CN109569344B - Method for preparing suspended micro-droplets by using micro-fluidic device - Google Patents
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
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- B01F23/4105—Methods of emulsifying
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- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
Abstract
The invention discloses a method for preparing suspended micro-droplets by using a microfluidic device. At the outlet of the internal phase channel of the microfluidic device, the internal phase solution forms micro-droplets under the combined action of the surface tension and the viscous force of the two-phase solution. Then, a solution for gelling the external phase is introduced at the outlet of the external phase channel of the microfluidic device. The gelled external phase solution has shear thinning properties. Under the condition of no shearing force, the inner phase liquid drop can be kept at a fixed position in the three-dimensional space of the outer phase for a long time, so that suspended micro-liquid drops with controllable particle size, uniform size and uniform dispersion are obtained. The technology realizes continuous production through a continuous injection pump and realizes batch production through parallel amplification of the microfluidic device, thereby realizing industrial mass production. The technology has simple preparation process and wide application value in the fields of food, cosmetics and medicines.
Description
Technical Field
The invention relates to the field of liquid drops, in particular to a method for preparing a suspended liquid drop by using a microfluidic device.
Background
Suspended micro-droplets are widely used in various fields. The traditional emulsification technology for preparing suspended micro-droplets generally adopts a high-speed shearing method, and oil, water, a surfactant and a thickening agent are placed in a container together, and a high-shear emulsifying machine is used for emulsification. Wherein surfactants are used to stabilize the emulsion and thickeners are used to impart shear-thinning properties to the solution. Without shear forces, the microdroplets can be suspended in fixed positions in three-dimensional space of the solution for extended periods of time. The high-speed shearing generated by the high-speed rotation of the rotor of the emulsifying machine ensures that the oil phase is quickly emulsified in the water phase. However, the high-speed shearing emulsification method cannot accurately control the emulsification process, phase separation is easy to form, the oil phase ratio is difficult to increase, and the particle size of micro-droplets is uneven.
Compared with the traditional shearing emulsification technology, the technology for preparing the liquid drops by microfluidics has obvious advantages. The prepared liquid drops are uniform and controllable in size, and can be parallelly amplified through a microfluidic device to realize batch production. However, the microdroplet suspension prepared simply by shearing the internal phase through the gelled external phase is unstable. This is because the gelled outer phase is subjected to strong high-speed shear when the inner phase is sheared in the microtube, and the shear thinning property is deteriorated, so that the droplet suspension is unstable and phase separation is easily caused. The present invention designs a microfluidic device that achieves long-term stabilization of a droplet suspension by first emulsifying the inner phase with an ungelled outer phase and then gelling the outer phase in a two-step process. The invention can also realize the accurate control of the oil-water ratio, the size of the prepared suspended micro-droplets is uniform and controllable, and the mass production can be realized by the parallel amplification of the micro-fluidic device. The technology can improve the overall attractiveness of the product, realize the function visualization effect and have important application value in various fields such as cosmetics, medicines, foods and the like.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a method for preparing stable suspended liquid drops based on a glass capillary microfluidic device. The method is simple and feasible, can obtain stable liquid drop suspension with uniform size and uniform distribution, and can be widely applied to various fields such as cosmetics, medicines, foods and the like.
To achieve the above object, the present invention provides the following solutions:
the invention provides a method for preparing micro-droplets based on a glass capillary microfluidic device, which comprises the following steps:
a method for preparing micro-droplets by utilizing a glass capillary microfluidic device is characterized by comprising the following steps:
(1) any one or a mixture of a plurality of oily substances is selected as the inner phase,
(2) the surfactant and thickener are dissolved in a solvent to give a shear-thinning solution as the external phase.
