CN1205034A - Improved method for determining lipase - Google Patents
Improved method for determining lipase Download PDFInfo
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- CN1205034A CN1205034A CN 97191263 CN97191263A CN1205034A CN 1205034 A CN1205034 A CN 1205034A CN 97191263 CN97191263 CN 97191263 CN 97191263 A CN97191263 A CN 97191263A CN 1205034 A CN1205034 A CN 1205034A
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Abstract
The invention relates to a method and reagent for determining lipase in the presence of chromogenic substrate and at least one N-substituted carboxylic acid amide derivatives, or a compound of formula (I) or (Ia), wherein, R<1> and R<2> are hydrogen, or a hydrocarbon residue which is saturated or unsaturated, substituted or unsubstituted, and comprises 2-24 carbon atoms, Z is a hydrocarbon residue which is saturated or unsaturated, substituted or unsubstituted, ring or chain, and comprises 1-10 carbon atoms, X is a atom or atomic group with positive charges, n is 1-3. N-(1,2-dicarboxy ethyl)-N-alkyl sulfo succinamide tetra-sodium or a mixture comprising the compound can avoid the nonspecific reaction in the mensuration of lipase in biological samples.
Description
The invention relates to method and the reagent of measuring lipase in the biological fluid, and the application in a kind of nonspecific reaction of carboxylic acid amide derivative in eliminating enzymatic determination of low molecule N-replacement.Particularly, now proving, is favourable when N-(1,2-dicarboxyl ethyl)-N-alkyl-sulfosuccinic acid amides or N-(1,2-dicarboxyl ethyl)-N-alkaryl-sulfosuccinic amide derivatives exist with the concentration of about 0.001%-2.0% (w/v).
For a long time, measure lipase, particularly human pancreas's lipase (E.C.3.1.1.3), the diagnosis and the course of disease calibrating of pancreatic disease had apodeictic importance (W.Steinberg etal, Annals of Internal Medicine 102 (1985), 576; M.Panteghini et al, Clin.Biochem.24 (1991), 497, N.W.Tietz and D.F.Shuey, Clin.Chem.39/5 (1993), 746).For example, when acute pancreatitis, very huge increase took place in lipase sometimes in the serum in several hours.
In vivo, the function of lipase mainly is the α ester bond that preferentially disconnects triglyceride, and this triglyceride contains long chain fatty acid ester, is broken into to be Diglyceride part and free fatty acids.And then change into monoglyceride, but this process is quite slow.
Usually, enzymic catalytic reaction carries out at aqueous phase, and still, because its physiological importance, lipase only plays a role in oil/water termination.In this respect, lipase significantly is different from esterase.And lipase can only be attached to by on the sex change bile acide emulsive interface not, and disconnects triglyceride under the help of colipase.For this reason, its reaction kinetics also mainly is the influence that is offered the interfacial property of enzyme except being subjected to chemical parameters influences.
At present, known have several different methods to can be used for measuring lipase.These methods mainly are based on titrimetry, turbid setting analysis and immunological testing principle.In titration measuring, at first to add the excess fats enzyme substrates, for example sweet oil by using indicator, carries out titration with alkali then, perhaps by the corresponding mantoquita of extraction, detects the lipid acid amount that is discharged from it by lipase.Volumetry only adopts in the routine clinical chemical laboratory at present once in a while, grasp because be difficult to sometimes, and need very long reaction times and a large amount of sample (W.Junge in Methods ofEnzymatic Analysis, Weinheim VCH.U.Bergmeyer ed.vol.4 (1984), 15).
Detect the clarifying nephelometry lipase measurement of triglyceride/aqueous emulsion turbidity by means of photometer, be method (the W.Rick andM.Hockeburn that in the routine clinical chemical laboratory, generally adopts at present, J.Clin.Chem.Biochem.20 (1982), 735; J.Ziegenhorn et al, " Medica Sonderheft " 11 (1980).The problem of tuurbidimetry is, indivedual serum samples do not show that detection signal weakens linearly in the photometer detection window, but even demonstrates turbidity and increase.Another difficulty of this measuring method is to be difficult to repeatedly produce the emulsion that always has identical sized particles reliably.
