operation instruction manual

UV/Visible Spectrophotometer

T8DCS

Op

era

tion

Instru

ction

Man

ual

- 2 -

Chapter 1 - IntroducƟŽŶ Packaging Contents 2UV-Vis Spectroscopy 2UV-Vis Spectrophotometer 2Double Beam Spectroscopy 2Performing Measurements with a Double Beam Instrument 2Measurement CondiƟons 3

Choice of Wavelength 3Absorbance Range 3Bandwidth SelecƟŽŶ 3

Instrument DescripƟon 4Front View 4Rear View 5Side View 5Cell Holder 5

Instrument ^ƉĞĐŝĮĐĂƟŽŶ 6

Chapter 2 - Instrument InstallaƟŽŶ Environmental ConsiĚĞƌĂƟŽŶƐ 7Computer PreparaƟŽŶ and ^ŽŌǁĂƌĞ /ŶƐƚĂůůĂƟŽŶ 7Instrument /ŶƐƚĂůůĂƟŽŶ 7^ƚĂƌƟŶŐ The Instrument and ^ŽŌǁĂƌĞ 8Before Analysing any Samples 9

Chapter 3 - ^ŽŌǁĂƌĞ InsƚĂůůĂƟŽn and ŽŶĮŐƵƌĂƟŽŶ ^ŽŌǁĂƌĞ /ŶƐƚĂůůĂƟŽŶ 10^ŽŌǁĂƌĞ ŽŶĮŐƵƌĂƟon 11

Chapter 4 - UVWin SoŌǁare Chapter 5 - ApplicĂƟŽn Examples

Example of a Photometric Measurement 13Example of a Spectral Measurement 14Example of a YƵĂŶƟƚĂƟve Measurement 16

Chapter 6 - OƉƟŽŶĂl Accessories CH19-1 TheƌŵŽƐƚĂƟĐ Cell Holder 19DS19-1 Angle Adjustable Solid Sample Holder 20IS9-1 InteŐƌĂƟŶŐ Sphere 21LS19-1 Long Pathlength Cell Holder 22MH19-1 Micro Cell Holder 23MH19-2 Ultra Micro Cell Holder 24MR19-1 Specula ReŇecƟŽŶ Accessory 25PS19-2 Sipper Pump 26S19-1 Solid Sample Holder 27SS19-1 Short Pathlength Cell Holder 28TR19-1 Test Tube Holder 29

T8DCS UV-Vis Spectrophotometer User Manual

- 3 -

Chapter 7 - Maintenance Replacement of the W (Tungsten) Lamp 30Replacement of the D2 (Deuterium) Lamp 32Fuse Replacement 33


- 4 -

Chapter 1 - IntroducƟŽŶ Packaging Contents On receipt of your PG Instruments Spectrophotometer please ensure that the box contents are thoroughly checked against the packing list to ensure completeness. In the event of the instrument being damaged during shipment or any of the box contents missing please contact your local distriďƵƟŽŶ agent immediately. UV-Vis Spectroscopy UV-Vis spectroscopy is an analyƟcal method used to measure the absorbance of ultra-violet or visible radiaƟon through an analayte. The molecular ĂďƐŽƌƉƟŽn of the analayte corresponds to both excitaƟon of valence electrons and excitaƟon of electrons in diīerent atomic orbitals. UV-Vis Spectroscopy is an eīecƟve technique for both qualitaƟve and quanƟtaƟve analysis of organic and inorganic compounds. UV-Vis spectroscopy is based on the Lambert-Beer principle which states that the Absorbance of a ƐŽůƵƟŽn (A) is directly prŽƉŽƌƟŽŶĂů to its pathlength (l) and its concentraƟon (c) when the wavelength of the incidence light remains Įxed. This is summarized in the following equaƟŽŶ whereɸis the molar absorbƟvity.

ClA ..H UV-Vis Spectrophotometer

The UV-Vis Spectrophotometer is the analyƟcal instrument used for the UV-Vis spectroscopic analysis. Spectrophotometers are available in diīerent ĐŽŶĮŐƵraƟŽŶs however most can be categorized into either single beam or double beam types depending on the design of their opƟcal system. Such types of instrument comprise the following components in their construcƟŽŶƐ

x Light Source x Monochromator x Cell Compartment x Detector x Signal Processing System

Double Beam Spectroscopy The double beam approach to UV-Vis spectroscopy requires two beams of light both having the same intensity to measure the Absorbance through sample and reference ƉŽƐŝƟŽŶƐ simultaneously. The Sample ƉŽƐŝƟŽn is used for measurement of the analyte whereas the reference posiƟon is used for the correcƟŽn against a blank ƐŽůƵƟŽn or sample matrix. A clear advantage of the double beam opƟcal system is the improvement in measurement stability and precision as a result of having a real-Ɵme feedback of both the reference and sample signals. The opƟcal schemaƟc shown in Įgure 1 clearly demonstrates the diīerence between and single beam and double beam instrumenƚ in that the sample compartment accommodates both the sample and reference ƐŽůƵƟŽŶƐ allowing a direct measurement to be performed.

Performing Measurements with a Double Beam Instrument Due to the diīerent opƟcal design of a double beam instrument a slightly diīerent approach can be taken when performing sample measurements. When using a single (or split) beam instrument it is necessary to Įrst perform an Autozero by means of placing your blank soluƟon in the sample beĂŵ the blank soluƟon can then be replaced by the sample soluƟon and the measurement can be performed. Double beam instruments allow for measurement of both the sample and reference ƐŽůƵƟŽŶs simultaneously hence it is possible to place the sample in the sample beĂŵ the blank in the reference beam and measure directly. It is considered good in the interest of ensuring that you obtain an accurate Autozero that a blank ƐŽůƵƟŽn is placed in both the sample and reference beam and an Autozero be performed prior to performing sample measurements. Figure 2 shows the sample compartment of the T92+ spectrophotometer clearly showing the two ƉŽƐŝƟŽŶƐ for sample and reference.