(3) Injecting the internal phase prepared in the step (1) and the external phase prepared in the step (2) into an internal phase inlet and an external phase inlet of the microfluidic device respectively through a syringe; at the outlet of an internal phase channel of the microfluidic device, an internal phase solution forms micro-droplets under the combined action of surface tension and viscous force of a two-phase solution, then a solution for gelatinizing an external phase is introduced at the outlet of an external phase channel of the microfluidic device, the gelatinized external phase solution has the shear thinning property, and the internal phase droplets can be kept at fixed positions in a three-dimensional space of the external phase for a long time under the condition of no shear force, so that suspended micro-droplets dispersed in the external phase are obtained.
Preferably, the oily substance may be one or more of rosemary, perfume, lavender essential oil or vitamin E.
Preferably, in step (2), the surfactant in the external phase is polyvinyl alcohol or sodium lauryl sulfate, etc.
Preferably, in step (2), the solvent is water.
Preferably, in step (2), the thickener in the external phase may be carbomer or xanthan gum.
Preferably, in the step (2), the mass percentage of the surfactant in the external phase is 1% -10% in the water phase when the surfactant is polyvinyl alcohol, and the mass percentage of the surfactant in the water phase when the surfactant is sodium dodecyl sulfate is 0.5% -1%.
Preferably, in the step (3), the solution for gelling the external phase is an aqueous sodium hydroxide solution, and the aqueous sodium hydroxide solution has a mass percent solubility of 0.01% to 50%.
Preferably, the glass capillary microfluidic device is prepared by the following steps:
(1) preparing a glass tube: preparing a round glass capillary tube and a square glass capillary tube, ultrasonically cleaning the glass capillary tube and drying;
(2) stretching the glass tube: processing one end of a round glass capillary tube into a taper shape by using a stretcher;
(3) treating the capillary: rubbing one end of a conical tip of the round glass capillary tube by using abrasive paper to flatten an end opening;
(4) preparing a device: and (3) placing one end of the processed pointed cone of the round glass capillary into the square glass capillary, and then fixing all the glass capillaries on the glass sheet by using glue.
The invention has the following beneficial effects:
1) the suspended micro-droplets obtained by the traditional emulsification technology are generally subjected to a high-speed shearing method, and the particle size of the micro-droplets in the emulsion obtained by the method is difficult to accurately control and has uneven size. The invention provides a method for preparing suspended micro-droplets by using a glass capillary microfluidic device, the droplets prepared by the method have controllable particle size and uniform size, and the problems of difficult particle size control and uneven size in the traditional emulsification method are solved.
2) The traditional shearing emulsification method cannot accurately control the dispersion process, and is easy to form phase separation, so that the oil phase ratio is difficult to improve.
3) The microdroplet suspension prepared by simply shearing the internal phase through the gelled external phase is unstable. This is because the gelled outer phase is subjected to strong high-speed shear when the inner phase is sheared in the microtube, and the shear thinning property is deteriorated, so that the droplet suspension is unstable and phase separation is easily caused. The present invention designs a microfluidic device that achieves long-term stabilization of a droplet suspension by first emulsifying the inner phase with an ungelled outer phase and then gelling the outer phase in a two-step process.
4) The suspended micro-droplets prepared by the glass capillary microfluidic device have uniform particle size, and compared with products prepared by traditional emulsification, the suspended micro-droplets prepared by the glass capillary microfluidic device have the advantages that the overall attractiveness of the products can be improved, the function visualization effect is realized, and the attraction of the products is improved.
5) In the invention, the single emulsion is used as a template to prepare suspended micro-droplets, continuous production is realized through a continuous injection pump, and batch production is realized through parallel amplification of a micro-fluidic device, so that industrial mass production is realized, and the method has wide application prospects in the fields of cosmetics, medicines, foods and the like.