A concrete shortcoming of immunological method is that what to measure in this method is quality, rather than enzymic activity (W.UhL et al, Internat, J.Pancreatology 12/3 (1992), and 253, G.E.Hoffann et al, _ rztl.Lab.30 (1984), 193, H.Herden and K.Walter, Klin.Lab, 38 (1992), 89).
Thereby having developed the assay method based on colour developing, is the main at present method that adopts.For example, with 1, the 2-Diglyceride is made substrate, and it is degraded into the 2-monoglyceride by lipase, and is broken into glycerine by the 2-monoglyceride lipase that sample is added subsequently.The free glycerol of Xing Chenging in this way, by means of GPO, degraded forms otan and hydrogen peroxide (H
2O
2).Can measure formed H by suitable colour developing indication mechanism
2O
2(P.Fossatiet al.Clin.Chem.38/2 (1992), 211).
Also may directly transform chromogenic substrate now and be used for lipase measurement (EP 0207252).In this method, because the activity of people's steapsase, directly discharging from this substrate can photometric dyestuff, thereby has avoided to producing the complicated enzyme cascade process of dyestuff.
And, know that for a long time the ester bond of used chromogenic substrate is because non-specific hydrolysis may produce too high signal.For example, chromogenic substrate acetate just is a problem (W.Downey and P.Andrews, Biochem.J.96 (1965) 21) to the mononitro-benzene ester by albumin and the hydrolysis of r-sphaeroprotein very much.This class reaction can produce false measured value, with under the situation of serum sample, may cause clinical misdiagnosis especially.
Therefore, the objective of the invention is to eliminate the nonspecific reaction in the lipase measurement, thereby improve precision of analysis.
By adding lipase chromogenic substrate and a kind of low molecule N-substituted carboxylic acid amide derivatives, the dyestuff that discharges from substrate with spectrphotometric method for measuring is measured lipase by means of this method, thereby has been reached purpose then.Prove, have general formula (I) or (Ia) compound of structure be particularly suitable for according to the present invention as this carboxylic acid amide derivative.
R wherein
1And R
2Represent hydrogen independently of each other, perhaps one saturated or undersaturated, be substituted or unsubstituted, by any hydrocarbon residue carboxylation, that have 2-24 carbon atom,
Z represent one saturated or undersaturated, be substituted or hydrocarbon residue unsubstituted, ring-like or straight chain, that have 1-10 carbon atom,
X represents the atom or the atomic group that have positive charge, and n is the numeral of 1-3.
According to the present invention, general formula (I) or carboxylic acid amide compounds (Ia) they are preferred, wherein:
Methylene radical and/or ethylene that Z is made up of 1-10 C atom, it is randomly replaced by following electron-withdrawing group: carboxyl, alkylsulfonyl, phosphate-based, phosphonate group, nitro, nitrous acid ester group or itrate group, halogen or alkoxyl group,
R
1And R
2Represent hydrogen independently of each other, the straight or branched that is optionally substituted, saturated or undersaturated alkyl, aryl, alkaryl or alkylidene group, it is made up of 3-24 carbon atom, and wherein carboxyalkyl or the dicarboxyl alkyl residue of being made up of 2-24 carbon atom is particularly preferred.
Proved that X represents a positively charged atom or atomic group, on behalf of numeral 1 or 2, n be fit to.
