System for flushing out clogged particles from an HPLC column

Abstract

The invention relates to a HPLC workstation (500) for identification and quantification of components of an eluent. The HPLC workstation comprises a HPLC column (400, 500) configured for separation of the components of the eluent flowing through it. The HPLC column comprises a first side (402), wherein the first side is configurable as an inlet and an outlet for the eluent; a second side (404), wherein the second side is configurable as the inlet and the outlet for the eluent; and a flow direction (406), wherein the flow direction configures one of the first side and the second side as the inlet and the other side as the outlet for flowing of the eluent through the column; an injection valve (504) connected to the HPLC column through both the first side and the second side, wherein the injection valve is configured for reversing the flow direction in the HPLC column through a switching mechanism; a pump (516) configured to create a mobile phase in the column by flowing the eluent through the HPLC column at a specific flow rate and increasing the flow rate in a predetermined order and at a predetermined interval; wherein the eluent flow rate is increased by the pump to flush out the eluent particles clogged on one of the first and the second side of the HPLC column.

Description

[0001] SYSTEM FOR FLUSHING OUT CLOGGED PARTICLES FROM AN HPLC COLUMN

[0002] FIELD OF THE INVENTION

[0003] The disclosure herein relates to a system for the improvement of HPLC workstation. In particular, a system for enhancing the HPLC column lifetime while minimizing the maintenance time and cost is described.

[0004] BACKGROUND OF THE INVENTION

[0005] High-performance liquid chromatography (HPLC) is an analytical chemistry technique used to separate compounds in a chemical mixture. HPLC helps to identify each component of the chemical mixture and quantify each component. The separation technique utilizes a pressure-driven flow of a mobile phase through a column packed with a stationary phase.

[0006] A schematic overview of the HPLC system 100 is illustrated in Fig. 1A. The HPLC system comprises a solvent 102 needed to create a mobile phase which is needed to transport the samples through the HPLC system. The mobile phase is a phase in motion and composed of solvent or eluents flowing from injection to detection. The mobile phase is created through a pump 104 which moves the solvent continuously from the solvent flask 102 to a waste can 112. An injection system injects the samples into the mobile phase. The injection system comprises an injection valve 106 into which the samples are injected by an injection needle 108.

[0007] The separation of the components happens in a column 110. When the sample reaches the column 110, it leaves the mobile phase and enters a stationary phase. The stationary phase is the substance fixed in place in the column 110 for the chromatography procedure. The components or analytes separate in the column 110 due to varying degrees of interaction with the stationary phase. Fig. 1 B illustrates a column 100’ with the separated components 120, 122 and 124 of a sample.

[0008] The separated components are transmitted through a detector 112 after leaving the stationary phase in the column 110. The detector 112 detects the components and sends the signals to a software on a computing device 116 which converts it in a chromatogram 118.The software used for process is generally known as chromatography data system (CDS) which produces the chromatogram 118 from the detected signals.

[0009] A chromatogram 118 comprises an x-axis which represents a measure of time and a y-axis which measures a specific signal generated by the detector 112. A sample chromatogram 200 of Enthone SnAg bath is illustrated in Fig. 2A. The chromatogram 200 shows the quantities of different components of the Enthone SnAg bath as “Inorganics”, “Complexer”, “Leveler”, “Antioxidant” and “Refiner”. Another sample chromatogram 200’ of Mitsubishi SnAg is illustrated in Fig. 2B. The chromatogram 200’ illustrates quantities of various components “Antioxidant”, “TS-140AD” and “TS-SLG Polymeric group”.

[0010] HPLC provides the following benefits:

[0011] • Ability to analyze / quantify multiple components / compounds in a single run.

[0012] • Direct analysis / quantification of the component(s) of interest.

[0013] • Fast analysis - single run often takes <30min.