- 5 -

Figure 1

Figure 2

Measurement

Choice of Wavelength

When choosing a suitable wavelength for analyzing the sample, considera ons of the nature of the sample must be made. It is

common prac ce to use the strongest abso on ( maxO ) line for quan ta on. This concept is p cularly well suited to samples

with low concentra on. When measuring samples with high concentra on, in order to ensure that measurements are performed within a linear working range, it may be more appropriate to perform the measurement at a less sensi ve n peak.

Absorbance Range

In order to obtain accurate and stable measurement results it is important to ensure that measurements are performed within a linear working range. The nature of the rela onship between absorbance and transmission is such that at high absorbance values, linearity is decreased. Care should also be taken when measuring at p cularly low Absorbance values. In prac ce steps, such as sample di and pathlength adjustment, can be taken to ensure that the sample is measured within a linear working range.

Bandwidth Sel on

In cases were sample spectra show adjacent peaks it may be advantageous to reduce the instrument bandwidth in order to increase the spectral re n power. When selec g instrument bandwidth other considera ons need to be made. Wide instrument bandwidths will reduce instrument sensi vity and re . Narrow bandwidths will eīec vely decrease the signal


- 6 -

intensity to which the sample is exposed which could potenƟally deteriorate the signal to noise raƟo. SelecƟŽn of instrument bandwidth needs to be a compromise between achieving the required resoluƟon power whilst ensuring no other measurements characterisƟĐƐ are jeopardized.

The T92+ series spectrophotometer features the “ConƟŶƵŽƵƐůy variable bandwidth feature”. This diīers from convenƟŽnal variable bandwidth spectrophotometers in as much as they have pre-deĮned bandwidth seƫŶŐƐ. The conƟŶƵously variable nature of the bandwidth seleĐƟŽn on the T92+ allows for sĞůĞĐƟŶŐ the bandwidth anywhere in the range 0.1nm to 5nm with an interval of 0.1nm.

Instrument DescripƟŽŶ A descrŝƉƟŽn of the instrument appearance is as follows and as indicated in Įgure 3:

Figure 3

FrŽnt View

A summary of the front view of the instrument is as follows, and image of which can be seen in Įgure 4:

1. Instrument casework – produced from a high intensity reinforced Įberglass.

2. Sample compartment front baŋe – to be removed with the use of certain instrument accessories.

3. Instrument Chassis – Highly strengthened steel ensures opƟcal stability and measurement precision.

4. Instrument model number label.

5. Sample compartment cover –giving access to the sample compartment and any opƟonal accessories that might be installed.

Figure 4

1

2

3

4

5


- 7 -

Rear View A summary of the rear view of the instrument is as follows, and image of which can be seen in ĮŐƵre 5:

1. VenƟlaƟon Fan.

2. USB Interface for communicaƟŽn with PC.

3. Mains Inlet – 115/230V AC @50Hz.

4. Fuse 1.

5. Fuse 2.

Figure 5

Side View A summary of the side view of the instrument is as follows, and image of which can be seen in ĮŐƵre 6:

1. Lamp Compartment.

2. Power Switch.

Figure 6

Cell Holder The cell holder should be cŽŶĮgured to suit the accessory ĮƩed in the sample compartment. The Standard cell holder for The T92+ series spectrophotometer has Įxed posiƟŽŶs for both Sample and Reference beams, however diīerent accessories are available and will require appropriate ƐŽŌware ĐŽŶĮguraƟŽn to be made. Figure 7 shows the standard 10mm pathlength cell accessory for the T92+.

Figure 7

2

1

4

3

5

2

1


- 8 -

Instrument SpeciĮcaƟŽŶ

Parameter ^ƉĞĐŝĮĐaƟŽn Wavelength Range 900nm190nm Wavelength Accuracy ±0.3nm Wavelength ReprŽĚƵĐibility ч0.1nm Spectral ĂŶĚǁŝĚƚŚ 0.1nm – 5nm (0.1nm increments) PŚŽtŽŵetric Accuracy ±0.3%t (0100%T) PŚŽtŽŵetric ReprŽĚƵĐibility ч0.1 %t PŚŽtŽŵetric ƌŝŌ ч0.0004 Abs/h (2hr ǁarm-up, kineƟĐ scan at 500nm)

Table 1 The speciĮcaƟŽŶƐ quŽteĚ in Table 1 are referenceĚ tŽ measurements perfŽrmeĚ ǁith a 2nm spectral baŶĚǁŝĚth. SpeciĮcaƟŽŶƐ may vary at Žther baŶĚǁŝĚth seƫŶŐƐ. These specŝĮcaƟŽŶs are alsŽ subject tŽ change ǁŝƚŚŽƵƚ nŽƟĮcaƟŽŶ


- 9 -

Chapter 2 - Instrument InstĂůůĂƟŽŶ EŽte: For detailed installaƟŽn insƚƌƵĐƟŽŶƐ in regulated laboratories please refer to the T92+ series ƋƵĂůŝĮcaƟon workbook. The

user manual provides a brief overview to the installaƟŽn only.

EnvirŽnmental ŽŶƐŝĚĞraƟŽŶƐ The T92+ series spectrophotometer (hereaŌer referred to as the T92 or the Instrument) is a versaƟle high-precision laboratory instrument. In order to ensure proper performance and longevity of its operaƟon environmental consideraƟŽŶƐ should be made prior to installaƟon. As summary of the most important environmental condiƟons are as follows:

x Temperature…..15 - 35C x RelaƟve Humidity….. ч70% x Power Supply…..115 / 230V AC @50Hz. x The instrument should be placed on a stable and Ňat work surface free from vibraƟŽŶ x The instrument should not be exposed to toxic or causƟc liquids or gases. x The instrument should not be exposed to electromagneƟĐ radiaƟŽŶ

ŽŵƉƵter PreparaƟŽŶ anĚ ^ŽŌware InstallaƟŽŶ As the T92+ is considered to be a high speciĮcaƟŽn spectrophotometer amongst the PG Instruments product range it is supplied as standard with the UVWin soŌware package for instrument control and data acquisiƟŽŶ.