In special those compounds considered of the present invention, residue Z carries carboxyl and/or alkylsulfonyl, but also comprises wherein R
1Or R
2The compound of the following low alkyl group of one of residue representative: methyl, ethyl, propyl group, butyl, amyl group, vinyl or propenyl, perhaps C
10-C
18Alkyl.Can realize that randomly suitable alkyl or alkylidene group replace in accordance with known methods.And proved that following situation is favourable, if R for example
1Represent one based on propanedioic acid, pentanedioic acid, hexanodioic acid, pimelic acid, nonane diacid and sebacic acid are particularly based on the dicarboxylic acid residue of succsinic acid.Can be used for compound of the present invention and preferably have the daltonian molecular weight of about 200-1000, still, prove, also be fit to until 3000 daltonian compounds.The compound or the salt that very advantageously have at least two acid groups have been proved, for example based on having low alkyl group (1-6 C atom) and/or having the N-(1 of senior alkyl (12-18 C atom), 2-dicarboxyl-ethyl)-N-alkyl-sulfosuccinic acid amides, the perhaps derivative that obtains based on N-(1,2-dicarboxyl ethyl)-N-alkaryl-sulfosuccinic acid amides or by it and the tetra-na salt of respective mixtures.
In principle,, all be used for, have mensuration lipase down as general formula (I) or compound (Ia) at compound of the present invention as all compounds that substrate is discerned and transformed by lipase.Proved that a kind of method of measuring lipase is superior especially, wherein be as the lipase substrate with the compound of EP 0207252, for example, 1,2-O-octyl group-racemize-glyceryl-3-nonane diacid, 1,2-O-didecyl-racemize-glyceryl-3-pimelic acid or 1,2-O-dilauryl-racemize-glyceryl-3-pentanedioic acid resorufin ester, perhaps 1,2-O-dilauryl-racemize-glyceryl-3-pentanedioic acid-(4 '-or 6 '-the methyl resorufin) ester.Be surprisingly found out that in this respect these materials of the present invention can prevent from non-specific hydrolysis takes place, and don't can suppress the activity of lipid acid in containing the emulsion of corresponding chromogenic substrate.
With general formula of the present invention (I) or (Ia) (its R
1, R
2, Z, X and n have aforementioned implication) compound, perhaps the mixture of several compounds is added in the lipase measurement reaction solution, its concentration (final concn) is approximately 0.001-2.0% (w/v), is preferably about 0.01-1.0% (w/v).
The further condition of lipase measurement and additive are that one of skill in the art knows; for example washing agent is (as the cow-bezoar deoxycholate salt; Sodium desoxycholate polydocanol), has the material (sodium azide of sterilization or fungicidal action; methyl-different-thiazolone etc.); stablizer (as DMSO), cofactor, emulsifying agent (as propyl alcohol); activator is perhaps avoided the measure of unwanted side reaction.Prove that particularly additive and technical qualification described in DE 2904305 or the EP 0207252 are fit to this situation.Lipase can be measured in deriving from (for example pig pancreas lipase) sample of humans and animals, for example at blood, in serum or the tissue.
General formula of the present invention (I) or N-substituted carboxylic acid amide compound (Ia) or its salt can prepare by the method that this area professional knows.In addition, compound of the present invention, for example N-(1,2-dicarboxyl ethyl)-N-alkyl sulfosuccinic acid amides four sodium (REWOPOLB 2003) can be from known company such as Witco Surfactants GmbH Company, and Steinau. buys.
Another theme of the present invention is reagent or a test kit of measuring lipase, mainly be grouped into: (a) a kind of chromogenic substrate of lipase by following several one-tenth, (b) a kind of suitable buffer material, and (c) be included in composition-a kind of low molecule N-substituted carboxylic acid amide derivatives in any required reagent part, promptly for example a kind of general formula (I) or compound (Ia).Any known buffer reagent that can its pH value be adjusted in 6.0-10.5 in reagent of the present invention all is suitable as this buffer material.Preferred pH value scope is 7.0-9.5.Suitable buffer reagent example has the diethanolamine buffer reagent, the trolamine buffer reagent, Tris or tartrate buffer reagent, perhaps so-called Good ' s buffer reagent, as the Hepes buffer reagent, N, N-two (hydroxyethyl) glycine buffer, Taps buffer reagent and CHES buffer reagent (2-(hexamethylene amino)-ethylsulfonic acid buffer reagent).Particularly preferably be N, N-two (hydroxyethyl) glycine buffer.In the case, the concentration of this buffer reagent preferably at 30-100mM, and particularly preferably is about 50mM usually at 5-200mM.
