SY-8100 Manual operacion

8/9/2019 SY-8100 Manual operacion

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Users’ HandbookFor

SY-8100 HPLC System

and

software of workstation


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SY-8100 HPLC Chromatograph

CONTENT

CHAPTER ONE PREFACE

SOP’s in this manual...............................................................................................1

Unpacking...............................................................................................................1

Packing list..............................................................................................................1

CHAPTER TWO HPLC PUMP

Brief descriptions of HPLC Pump ..........................................................................2

1. Front view of HPLC Pump ..............................................................................2

2. Rear panel view of the HPLC pump................................................................3

Operation of HPLC pump.......................................................................................4

1. Connection of power supply ............................................................................4

2. User’s page characteristics and operation........................................................4

Simple Maintenance: ..............................................................................................7

1. Replacing and purging the pump head.............................................................7

HPLC pump technical parameters ........................................................................10

CHAPTER THREE UV DETECTOR

Brief descriptions of UV Detector ........................................................................11

1. General description ........................................................................................11

2. Optical path of the UV detector .....................................................................11

Preparation of UV detector before the operation:.................................................12

1. Front view of UV detector .............................................................................12

2. Rear view of UV detector ..............................................................................14

3. Power supply, ON/OFF, and self-test.............................................................14

Operation of UV detector......................................................................................15

1. Internal software structure and menu explanation .........................................15

2. Installation of flow cell ..................................................................................17

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Simple Maintenance..............................................................................................18

1. Lamp maintenance .........................................................................................18

2. Cleaning the flow cell ....................................................................................18

3. Flow cell.........................................................................................................19

Technical data of UV detector ..............................................................................20

CHAPTER FOUR CONNECTION

Connection of detector to HPLC system ..............................................................21

Connection of UV detector with computer...........................................................22

CHAPTER FIVE WORK STATION controlled instrument

Brief instructions of work station .........................................................................23

1. Characteristics of work station.......................................................................23

2. Running environment of work station software.............................................23

3. Integral environment of work station.............................................................24

Operation of work station .....................................................................................25

1. Preparation of work station before the operation...........................................25

2. Operation procedures of work station............................................................27

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CHAPTER ONE PREFACE

Thanks for purchasing Model SY-8100 Liquid Chromatograph developed by our company.

This device includes HPLC Pump,UV detector and software of work station. Read this manual

carefully before the operation.

SOP’s in this manual

The Standard Operation Procedures (SOP) provided with this manual offer a convenient way of

structuring complex tasks in the operation of your HPLC pump and UV detector. They include

step-by-step instructions leading the user through all routine tasks during the operation. They can

be used for documentation purpose and be copied, applied signed, and filed in order to document

the performance of the instrument.

Please operate the instrument and all accessories according to instructions and SOP’s in this

manual. This ensures proper results and longevity of the instrument.

Unpacking

Unpack the instrument and check the device and accessories thoroughly for any damage that may

have occurred during transport. If necessary, put forward any claim for damages to the carrier

and inform of your local dealer immediately.

Use packing list to check the device delivery is complete. Please contact your local dealer if you

miss something or if you need support. Please fill out the guarantee registration card and return it

to the local dealer immediately.

Remove the transport protections from the display.

Packing list

Packing list (Standard delivery) Quantity

HPLC Pump 1Set

UV Detector 1Set

Standard flow cell 1Pc.

Start-up kit,analytical 1Set

Operation manual 1Copy

Power cable 220V/10A 2Pcs.

RS232 Cable 1Pcs.

Software of work station 1Set

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CHAPTER TWO HPLC PUMP

Brief descriptions of HPLC Pump

Front view of HPLC Pump

Fig. 1 Front view of Pump

1-Display information area 2- Foil key area 3-Pump front door

Display area and key area (1 and 2 in Fig.1)

Fig 2 Display area and key area

1-Display information area 2.1 Pump START/STOP

2.2-Purge START/STOP 2.3-Numeric key pad

2.4-Arrow keys and ENTER key

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Operation of HPLC pump

HPLC pump is controlled by keyboard and work station of the computer control. The following

describes the operation of the keyboard. The work station of the computer control is described in

the Chapter four.

Connection of power supply

HPLC Pump is equipped with a universal power supply for input voltage 220±20V Ac,

frequency 50 ±0.5Hz. 1KVA stabilizer is required. Connect the power cable to the socket on

the rear panel of the instrument and switch on the device with the ON/OFF key, see (4.1) and

(4.3) in Fig.4.

