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The sensitivity of this lipase test is especially influenced by the extinction of the substrate solution, the concentration of taurodesoxycholate, the pH value, and the molarity of the Tris buffer.

Final test concentration of the substrate solution:
chrom. Lipase Substrate: 0.24 mmol/L (=180 mg/L)
colipase: 0.98 mg/L
taurodesoxycholate *(see buffer solution)
CaCl2 :0.1 mol/L
tartrate buffer, pH 4.0: 1.6 mmol/L
stabilizers
The lipase substrate has to be dissolved in a small quantity of an organic solvent (e.g. n-propanol) first. Under vigorous stirring this organic solution should be injected into the tartare buffered aqueous solution with a thin beam. (The lipase substrate starts to hydrolyze at alkaline pH values.) The lipase substrate containing solution should be a micro-emulsion with
an extinction of about 0.5 E. (Lower extinction of the reagent results in measurement of nonspecific serum esterases.) Stabilizers like mannitol, polywax 4000 and co-emulsifiers like lecithin, phosphoryl choline or dilauryl-glycerol-sulfate improve the stability of the micro-emulsion.

Final test concentration of the buffer solution:
Tris-HCl, pH 8.4: 41 mmol/L
taurodesoxycholate: *7.2 mmol/L (total)
desoxycholate: 1.77 mmol/L
Wavelength: 578 nm or 580 nm
Temperature: +25°C, +30°C or +37°C, respective
Buffer solution: 1 mL
Substrate solution: 0.2 mL
Sample volume: 0.02 mL
Assay time: 2 to 10 minutes
Start of reaction: with substrate", "Name": "Background Information" }, { "Language": "en", "Value": "Nomenclature: 1,2-O-Dilauryl-rac-glycero-3-glutaric acid-(6'-methylresorufin)ester
CAS Number: 195833-46-6
Formula: C45H69NO8
Molecular weight: 752.05 D
λmax.substrate: 470 nm (Tris-HCI, pH 8.4)
ε470: 57.94 [L x mmol-1 x cm-1]
λmax.methylresorufin: 581 nm (Tris-HCI, pH 8.4)
Melting range: +29 to +31°C
pH optimum: 7.0-9.5
Solubility: Soluble in polar organic solvents, e.g., n-propanol, ethyl acetate, dioxane, methanol, dimethyl sulfoxide. The limit of solubility in n-propanol is 42.9 mg/mL.", "Name": "Properties" }, { "Language": "en", "Value": "Appearance: Red, smear substance
Chromogenic Lipase Substrate (from C): ≥95%
Chromogenic Lipase Substrate (HPLC): ≥95 area%
C (elementary analysis): 68.2-72.5%
H (elementary analysis): 8.7-9.7%
N (elementary analysis): 1.4-2.4%
Methylresorufin, free (HPLC): ≤0.5 area%
Isomer (HPLC): ≤2 area%
Stability: At +2 to +8°C within specification range for 36 months.", "Name": "Specification" } ] } } ] }

Chromogenic Substrate for Lipase

1,2-O-Dilauryl-rac-glycero-3-glutaric acid-(6'-methylresorufin)ester 

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