Prove, is favourable when this reagent is become by two part reagent when being grouped into, wherein except other additive well-known to those having ordinary skill in the art, in first part, also have carboxylic acid amide derivative of the present invention, and the lipase chromogenic substrate is included in the second section reagent.In the case importantly, before measuring, carboxylic acid amide compounds of the present invention is kept in the weak base buffered matrix, and the lipase chromogenic substrate is kept under the acid ph value.In an especially preferred embodiment, reagent of the present invention is made up of two part reagent, wherein first part has weak base buffered pH value (pH7.5-9.0, preferably about pH8.0), and contain general formula of the present invention (I) or one or more carboxylic acid amide compounds (Ia), and other salt, sanitas and washing agent.Second section reagent contains a kind of being buffered in basically in the 2.0-5.0pH value scope, the material of about pH4 preferably, and a kind of suitable lipase substrate, and preferably contain other auxiliary materials.For this second section reagent, proved by adding emulsifying agent for example cholate compound and/or Thesits under the pressure of spray technology, be particularly advantageous.Preferred oil phase promptly can be contained the reagent R2 of whole compositions for this reason, enter the aqueous phase that is at first added by a very thin conduit spray.As a result,, obtained a kind of the limpid of the stability improved that have, do not had muddy emulsion/suspension by described method.And this reagent can be owing to for example existing non-specific enzyme (esterase of non-lipase) that unwanted side reaction takes place.A kind of like this particle size with unanimity is the lipase substrate reagent of feature, can never degenerate about 2-8 ℃ of storage 12-18 month.Guarantee to make complete reagent that adequate stability is arranged so again.The preferably about 100nm of the distribution of sizes scope of particulate maximum is equivalent to the turbidity less than 300NTU (turbidimetric assay turbidity unit).
Description of drawings: Fig. 1: IgG progressive concentration 0-3g/dl, R1 (reagent 1) does not add compound of the present invention,
Sample: 5 μ l,
-NaCl blank,---IgG 0 ... IgG 0.5g/dl,
……IgG??1．0g／dl，……IgG??1．5g／dl，
-IgG??2．0g／dl，---IgG??2．5g／dl，
……IgG??3．0g／dl。Fig. 2: IgG progressive concentration 0-3g/dl, R1 contain 0.27% (w/v) N-(1,2-dicarboxyl ethyl)-N-alkyl-sulfosuccinic acid amides four sodium, and sample: 10 μ l, other details is with the explanation among Fig. 1.Fig. 3: lipase, the colorimetric estimation of turbidity, R1/R2 does not add compound of the present invention, calibrator: Cfas (=calibration automatic operation system).
Numerical value is to quantity: 25
-BaPa?y=57．8108+1．0315
*x(r=0．9828)
Y=x is equal to straight line,
X human serum Fig. 4: the colorimetric estimation of lipase turbidity, R1 does not add, and R2 has added as the The compounds of this invention among Fig. 2, the same explanation of other details to Fig. 3,
Numerical value is to quantity: 25
-BaPa??y=12．6916+1．3995
*x(r=0．9944)
Hour Hk y=9.4818=1.4178
*x
---y=x is equal to straight line,
The x human serum
Will the present invention is further elaborated by the following examples: embodiment 1
IgG with various concentration makes sample
The reagent of lipase colorimetric estimation forms: reagent 1 (R1): 4.55mmol/l taurodeoxycholate 1.77mmol/l NaTDC 50.00mmol/l N; N-two (ethoxy) glycine buffer 0.98mg/l colipase 5.00mmol/l calcium chloride anticorrisive agent reagent 2:(R2): 8.1mmol/l taurodeoxycholate 0.24mmol/l chromogenic substrate (1,2-O-dilauryl-racemic-glyceryl-3-glutaric acid-(6 '-the methyl resorufin) ester) 1.60mmol/l tartrate buffer 0.10mmol/l calcium chloride anticorrisive agent/surfactant
Sample: the NaCl that contains different concns IgG (0-3g/l).On BM/Hitachi 717 automatic analysers, carry out colorimetric estimation according to following schedule of operation:
Temperature: 37 ℃
Measuring and calculating: calculate by reading numerical values from working curve, this curve also is under above-mentioned instrument condition, by measuring from the O standard of Boehringer Mennheim Co. and lipase modular system (Cfas).