Prior to instrument setup it is recommended to setup the PC intended for use with the UVWin soŌware. For instrucƟons on the setup of your PC please refer to manufacturer’s documentaƟŽŶ

Once successful compleƟon of the PC installaƟŽŶ, it is recommended to install the UVWin soŌware. The process of installing the UVWin soŌware is extremely simple and follows a typical Windows installaƟŽn Wizard.

Run Setup from the installaƟon directory and follow the instrucƟons indicated in the installaƟŽn wizard.

During the installaƟŽn the wizard will ask you to enter a soŌware serial number. The is a Hex-Decimal code generated by PG Instrument Ltd to control to distriďƵƟŽŶ and ĂĐƟvaƟŽŶ of the UVWin soŌware.

The soŌware serial number is ƐƉĞĐŝĮĐ to a parƟcular instrument serial number and therefore can only be used with a ƐƉĞĐŝĮĐ instrument.

Please refer to the documentaƟŽn supplied with your instrument for the soŌware serial number.

Please refer to the soŌware manual for further informaƟon on installing the ƐŽŌware.

On successful installaƟŽn of the ƐŽŌware please progress to the instrument installaƟon.

Instrument InstallaƟŽŶ

x Place the instrument on a stable and horizontal workbench suĸĐŝĞnt in strength to support its weight and close enough to the PC to allow for connecƟons to be made.

x Connect the Mains Cable to a suitable power supply and also to the instrument.

x Connect the USB cable to the USB port on the instrument and also to an available USB port on the PC.

x It is recommended that the USB cable not be removed during operaƟon or whilst the instrument is powered up as this could result in damage to the instrument electronics.

x A representaƟon of the instrument and PC connecƟŽns can be clearly seen from the image in Figure 8.


PC RS 232PO RT

USB

pow er cord PC pow ercord

m ain unit

PC parallel

Figure 8

St the Instrument and So ware Now that all of the required connec have been made it is possible to start the instrument and the UVWin ware.

To start the instrument make the following checks and follow steps as follows:

x Ensure that there is no cell in the sample compartment or any other object which could poten ally block the beam.

x Ensure that the sample compartment is closed.

x Ensure that the PC is started and that the UVWin ware has been successfully installed.

x Power up the instrument from the power switch to the right hand side.

x Start UVWin from the PC

x The PC will then empt to connect to the instrument.

x On successful conne the za on process will start.

x Allow several minutes for the instrument in za on to complete.

x The screenshot in Figure 9 shows the ini aliza on window.

Figure 9

Note: Error messages can occur during za on if the instrument conĮgura on is not set correctly, or if the cells or any

other such objects are le in the sample compartment. If the error message indicates that the PC cannot communicate with the instrument ensure that all the connec on are present and correct then repeat the ini aliza on.

- 01 -


- 11 -

Before Analyzing any Samples AŌer the instrument has completed its iniƟalizaƟon self tests the UVWin soŌware will show each of the measurement workspaces as seen in Figure 10. It is recommended that, in order to obtain accurate and reliable results, the instrument be leŌ for 60 minutes warm-up Ɵme prior to performing any analysis.

Figure 10

When you have ĮŶŝƐŚed performing the required analysis the UVWin soŌware should be closed, ensuring all of the required data Įles are saved, then the spectrophotometer should be switched oī from the mains and ĮŶally the PC can be shutdown from the windows operaƟŶŐ system.


l

- 21 -

Chapter 3 – So ware Installa on and ŽŶĮŐƵra This on of the manual will describe how to correctly install the UVWin so ware then make the correct cŽŶĮgura s and se g for your instrument, taking into considera n any a accessories that you may have. So ware Installa UVWin version 6 is designed to run on the Microso Windows XP Professional and Windows 7 Professional opera systems. Minimum PC speciĮca ons are as follows:

x Pen um 1.4GHz Processor x 512MB RAM x 32MB Graphic Capability x CD/DVD Op cal Drive x Minimum 100MB Free Drive Space for Installa n

The installa n of UVWin is simple and can be performed quickly. Locate the directory in which the UVWin installa n Įles are stored and run Setup.exe. This will start the installa n wizard as shown in Figure 11.

FiŐƵre 11

Select Next to move through the installa n Wizard specifying a company name when prompted.

FiŐƵre 12

Once the installa n is complete and all the necessary Įles have been copied across to the computer a shortcut icon will be


- 31 -

created on the Windows desktop and a new program group will be added to the Start Menu. Included in this Program Group are op ons for the following as can be seen from Figure 13:

x Run UVWin So ware x Start UVWin OŋŝŶĞ – This allows you to run UVWin without emp ng to connect to and ini alize an instrument. x Connect to another Machine – This feature should be used when two or more spectrophotometers are being run from

the same PC and UVWin installa on. x UVWin ConĮgura – Allows for the se ng of communica n port, bandwidth, accessory op ons etc. x Uninstall UVWin S ware

Figure 13

When UVWin is installed a new directory is created for both the installa n Įles and also the data Įles. The default directory if not changed is: C:\Program Files\UV-Vis\UVWin6 – for the installa n Įles and C:\Program Files\UV-Vis\UVWin6\Data – for the Data ConĮŐƵra In order to set some ConĮgura n parameters for UVWin run the UVWin ConĮg program from the UVWin program Group in the Start Menu. First set the correct COM Port by making your selec on from the drop down menu. As show in Figure 14.

Figure 14

Next set the correct accessory op on by selec on the appropriate radio bu on as shown in Figure 15. The standard conĮgura n for the T92+ is Fixed Cells, however op onal accessories are available which would require a diīerent se ng to be ƐƉĞĐŝĮed.