[0014] Inspite of the abovementioned advantages, HPLC has its own disadvantages. The HPLC system uses expensive hardware, especially the column. Further, it has a complicated methodology and any issue raised is difficult to troubleshoot for the operators. For this, advanced knowledge of the hardware and operating principles is required. Moreover, the components of interest are often proprietary involving difficult development methods. Most importantly, the problem is caused by the use of dirty samples. The turbid and particulate filled samples cause hardware failure requiring frequent maintenance.

[0015] HPLC column 110 is the most important part of the system. It comprises the following features:

[0016] • Different manufacturers and prices

[0017] • Different dimensions (i.e. ID / OD, length, etc.)

[0018] • Different selectivity (i.e. C8 / C18 / Phenyl / etc.)

[0019] • Different base structure (silica based / monolithic / etc.)

[0020] • The component(s) of interest directs the column selection and application• The method / application development optimizes the interactions between the stationary phase (column packing) and the mobile phase (eluent - water / solvent), to yield good separation / resolution between the components of interest.

[0021] • The job of the column is to retain / release the component(s) of interest separately.

[0022] • The column (and detection method) selection and optimization are paramount.

[0023] A standard HPLC column 302 is illustrated in Fig. 3A. The column 302 comprises an inlet 304 and an outlet 306. From a hardware point of view there is no difference between the column inlet 304 and the outlet 306. All manufacturers specify a flow direction on columns. The column 302 has the flow direction 308 marked by the manufacturer. The flow and analysis can be run in reverse direction with proper and complete flushing during / after analysis. By running the next analysis in reverse, the flow and gradient itself also flushes out any accumulation of particulates at the column inlet.

[0024] There are two main causes for column damage or degradation. Firstly, clogged column due to particles leading to pressure increase. Secondly, material fatigue & wear of the column separation material leading to peak broadening.

[0025] A conventional method to increase the column lifetime is illustrated in Figs. 3B and 3C. A basic structure of the column 300’ is illustrated in Fig. 3B. The column 300’ comprises a main column 310 and a guard column 312. The flow direction 314 is also indicated. The guard column 312 protects the main column 310 from particles mixed in the solvent as shown in Fig. 3C. The particles 316 are filtered by the guard column 312 while a small amount of particles 318 reaches the main column. If the pressure is too high the guard column 312 can be replaced. This saves money and increases the main column 310 lifetime. However, there are sample types where the particles are so small that these cannot be filtered by the guard column 312. The exemplary sample types can be galvanic tin, tin / silver palladium, nickel, lead, copper electroplating baths, etc. These particles can clog the expensive column over time, so it needs to be replaced frequently.There is a need for a method or a system which can remove particles to minimize pressure increase within the column and increase the column lifetime, especially with turbid samples. Further, the system should eliminate the need for costly pre-column or guard column to lower the HPLC system cost. The invention described herein addresses the above-described needs.SUMMARY OF THE EMBODIMENTS

[0026] In one aspect of the invention, an HPLC workstation with flow reversal and column backflush functionality is disclosed. The workstation reverses the eluent flow and flush out the particles clogged at the inlet of the HPLC column to mitigate / minimize pressure increase and to increase the column lifetime, especially with turbid samples. This enables low operating cost and lower frequency of maintenance of the system.

[0027] In another aspect of the invention, an HPLC workstation for identification and quantification of components of an eluent is disclosed. The HPLC workstation comprises an HPLC column configured for separation of the components of the eluent flowing through it. The HPLC column a first side and a second side configurable as an inlet and an outlet for the eluent flowing through the column and a flow direction which configures one of the first side and the second side as the inlet and the other side as the outlet for flowing of the eluent through the column. The HPLC workstation further comprises an injection valve connected to the HPLC column through both the first side and the second side, wherein the injection valve is configured for reversing the flow direction in the HPLC column through a switching mechanism. The workstation also comprises a pump configured to create a mobile phase in the column by flowing the eluent through the HPLC column at a specific flow rate and increasing the flow rate in a predetermined order and at a predetermined interval. The eluent flow rate is increased by the pump to flush out the eluent particles clogged on one of the first and the second side of the HPLC column.