The result: find not contain in emulsion under the situation of The compounds of this invention, the IgG concentration (referring to Fig. 1) of adding is obviously depended in chromogenic substrate generation hydrolysis.Embodiment 2
The additive of different concns of the present invention (interference eliminated agent)
The reagent of lipase colorimetric estimation is formed and is equivalent to reagent R1 and the R2 described in the embodiment 1, and different is, and R1 also contains 0.27% has C
12-C
18The N-of alkyl (1,2-dicarboxyl ethyl)-N-alkyl-sulfosuccinic acid amides four sodium mixtures.
Sample: the NaCl that contains different concns IgG (0-3g/l).
According to method and the detailed description described in the embodiment 1, on BM/Hitachi 717 automatic analysers, carry out colorimetric estimation and measuring and calculating.
Result: under predetermined concentration, eliminated nonspecific nonenzymic hydrolysis effect (referring to Fig. 2).Embodiment 3
The preparation of complete reagent
Reagent 1: described in embodiment 1.
Reagent 2:
0.6g lipase chromogenic substrate (for example 1,2-O-dilauryl-racemize-glyceryl-3-pentanedioic acid-(6 '-methyl resorufin) ester) is dissolved in the suitable alcohol of 9ml, in ethanol.This solution is added the 1g emulsifying agent, as Brij 35 or Triton X-114.Formed oil phase is drawn in the injection needle, and high pressure is pushed then, makes it to inject the aqueous solution by fine duct (internal diameter 0.15-1.0mm), stirs simultaneously.This aqueous solution for example contains, 0.15g tartrate buffer reagent (pH5.0), and 0.009g calcium chloride, and 0.5g taurodeoxycholate are dissolved in the 100ml water.In addition, this aqueous solution also may contain one or several sanitas and other any co-emulsifier composition.Embodiment 4
The comparison of nephelometry and colorimetric method when not adding material of the present invention
The reagent of lipase colorimetric estimation is formed and is equivalent to the component described in the embodiment 1.
The reagent of " lipase nephelometry " is composed as follows: the R1:19.0mmol/l Sodium desoxycholate
26.0mmol/l Tris damping fluid
3.0mg/ml colipase
0.1mmol/l calcium chloride
0.30mmol/l triolein sanitas
Sample: human serum
According to following program, on BM/Hitachi 717 automatic analysers, carry out different lipase measurements:
The colorimetric estimation value
Temperature: 37 ℃
| Program 2 | Chemical parameters |
| The determinand detection of code, (speed A) sample volume, (μ l) R1 volume, (μ l) R2 volume, (μ l) wavelength, (nm) calibration steps standard 1 CONC.POS standard 2 CONC.POS standards 3 CONC.POS standards 4 CONC.POS standards 5 CONC.POS. standards 6 CONC.POS. SD limit iteration limit sensitivity limit value ABS scopes, (INC/DEC) prozone scope desired value, (mg/dl) emergent value, (mg/dl) instrument factor | Lipase 5-32-39 7-3 250-100-0 50-50-0 800-570 2-0-0 0-1 designated value-2 0-0 0-0 0-0 0-0 0.1 100 0 32000-0 0-0 0-190 ... 1.0 |
The turbidimetric assay value
Temperature: 37 ℃
| Program 2 | Chemical parameters |
| The determinand detection of code, (dynamics A) sample volume, (pl) R1 volume, (μ l) R2 volume, (μ l) wavelength, (nm) calibration criterion 1 CONC.POS., (U/1) standard 2 CONC.PO S. standards 3 CONC.POS. standards 4 CONC.POS. standards 5 CONC.POS. standards 6 CONC.POS. SD limit iteration limit sensitivity limit value ABS scopes, (INC/DEC) prozone scope normal range (NR), (the emergent value of μ/l), (U/1 or μ g/cat/l) instrument factor | Lipase 5-30-50 10-2 250-20-0 0-20-0 660-340 1-0-0 0-1 (μ cat/l) 0.00-1 designated value-2 0-0 0-0 0-0 0-0 0.1 100 0 10000-0 0-0 0-190 (μ g/cat/l) 0.00-3.17 ... 1.0 |
Calculate with the method that is similar to embodiment 1.
