Solid-liquid separation method, solid-liquid separation apparatus, and kit to be used therefor

Patent metadata

Field	Value
Publication number	US-10702864-B2
Application number	US-201815993013-A
Country	US
Kind code	B2
Filing date	May 30, 2018
Priority date	May 31, 2017
Publication date	Jul 7, 2020
Grant date	Jul 7, 2020

Abstract

Disclosed is a solid-liquid separation method including: sucking a suspension containing particles, through an opening formed at a leading end of a suction nozzle, into the suction nozzle; allowing at least a part of the particles contained in the sucked suspension, to sediment to be clogged in the suction nozzle; and ejecting a liquid inside the suction nozzle, through the opening of the suction nozzle clogged with the particles.

First claim

What is claimed is:

1. A solid-liquid separation method comprising:
sucking a suspension containing particles, through an opening formed at a leading end of a suction nozzle, into the suction nozzle;
allowing at least a part of the particles contained in the sucked suspension, to sediment by gravity to be clogged in the suction nozzle; and
ejecting a liquid inside the suction nozzle, through the opening of the suction nozzle clogged with the particles, wherein
a minimum opening diameter of the suction nozzle is greater than a maximum diameter of the particles, and is smaller than six times the maximum diameter of the particles,
with respect to a particle diameter and a particle diameter distribution, the particles have D 90 -D 10 being not smaller than 120 μm and D 50 being not smaller than 170 μm,
an inner diameter of the suction nozzle is gradually reduced toward the leading end.

2. The solid-liquid separation method of claim 1 , wherein
the ejecting the liquid is performed after a difference in particle concentration due to sedimentation has occurred between: a vicinity of the leading end opening in the suspension having the sucked particles substantially uniformly dispersed therein; and a farthest portion from the leading end opening in the suspension, and
the ejecting the liquid is ejecting a liquid that is capable of passing through the suction nozzle clogged with the particles, in association with the ejection.

3. The solid-liquid separation method of claim 2 , wherein
the suspension is obtained by mixing a sample liquid containing a target substance, and particles for adsorbing the target substance, and
the ejecting the liquid is causing the particles having the target substance adsorbed thereon to remain at a near side of the opening.

4. The solid-liquid separation method of claim 3 , wherein
the target substance is a desired peptide or nucleic acid, and
the liquid contains a peptide and/or a nucleic acid and a salt other than the desired peptide or nucleic acid.

5. The solid-liquid separation method of claim 3 , wherein
the sample liquid is whole blood, serum, or plasma.

6. The solid-liquid separation method of claim 3 , wherein
the sample liquid is a supernatant obtained by heating whole blood, serum, or plasma by means of a microwave oven.

7. The solid-liquid separation method of claim 3 , wherein
the sample liquid is a desalted liquid of a supernatant obtained by heating whole blood, serum, or plasma by means of a microwave oven.

8. The solid-liquid separation method of claim 3 , wherein
the particles are particles of a porous material for adsorbing the target substance.

9. The solid-liquid separation method of claim 3 , wherein
the particles are made from silica, alumina, zirconia, or polystyrene.

10. The solid-liquid separation method of claim 3 , wherein
after the liquid is ejected, an elution liquid for eluting the target substance from the particles having the target substance adsorbed thereon is sucked into the suction nozzle,
at least a part of the particles mixed with the sucked elution liquid is allowed to sediment at the near side of the opening, and
the elution liquid and the eluted target substance are ejected through the leading end opening.

11. The solid-liquid separation method of claim 1 , wherein
the sucking the suspension is immersing the leading end of the suction nozzle in the suspension and causing an air pressure in the suction nozzle to be a negative pressure relative to an external air pressure,
the allowing the particles to sediment is keeping the air pressure in the suction nozzle constant, and
the ejecting the liquid is causing the air pressure in the suction nozzle to be a positive pressure relative to the external air pressure.

12. The solid-liquid separation method of claim 11 , wherein
controlling the air pressure in the suction nozzle is controlling a position of a plunger in a state where a syringe including the plunger is connected to a rear end of the suction nozzle,
the sucking the suspension is increasing, by moving the plunger, a volume inside the syringe including the suction nozzle,
the allowing the particles to sediment is causing the plunger to rest and keep a same position, and
the ejecting the liquid is decreasing, by moving the plunger, the volume inside the syringe including the suction nozzle, and then causing the plunger to rest and keep a same position.

13. The solid-liquid separation method of claim 1 to be used in BF separation.

14. The solid-liquid separation method of claim 1 , wherein
a concentration of the particles in the suspension is 10 to 100 mg/mL.

15. The solid-liquid separation method of claim 1 , wherein
an inner diameter of the suction nozzle is gradually reduced toward the leading end.

16. A solid-liquid separation apparatus comprising:
a suction unit to which a suction nozzle having an opening formed at a leading end thereof is attached;
a controller programmed to control operation of the suction unit; and
a container supporting body configured to support a container storing a sample liquid that contains a target substance, and particles configured to absorb the target substance thereon, wherein
the controller is programmed to control the suction unit so as to
suck at least a part of a suspension of the sample liquid containing the particles, through the opening into the suction nozzle,
allow at least a part of the particles contained in the sucked suspension, to sediment by gravity to be clogged in the suction nozzle, and
eject a sample liquid inside the suction nozzle, through the opening of the suction nozzle clogged with the particles, thereby causing the particles having the target substance absorbed thereon, to remain in the suction nozzle, wherein

a minimum opening diameter of the suction nozzle is greater than a maximum diameter of the particles, and is smaller than six times the maximum diameter of the particles,
with respect to a particle diameter and a particle diameter distribution, the particles have D 90 -D 10 being not smaller than 120 μm and D 50 being not smaller than 170 μm,
an inner diameter of the suction nozzle is gradually reduced toward the leading end.

17. The solid-liquid separation apparatus of claim 16 , wherein the controller is programmed to control the suction unit such that
in a resting period determined in advance on the basis of a period after the suction of the suspension until a difference in particle concentration due to sedimentation occurs between:
a vicinity of the leading end opening in the suspension having the particles substantially uniformly dispersed in the suction nozzle; and
a farthest portion from the leading end opening in the suspension, the suction unit stops the suction of the suspension to allow at least a part of the particles to sediment, and then, after a lapse of the resting period, the suction unit ejects the sample liquid inside the suction nozzle through the opening of the suction nozzle clogged with the particles.

18. A solid-liquid separation kit comprising:
a pipette tip as a suction nozzle that is used in the method of claim 1 and that does not have a filter in a vicinity of the leading end opening.

Assignees

• Sysmex Corp
• Mitsubishi Chem Corp

Inventors

• Kishimoto Satomi
• Kabata Hiroyuki
• Katsuki Takanobu

Classifications

• B01L2200/0647

  Handling flowable solids, e.g. microscopic beads, cells, particles · CPC title

• B01L3/0293

  for liquids · CPC title

• B01L2200/0631

  Purification arrangements, e.g. solid phase extraction [SPE] · CPC title

• B01L2300/0681

  Filter · CPC title

• G01N2035/1053

  for separating part of the liquid, e.g. filters, extraction phase · CPC title