Figure 15

Finally select the instrument Bandwidth at which you would like to perform your analysis. This can be changed within the UVWin so ware once the instrument has been successfully ini alized.


- 41 -

Chapter 4 – UVWin SoŌware UVWin is a powerful and intuiƟve soŌware tool for the control of PG Instrument spectrophotometers and the acquisiƟŽŶ and interpretaƟon of spectrophotometric data. For a more in depth descrŝƉƟŽn of the usage and feature of the UVWin soŌware please refer to the UVWin User Manual.


- 51 -

Chapter 5 – ApplicaƟŽŶ Examples Example Žf a WŚŽtŽmetric MeasuremeŶt

Vitamin B1 can promote the metabolizability of carbohydrate and fat, provide energy for the nervous Ɵssue, prevent the nervous

Ɵssue from atrophy and degeneraƟŽn, prevent and cure beriberi. The quality determinaƟŽn method of Vitamin B1 for the

pharmaceuƟcal industry is to measure the absorbance at 400nm; The conforming value should be less than 0.020.

We need to select the photometric measurement funcƟon to determine the quality of Vitamin B1 by UVWin soŌware, the

detailed operaƟon steps follow:

PreparaƟŽŶ Žf ReageŶts

Weight 1g Vitamin B1 in a precision microbalance. Put the 1g of Vitamin into a 10ml volumetric Ňask. Add adequate disƟlled

water to dissolve the Vitamin B1. Select two clean matched quartz cells to Įll with disƟlled water as the reference soluƟŽŶ

Start the IŶstrumeŶt ĂŶd UVWŝŶ ^ŽŌware

Power up the PC which is connected with the T90+, enter into the Windows operaƟŶg system. Be sure there is nothing blocking

the beam of the sample cell holder, and power on the instrument. Start UVWin soŌware and allow the instrument to iniƟalize.

Enter the main working program. The measurement can only be performed aŌer the preheat Ɵme of 60 minutes aŌer switching

on the instrument.

Set the MeasuremeŶt Parameters

Select the “Photometry” funcƟon and enter into the parameter seƫŶg interface. Set the parameters as shown in Table 2:

Measurement parameters InstructionPhotometric mode Abs Measurement wl 400nm

Instrument parameter InstructionSpectral bandwidth 2.0nm Fixed or variable

Table 2

PerfŽrm a Dark CurreŶt ŽƌrĞĐƟŽŶ

Insert the black block into the sample cell holder, to carry out the dark current correcƟŽn using the dark current correcƟŽn

ĨƵŶĐƟŽŶ of the UVWin soŌware. Take out the black block aŌer correcƟŽn.

EŽteφ Dark curreŶt cŽrrecƟŽŶ caŶ eliŵŝŶate ƐŽme ŶŽŝƐe Žf the iŶstrumeŶt, ĂŶd iŶsure the precisiŽŶ Žf result. This dŽes ŶŽt

have tŽ be carried Žut fŽr every measuremeŶt, ŽŶly wheŶ the eŶvirŽŶmeŶt has beeŶ chaŶged, such as the temperature, the

ůŽcaƟŽŶ, measurŝŶg high absŽrbaŶce sample ĂŶd sŽ ŽŶ. (we advise that yŽu perfŽƌŵ the dark curreŶt cŽrrĞĐƟŽŶ aŌer

preheaƟŶg ĂŶd befŽre measuremeŶt.)

PerfŽrm the AutŽ ZerŽ

Put the two matched quartz cells which are Įlled with disƟlled water into the sample light beam cell holder and reference light

beam cell holder. Cover the sample compartment and press the “Zero” buƩon of the UVWin soŌware. The absorbance shown on

the top of the main page should be 0.000 Abs.


- 61 -

Perform the Sample Measurement

Take the cell out of sample cell holder, and replace the disƟlled water with the Vitamin B1 ƐŽůƵƟŽŶ Be sure to wash out the cells

between samples Put the cell into the sample cell holder and cover the sample compartment Press the “Start” buƩon of the

UVWin ƐŽŌware to perform the measuremenƚ AŌer ĮŶŝshing the measurement, press “Start” again to measure the next datĂ

The result can be made more accurate by the averaging of several measurements

Analysis of Results

The result can be saved as a special Įle format of UVWin soŌware for checking and prinƟŶŐ later

Example of a Spectral Measurement

Vitamin B12’s characterisƟĐ ĂďƐŽƌƉƟŽn peak is at 361nm, so the quality determinaƟŽn method of Vitamin B12 is to measure the

absorbance at 36ϭŶŵ The detailed operaƟon steps of determining the absorpƟŽn peak of Vitamin B12 by T90+ is as follows:

PreparaƟŽŶ of Reagents

The Vitamin B12 ƐŽůƵƟŽn can be mixed with disƟlled water according to the prŽƉŽƌƟon of 1:10 to obtain a 10ml diluted ƐŽůƵƟŽŶ

Prepare two clean matched quartz cells to Įll with disƟlled water as the reference ƐŽůƵƟŽŶ

Start the Instrument and UVWin SoŌware

Power up the PC which is connected with the T90+, enter into the Windows operaƟŶg system Be sure there is nothing blocking

the beam of the sample cell holder, and power on the instrumenƚ Start UVWin soŌware and allow the instrument to iniƟalize

Enter the main working progrĂŵ The measurement can only be performed aŌer the preheat Ɵme of 60 minutes aŌer switching

on the instrumenƚ

Set the Measurement Parameters

Select the “Photometry” funcƟon and enter into the parameter seƫŶg interfĂĐĞ Set the parameters as shown in Table 3:

Measurement parameters Comments Photometric mode Abs Scanning speed Medium Scanning interval 1nm

Wavelength range 220660 nm

Vertical coordinate range

0.0002.000

Instrument parameters Comments Spectral bandwidth 2.0nm Fixed or variable

Table 3


- 71 -

Perform a Dark Current CorrĞĐƟŽn

Insert the black block into the sample cell holder, to carry out the dark current correcƟŽn using the dark current correcƟŽn

ĨƵŶĐƟŽŶ of the UVWin soŌware. Take out the black block aŌer correcƟŽn.