CAUTION: Make sure that the main power supply is properly grounded and a

correct three-pronged power cable is used. Connecting the instrument to faulty power

sockets can cause damages.

2 User’s page characteristics and operation

Press the key in 2.2 of Fig.2 at any time to make system purge, again press this key to stop

purging.

CAUTION: If the purge is required, first loose “PEEK screw by hands”, see 4.2 in

Fig3, otherwise chromatographic column will be damaged!

Self-test page

After switching on the instrument, firstly enter into the self-test status, display the self-test page

as shown the follows:

Page of switching on

BFRL SY-8100 HPLC PUMP

RAM TEST OK

BFRL SY-8100 HPLC PUMP

EEPROM TEST OK

FLOW►0.00ml P.max 19.0 MPa A

PRES 0.0mPa P.min 0.0 MPa STOP

Turn on the switch of the pump on this page. After completing the self-test of the pump, the

above page is appeared. Press 2.1 key in Fig.2 to make the pump START/STOP.

CAUTION: Never run the HPLC Pump without liquid in the pump head or in the piston

backflushing compartment. Operating the pump without solvent may lead to damage of the

pump seals

SOP 1:

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Press “ENTER” key, move the triangle cursor among “Flow”, “P. max.” and “P. min.”

As shown in Fig., the triangle cursor points “Flow”, use the numeric key for flow input, again

press “ENTER” key to confirm the setting. Therefore, the system flow is transferred to the flow

value of setting.

As the same operation, press “ENTER” key to move the cursor to “P. max.” and “P. min”.

The data value enables to be renewed with the same operation.

Press the arrow “▲”and“▼”keys to enter into the other pages.

Vocabulary explanation:

Flow － Current flow of pump

P. max. － Maximum pressure allowable for the system

A Pump － Series number of Pump

Pressure －Current pressure of pump

P. min. －Minimum pressure of system

Stop －Running status of pump（When pump stops, display“Stop”, when

running, display“Run”）

Gradient mode page

GE MODE: High-Pressure Gradient A

1: HP. GE A 2: HP. GE B 3: LP. GE

This page is the selection page of the gradient mode of the current pump

Press the numerical keys 1, 2 or 3, again press ENTER key to select the corresponding gradient

mode.

SOP 2

Press 1, 2 or 3 keys, again press “ENTER” key to select the corresponding gradient mode.

Press“▲” and“▼”keys to enter into the other pages.


HP. GE A － Pump located in A pump system of binary high pressure system

HP. GE B － B pump in binary high pressure system

LP. GE － Pump in quarter low pressure gradient system.

During the operation of high pressure gradient and low pressure gradient, the corresponding

setting is required in “SY8000 chromatograph work station to realize the work station to fully

control the instrument.（See the control section in the operation manual of data work station of

chromatograph. Pressure zeroing page

Pressure zero Adj

ENTER to zero adj

This page is the pressure zeroing page. There is no need for the user to operate it. User can press

“ENTER” key to adjust the pressure zeroing unless “PEEK screw is loosen and the pressure

value does not display the zero in the page of switching on the instrument.

SOP 3

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Press “ENTER” key to adjust the pressure to be the zero.

Press“▲”and“▼”keys to enter into the other pages.


Pressure zero Adj － Current pressure display value in the system is adjusted

to be the zero.

Status page

This page is a using status page of current pump, the user can not operate it any more.

SOP 4

Press“▲”and“▼”keys to enter into the other pages.


SN － Production series of the instrument, not series No. of the delivery

Total time － Accumulated using time of current pump.

Total volume － Accumulated transfusion volume of current pump.

Remark page

SN: 815299 Total time 0.1 (H)

Total volume 0.1 (L)

BFRL HPLC PUMP

SN. 815299 Rev: 1.7

This page shows Model of HPLC Pump, Series No and Version No. It is not necessary for

the user to operate it.

SOP 5

Press“▲”and“▼”Keys to enter into the other pages.


BFRL HPLC Pump －Model of instrument

SN. 815299 －Production number of instrument

Rev: 1.7 －Software version No.

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Simple Maintenance:

Replacing and purging the pump head

1.1 Disassembling the pump head

CAUTION: If the organic residential is remained in the pump head, it may cause the

skin irritation..

