Method for determining the volume of a liquid sample
A method, a device, and a test kit for performing this method for determining the volume of a sample of the liquid. In the method, a specific concentration of a chromophoric indicator is provided in this liquid, a sample is separated from the liquid, the optical absorption of the separated sample is measured, and the volume of the separated sample is determined by correlation of the measured optical absorption with the concentration of indicator in this liquid. Ions that generate a color in the sample by complexing with a specific ligand are used as the indicator to stain the liquid.
1. A method for determining the volume of a dispensed liquid sample, wherein the sample liquid comprises a fixed concentration of indicator ions comprising the steps of:
providing a diluent in a container, the diluent comprising polydentate chromogenic ligands of one type;
dispensing a volume of the sample liquid into the diluent, thereby forming complexes of the polydentate chromogenic ligands with the indicator ions, which complexes are chromophoric indicators for staining the resulting mixture;
measuring the optical absorption of the stained mixture in the container; and
determining the volume of the dispensed liquid sample by correlating the measured optical absorption of the stained mixture with the optical absorption of a test sample that has an exactly defined concentration of the same chromophoric indicator, and wherein the chromophoric indicator is a colored complex that has a three-dimensional coordination geometry, which greatly hinders adsorption of this type of molecule to apolar surfaces.
2. A method for determining a residual volume of a liquid in a sample holder comprising the steps of:
providing a liquid in a sample holder comprising a fixed concentration of indicator ions;
removing a volume of the liquid from the sample holder;
adding a diluent comprising polydentate chromogenic ligands of one type to the liquid remaining in the sample holder to form the complexes of the polydentate chromogenic ligands with the indicator ions, which complexes are chromophoric indicators for staining the resulting mixture;
measuring the optical absorption of the stained mixture in the sample holder; and
determining the residual volume of the liquid by correlating the measured optical absorption of the stained mixture in the sample holder with the optical absorption of a test sample that has an exactly defined concentration of the same chromophoric indicator, and wherein the chromophoric indicator is a colored complex that has a three-dimensional coordination geometry, which greatly hinders adsorption of this type of molecule to apolar surfaces.
3. A method for determining the volume of a dispensed liquid sample, comprising the steps of:
providing a diluent in a container;
dispensing a volume of a sample liquid comprising a chromophoric indicator that stains the sample liquid to form a mixture in the container;
measuring the optical absorption of the stained mixture in the container; and
determining the volume of the dispensed liquid sample by correlating the measured optical absorption of the stained mixture with the optical absorption of a test sample that has an exactly defined concentration of the same chromophoric indicator, wherein the chromophoric indicator to stain the sample liquid is a colored complex that is formed by complexing indicator ions with specific polydentate chromogenic ligands with a three-dimensional coordination geometry which greatly hinders adsorption of this type of molecule to apolar surfaces.
4. A method for determining a residual volume of a liquid in a sample holder comprising the steps of:
providing a liquid in a sample holder comprising a chromophoric indicator;
removing a part of the liquid in the sample holder;
adding a diluent to the residual volume of the liquid in the sample holder;
measuring the optical absorption of the diluted residual volume of the liquid; and
determining the residual volume of the liquid in the sample holder by correlating the measured optical absorption of the mixture with the optical absorption of a test sample that has an exactly defined concentration of the same chromophoric indicator, wherein the chromophoric indicator to stain the liquid is formed by complexing indicator ions with specific polydentate chromogenic ligands with a three-dimensional coordination geometry, which greatly hinders adsorption of this type of molecule to apolar surfaces.
5. The method according to claim 1 or 2 , wherein, prior to dispensing the liquid sample, a compensating volume is provided in the container as part of the diluent.
6. The method of claim 1 wherein the solution from which liquid is dispensed further comprises complexes composed of one type of auxiliary ligands and indicator ions for improving the solubility of the indicator ions in the solution.
7. The method of claim 2 wherein the liquid in the sample holder further comprises complexes composed of one type of auxiliary ligands and indicator ions for improving the solubility of the indicator ions in the liquid.
8. The method according to claim 1 or 2 , wherein the polydentate chromogenic ligand is added to the diluent in excess.
9. The method according to claim 1 or 3 , wherein after dispensing the liquid sample into the container, a supplementary volume is added to this container as part of the diluent.
10. The method according to claim 1 , 2 , 3 , or 4 , wherein the indicator ions for complexing with the polydentate chromogenic ligands comprise metal ions.
11. The method according to claim 10 , wherein the metal ions are Fe ++ , Fe +++ , mixtures of Fe ++ and Fe +++ , or Cu ++ .
12. The method according to claim 1 , 2 , 3 , or 4 , wherein indicator ions for complexing with the polydentate chromogenic ligands are anions.
13. The method according to claim 12 , wherein the anions are F − , Cl − , or H 2 PO 4 − .
14. The method according to claim 10 , wherein metal ions which cannot be quantitatively complexed with the polydentate chromogenic ligands are reduced or oxidized to indicator ions prior to complexing with the polydentate chromogenic ligands.
15. The method according to claim 14 , wherein the metal ions are reduced with hydroxyl amine hydrochloride, a tartrate salt, or ascorbic acid or the metal ions are oxidized with hexacyanoferrate or elementary bromine.
16. The method of claim 1 , 2 , 3 , or 4 , wherein the polydentate chromogenic ligand is FerroZine®, bathophenanthroline-disulfonic acid disodium, bathocuproine-disulfonic acid disodium or Chromazurol S.
17. The method according to claim 6 or 7 , wherein the auxiliary ligands are β-diketones.
18. The method of claim 17 , wherein the β-diketones are acetyl acetonate or pentane-2,4-dione-1,5-diol.
19. The method of claim 12 , wherein the chromogenic ligands are anthraquinone functionalized systems covalently bonded at the 62 position.
20. The method of claim 19 , wherein the anthraquinone functionalized system covalently bonded at the β position is calix[4]pyrrole-anthraquinone.
21. The method according to claim 16 , wherein the chromophoric indicator comprises substituted ionic groups that further amplify the hydrophilic properties of the chromophoric indicator.
22. The method according to claim 1 or 2 , wherein metal ions, which cannot be quantitatively complexed with the polydentate chromogenic ligands, are complexed with auxiliary ligands thereby improving their solubility in a liquid, a sample of the liquid is dispensed into the diluent, wherein the diluent is an existing reaction solution that comprises polydentate chromogenic ligands and a reducing or oxidizing agent, wherein a reducing or oxidizing agent is reducing or oxidizing the metal ions to indicator ions which are then complexed with the polydentate chromogenic ligands under color development.
23. The method according to claim 6 or 7 , in which the metal ions that have been complexed with auxiliary ligands are mixed with the diluent containing polydentate chromogenic ligands, wherein the indicator ions are then complexed with the polydentate chromogenic ligands under color development while suppressing the auxiliary ligands.