Patent ID: 11879880
Assignee: RESEARCH CENTER FOR ECO-ENVIRONMENTAL SCIENCES, CHINESE ACADEMY OF SCIENCES
Field: Measurement (Instruments)
Classification: CPC G  B  H | IPC B  G  H

Claim 3:
4. A method for performing fully-automated biological evaluation and chemical analysis based on the integrated machine for performing fully-automated biological evaluation and chemical analysis of claim 3, characterized in that it the method comprises the steps of:
step 1: enriching, concentrating, and purifying the sample to be analyzed in the pretreatment module to produce a mother liquor to be analyzed;
step 2: the mother liquor to be analyzed enters the component separation module through the high-pressure liquid pump and the sample injector, and then after crude separation through an analysis of a chromatographic column or a preparation of a chromatographic column, a liquid effluent is split into two paths in the three-way diverter valve in a proportion, and enters the monitoring and identifying module and the component collection module, respectively;
step 3: the liquid effluent entering the monitoring and identifying module sequentially passes through the ultraviolet detector and the high-resolution mass spectrometer detector to provide a spectrogram of chromatography-mass spectrometry of the liquid effluent, and the data processing and automated control module acquires the spectrogram of chromatography-mass spectrometry of the liquid effluent;
step 4: the mechanical arm in the component collection module grabs a first plate in the plate storing scaffold and places the first plate on the fraction collector, and the liquid effluent entering the component collection module after being split is collected into the first plate via the fraction collector; the mechanical arm transfers the first plate to the nitrogen blower for nitrogen blowing the liquid effluent in the first plate to dryness, and then transfers the first plate to the automated liquid distributor for redissolution by distributing a constant volume of organic solvent into the first plate, to produce a redissolved solution; after distributing an appropriate amount of the redissolved solution into a second plate which is different from the first plate and nitrogen blowing the second plate to dryness, redissolution is performed with a biocompatible solvent to provide a mother liquor for administration, and a remaining redissolved solution is preserved in the refrigerator; an appropriate amount of positive control mother liquor taken from outside of the integrated machine and an appropriate amount of mother liquor for administration each are formulated with an appropriate amount of complete culture media suitable for cell growth into a positive control exposure liquid and a sample exposure liquid, respectively, at the pipetting workstation, and sample application of the sample exposure liquid and the positive control exposure liquid to a plate for administration is performed;
step 5: the plate for administration after the sample application is transferred to the CO2 incubator via the mechanical arm and then is exposed within the CO2 incubator; the plate for administration is transferred to the biological evaluation detector for biological effect signal detection after exposure is completed; the biological effect signal of each cell in the plate for administration is acquired and a biological assay effect spectrogram is reconstructed by the data processing and automated control module;
step 6: the spectrogram of chromatography-mass spectrometry is overlapped with the biological assay effect spectrogram by the data processing and automated control module to complete a primary screening of the biological effect activity of the mother liquor to be analyzed and determination of biological effect active components;
step 7: if the biological effect active components are still a complex mixture, steps 2-6 are repeated using the remaining redissolved solution in step 4 as a mother liquor to be analyzed, for the purpose of a further subdivision of the biological effect active components;
step 8: after the subdivided biological effect active components are determined, the remaining redissolved solution in step 4 is taken and injected into the monitoring and identifying module for identification by high-resolution mass spectrometry, and accurate mass-to-charge ratios and secondary fragment information of suspicious effectors are acquired; finally, alignment and screening of a chromatographic library of the suspicious effectors are carried out by the data processing and automated control module and a list of the suspicious effectors is obtained.