Use proper cleaning reagent to purge the pump head, and then to purge it with the distilled water.

After so, `the replacement for the pump head is made.

SOP 6:

1. Loosen“Eluent outlet”, see Fig.3（3.5），Loosen inlet valve housing, see Fig. 3(3.4).

2. Use 3#（3mm）hexagonal spanner to loose “fixing screws of pump head “on the two

slanting angles, see Fig. 3 (3.3). Remove them.

3. Carefully loosen the rest two screws about half a turn, alternating from one to the other

to avoid the damaging the pump head. Unscrew the two screws strictly alternating due

to strong force of springs, it may be help to either clamp the pump head or to press one

of its side surfaces against a table with one hand and remove it.

4. Carefully remove the pump head

Disassembling and Check piston

SOP 7

1. Detach the pump head.

2. The piston is shown as Fig.5（11）. A clipper (nipper) is available for detaching it.

Use the clipper to clamp the metal material part of the piston end and carefully pull it out along

on the straight line.

CAUTION: If the piston is broken the pump head must be thoroughly checked for the damage.

1.2 Detaching the pump head

1) Detach the pump head.

2) Detach the piston.

3) Loosen the two “retaining plate screws” as shown in Fig.5（20），half a turn,

alternating from one to the other to avoid damaging the “retaining plate”.

4) Because the two screws are very tight, it may be help to either clamp the pump head or

to press one of its side surfaces against a table with one hand while loosening the

screws.

5) Unscrew the screws strictly alternating due to strong force of the spring behind the plate

as shown in Fig. 5（10）and remove them.

6) Dismount the retaining plate, see Fig.5（12）.

7) The moment, remove “spring guide sleeve，see Fig.5（19）and “spring:”.

8) Use the spinner to loosen “ retaining nut” as shown in Fig.5（9）, half a turn alternately.

9) Remove “Rear pump head” as shown in Fig 5（17），and take out（15） and（16）.

10)Insert the piston into “high pressure seal ring”, and then pull it out. The moment, the

seal ring is adhered on the piston and take it out.

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图 5 Disassemble diagram of the pump head and parts sequence.

Ser. No Part’s Name

1 Retaining screw of pump head

2 Front pump head

3 Front connection tube

4 High pressure seal ring

5 Stainless steel block ring

6 O- ring

7 Rear purge tube

8 Low pressure seal ring

9 Retaining bolt

10 Spring

11 Piston

12 Retaining plate

13 Valve sleeve

14 Single way valve

15 Plastic baffle ring

16 Rear purge tube

17 Rear pump head

18 Bushing

19 Guide for spring

20 Screw of retaining plate

Table 1 Pump head

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1.3 Assembling the pump head

All positions of components of the pump head refer to Fig 5

SOP 8

1. Detach the pump head to check it. After solving the problems, mount it into (4），（15），

and（5）at the front pump head in regular sequence.

2. After mounting the pump head, piston the head in a straight line onto the pump

housing (9），and then take the “piston” out .

3. Mount “spring guide sleeve and spring”, see Fig.5（19）and (10).

4. Install “retaining plate”, see Fig.5（12）.

5. Insert and tighten the two screws strictly alternating due to strong force of the spring

behind the plate.

6. Alternating from one to the next, tighten two screws half a turn at a time on the

retaining board so as not to damage it.

7. Carefully insert the piston rod, see Fig.5（11）without bending or quenching the rod..

8. Both “the spacing bolts” and “retaining board screws “must be tightened securely

as before.

1.4 Installing the pump head

SOP 9:

1. Make sure that the pump head has been correctly assembled.

2. Piston the head in a straight line onto the pump housing..

3. Tighten all four pump head set screws a few turns by hand, see Fig.5（10）.

4. Alternating from one to the next, tighten two diagonally opposed screws half a turn at a

Time until the pump head is correctly seated

5. Tighten the two remaining screws. Make sure that all four pump head set screws are

Securely tightened, see Fig. 5 (1).

6. Mount the capillary connection between the pump head outlet and the pressure

transducer.

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HPLC pump technical parameters

Flow rate range: 0.01～9.99 ml/min

Structure of Pump: Dual piston rod, series type oscillating

displacement pump

Flow accuracy


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CHAPTER THREE UV Detector

Brief descriptions of UV Detector

General description

BFRL UV detector is a valuable new type detector ，especially developed for the routine analysis.

