility
To secure specificity for the pancreatic lipase, sodium
In order to assess whether the degree of variation of data
dodecyl benzene sulfonate was prescribed in addition to
provided by repeated assays with the FDC v-LIP-P slide is
colipase and bile salt. By prescribing these materials, a good
in the acceptable range, simultaneous reproducibility assays
correlation could be achieved in canine test samples.
were conducted. In Table 3, are provided the results of
I n addition, improvement of reactivity of the method
assays performed 20 times repeatedly for each of 3 canine
for the pancreatic lipase could be attained by emulsifying
serum samples with different lipase activities. Since a
and dispersing triolein, a substrate, and by prescribing the
coefficient of variation (CV) as a measure of variation was
emulsified substrate on the slide.
lower than 4% at each activity level, it was confirmed that
A very good correlation was observed between the PLI
the reproducibility of assay with the FDC v-LIP-P slide was
method and the FDC v-LIP-P slide, into which the above
in the acceptable range for diagnosis of pancreatitis.
measures were introduced, with the use of canine test
Table 3 R
 eproducibility in measurement with FDC v-LIP-P Slide samples. A very good correlation was observed additionally
for dog serums. between the FDC v-LIP-P slide and the DGGR method,
dog serum 1 dog serum 2 dog serum 3 which showed also a very good correlation with the PLI
1 37 678 412 method. It was also confirmed that the degree of variation
2 36 706 406 of data provided by the FDC method was in the acceptable
3 38 671 392
4 37 707 396
range.
5 37 723 375 The PLI method, only one effective diagnosis method that
6 37 686 382 is said to be available today for pancreatitis in dogs and cats,
7 38 719 386
is not an easy and rapid procedure with taking a few days to
8 35 680 393
9 36 695 377 get measurement results and has a disadvantages that it do
10 37 713 386 not lead to rapid diagnosis. Since the FDC v-LIP-P Slide
11 37 725 394
that was developed this time has high specificity for the
12 35 700 399
13 35 722 399 pancreatic lipase and can be packed onto the easy and rapid
14 34 749 397 FDC system which has been used hitherto in the veterinarian
15 38 730 397 medical care, the slide is expected to be a very effective
16 34 704 400
17 36 708 397
diagnostic product for pancreatitis in the veterinarian medical
18 36 683 388 care.
19 37 676 390
20 40 685 394 5. Acknowledgment
average (U/L) 36.5 703.0 393.0
SD (U/L) 1.47 21.1 9.0 The authors express their deep gratit ude to Mr.
CV (%) 4.0 3.0 2.3 Jun Arakawa of Pharmaceutical & Healthcare Research
Laboratories for his cooperation for assessing substrates and
subsequent manufacturing/introducing an emulsifier. The
4. Conclusion authors are also grateful to everybody of FUJIFILM Medical
Co., Ltd. who kindly provided us with animal samples.
We assessed FDC v-LIP-P slide that was developed for
enabling rapid and easy screening of pancreatitis in the
veterinarian medical care.
The DGGR substrate method or the triglyceride method
is suitable for screening of pancreatitis as a measurement
principle because these methods have high specificity for

10 Development of FUJI DRI-CHEM v-LIP-P Slide that Has the High Specificity to Pancreatic Lipase
 References 14) Baginsky, M.L.; Brown, W.V. Differential characteristics
of purified hepatic triglyceride lipase and lipoprotein
1) Tomoda, Isamu. Veterinary clinical science series (14),
lipase from human postheparin plasma. J. Lipid Res. 18
Clinical blood chemistry test V - Guideline for clinical
(4), 423-437 (1977).
hematology test in small animals -, 77-99 (1996).
15) Gargouri, Y.; Pieroni, G.; LOWE, P.A.; Sarda, L.; Verger,
2) Steiner, J.M.; Broissard, J, Mansfield C.S.; Gumminger,
R. Human gastric lipase. Eur. J. Biochem. 156 (2), 305-310
S.R.; Williams D.A. Serum canine pancreatic lipase
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