[0028] As appropriate, during a first run, the first side is configured as the inlet and the second side is configured as the outlet of the HPLC column, wherein the eluent particles are clogged at the inlet of the HPLC column.

[0029] As appropriate, during a second run, the injection valve reverses the HPLC column direction by switching the inlet and the outlet of the HPLC column and the pump increases the eluent flow rate to flush out the clogged particles from the HPLC column.

[0030] As appropriate, the flushing out of the clogged particles mitigates or minimizes the pressure increase within the column.As appropriate, the flushing out of the clogged particles reduces maintenance and increases the column lifetime.

[0031] As appropriate, the eluent can be one or more of galvanic tin, tin / silver palladium, nickel, lead, copper electroplating baths, or a combination thereof.

[0032] In another aspect, the HPLC workstation further comprises a pressure detector or a transducer configured to monitor the pressure in the HPLC column.

[0033] In a further aspect, the HPLC workstation further comprises a flow detector configured to monitor the flow limit in the HPLC column.

[0034] In yet another aspect, the HPLC workstation further comprises a UV detector configured to monitor the flushed particles from the HPLC column.

[0035] As appropriate, the eluent flow rate is increased by the pump until a pressure limit or a flow limit is reached in the HPLC column.

[0036] As appropriate, the eluent flow rate is stepwise increased by the pump.

[0037] As appropriate, the steps are same or different for every increase in the eluent flow rate.

[0038] As appropriate, the eluent flow rate is linearly or non-linearly increased by the pump.

[0039] As appropriate, the eluent flow rate is constant for a predetermined period and then increases stepwise, linearly or non-linearly.

[0040] As appropriate, the HPLC column is a Monolithic column.

[0041] According to another aspect of the invention, An HPLC column of an HPLC workstation for flushing out the clogged particles of an eluent flowing through it is disclosed. The HPLC column comprises a first side and a second side configurable as an inlet and an outlet for the eluent and a flow direction, wherein the flow direction configuresone of the first side and the second side as the inlet and the other side as the outlet for flowing of the eluent through the column.

[0042] As appropriate, during a first run of the eluent through the HPLC column, the first side is configured as the inlet and the second side is configured as the outlet for the eluent and the eluent particles are clogged at the inlet of the HPLC column

[0043] As appropriate, during a second run, the flow direction in the HPLC column is reversed by an injection valve through a switching mechanism.

[0044] As appropriate, an eluent flow rate in the HPLC column is increased in a predetermined order and at a predetermined interval by a pump to flush out the clogged particles from the HPLC column.

[0045] As appropriate, the HPLC column is configured for separation of the components of the eluent flowing through it.

[0046] As appropriate, the flushing out of the clogged particles mitigates or minimizes the pressure increase within the column.

[0047] As appropriate, the flushing out of the clogged particles reduces maintenance and increases the column lifetime.

[0048] As appropriate, the eluent can be one or more of galvanic tin, tin / silver palladium, nickel, lead, copper electroplating baths, or a combination thereof.

[0049] As appropriate, the eluent flow rate is increased by the pump until a pressure limit or a flow limit is reached in the HPLC column.

[0050] As appropriate, the pressure in the HPLC column is monitored by a pressure detector or a transducer.

[0051] As appropriate, the flow limit in the HPLC column is monitored by a flow detector.

[0052] As appropriate, the flushed particles from the HPLC column are monitored by a UV detector.As appropriate, the eluent flow rate is stepwise increased by the pump.

[0053] As appropriate, the steps are same or different for every increase in the eluent flow rate.

[0054] As appropriate, the eluent flow rate is linearly or non-linearly increased by the pump.

[0055] As appropriate, the eluent flow rate is constant for a predetermined period and then increases stepwise, linearly or non-linearly.

[0056] As appropriate, the HPLC column is a Monolithic column.