Noteφ Dark current correcƟon can eliminate some noise of the instrument, and insure the precision of result. This does not

have to be carried out for every measurement, only when the environment has been changed, such as the temperature, the

locaƟŽŶ, measuring high absorbance sample and so on. (we advise that you perform the dark current corrĞĐƟon aŌer

preheaƟŶg and before measurement.)

Perform a Baseline CorrecƟŽn

Place the two matched quartz cells which are Įlled with disƟlled water into the sample cell holder and reference cell holder.

Cover the sample compartment and select the “Baseline” buƩon of the UVWin soŌware. The result of the baseline will be

shown on the graph. Please wait for it to Įnish automaƟcally, it may take several minutes. During this process no other operaƟon

of the soŌware can be performed but it can be stopped by pressing the “Cancel” buƩon, and the data of correcƟŽn will be lost.


Remove the cell from the sample cell holder and replace the disƟlled water with the Vitamin B12 ƐŽůƵƟŽŶ Be sure to wash the

cells between samples. insert the cell which is Įlled with sample soluƟŽn into sample cell holder and cover the sample

compartment .Press the “Start” buƩon of the UVWin ƐŽŌware to begin the spectrum scanning. AŌer ĮŶŝƐŚŝŶŐ measurement

automaƟĐĂůůy, it will take several minutes, press the “Start” again to measure the next data. The result can be made more

accurate by the averaging of several measurements. During this process no other operaƟŽŶ of the soŌware can be performed. if

it is necessary to stop the operaƟŽn please press the “Cancel” buƩon and the scanning spectrum will be saved.

Note: Please pay ĂƩenƟon to the posŝƟŽŶ of the cell when taking it out as it needs to be consistent to insure minimum error.

Analysis of Results You can use the “Peak-picking” ĨƵŶĐƟon of soŌware to analyze the spectrum aŌer ĮŶŝshing the measurement. (By the shortcut of “Peak-picking” or under the “Graph” menu) You can search for the peak by this peak picking funcƟon although it may be necessary to re-set the threshold. The soŌware will list the result of measurements corresponding to the peaks. Output of Results You can save the result of the spectrum and the peak-picking as a special Įle format of UVWin ƐŽŌware for checking and prinƟŶŐ later.


- 81 -

Example of a QuanƟtaƟve Measurement

[Cr(VI)] detecƟŽŶ is ĂŶ ŝŵƉŽƌtaŶt item fŽr the deteĐƟŽŶ fŽƌ electrŽŶic ĂŶd electrical prŽĚucts. Take DipheŶylcarbazide. The

UV/Vis spectrŽphŽtŽmeter methŽĚ cĂŶ detect the cŽŶceŶtraƟŽŶ Žf [Cr(VI)] iŶ electrŽŶic ĂŶĚ electrical prŽĚucts.

The detailed ŽperaƟŽŶ Žf deterŵŝŶŝŶg the cŽŶceŶtraƟŽŶ Žf [Cr(VI)] by T90+ is as fŽůůŽws:

PreparaƟŽŶ of Reagents:

First prepare the fŽllŽǁiŶg ƐŽůƵƟŽŶ Water used fŽƌ these ƐŽluƟŽŶ shŽuld be disƟlled water, ĂŶd the reageŶt ƐŚŽƵůd be aŶalyƟcal

pure.

x Sulphuric sŽůƵƟŽŶ (1+9): sampliŶg 10 mL sulphuric˄GR˅ ĂŶd mix it ŝŶƚŽ 90mL Žf water ƐůŽwly.

x Nitric acid ƐŽůƵƟŽŶ (5 mol/L): samƉůŝŶg 31mL Ŷŝtric acid (GR) aŶd add it iŶtŽ 69mL water..

x Leaching soluƟŽŶ: weight 20.0g ƐŽĚŝƵŵ hydrŽxide aŶd 30.0g aŶhydrŽus ƐŽĚŝƵŵ ĐĂƌďŽŶate. ŝƐƐŽůve them iŶ disƟůůed

water ĂŶĚ remŽve iŶtŽ 10ml vŽlumetric Ňask. RemŽve iŶtŽ plasƟĐ bŽƩůĞ fŽƌ preservaƟŽŶ

x Buīer ƐŽůƵƟŽn: ĚŝƐƐŽůve 87.09g Žf dipŽtassium ƉŚŽƐƉŚate ĂŶĚ 68.04g ƉŽtassium dihydrŽgeŶ ƉŚŽsphate iŶ disƟlled

water, ĂŶĚ remŽve iŶtŽ 1000mL vŽlumetric Ňask.(pH=7).

x [Cr(VI)] stock standard soluƟon 100mg/L: measure 0.2828g ƉŽtassium dichrŽŵate exactly ĂŶd dry ƵŶĚer 120C fŽr 2

ŚŽƵrs. ŝƐƐŽůve it with disƟlled water ĂŶd remŽve iŶtŽ 1000 mL vŽlumetric Ňask.

x [Cr( )] standard soluƟon 5.0 mg/L: measure 5.0ml stŽck staŶĚard ƐŽůƵƟŽŶ iŶtŽ 100 ml vŽlumetric Ňask. ŝƐƐŽůved it iŶ

disƟlled water ĂŶĚ mix.

x Diphenylcarbazide (DPC) chromogenic agent: weight 0.5g DPC iŶtŽ 100 mL acetŽŶe, preserve iŶ brŽwŶ bŽƩůĞ If the

ƐŽůƵƟŽŶ’s cŽůŽr fades, re-prepare it.