The deuterium lamp is used as the light source of the instrument.

UV detector selects the flow cell of standard type（Optical path:10mm，volume:10µL，the flow

rate range of eluent phase of UV detector is available from 0.001～9.999mL/min.

After powering up, the UV detector performs the self test and calibration automatically.

The internal software includes an auto-calibration of the wavelength scale. The

position of the grating reflection (0. order) will be located automatically.

The deuterium lamp with the wavelength of 656nm is used to check and to calibrate

the display.

The distance of the wavelength passed through the monochromator should be 1nm for per

step, arbitrary setting can be made in the range of 190nm to 740nm. Accuracy is ±2nm.

In addition, UV detector still have the function of the whole wavelength or the wavelength

range set by user to process stopping the flow scanning.

UV detector provides with very low noise（2×10-5

AU）and shift（15×10-5

AU/h），available

for auto-zeroing of the full scale.

Key pad is clear, clean film, convenient for completing the operation.

The control and data acquisition of the UV detector is digitalization to secure running

without the trouble.

2 Optical path of the UV detector

1. Deuterium lamp

2. Slit

3. Filter

4. Concave mirror

5. Plane grating monochromator

6. Half transparent mirror

7. Flow cell

8. Photodiode (Sample)

9. Photodiode (Reference)

9. 参考光电二极管

图 6 Optical Path of UV detector

Instructions for optical path

A beam of emitted light from the deuterium lamp（1）is faded out by a slit（2）. After passing a

filter （3） it is focused by a concave mirror （4）on a plane grating monochromator （5），so

as to obtain a beam light of characteristic wavelength. The bream is then split by a half

transparent mirror （6）. One part beam delivers the reference signal（9），and the other after

pathing the flow cell（7）irradiates to photodiode (8) to produce the measurement signal.

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Preparation of UV detector before the operation:

Front view of UV detector

1.Display information area 2.Foil key pad 3.UV detcector front door

Fig.7 Front panel of UV detector and flow cell

Display area and foil key pad

Fig. 8 Display area and foil key pad

1. Display area 2. Control function key

3.Input key 4. Arrow key and ENTER key

Information display area

ABS－0.0001 AU LAMP IS LIT

WAVE 254 nm

Fig. 9 Display information area

Information display area displays real time parameters and result data of UV detector.

Foil key pad area

Foil key pad includes control function key（Auto-zeroing key and running program key）,Data

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input key, arrows keys and ENTER key.

Auto-zeroing

Press this key to adjust the baseline of the instrument（ baseline returns to zero）.

Normally press this key before the analysis. Use sampling valve of signal cable

(7725i) equipped with the instrument for sampling, the work station will

automatically control to process the zeroing adjustment for once at the same time

the valve is changed over. The moment, there is no need for user to press this key

again.

Running program

Press this key to run the wavelength scanning program set by user.

Auto

Zero

Prog

Data input key

As shown in Fig.8－3. Data input key is used to modify the operation parameters.

At the same time, CE key is set at the key area（CANCEL key）. It is used to

cancel error parameters being input.

Arrow keys and ENTER key

As shown in Fig.8－4. Arrow keys are used to turn over the pages. “Enter” key is

used to confirm currently entered or selected parameters.

Flow cell area

Fig. 10 Flow cell area and flow cell

1. Knurled cell screw 2. Flow cell cradle

3. Inlet liquid 4. Cell body

5. Outlet liquid

Inlet liquid of the flow cell is connected to column, outlet liquid is connected to waste liquid

bottle or collecting device. Every constituent separated by the column is entered into the flow

cell to make the determination.

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2 Rear view of UV detector

Fig. 11 Rear view of UV detector

1.RS—232 interface 2.Remote terminal

3. Analog output 4. 24V ventilator

5.12V ventilator 6. Power switch

7., power socket 8. Slow melting fuse（1A）

3 Power supply, ON/OFF, and self-test

UV detector operates with 220±20V AC of 50±0.5Hz. 1A slow melting fuse is used. The

setting is done by the manufacture on custom request. The actual setting is indicated on the

rear panel of the instrument (Fig. 12).

Caution:

Cut off power supp ly before opening cover.

220±20V 50±0.5 Hz

100VA

FUSE: 1A SLOW

Fig. 12 Indication of the input voltage

CAUTION: Make certain that the correct voltage has been set on the rear panel of the

instrument, the power supply is reliably grounded and a corresponding 3-pole power cable

is used.