[0057] According to yet another aspect of the invention, a method for flushing out the clogged particles of an eluent flowing through an HPLC column of an HPLC workstation is disclosed. The method comprises configuring a flow direction of the HPLC column by configuring a first side as an inlet and a second side as the outlet for flowing the eluent through it and running the analysis with the configured flow direction at a first flow rate, wherein the eluent particles are clogged at the inlet during the analysis. The method further comprises reversing the flow direction by switching an injection valve and running the analysis in the reverse direction at a second flow rate. The method also comprises increasing the second flow rate in the HPLC column in a predetermined order and at a predetermined interval by a pump to flush out the clogged particles from the HPLC column.

[0058] As appropriate, the second flow rate is same as the first flow rate.

[0059] As appropriate, increasing the flow rate further comprises determining if a pressure limit or a flow limit is reached in the HPLC column.

[0060] As appropriate, increasing the flow rate comprises stepwise increase in the flow rate.

[0061] As appropriate, increasing the flow rate comprises linear or non-linear increase in the flow rate.As appropriate, the method further comprises measuring a column pressure and UV absorbance of the eluent.

[0062] BRIEF DESCRIPTION OF THE FIGURES

[0063] For a better understanding of the embodiments and to show how it may be carried into effect, reference will now be made, purely by way of example, to the accompanying drawings.

[0064] With specific reference now to the drawings in detail, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of selected embodiments only and are presented in the cause of providing what is believed to be the most useful and readily understood description of the principles and conceptual aspects. In this regard, no attempt is made to show structural details in more detail than is necessary for a fundamental understanding; the description taken with the drawings making apparent to those skilled in the art how the various selected embodiments may be put into practice. In the accompanying drawings:

[0065] Fig. 1A illustrates a schematic overview of the HPLC system 100 as known in the art;

[0066] Fig. IB illustrates a standard HPLC column 100’ with separated components of a sample as known in the art;

[0067] Fig. 2A illustrates a sample chromatogram 200 of Enthone SnAg bath;

[0068] Fig. 2B illustrates a sample chromatogram 200’ of Mitsubishi SnAg;

[0069] Fig. 3 A illustrates a standard HPLC column 300 with a predefined flow direction; Fig. 3B illustrates basic structure of a column 300’ with a main column and a guard column as known in the art;

[0070] Fig. 3C illustrates the column 300” with the mixed particles filtered by the guard column as known in the art;

[0071] Fig.4A illustrates a column 400 with one end configured as an inlet 402 and another end configured as an outlet 404 according to an aspect of the invention;Fig. 4B illustrates a column 400’ with the eluent flow direction reversed;

[0072] Fig. 4C illustrates a column 400” with the clogged particles back flushed from the main column;

[0073] Fig. 5A illustrates an HPLC workstation 500 according to an aspect of the invention;

[0074] Fig. 5B illustrates an HPLC workstation 500 with the flow direction in the column in a forward direction;

[0075] Fig. 5C illustrates an HPLC workstation 500 with the flow direction in the column in a reverse direction;

[0076] Fig. 6 illustrates a flowchart showing method steps for reverse flow backflush according to an aspect of the invention;

[0077] Fig. 7 A illustrates the graphical analysis 700 of pressure vs time for standard and slip streams for different HPLC columns; and

[0078] Fig. 7B illustrates the graphical analysis 700’ of the flow reversal test.DESCRIPTION OF THE SELECTED EMBODIMENTS

[0079] Aspects of the present disclosure relate to a method and a system for the improvement of HPLC workstation. The system reverses the eluent flow and flush out the particles at the inlet of the column to mitigate / minimize pressure increase and to increase the HPLC column lifetime. An additional injection valve is installed in the standard HPLC hardware to reverse the flow direction. The flow is increased stepwise from the reverse direction to flush out the clogged particles. The flow is increased until a pressure limit or a flow limit is reached.

[0080] In particular embodiments of the system, the UV absorbance of the eluent is measured to monitor the flushed particles and contaminants. The pressure within the column is detected to check the pressure dip when the particles are flushed out and to avoid exceeding a certain pressure limit given by the system. A detector is also used to detect the flushing out of particles.