PreparaƟŽŶ of the Sample

ŝƐƉŽƐĂů Žf the electrŽŶic aŶd electrical prŽĚƵcts have ďĞĞŶ split iŶtŽ strips Žƌ pŽwder samples which have a diameter Žf ŶŽ

ŵŽre thaŶ 1 mm ĂŶĚ a ůĞŶgth Žf ŶŽ ŵŽre thaŶ 5 mm by a crusher.

Weight (0.5~2.5)g˄precŝƐŝŽŶ tŽ 1mg˅sample exactly iŶtŽ a cŽŶical Ňask ĂŶd add iŶtŽ the 25.0 mL leĂĐŚŝŶg ƐŽůƵƟŽŶ. Add alsŽ tŽ

the 0.5 mL buīer ƐŽůƵƟŽŶ The sample ƐŚŽƵůd be submerged iŶ the leacŚŝŶŐ ƐŽůƵƟŽŶ, ĂŶĚ mixed rŽuŶd iŶ a(90~95)C water bath

;Žƌ shakiŶg bath ) fŽƌ 1.5 hŽurs. RemŽve the sample aŶd ĐŽŽl it dŽǁŶ tŽ rŽŽŵ temperature. Filtrate ĂŶĚ wash the cŽŶical Ňask

with water ŽůůĞĐƚ the Įltrate ĂŶĚ cleaŶŝŶg ƐŽluƟŽŶ iŶtŽ a beaker aŶd add 5 mŽů/L Ŷŝtric acid iŶtŽ it, ŽŶtrŽů the pH value Žf

ƐŽluƟŽŶ iŶ the raŶge Žf 7 tŽ 8 measurŝŶg with a pH meter. Filtrate further if required, keepiŶg the Įltrate.

Start the Instrument and UVWin SoŌware

PŽwer up the PC which is cŽŶŶĞĐted with the T90+, eŶter iŶtŽ the WiŶdŽws ŽƉeraƟŶg system. Be sure there is ŶŽthŝŶŐ blŽckiŶg

the beam Žf the sample cell ŚŽůĚĞr, ĂŶĚ ƉŽwer ŽŶ the ŝŶstrumeŶt. Start UVWiŶ ƐŽŌware ĂŶĚ allŽw the ŝŶstrumeŶt tŽ ŝŶŝƟalize.

EŶter the maiŶ wŽƌŬiŶg prŽgram. The measuremeŶt caŶ ŽŶly be perfŽƌmed aŌer the preheat Ɵme Žf 60 mŝŶƵtes aŌer switĐŚŝŶŐ

ŽŶ the ŝŶstrumeŶt.


- 91 -

Set the Measurement Parameters

Select the “Photometry” funcƟon and enter into the parameter seƫŶg interface. Set the parameters as shown in Table 4:

Measurement parameters Comments Measure method Single wavelength Main WL 540nm Calibration curve Abs=f(c) Equation Order 1st Conc. Unit mg/L Calibration method

Concentration method

Instrument parameters Comments Spectral bandwidth 2.0nm Fixed or variable

Table 4

Color producing reaĐƟon

Standard soluƟon: sampling 0.0, 2.0, 4.0, 6.0, 8.0, 10.0, 12.0mL [Cr(VI)] standard soluƟon (5.0 mg/L) and pipeƩe it into 100mL

volumetric Ňask. Add 50mL water into it, and add 2.0mL Diphenylcarbazide color producing reagent. Mix. Drop 2 beads of vitriol

soluƟon (1+9) and make the pH value of the Įltrate in the range of 1.5 to 2.5, make up to volume with water. The concentraƟŽŶƐ

of [Cr( )] in these calibraƟŽŶ soluƟon are 0.0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6mg/L. Mix, shake and leave to seƩle for half an hour.

Sample ƐŽůƵƟŽŶ: Add 2.0mL Diphenylcarbazide color producing reagent into sample Įltrate. Mix and add drop vitriol soluƟon

(1+9). Make the pH value of Įltrate in the range of 1.5 to 2.5, and move all the Įltrate into a 100mL volumetric Ňask. Make up

volume with water. Mix, shake and leave to seƩle for half an hour

Establish the calibraƟŽn curve

Input the ID and concentraƟŽn of the standard ƐŽůƵƟŽŶƐ from 1 to 7 in the “Standard” table. The concentraƟons are 0.0, 0.1, 0.2,

0.3, 0.4, 0.5, 0.6˄mg/L˅.

Put the two matched quartz cells which are Įlled with blank reagent soluƟon into the sample cell holder and reference cell

holder. Cover the sample compartment and press the “Zero” buƩon of the UVWin soŌware. The absorbance shown on the top

of the main screen should be 0.000 Abs or. Replace the soluƟŽn in the sample cell with the [Cr(VI)] standard soluƟŽŶs from NO.1

to NO.7 and press “Start” to measure the absorbance. The ƐŽŌware will plot the calibraƟŽn curve between ConcentraƟŽn and

Abs in real Ɵme (as shown in Figure 16).

Figure 16


- 02 -

The parameters of calibraƟon curve are: Abs = K1*(C) + K0ˈK0=0.0014ˈK1=0.68847ˈR2=0.9999


Input the ID of the sample in the “Sample” table using the same condiƟŽŶs and measure the Abs of sample soluƟon and blank

ƐŽůƵƟŽŶ The soŌware will calculate the concentraƟon of [Cr(VI)] by using the calibraƟon curve, expressed as A and B.