Connect the detector to the power supply. Switch on the instrument. Power switch is on the rear

part of the instrument.

After switching on, the instrument first performs the self-test as follows:

BFRL UV/VIS DETECTOR

RAM TEST OK

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Afterwards, display model of instrument and simultaneously the deuterium lamp is lit.

Power on the instrument a time, the wavelength should be calibrated for once, display as

follows:

After a certain period of zeroing calibration（about one minute）, display as follows：

The detector enters into the following page afterward. This page is also a acquiesce page of

UV detector ：

15 minutes later, UV detector is ready for the analytical determination.

Operation of UV detector

Internal software structure and menu explanation

The software is divided into several menus. Settings and operation modes are made on every

menu. Use“▲”and “▼”keys to enter into each menu as shown in Fig.13.


EEPROM TEST OK >


Pre heating ＞

ABS －0.0001 AU LAMP IS LIT

WAVE 254 nm

ABS －0.0001 AU LAMP IS LIT

WAVE 254 nm

RESPONSE 1.00 （0.05－10）S

Press [ENTER] TO Change


Check Home Pos ＞


Wave calib ＞

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D2 LAMP ENERGY SMPL 0.794583

USED TIME 50.3 h REF. 0.401205

SERIAL 825365 Option

1:D2 OFF 2：WAVE CALIB [Enter]


SN. 825282 Rev: 1.70

Fig. 13 Menu sequence of UV detector

Describe every menu in details as follows. When the parameter on the menu is required to

modify, use the data input key to modify the setting of the corresponding parameter. Thus press

the “ENTER” key，the moment the value just input is confirmed. If error parameter is input, use

“CE”key to delete it, then input the correct one according to the procedures mentioned above.

Signal menu

ABS -0.0001AU LAMP IS LIT

WAVE 254nm

Signal menu（or main menu）includes: absorbance（signal intensity），D2 lamp status and

wavelength. Absorbance is in unit of “AU”. D2 lamp status shows “ON” and “OFF”. The

wavelength is in unit of “nm”.

The signal value for real measurement displays in four digits. The wavelength range is from 190

to 740nm for arbitrary setting.

Response time menu

RESPONCE 1.00 （0.05－10）S

Press [Enter] TO Change

Use the response time (time constant) to make the signal smooth. Press ENTER key to select the

input. Respectively set: 0.05，0.10，0.15，0.20，0.25，0.30，0.35，0.40，0.45，0.50，0.55，

0.60，0.65，0.70，0.75，0.80，0.85，0.90，0.95，1.00，1.50，2.00，2.50，3.00，3.50，4.00，

4.50，5.00，6.00，7.00，8.00，9.00，10.00sec. The value is more bigger, the signal is more smooth.

Time constant of 1second. is satisfied with the most of the analysis requirements.

D2 lamp energy and using time menu

D2 lamp energy menu displayed is the energy values of ultraviolet light received from the

D2 LAMP ENERGY SMPL 0.794583

USED TIME 50.3 h REF 0.401205

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photodiode on the reference signal board and the sample of the detector. These two energy

values can be directly known the energy of the deuterium lamp. If these energy values show too

small, replace D2 lamp on time.

The used time means the ignition time of D2 lamp.

Series No. and operation function menu

Series No. means production series No. of the detector, not delivery series No.

Operation function menu includes:“1：OFF lamp”,“2：Wavelength calibration”. Respectively

input the numeric number corresponded to the operation function under this menu. Press

“Enter ”key to complete the corresponding function. For example: Complete the operation of

switching off the lamp, press the corresponding digit“1”under this menu and then press“Enter ”

key to finish the operation of OFF lamp. As the same, complete the operation of the wavelength

calibration, press the corresponding digit“2”on this menu，after so, press“Enter ”key to finish

the operation of the wavelength calibration.

Identification menu

SERIAL 825365 Option

1：D2 OFF 2：WAVE CALB [Enter]


SN. 825282 Rev: 170

Identification menu displays model and name of the instrument，simultaneously display

production series number of the instrument and software version No. It is not necessary for the

user to modify any item in the identification menu.

2 Installation of flow cell

The flow cell of UV detector has not been delivered during transport. There is a dummy flow cell

without any of optics equipped with the instrument. Before the detector is used, a proper

standard flow cell must be installed.