[0081] As required, the detailed embodiments of the present invention are disclosed herein; however, it is to be understood that the disclosed embodiments are merely examples of the invention that may be embodied in various and alternative forms. The figures are not necessarily to scale; some features may be exaggerated or minimized to show details of particular components. Therefore, specific structural and functional details disclosed herein are not to be interpreted as limiting, but merely as a representative basis for teaching one skilled in the art to variously employ the present invention.

[0082] It is particularly noted that the systems and methods of the disclosure herein may not be limited in its application to the details of construction and the arrangement of the components or methods set forth in the description or illustrated in the drawings and examples. The systems and methods of the disclosure may be capable of other embodiments, or of being practiced and carried out in various ways and technologies.

[0083] Alternative methods and materials similar or equivalent to those described herein may be used in the practice or testing of embodiments of the disclosure. Nevertheless, particular methods and materials described herein for illustrative purposes only. The materials, methods, and examples are not intended to be necessarily limiting. Accordingly,various embodiments may omit, substitute, or add various procedures or components as appropriate. For instance, the methods may be performed in an order different from described, and that various steps may be added, omitted or combined. In addition, aspects and components described with respect to certain embodiments may be combined in various other embodiments.

[0084] Reference is now made to Fig. 4A, which illustrates a column 400 with one end configured as an inlet 402 and another end configured as an outlet 404 according to an aspect of the invention. From a hardware point of view there is no difference between the column inlet and outlet. The inlet and the outlet are defined by the flow direction. The flow direction 406 is shown from the inlet 402 towards the outlet 404. The column is clogged with particles 408 at the inlet 402. The particles are from the samples which can be galvanic tin, tin / silver palladium, nickel, lead, copper electroplating baths, etc. It should be noted that any other metallic, non-metallic or alloy-based eluent can be used as a sample in the HPLC analysis without limiting the scope of the invention.

[0085] Fig. 4B illustrates the column 400’ with the eluent flow direction reversed. The flow direction 406’ configures the column end 404 as the inlet and the other end 402 as the outlet. The reverse flow 410 from the inlet 404 towards the outlet 402 backflushes the outlet 402 and flushes out the most of the clogged particles 408 as shown in Fig. 4C.

[0086] The flow direction is reversed by using an additional injection valve installed in the standard HPLC hardware as shown in Fig. 5 A which illustrates an HPLC workstation 500 according to an aspect of the invention. The HPLC workstation 500 comprises an additional injection valve 504 connected to the HPLC column 502 through both sides. The column 502 is configured to support flow direction in both forward and reverse directions. The flow direction in the column 502 reversed by switching the additional injection valve 504. Fig. 5B illustrates the flow direction in the column 502 in a forward direction through the injection valve 504. Fig. 5C illustrates the flow direction in the column 502 in the reverse direction through the injection valve 504.

[0087] Referring to Fig. 6 which shows a flowchart illustrating method steps for reverse flow backflush. The process starts at step 602 and an HPLC workstation 500 is provided with an HPLC column 502 with a defined flow direction at step 604. The column 502 isconfigured to support flow direction in both forward and reverse directions. At step 606, the analysis is run with the flow direction by creating a mobile phase in the column 502 through a pump 516 which moves the solvent continuously from the solvent flask 506 to a waste can 518 as shown in Fig. 5B. The additional injection valve 504 is switched in a forward direction and the samples are run through the column 502 in the forward direction. The sample particles are clogged in the column 502 in the inlet part at step 608 as shown in the Fig. 4 A. At step 610, the column 502 flow is reversed by switching the additional injection valve 504 and the samples are run through the column 502 in the reverse direction through the pump 516 as shown in Fig. 5C. The gradient is switched to the aqueous eluent. This saves solvent usage and increases the pressure due to higher eluent viscosity. The gradient is switched by a gradient mixture 508 which performs the process of gradient elution. Gradient elution refers to a technique of altering the composition of the mobile phase during the course of the chromatographic run. Gradient elution is used when a mixture of eluents with a wide range of retention factors is to be separated. The analysis is run through the HPLC workstation 500 in the reverse direction at step 612. The pressure and the UV absorbance of the eluent is measured at step 614. The pressure in the HPLC column is monitored by a pressure transducer 510. The UV absorbance of the eluent is measured by a UV detector 512 to monitor the flushed particles and contaminants from the column 502.