Analysis of Results

AŌer the measurement, calculate the mass fracƟon W, units shown as mg/kg. The concentraƟŽn of [Cr(VI)] is 704.76 mg/kg by

the formula below:

mNVBAW **)(

W - ConcentraƟon of [Cr(VI)] in the sample, the unit is mg/kg A - ConcentraƟon of the leaching ƐŽůƵƟŽn from the calibraƟon curve. The unit is mg/L B - ConcentraƟŽŶ of the blank soluƟon from the calibraƟŽn curve. The unit is mg/L

V - Constant volume of leaching ƐŽůƵƟŽn The unit is ml N - DiůƵƟŽŶ mulƟƉůe of leaching ƐŽůƵƟŽŶ. M - Mass of sample, the unit is gram (g)


- 12 -

Chapter 6 – OpƟŽnal AcceƐƐŽries CH19-1 ThermŽstaƟc Cell ,ŽůĚĞƌ The thermostaƟc cell holder is used together with a constant-temperature water bath and water pump. It is designed to measure samples at a constant and accurate temperature.

The ThermostaƟc Cell Holder can be seen in Figure 17, along with the speciĮcaƟon in Table 5.

Figure 17

NO. Item Specifications Remark

1 Product model CH19-1

2 Connection tube Inner diameter ĭ4mm

3 Wavelength range 190ˉ900nm

4 Path length 10mm

5 Cell holder NO. 2 Sample beam and

reference beam sample holder

Table 5


- 22 -

DS19-1 Angle Adjustable Solid Sample Holder Sliced solid samples can be installed onto the angle-changeable solid sample holder. The angles can be adjusted within ±45° and it is easy to measure the absorbency for solid samples.

The Angle Adjustable Solid Sample Holder can be seen in Figure 18, along with the speciĮcaƟon in Table 6.

Figure 18


1 Product model DS19-1


3 Changeable angle range ±45o

4 Min. sample size ĳ4mm

5 Max. sample size 80x55x5mm

Table 6


- 32 -

IS19-1 IntegraƟŶg Sphere

The integraƟŶg sphere is used to measure the diīƵƐĞ reŇectance of solid sample and powder, as well as measure the transmiƩance of glass and membrane. The integraƟng sphere can analyze the chrominance, color diīerence and whiteness.


Figure 19


1 1-91SI ledom tcudorP

2 Incident angle Sample light beam 0°ˈReference light

beam8°

3 Min. sample dimensions

Width 15mm×Height25mm Diffuse reflectance

4 Min. sample dimensions Diameter 20mm Transmittance

5 Wavelength range 230nm-850nm Slit 5nm

6 Sphere diameter mm06

Table 7


- 42 -

LS19-1 Long Pathlength Cell Holder

The long pathlength cell holder can be used with 5-50mm wide cells making it is easy for the measurement of samples.


Figure 20


1 Product model LS19-1

2 Wavelength range 190900nm

3 Pathlength 5mmǃ10

mmǃ20mm,30 mmǃ40 mmǃ50 mmǃ

Adjusted according to requirements

4 Cell NO. 2 Sample light beam and reference light beam

Table 8


- 52 -

MH19-1 Micro Cell Holder

Micro cuveƩĞs can be used in the micro cell holder. The bases can be carefully adjusted to enable the light path to pass through the cell windows giving maximum light throughput and improve measurement precision for micro samples.

The Micro Cell Holder can be seen in Figure 21, along with the speciĮcaƟŽn in Table 9.

Figure 21


1 1-91HM ledom tcudorP


3 Cell NO. 2

Table 9


- 62 -

MH19-2 Ultra Micro Cell Holder Ultra-micro cuveƩĞs need to be used with the ultra-micro cell holder due to the clear aperture on the cuveƩĞs being very small. Using a standard cell holder for the measurement would block some light causing the energy to be lost and the instrument precision reduced.

Two quartz lens are used to shrink the imaging of the beam in the sample and reference opƟcal circuits through the ultra-micro cell holders. AŌer this the beams are magniĮed by two more quartz lenses which collect energy and return back to normal state to be read by the detector. By using this accessory the signal-noise raƟo and measurement precision of the instrument will be greatly improved.

The Ultra Micro Cell Holder can be seen in Figure 22, along with the speciĮcaƟon in Table 10.

Figure 22


1 Product model MH19-2


3 Cell NO. 2

Table 10


- 72 -

MR19-1 Specula ReŇecƟon Accessory

The specula reŇecƟon accessory can measure the reŇectance spectrum of opƟcal surfaces, crystals and coated samples. If the reŇecƟng mirror on the reference beam is the “standard reŇecƟng mirror”, the user can calculate the absolute reŇecƟvity of the measured surface.

The Specula ReŇecƟon Accessory can be seen in Figure 23, along with the speciĮcaƟon in Table 11.

Figure 23


1 Product model MR19-1

2 o5 elgna ecnedicnI

3 Measured sample

area 11×9mm to 60×40mm

4 Wavelength range 190nm-900nm

Table 11


- 82 -

PS19-2 Sipper Pump The sipper pump accessory consists of a sipper pump, front baŋĞ and connecƟŽŶ tubes.

The sipper pump is a soŌ-tube pump that can transmit liquid in dual dirĞĐƟŽŶƐ It is widely used in research and prŽĚƵĐƟon for reĮne chemistry, biophysics, biochemistry, environmental protĞĐƟŽn, pharmaceuƟcals and other industries. It is an ideal device for transmiƫŶg sƟcky and corrosive liquids. The PS181-2 sipper pump accessory has the following features:

1. LCD displays liquid sampling or motor rotaƟon speed. All ĨƵŶĐƟŽŶƐ and parameters can be set from six buƩons.

2. It can store data if the power supply is lost. The device can automaƟcally store the sampling volume or the rotaƟon speed of the motor. The LCD displays the value that is set in the last operaƟŽŶ when it is turned on.

3. It has a new control system and quanƟtaƟve sampling system. There are two working modes, conƟŶƵĂů and quanƟtaƟve. It can be used for conƟnuous sampling and disconƟnuous quanƟtaƟve sampling.

4. It has an auto-calibraƟŽn funcƟŽn. Users can calibrate the sampling volume through the calibraƟŽn funcƟŽŶ which will avoid errors with prolonged running.

