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I. The basic principle of centrifugation technology centrifugation technology is a commonly used technology in the laboratory, mainly by using centrifugal force to quickly sink suspended particles in suspension, so as to separate the various material components with different proportions.
sedimentable particles in the separated liquid, the sinking force under the interaction of gravity and buoyant force has the following relationship: . F
-gravity-buoyant,
VPg
-
V-deltaVg
(
p
-
-delta;
F
- sinking force; V
- particle volume; P
- particle density;s delta;
- media density; g
- gravity acceleration.
be
. The
>
the difference
between
P
and
and delta
the faster the particles sink when the P
delta;
is found in P-delta; Therefore, increasing
centrifuges
speed can speed up the sendication of centrifuges.II, the basic principle of preparation centrifugation technology preparation centrifugation technology is to separate, purify, prepare a biological molecule, subcellular particles, cysts and cells for the purpose of centrifugation technology. According to the principle, the preparation centrifugation technology can be divided into differential centrifugation technology and density gradient centrifugation technology.
differential centrifugation technology refers to the particles in the suspension according to the unealance, different particles have different seboding speed, usually large particles sethering speed. In order to separate the different particle components, you can choose different centrifugal speed, you can be from low speed to high-speed phased centrifugation, or the use of high speed and low speed alternate recurring centrifugation, so that particles with different semosis speed in different centrifugal speed and different centrifugal time in batches of precipitation, while getting the required components.
density gradient centrifugation technology refers to the prior in the
centrifuge tube
into the different density of centrifugal medium, centrifugal tube from top to bottom, medium density from low to high, forming a density gradient centrifugal medium. The maximum density of the medium needs to be less than the density of the smallest particles in the sample, thus ensuring that all particles in the sample are sethed at a rate greater than zero. The sample is added to the density gradient centrifugal medium, during which the particles 2 000 d'or float up and move along the gradient until a zone is formed at a location equal to its density. Particles at iso-density do not float because they dissolve in the density gradient medium without weight. The effective separation of different particles depends on the difference in buoyant density of the particles, the greater the difference in density, the better the separation effect. The shape and position of the zone strip are not affected by centrifuge time.third, centrifuge classification(1) according to the speed classification
1 . The normal-speed centrifuge
refers to centrifuges with a speed of less than 5,000r/min, which are mainly used for the separation of precipitated particles in general liquids, or for the preparation of general
tissue
homogenous slurry.
2 . High-speed centrifuges
refer to centrifuges with speeds between 5,000 and 25,000 r/min and are mainly used for cell separation and preparation of biometric molecules.
3 . The superstat centrifuge
refers to centrifuges with a speed of more than 25,000 r/min and are used for the separation of subcellular structures and the preparation of biomosulations. Overspeed centrifuges currently have speeds of more than 85,000 r/min. Under the effect of super-fast centrifugal force field, the centrifugal force is greater than the diffusion force of the molecule, the biomolelees gradually sink, and the rate of their subsidon is different with the molecular weight and shape of the biomolees, so that different biological large molecules separate from each other.
(ii) Classification by temperature
1 . The temperature centrifuge
refers to centrifuges that are centrifuged at room temperature (25 degrees C). In general, when centrifuging at normal speed, use a room temperature centrifuge.
2 . A low-temperature centrifuge
a centrifuge that is centrifuged at 4 degrees C. In general, low-temperature centrifuges are used when centrifuges are at high speed.
3 . A frozen centrifuge
a centrifuge that is centrifuged at 0 to 25 degrees C. In general, when overspeeding centrifuges, use a frozen centrifuge. .