[0088] At step 616, the analysis is run at a constant flow rate for a predefined time period. In a particular embodiment, the reverse flow rate is kept the same as the forward flow rate. In an exemplary embodiment, initially, the flow is pumped at the normal analysis flow rate of 1 ml / min and the pressure within the column is measured. Any lower or higher flow can also be taken as the starting point. The flow is kept constant for a predefined time interval, for example, 30 seconds. Any shorter or longer time interval can also be selected for analysis. At step 618, the flow rate is increased in a particular order, for example, the flow rate is increased stepwise, linearly, non-linearly, and so on.

[0089] In a particular embodiment of the present invention, the flow is increased stepwise to increase the effectiveness of backflush and flush out all the clogged particles. The flow is increased at a particular step, for example, 0.5 ml / min. Any lower or higher step can also be selected for analysis. The flow is increased stepwise until a pressure limit or a flow limitis reached. Alternatively, in stepwise flow increase, the flow gradient need not be constant and can increase linearly. For example, the flow is increased at 0.5 ml / min during the first cycle, at 1 ml / min in the next cycle, at 1.5 ml / min in the next cycle and so on until the pressure limit or the flow limit is reached. Further alternatively, the flow gradient might be kept constant for certain flow cycles and then increased for the subsequent cycles. For example, the flow is increased at 0.5 ml / min for the first three cycles and then increased at 1 ml / min for the next three cycles and so on.

[0090] In alternative to the stepwise flow increase, a linear or non-linear flow change is also possible. For example, the flow is increased at the constant rate of 0.5 ml / min until the pressure limit or the flow limit is reached. Alternatively, the flow is increased non-linearly at 0.5 ml / min for first one minute, then at 1 ml / min for the second minute, then at 1.5 ml / min for the third minute, and so on until the pressure limit or the flow limit is reached.

[0091] It should be noted that the above-mentioned methods for flow increase are exemplary in nature and should not limit the scope of the invention. The flow can be increased or decreased using any suitable methods.

[0092] At step 620, the workstation 500 checks if the pressure limit or flow limit is reached. The pressure limit is checked by the pressure transducer 510 and the flow limit is checked by the flow detector 514. If the pressure limit or flow limit is reached, the system 500 checks if HPLC analysis is completed at step 622. If the HPLC analysis is completed, the process stops at step 628. If the pressure limit and the flow limit is not reached at step 620, the process goes back to step 612. In case the analysis is not completed at step 622, the next analysis is started at step 624. The analysis is run either in the same flow direction as above or in the opposite direction by switching the additional injection valve 504 at step 626. The process then goes back to step 614.

[0093] In a preferred aspect of the present invention, a monolithic HPLC column is used. The monolithic HPLC column is used in high-performance liquid chromatography (HPLC). The internal structure of the monolithic column is created in such a way that many channels form inside the column. The material inside the column which separates the channels can be porous and functionalized. Monolithic columns can be broken down into two categories, silica-based and polymer-based monoliths. Silica-based monoliths areknown for their efficiency in separating smaller molecules while, polymer-based are known for separating large protein molecules. The use of Monolithic column in flow reversal and column backflush is more effective as the column is not packed with small particles, and the package inside the column if fixed and cannot move or build voids due to the forward / backward flow direction.

[0094] Referring now to Fig. 7 A which illustrates the graphical analysis 700 of pressure vs time for standard and slip streams for different HPLC columns. As evident from the graph, the column pressure is higher before the start of the backflush function 702 in the column. After the start of the backflush function 702, the pressure remains unstable for a short duration and then becomes lower and stable in the column. The pressure increase within the column is mitigated or minimized due to flush out of the particles at the inlet of the column as a result of the backflush function. This significantly increases the column lifetime, especially with turbid samples.