5. The mixing stepper-motor runs smoothly and accurately with large torque and low noise through a high-subdivision driver.

6. The membrane touch-switches are easy to operate and the tube clips are easy to install and dismantle.

7. It is easy for maintenance. For diīerent Ňuids it is only necessary to replace the soŌ tube. There is no ƉŽůůƵƟŽŶ from the valves or sealing parts.

8. It can transmit liquids from dual-dirĞĐƟon, have accurate quanƟtaƟve sampling, have stable Ňow rate and be used with an outside interface control.

The speciĮcaƟŽŶ for the Sipper Pump can be seen in Table 12.


1 2-91SP ledom tcudorP

2 Rotation speed range 0.1-250rpm Clockwise & counterclockwise

3 Speed resolution 0.1rpm: lower than 30rpm 1 rpm: higher than30rpm

4 Display mode 3 1/2 LCD display

5 Sampling accuracy 1 mL

6 Power supply AC85v-264v, 50-60Hz

7 Working environment Temp: 0-400C,

Relativity humidity<80%

8 Driver size (L*W*H)370*210*500(mm)

9 Driver weight 10kg

Table 12


- 92 -

S19-1 Solid Sample Holder Sliced solid samples can be put in the solid sample holder which makes it easy to measure the absorbency.

The Solid Sample Holder can be seen in Figure 24, along with the speciĮcaƟon in Table 13.

Figure 24


1 Product model S19-1


3 Cell NO. 2

4 Max. sample size 80x55x5mm

Table 13


- 03 -

SS19-1 Short Pathlength Cell Holder The short pathlength cell holder is inserted into the standard cell holder making is easy to measure samples using short pathlength cells. The short pathlength cell holder can be used in the following holders: standard Įxed cell holder, 8-cell holder, micro cell holder and ultra-micro cell holder.

The Short Pathlength Cell Holder can be seen in Figure 25, along with the speciĮĐĂƟon in Table 14.

Figure 25


1 Product model SS19-1


3 Path length 1mmǃ2mmǃ5mm

Table 14


- 13 -

TR19-1 Test Tube Holder The test tube holder is used for the measurement of liquids contained in test tubes. The measurement results are dependent on the material and quality of the test tubes. Therefore if precise measurement is required, it is recommended that the related accessory for cuveƩĞs be used. The Short Pathlength Cell Holder can be seen in Figure 26, along with the speciĮĐĂƟon in Table 15.

Figure 26


1 Product model TR19-1


3 Test tube range Diameter: 1525mm Height: 90120mm

4 Cell NO. 2

Table 15


- 2 3 -

Chapter 7 – Maintenance Replacement of the W (Tungsten) Lamp

The tungsten lamp is plugged into a socket and sleeved by a plas c holder. It is well adjusted before delivery. If you need to

replace the tungsten lamp, please observe the following procedures. The Tungsten Lamp Assembly can be seen in Figure 27.

Figure 27

Remove the two screws which secure the lamp housing cover in place. Gain access to the Lamp housing by removing the cover as

shown in Figure 28.

Figure 28

Loosen the two screws which secure the tungsten lamp sleeve into the lamp housing as seen in Figure 29. The Lamp assembly including both the lamp and sleeve can now be removed from the lamp housing.

Ceramic sieeve

Metal foot

Adjuster screw

Tungsten lamp

Fixed screw


- 33 -

Figure 29

Remove the lamp from the sleeve by Įrst loosening the two screws which secure it. Insert a new lamp taking care not to get any ĮŶgerprints on the opƟcal surface Ensure that the orientaƟŽn of the lamp is correct as shown in Figure 31. The lamp can be gently cleaned using a mild alcohol to ensure it is clean. Secure the new lamp in the sleeve by fastening the screws then install it back in the lamp housing again fastening the respecƟve screws - Figure 30.

Figure 30

If the performance of the instrument changes greatly aŌer the replacement, the height of tungsten lamp should be adjusted (The

horizontal posiƟon of tungsten lamp is adjusted automaƟcally when the self-tesƟng is complete aŌer power-up). First, remove

the tungsten lamp holder, loosen the fastening screws on the tungsten lamp sleeve with a model screwdriver and adjust the

height of tungsten lamp. The distance between the center of the Įlament and the boƩom of the sleeve should be 40mm,

however slight adjustment may be required. Tighten the fastening screws and insert the W lamp sleeve into the socket.


- 4 3 -

Replacement of the D2 (Deuterium) Lamp The D2 lamp is ng into an Octal base socket inside the Lamp Housing. When replacement is made, directly and securely insert

a new one into the socket. The procedure is as follows:

x Open the Lamp Housing, the same in the Įrst step of the tungsten lamp replacement.

x Remove the deuterium lamp, pull it out upwards, you can shake it when drawing, because the D2 lamp is installed

htly. This can be seen in Figure 31.

Figure 31

Clean the surface of D2 lamp with a clean cloth socked with alcohol (especially the window of it). Insert the D2 lamp into its

socket. The keyway of the deuterium lamp base should align to the recess in the lamp socket. and the dent of the holder

should be rightly overlapped (the light beam window should face to the convergent lens), as shown in Įgure 32.

Figure 32

Note: x The Deuterium Lamp will become extremely hot after operation; take care

when accessing the lamp compartment.

x The Deuterium Lamp emits high intensity UV radiation, take the necessary precautions when exposing to eyes or skin.

Dent of D2 lamp holder

Bulge of D2 lamp



- 5 3 -

Fuse Replacement

This chapter introduces the user how to replace the fuse. If you suspect the fuse is brown, please replace it according to the

following procedures.

x Switch oī the instrument.

x Remove the fuse holder using a posi-drive screwdriver, as shown in Įgure 33.

Figure 33

Figure 34

Remove the old fuse, and replace it with a new one. Re-install the fuse holder. The fuse replacement is now complete.

Plus screwdriver

Fuse

Fuse holder

operation instruction manual

Documents