CAUTION: There is no need to change the status of the instrument and the lamp

during the replacement of the flow cell.

SOP 1 Installation of flow cell

This instruction refers to UV detector as shown in Fig10 flow cell area and flow cell (see the

preparation of UV detector before the operation in the concerned section).

a) Loosen the two knurled cell screws (1) by hand and remove them.

b) Pull out the flow cell cradle (2).

c) Take the cell or dummy cell with two figures and remove it upward.

d) Insert the new flow cell and make sure engraved specification towards the user

(….can be read) and the fixing hole on the back side of the cell meets the

corresponding metal pin.

e) Now push the complete system towards the housing, insert the two screws and

tighten them manually.

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Simple Maintenance

Lamp maintenance

The deuterium used with UV detector has an extended lifetime to ensure long-time functionality

and reliable measurements with low noise and baseline drift as well as high sensitivity. (life time

about 2000 hours). The actual using time of the deuterium lamp depends on different factors, like

the number of lamp starts, the average burning time and your requirements concerning noise and

sensitivity.

To check the functionality of the lamp, the two intensity values of sample and reference（see“D2

lamp energy and used time” menu），the help information for D2 lamp can be found there. The

reference value refers to the intensity of the light measured in the reference channel and be used

for checking the quality of the lamp.

We recommend to check the reference value at regular intervals under the conditions mentioned

above（dummy cell,λ= 240nm）. This applies especially in case higher noise levels or decreased

sensitivity is observed on working with UV detector. In these observations coincide with a

reference value of approx. 0.1 or less, or used time is over 2000h, a new deuterium lamp should

be installed.

2 Cleaning the flow cell

Noisy baselines and low sensitivities may be due to a dirty flow cell. This may also be indicated

by a low absorbance value in the signal menu when flushing the cell with pure solvent. That

shows the flow cell has been polluted. In most cases it is sufficient to purge the flow cell

according to the following SOP.

SOP 1 Purging the flow cell

This instruction applies to the flow cell delivered with UV detector

a) Purge the flow cell using one of following solvents: sodium dodecyle sulfate

(SDS), 1m HCL, 1m NaOH, ethanol, or acetone.

b) Run the solvent through the flow cell using a syring and leave for approximately 5

minutes.

c) Rinse extensively with water and the blow dry using a gentle stream of pure

nitrogen.

CAUTION: Never dry with compressed air to blow the flow cell, this will contain

microdroplets of oil that will coat the cell.

CAUTION: When the optics module is not in use, disconnect the flow cell and clean

outer traces of salt and protein with a syringe filled with distilled water.

Before storing the flow cell inject a dilute solution (10-25%) of ethanol or

i-propanol to prevent microbial growth.

CAUTION: In case the flow cell purging do not provide sufficient success, all flow cells

can easily be disassembled for cleaning the lenses.

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3 Flow cell

Fig.17 Sectional view of flow cell

SOP 2 Cleaning the flow cell

This instruction applies to the flow cells enclosed in UV detector.

a) Unscrew the outer threads with 3mm hexagonal spanner enclosed in the flow

cell’s delivery.

b) Remove the black gasket that carries the lenses with a pair of tweezers or by

gently tapping it on a clean surface. The lens is embedded in the gasket and sealed

against the flow path with PTFE seal. This seal should be changed every time

when assembling the flow cell.

c) Take out the lenses and clean them by wiping them with a soft cloth or with an

appropriate solvent in an ultrasonic bath. Be careful 取 not to touch the lenses

with the figures.

d) Reassemble the cell in the reverse manner, making sure that PTFE seal does not

block the light path.

e) Tighten the outer threads carefully with the spanner in order not to damage the

lenses.

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CHAPTER FOUR Connection

Connection of detector to HPLC system

CAUTION：Before taking a measurement cell filled with fluid into operation, please

make certain that the reagent used is miscible with that one used

previously. Or purge the flow cell with a medium miscible and two

miscible reagents.

The connection of the fluid piping in the isocratic system is shown in the following Fig.

Convenient for the instruction, the pump head represents HPLC pump and UV detector shows as

flow cell.