The result: method comparison shows that, colorimetric estimation demonstrates and coordinate axis intersects (axisintercept), this be since chromogenic substrate by (Fig. 3) embodiment 5 due to the non-specific hydrolysis of serum composition
The comparison of nephelometry and colorimetric method when adding material of the present invention
Reagent composition and measuring method are as described in example 4 above.In addition, also added N-(1,2-dicarboxyl ethyl)-N-octadecyl-sulfosuccinic acid amides four sodium as material of the present invention in the reagent 1 (R1), or contained the suitable mixture of this material, concentration is 0.27% (w/v).
Sample: human serum
According to the method described in the embodiment 4, on BM/Hitachi 717 automatic analysers, carry out different lipase measurements.Measuring method also is similar to embodiment 4 and 1.
The result: removed nonspecific reaction owing to adding material of the present invention, and therefore eliminated be similar to nephelometry intersect (Fig. 4) with coordinate axis.
Claims (14)
1. by adding the lipase chromogenic substrate, then the dyestuff that discharges from this substrate being carried out lipase detection method the photometric biological sample, wherein is to carry out this mensuration under the situation that has a kind of low molecule N-substituted carboxylic acid amide derivatives.
2. the process of claim 1 wherein it is when having following general formula (I) or compound (Ia), to measure:
R in the structural formula
1And R
2Represent hydrogen independently of each other, perhaps one saturated or undersaturated, be substituted or the hydrocarbon residue of the unsubstituted 2-24 of a having carbon atom,
Z represent one saturated or undersaturated, be substituted or the hydrocarbon residue with 1-10 carbon atom of unsubstituted, ring-like or straight chain,
X represents the atom or the atomic group that have positive charge, and n is the numeral of 1-3.
3. the method for claim 2, wherein R
1Or R
2Represent the alkyl that is optionally substituted, aryl, alkaryl or an alkylidene group, it is made up of 3-24 carbon atom,
Z has the methylene radical and/or an ethylene of 10 C atoms at the most, and it is randomly replaced by one or several following group: carboxyl, alkylsulfonyl, phosphate-based, phosphonate group, nitro, nitrous acid ester group or itrate group, halogen or alkoxyl group.
4. claim 2 or one of 3 method wherein add general formula (I) or compound or its salt (Ia), and wherein Z carries a carboxyl or alkylsulfonyl.
5. the method for one of claim 2-4, wherein R
1Or R
2One of residue represent methylidene, ethyl, propyl group, butyl, amyl group, vinyl or propenyl, perhaps C
12-C
18Alkyl.
6. the method for one of claim 2-5, salt wherein is N-(1,2-dicarboxyl ethyl)-N-alkyl-sulfosuccinic acid amides four sodium, and N-(1,2-dicarboxyl ethyl)-N-alkaryl sulfosuccinic acid amides four sodium or the derivative that produces by their, or contain the respective mixtures of this compound.
7. the method for one of claim 1-6, carboxylic acid amide derivative wherein are that the concentration with 0.001-2.0% (w/v) is present in the sample liquid.