[0095] Fig. 7B illustrates the graphical analysis 700’ of the flow reversal test. During the first run 704 in the forward direction, the sample particles are clogged at the inlet of the column resulting in the end of lifetime of the column only after few subsequent runs 706. The flow direction is reversed in graph 708 by flipping the column inlet and outlet using the additional injection valve 504 resulting in almost doubling of the column lifetime in graph 710 by flushing out the clogged particles and reducing the column pressure.

[0096] The HPLC workstation of the present invention with Flow reversal and column backflush functionality can remove particles to minimize pressure increase and significantly increases the column lifetime, especially with turbid samples. Further, the system does not require the precolumn or the guard column reducing the workstation, cost.

[0097] The HPLC workstation of the present invention is also advantageous for the customers by lowering the system operating cost due to longer column lifetime. It also lowers the tool downtime and the frequency of maintenance. Consequently, this results in lower customer frustration generating more profitable business for him.

[0098] While the preferred embodiment of the present invention and its advantages has been disclosed in the above detailed description, the invention is not limited there to but only by the scope of the appended claim.As will be readily apparent to those skilled in the art, the present invention may easily be produced in other specific forms without departing from its essential characteristics. The present embodiments are, therefore, to be considered as merely illustrative and not restrictive, the scope of the invention being indicated by the claims rather than the foregoing description, and all changes which come within therefore intended to be embraced therein.

Claims

I / WE CLAIM:

1. An HPLC workstation (500) for identification and quantification of components of an eluent, the HPLC workstation comprising:an HPLC column (400, 500) configured for separation of the components of the eluent flowing through it, the HPLC column comprising:a first side (402), wherein the first side is configurable as an inlet and an outlet for the eluent;a second side (404), wherein the second side is configurable as the inlet and the outlet for the eluent; anda flow direction (406), wherein the flow direction configures one of the first side and the second side as the inlet and the other side as the outlet for flowing of the eluent through the column;an injection valve (504) connected to the HPLC column through both the first side and the second side, wherein the injection valve is configured for reversing the flow direction in the HPLC column through a switching mechanism;a pump (516) configured to create a mobile phase in the column by flowing the eluent through the HPLC column at a specific flow rate and increasing the flow rate in a predetermined order and at a predetermined interval; wherein the eluent flow rate is increased by the pump to flush out the eluent particles clogged on one of the first and the second side of the HPLC column.

2. The HPLC workstation of claim 1, wherein during a first run, the first side is configured as the inlet and the second side is configured as the outlet of the HPLC column, wherein the eluent particles are clogged at the inlet of the HPLC column.

3. The HPLC workstation of claim 2, wherein during a second run, the injection valve reverses the HPLC column direction by switching the inlet and the outlet of the HPLC column and the pump increases the eluent flow rate to flush out the clogged particles from the HPLC column.

4. The HPLC workstation of claim 3, wherein the flushing out of the clogged particles mitigates or minimizes the pressure increase within the column.

5. The HPLC workstation of claim 3, wherein the flushing out of the clogged particles reduces maintenance and increases the column lifetime.

6. The HPLC workstation of claim 1, wherein the eluent can be one or more of galvanic tin, tin / silver palladium, nickel, lead, copper electroplating baths, or a combination thereof.

7. The HPLC workstation of claim 1 further comprising a pressure detector or a transducer (510) configured to monitor the pressure in the HPLC column.

8. The HPLC workstation of claim 1 further comprising a flow detector (514) configured to monitor the flow limit in the HPLC column.

9. The HPLC workstation of claim 1 further comprising a UV detector (512) configured to monitor the flushed particles from the HPLC column.

10. The HPLC workstation of claim 1, wherein the eluent flow rate is increased by the pump until a pressure limit or a flow limit is reached in the HPLC column.