图 15 Isocratic HPLC system

Brief description of fluid system：

a) Flow path of fluid：As shown above the Fig, Fluid is aspirated from sampling inlet

by HPLC pump, then flow into the sampling valve (2) from the fluid outlet,

afterwards, from the sampling valve (3) to the column. The fluid is entered into

the flow cell of the detector coming from the column. At last enter into the waste

liquid bottle or collection device.

b) Flow path of sample：As shown in Fig 16, see the operation mode of the 7725i

sampling valve，The operation of 7725i sampling valve is divided into two parts:

preparation status and sampling status.

Preparation status Sampling status

Fig. 16 Operation mode of 7725i sampling valve

Preparation status: The pump is directly connected with the column（connection

of hole 2 with hole 3），and the needle hole is connected with the sample ring. Themoment, use the sampling needle to aspirate 2-3 times sample (or more, to

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achieve the higher accuracy) of sample ring volume to inject it into the sample

ring. The residual sample is drained off through the discharge tube which is

connected with the hole 6.

Sampling status: The pump is connected with the sample ring (hole 1 and hole

2)，Purge all of samples from the sample ring to the column. Needle port is

connected with the discharge tube (hole 5).

The sample under the sampling status is brought into the column by the fluid for

the separation.

Connection of UV detector with computer

UV detector is connected with computer by RS232 interface. The analytical result enables to

display in the software of HPLC chromatograph by this interface. The software of HPLC

chromatograph can be also realizing the modification of the detection parameters by this

interface..

RS-232 series interface is located on the back of the instrument as shown in Fig.11（see the

section described in the preparation of UV detector before the operation）. Use this interface to

connect the computer installed the software of HPLC chromatograph with UV detector.

The series interface of the computer is directly connected with this interface.

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Better 64M

One free series interface on the system board (COM1 and COM2)

VGA Color display (better use an accelerated card of graphics)

Optical drive

Mouth

Printer

Software: Better Windows 20000 operation system

3 Integral environment of work station

Double-click the graphic of chromatograph work station on the table to enter into the intergral

environmental page of work station.

Main menu Prompt button

Information line

Spectrogram display window

Running indicating lamp

Fig 1

The main page of the integral environment consists of five parts: main menu, graphic display

window, prompt button, controlling menu, running indicating lamp and information line.

Main menu

As same as the other application program, according to the different requirement of treating data,

all of the function commands are classified, after so, respectively put them into eight menus, so

every menu has own group commands.

Prompt button

Some common used functions in the main menu are marked on these buttons with display

presentation. Therefore, these functions in common use can be directly selected by pressing the

concerned button without selecting it from the menu in order to speed up the operation.

Spectrogram display window: including plotting area, time coordinate scale, level coordinate

scale and peak hold time.

Plotting area Actively display the real time spectrogram area, the sizes can be automatically

adjusted with the size change of the spectrogram display window. When the user makes thespectrogram display window shortening to a certain extent, the system automatically delete the

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plotting coordinate scale and display of the hold time to secure observing the graphics clearly

even in a small size window.

Time coordinate scale——It is used as X axle coordinate scale, in unit: Min. The initial value is

automatically adjusted with the change of the sampling time. Modify the full time difference on

“screen plotting parameter edition”, minimum time is 1min.

Level coordinate scale——It is used as Y axle coordinate scale of the spectrogram plotting, in

unit : Mv. Modify the full scale maximum value and minimum value on “screen plotting

parameter”.

Peak hold time——During starting sampling, automatically display the hold time after

appearing the chromatograph peak top. In the “screen plotting parameter edition”, select “no

display the hold time” to close it.

Running indicating lamp

The red lamp is flash when the data is acquiring, while the green lamp is lit under the state of

preparation

Information line

It displays currently inputting level, sampling time, current sample name and current time.

As the main page mentioned above, when you select some command in the main menu or

single-click some of prompt button, the corresponding window or dialogue block will be

appeared.

Operation of work station

Preparation of work station before the operation

Before operating the work station, SY-8000 series HPLC Chromatograph should be connected

with the computer installed the work station software. Power the instrument on. When

completing the self test of the instrument, the computer displays the following page, That shows

the connection of the computer is successful.

Fig 2Current setting flow Current pressure of

Success for the connection Current Wavelength

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After success for the connection of the instrument with the computer, the operation page of the

work station simultaneously display current setting flow speed and current pressure of pump and

current wavelength of UV detector.