8. the method that one of requires of aforesaid right, wherein this compound or corresponding compounds are that ultimate density with 0.01-1.0% (w/v) exists.
9. the method that one of requires of aforesaid right, wherein determined is people's steapsase, or from the steapsase of animal.
10. be used to measure the reagent of lipase, contain following composition:
(a) a kind of lipase substrate,
(b) a kind of buffer material and
(c) a kind of low molecule N-substituted carboxylic acid amide derivatives.
11. the reagent of claim 10 contains following composition:
(a) a kind of lipase chromogenic substrate,
(b) a kind of buffer material,
(c) a kind of general formula (I) or compound (Ia),
R wherein
1And R
2Represent hydrogen independently of each other, perhaps one saturated or undersaturated, be substituted or the hydrocarbon residue of the unsubstituted 2-24 of a having carbon atom,
Z represent one saturated or undersaturated, be substituted or the hydrocarbon residue with 1-10 carbon atom of unsubstituted, ring-like or straight chain,
X represents the atom or the atomic group that have positive charge, and n is the numeral of 1-3.
12. the reagent of claim 10 or 11, the buffer material (b) that wherein contains the about 7.5-9.0 of pH value in first part's reagent of this reagent, composition (c), washing agent and/or sanitas, and second section reagent contains the buffer material (b) of the about 2.0-5.0 of pH value, composition (a) and emulsifying agent.
13. claim 10, one of 11 or 12 reagent, wherein composition (c) is N-(1,2-dicarboxyl ethyl)-N-alkyl sulfosuccinic acid amides four sodium, N-(1,2-dicarboxyl ethyl)-N-alkaryl sulfosuccinic acid amides four sodium or corresponding derivative, or contain the suitable mixture of this compound.
14. a N-substituted carboxylic acid amide derivatives or following general formula (I) or (Ia) application in the nonspecific reaction of the lipase measurement of compound in eliminating the biological sample material,
R wherein
1And R
2Represent hydrogen independently of each other, perhaps one saturated or undersaturated, be substituted or the hydrocarbon residue of the unsubstituted 2-24 of a having carbon atom,
Z represent one saturated or undersaturated, be substituted or the hydrocarbon residue with 1-10 carbon atom of unsubstituted, ring-like or straight chain,
X represents the atom or the atomic group that have positive charge, and n is the numeral of 1-3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 97191263 CN1205034A (en) | 1996-09-19 | 1997-09-16 | Improved method for determining lipase |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19638271.8 | 1996-09-19 | ||
| DE19733309.5 | 1997-08-01 | ||
| CN 97191263 CN1205034A (en) | 1996-09-19 | 1997-09-16 | Improved method for determining lipase |
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|---|---|
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Family
ID=5178858
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104198474A (en) * | 2014-08-14 | 2014-12-10 | 上海睿康生物科技有限公司 | Detection kit for measuring content of lipase in serum by colorimetric method |
| CN104215632A (en) * | 2014-08-28 | 2014-12-17 | 宁波瑞源生物科技有限公司 | Stable lipase kit |
| CN107782680A (en) * | 2016-08-26 | 2018-03-09 | 山东博科生物产业有限公司 | A kind of antiheparin fat enzyme detection kit of stabilization |
-
1997
- 1997-09-16 CN CN 97191263 patent/CN1205034A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104198474A (en) * | 2014-08-14 | 2014-12-10 | 上海睿康生物科技有限公司 | Detection kit for measuring content of lipase in serum by colorimetric method |
| CN104215632A (en) * | 2014-08-28 | 2014-12-17 | 宁波瑞源生物科技有限公司 | Stable lipase kit |
| CN104215632B (en) * | 2014-08-28 | 2017-12-19 | 宁波瑞源生物科技有限公司 | A kind of fatty enzyme reagent kit of stabilization |
| CN107782680A (en) * | 2016-08-26 | 2018-03-09 | 山东博科生物产业有限公司 | A kind of antiheparin fat enzyme detection kit of stabilization |
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