11. The HPLC workstation of claim 1, wherein the eluent flow rate is stepwise increased by the pump.

12. The HPLC workstation of claim 11, wherein the steps are same for every increase in the eluent flow rate.

13. The HPLC workstation of claim 11, wherein the steps are different for every increase in the eluent flow rate.

14. The HPLC workstation of claim 1, wherein the eluent flow rate is linearly or non- linearly increased by the pump.

15. The HPLC workstation of claim 1, wherein the eluent flow rate is constant for a predetermined period and then increases stepwise, linearly or non-linearly.

16. The HPLC workstation of claim 1, wherein the HPLC column is a Monolithic column.

17. An HPLC column (400, 500) of an HPLC workstation (500) for flushing out the clogged particles of an eluent flowing through it, the HPLC column comprising: a first side (402), wherein the first side is configurable as an inlet and an outlet for the eluent;a second side (404), wherein the second side is configurable as the inlet and the outlet for the eluent; anda flow direction (406), wherein the flow direction configures one of the first side and the second side as the inlet and the other side as the outlet for flowing of the eluent through the column;wherein during a first run of the eluent through the HPLC column, the first side is configured as the inlet and the second side is configured as the outlet for the eluent and the eluent particles are clogged at the inlet of the HPLC column,wherein during a second run, the flow direction in the HPLC column is reversed by an injection valve (504) through a switching mechanism, andwherein an eluent flow rate in the HPLC column is increased in a predetermined order and at a predetermined interval by a pump (516) to flush out the clogged particles from the HPLC column.

18. The HPLC column of claim 17 is configured for separation of the components of the eluent flowing through it.

19. The HPLC column of claim 17, wherein the flushing out of the clogged particles mitigates or minimizes the pressure increase within the column.

20. The HPLC column of claim 17, wherein the flushing out of the clogged particles reduces maintenance and increases the column lifetime.

21. The HPLC column of claim 17, wherein the eluent can be one or more of galvanic tin, tin / silver palladium, nickel, lead, copper electroplating baths, or a combination thereof.

22. The HPLC column of claim 17, wherein the eluent flow rate is increased by the pump until a pressure limit or a flow limit is reached in the HPLC column.

23. The HPLC column of claim 22, wherein the pressure in the HPLC column is monitored by a pressure detector or a transducer (510).

24. The HPLC column of claim 22, wherein the flow limit in the HPLC column is monitored by a flow detector (514).

25. The HPLC column of claim 17, wherein the flushed particles from the HPLC column are monitored by a UV detector (512).

26. The HPLC column of claim 17, wherein the eluent flow rate is stepwise increased by the pump.

27. The HPLC column of claim 26, wherein the steps are same for every increase in the eluent flow rate.

28. The HPLC column of claim 26, wherein the steps are different for every increase in the eluent flow rate.

29. The HPLC column of claim 17, wherein the eluent flow rate is linearly or non-linearly increased by the pump.

30. The HPLC column of claim 17, wherein the eluent flow rate is constant for a predetermined period and then increases stepwise, linearly or non-linearly.

31. The HPLC column of claim 17, wherein the column is a Monolithic column.

32. A method for flushing out the clogged particles of an eluent flowing through an HPLC column of an HPLC workstation, the method comprising:configuring (604) a flow direction of the HPLC column by configuring a first side as an inlet and a second side as the outlet for flowing the eluent through it;running (606) the analysis with the configured flow direction at a first flow rate, wherein the eluent particles are clogged at the inlet during the analysis; reversing (610) the flow direction by switching an injection valve; running (612) the analysis in the reverse direction at a second flow rate; increasing (618) the second flow rate in the HPLC column in a predetermined order and at a predetermined interval by a pump to flush out the clogged particles from the HPLC column.

33. The method of claim 32, wherein the second flow rate is same as the first flow rate.

34. The method of claim 32, wherein increasing the flow rate further comprises determining (620) if a pressure limit or a flow limit is reached in the HPLC column.

35. The method of claim 32, wherein increasing the flow rate comprises stepwise increase in the flow rate.

36. The method of claim 32, wherein increasing the flow rate comprises linear or nonlinear increase in the flow rate.

37. The method of claim 32 further comprising measuring (614) a column pressure and UV absorbance of the eluent.

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