If the instrument failed to connect to the instrument, under “instrument control” menu, make

setting of communicating terminal and let the terminal number of the setting be in accord with

the terminal number connected with the instrument shown as the following figure:

Setting of communicating terminal

Fig. 3

After success for the connection of the instrument with work station, firstly click “Pump” or

“Detector” on the work station, check the setting of working parameters of the pump and

detector as shown in the following:

Setting of pump

Fig. 4

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Setting of detector

Fig. 5

The pump requires to set the control mode (control mode divides: single pump, double pump

running independently, double pump high pressure gradient and single pump low pressure

gradient), If using gradient table, pump flow and up limit and down limit of pressure; Detector

requires to set the wavelength and deuterium lamp status, if using the wavelength program table,

etc. The sensitivity of the detector is set by the acqueuiescence. The spectral scanning is set

according to the concerning conditions when it is required.

2 Operation procedures of work station

Use the work station software to do the experiment after the settings mentioned above. As

example for outer standard method as follows:

Operation procedures of outer standard method

1. Create a new method file parameter to do the experiment. Its procedures are as follows:

1) Start HPLC chromatograph and work station software;

Fig. 6

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2) Record parameters of editing data

Signal-click the graphic of “data record parameter” to appear the window of the method editing,

set the position of file storage.

Fig.7

3) Editing integral parameter

Single-click the graphic of “integral parameter” to appear the window of the method edition.

Where set the integral parameter value and edit “ time program table”, If it is not set any, only

press the acquiescent value.

Fig. 8

4) Edit “sample information” table

Click the graphic of “ sample information”, edit “sample information table, define the name of

the experimentalized data file as “sample name” as shown in the following:

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Fig.9

5) Standard sample anslysis

When running the chromatograph stable, input the standard sample. During valve sampling, at

the same time, the program automatically process the data acquisition to the spectragram, and

remark the hold time. When all of peaks are appeared, stop the program manually, or set “ stop

time” in “integral parameter ” table to stop it automatically.

6) Sample analysis

Input the unknown sample. As same as the procedures 5. (this step can be made at last.) Data file

obtained from the procedures 5 and 6 will be saved automatically by the system.. The data files

are automatically, continuously named. Such as: Test001.dat, Test002.dat, Test003.dat……..

7) Draft the calibration curve

a. Edit the quantitative parameter

Click the graphic of “ the quantitative parameter” to appear the window of the method

edition:

Fig. 10

Set well “quantitative mode”, “quantitative method (outer standard method), “ standard

( eight concentration standard sample, select “ eight points”), “standard concentration

unit” and “sample result unit”.

b. Filling in constituent table

Click the graphic of “constituent table” to appear the window of the method edition, fill in

the analytical sample constituent table name, hold time and series standard concentration.

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Fig.11

c. Definition calibration curve

a)

Click the graphic of “calibration curve” to appear the window of the method edition.

Fig.12

b) Definite spectrogram peak file required at every point. Firstly use the mouth to move

to the point number of the standard sample, then click the right key, this number is

covered by blue frame, again click the left key to select the menu:

Fig.13

c) Select “ Add” item (this operation shows at the same concentration point, add the data

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of several parallel experiment to calculate the mean value), the window of

“spectrogram selection” is appeared.

Fig.14

Here select the corresponding spectrogram file. “+” appears before the file of sequence

number point. After completing the definition foe every points, click “re-read peak file” to

display the calibration curve.

8) Check sample result

Enter “re-treatment” page, click the graphic of “open data file” to appear the window of

“spectrogram selection”, select the sample spectrogram file here as follows:

Fig.15

And then click the button of “re-calculate”, Again click the graphic of “screen report” to look at

the screen report table.

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Fig.16

9) Printing report

2. Recall the method file parameters originally saved to do the experiment.

1) Use the mode of “open method”

Select “open method” in “file” menu of work station, appear the window of “method file

selection”, select the method file here as shown as follows:

Fig.17

Sampling is directly made after recalling the method file without the operation procedures from

1-7. After the experiment, the step 8 is made directly to look at the sample content.

2) Use “mode of “open method from data”

Select “open method from data” in “file” menu of work station (it is another mode for recalling

the method file, only recall the method file with the date), appear the window of “spectrogram

selection”, the moment, select the method file here as shown as the follows:

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Fig.18

The descriptions as same as the method above mentioned, Sampling analysis can be made after

recalling the method file without the operation procedures from 1-7 steps. The procedure of step

8 is made after completing the experiment in order to directly obverse the sample content.

SY-8100 Manual operacion

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