Drawings
FIG. 1 is a schematic diagram of the emulsification of micro-droplets in a glass capillary microfluidic device;
FIG. 2 is a diagram of suspended micro-droplets of uniform size prepared in example 1 of the present invention;
FIG. 3 is a diagram of a droplet object with different particle sizes prepared by controlling the flow rate in example 2 of the present invention;
FIG. 4 is a diagram of the different oil-water ratios obtained in example 3 of the present invention;
FIG. 5 is a graph of elastic modulus, dissipation modulus, and viscosity versus frequency for different concentrations of carbomer in water;
FIG. 6 is a graph of the effect of different concentrations of aqueous solutions of carbomer on the suspension of oil droplets in example 5 of the present invention as a function of time;
FIG. 7 is a schematic view of an apparatus in example 6 of the present invention (1: a first set of microfluidic devices, 2: a second set of microfluidic devices, 3: a third set of microfluidic devices, 4: a fourth set of microfluidic devices, 5: an aqueous NaOH solution, 6: a water tank, 7: an oil tank, 8: a single valve, 9: a continuous injection pump, 10: a guide tube, 11: a three-way tube, 12: a four-way tube, and 13: a glass capillary microfluidic device);
FIG. 8 shows the results of twelve microfluidic devices of example 6, with 1-3 showing the results of the first set of droplets, 4-6 showing the results of the second set of droplets, 7-9 showing the results of the third set of droplets, and 10-12 showing the results of the fourth set of droplets;
FIG. 9 is a particle size histogram of the droplets obtained in FIG. 8;
figure 10 comparative results of two preparation suspensions of example 7.
Detailed Description
The present invention will be described with reference to examples, but the present invention is not limited to the examples.
Example 1: and preparing suspended micro-droplets with uniform particle size.
Referring to the attached figure 1, the method for preparing the suspended micro-droplets comprises the following specific steps:
(1) manufacturing the microfluidic device: cleaning and drying the capillary glass tube; then, drawing one end of the glass capillary tube into a taper shape by using a drawing instrument; grinding the capillary glass tube to a proper diameter by using sand paper; fixing the glass capillary tube and the glass square tube on a glass slide by using glue; and finally, sealing and fixing the needle head at the inlet of the glass tube by using glue. The glass capillary microfluidic device is shown in figure 1 and comprises an inner phase tube and a glass square tube communicated with an outer phase.
(2) Preparation of the internal and external phases: the inner phase is selected from isononyl isononanoate; 10mg of surfactant polyvinyl alcohol and 0.2mg of carbomer 940 were dissolved in 989.8mg of water to give an external phase.
(3) Preparation of micro-droplets: injecting the internal phase obtained in the step (2) into an internal phase inlet of a capillary glass tube microfluidic device through a syringe pump; and (3) injecting the external phase obtained in the step (2) into an external phase inlet of the microfluidic device through a syringe pump. At the outlet of the inner phase tube, the inner phase solution obtains micro-droplets with uniform size under the combined action of the surface tension and the viscous force of two phases, and the aqueous solution of sodium hydroxide is injected into the outlet of the outer phase of the micro-fluidic device through an injection pump to obtain suspended micro-droplets. (as shown in figure 2).
Example 2: suspended micro-droplets of different sizes were prepared by glass capillary microfluidic devices.
(1) A glass capillary microfluidic device was prepared as mentioned in example 1, the internal phase being selected from isononyl isononanoate; 10mg of surfactant polyvinyl alcohol and 0.15mg of carbomer 940 were dissolved in 989.85mg of water to give an external phase.
(2) Respectively introducing the internal phase and the external phase into an internal phase nozzle and an external phase nozzle of the glass capillary microfluidic device, wherein when the flow rate of the internal phase is 1ml/h and the flow rate of the external phase is 5ml/h, the size of the obtained liquid drop is about 675 um; when the flow rate of the external phase is 2ml/h and the flow rate of the external phase is 5ml/h, the size of the obtained liquid drop is about 834 um. So that droplets of different sizes can be obtained with different ratios of internal and external phase flow rates (as shown in figure 3).
Example 3: suspended micro-droplets with different oil-water ratios are prepared by a glass capillary microfluidic device.
(1) A glass capillary microfluidic device was prepared as mentioned in example 1, the internal phase being selected from isononyl isononanoate; 10mg of surfactant polyvinyl alcohol and 0.15mg of carbomer 940 were dissolved in 989.85mg of water to give an external phase.
(2) Respectively introducing the internal phase and the external phase into an internal phase pipe orifice and an external phase pipe orifice of the glass capillary microfluidic device, and obtaining a left suspended micro-droplet system in the figure 4(a) when the internal phase flow rate is 1ml/h and the external phase flow rate is 9 ml/h; when the flow rate of the external phase is 0.5ml/h and the flow rate of the external phase is 9.5ml/h, the system of the suspended micro-droplets on the right side of the graph 4(a) is obtained.
(3) The suspended micro-droplet system obtained above was put into a centrifuge and worked at 4000rpm for 10 minutes to obtain the results in fig. 4(b), and the oil phase ratio was measured to be 10% and 5%, respectively.
Example 4: different concentrations of carbomer 940 were dissolved to test its shear thinning properties.
(1) Dissolving 0.06g of carbomer 940 into 99.94g of water to obtain a carbomer aqueous solution with the mass fraction of 0.06%, and then adding 200ul of a sodium hydroxide aqueous solution with the mass fraction of 10% to adjust the pH value of the carbomer aqueous solution to about 7 to obtain clear hydrogel; similar dissolution of different mass fractions of carbomer 940 resulted in clear hydrogels of mass fractions of 0.1%, 0.14%, 0.2% and 0.25%.
(2) The above hydrogels were tested for changes in elastic modulus, dissipation modulus, and viscosity using a rheometer (DISCOVERY HR-2hybrid rheometer), with the lighter line in FIG. 5 being the change in viscosity with frequency for different mass fractions of carbomer hydrogels. As can be seen from the figure, the viscosity decreased with increasing frequency, indicating that the series of hydrogels had shear-thinning properties.
Example 5: the minimum carbomer 940 concentration was tested to stabilize suspended oil droplets for a long period of time.
(1) Dissolving 0.06g of carbomer 940 into 99.94g of water to obtain a carbomer aqueous solution with the mass fraction of 0.06%, and then adding 200ul of a sodium hydroxide aqueous solution with the mass fraction of 10% to adjust the pH value of the carbomer aqueous solution to about 7 to obtain clear hydrogel; preparing carbomer hydrogel with the mass fraction of 0.05 percent by the same method.
(2) The hydrogels with different mass fractions in (1) above were placed in a centrifuge tube, and then one drop of isononyl isononanoate, which dissolves a green dye, was injected into each tube, as shown in fig. 6, in the carbomer hydrogel with a mass fraction of 0.05%, the oily droplets began to float up after two hours, but the carbomer hydrogel with a mass fraction of 0.06% could keep not floating up for five months, indicating that when the mass fraction of carbomer was 0.06% or more, the corresponding hydrogels had good suspension stabilization effect.
Example 6: suspension droplets were prepared in bulk.
(1) Twelve identical glass capillary microfluidic devices were prepared as described in example 1, with inner phase outlet orifices having an inner diameter of about 200 um. Twelve micro-fluidic devices are divided into four groups, each group comprises three micro-fluidic devices, four injectors can be pushed onto one injection pump at the same time, one injector can averagely divide fluid into the other three outlets through a four-way interface, the three outlets are respectively connected onto one group of micro-fluidic devices, so that one injection pump can simultaneously control the injection of the equivalent fluid of the twelve micro-fluidic devices, and the laboratory needs three injection pumps to respectively control the injection of an oil phase, the injection of a water phase and the injection of a sodium hydroxide aqueous solution (as shown in fig. 7). The flow rate of the oil phase was 12ml/h, the flow rate of the aqueous phase was 80ml/h, and the flow rate of the aqueous sodium hydroxide solution was 8 ml/h.
(2) The size and distribution of the droplets obtained under the above experimental conditions are shown in fig. 8, and the result of counting the size and distribution of the droplets in the picture by using ImageJ is shown in fig. 9, so that it can be observed that the overall particle size of the droplets obtained by one set of microfluidic devices and different sets of microfluidic devices is about 1.1mm, and the droplets have good uniformity.
Example 7: the effect of one-time emulsification and first emulsification then gelation on the suspension stability of the droplets.
(1) A single-emulsion microfluidic device and a three-phase inlet microfluidic device were prepared as described in example 1, the single-emulsion microfluidic device being suitable for a one-shot emulsification method, and the three-phase inlet microfluidic device being suitable for a one-shot emulsification-then-gelation method, wherein in the one-shot emulsification method, an aqueous solution of carbomer was first gelled, the gelled aqueous phase was taken as the external phase, isononyl isononanoate was selected as the internal phase, and the external phase inlet orifice and the internal phase inlet orifice of the single-emulsion microfluidic device were respectively passed through to obtain suspended micro-droplets. In the process of using the emulsification-first gelation method, the procedure was divided into two steps, after obtaining the aqueous solution of carbomer, the aqueous solution of sodium hydroxide was not added, and the aqueous solution of sodium hydroxide was introduced into the microfluidic device as the third phase, as described in example 1, to obtain the suspension droplets.
(2) The suspension droplets obtained above were allowed to stand for two weeks, and the suspension of droplets obtained by the one-time emulsification method was remarkably floated and unstable in suspension performance, but the suspension of droplets obtained by the first emulsification and then gelation was stable and did not float (as shown in fig. 10).
Claims (9)
1. A method of making suspended micro-droplets using a microfluidic device, comprising the steps of:
(1) any one or a mixture of a plurality of oily substances is selected as the inner phase,
(2) dissolving surfactant and thickener in solvent to obtain shear-thinned solution as external phase,
(3) injecting the internal phase prepared in the step (1) and the external phase prepared in the step (2) into an internal phase inlet and an external phase inlet of the microfluidic device respectively through a syringe; at the outlet of an internal phase channel of the microfluidic device, an internal phase solution forms micro-droplets under the combined action of the surface tension and the viscous force of a two-phase solution, then a solution for gelatinizing an external phase is introduced at the outlet of an external phase channel of the microfluidic device, the gelatinized external phase solution has the property of shear thinning, and the internal phase droplets can be kept at fixed positions in a three-dimensional space of the external phase for a long time under the condition of no shear force, so that suspended micro-droplets dispersed in the external phase are obtained; the solution for gelling the external phase is an aqueous sodium hydroxide solution.
2. The method of claim 1, wherein the oily substance is one or more of silk oil, cashmere ester, lavender essential oil, or vitamin E.
3. The method for preparing suspended micro-droplets of claim 1, wherein in step (2), the solvent is water.
4. The method for preparing suspended micro-droplets of claim 1, wherein in step (2), the surfactant in the external phase is polyvinyl alcohol or sodium dodecyl sulfate.
5. The method of claim 1, wherein in step (2), the thickener in the external phase is carbomer, and is present in the external phase at a level of at least 0.06% by weight.
6. The method for preparing suspended micro-droplets according to claim 1 or 4, wherein in step (2), the surfactant in the external phase is 1 to 10% by mass in the external phase when the surfactant is polyvinyl alcohol, and 0.5 to 1% by mass in the external phase when the surfactant is sodium lauryl sulfate.
7. The method of claim 1, wherein the microfluidic device is a glass capillary microfluidic device prepared by the following steps:
(1) preparing a glass tube: preparing a round glass capillary tube and a square glass capillary tube, ultrasonically cleaning the glass capillary tube and drying;
(2) stretching the glass tube: processing one end of a round glass capillary tube into a taper shape by using a stretcher;
(3) treating the capillary: rubbing one end of a conical tip of the round glass capillary tube by using abrasive paper to flatten an end opening;
(4) preparing a device: and (3) placing one end of the processed pointed cone of the round glass capillary into the square glass capillary, and then fixing all the glass capillaries on the glass sheet by using glue.
8. Suspended micro-droplets produced by the method according to any one of claims 1 to 7.
9. The suspended micro-droplets of claim 8, wherein the suspended micro-droplets are of uniform and controllable size and are stably suspended in the aqueous phase for a prolonged period of time.
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