Page 26 - Oligonucleotide Therapeutics Solution Guide

Characteristic analysis
Quality Control

Quantitation & Spectrum Confirmation UV-1900i

Features
Quantitation of Double-stranded DNA
The UV-1900i spectrophotometer features a space-saving and ergonomic hard design. It is equipped with a color touch panel and adopts a user Modification
– Trace Measurement Using TrayCell and Nano Stick – interface (UI) that realizes "Easy to Operate, Obtain Answers Easily and Rapidly". In addition, Six types of measurement conditions are built in: 1. Target selection
Oligonucleotide quantification, 2. Lowry method, 3. BCA method, 4. CBB (Bradford method), 5. Biuret method, and 6. UV absorption method. It
click here also has a screen shot function on the operation panel, so you can easily extract measurement results without connecting to a PC. A 10 mm square
cell requires a sample volume of approximately 4 mL, but the TrayCell or Nano Stick can be used to measure trace amounts of about 2 μL to 4 μL.

• It enables spectrum acquisition with ultrafast scanning at a maximum speed of 29,000 nm/min.
• The use of TrayCell (manufactured by Hellma Analytics) and Nano Stick (manufactured

by SINCO) enables measurement from a sample volume of as little as 3 μL.
benefits
• The quantitative value of oligonucleotide concentration can be calculated by Excision
using the oligonucleotide quantification function in Biomethod. Unprotected Oligomer synthesis

Methods and Results

Sample Double-stranded DNA
Conc., Volume 1 ~ 300 ng/mL, 2 µL
Preparation Dilution in ultrapure water to the concentrations above.
Analytical As shown in Table 1
Conditions 1. The sample is placed in the central
Method for We prepared ds DNA and prepared standard samples at 27.5, silver region. 2. set the lid on
measuring 55, 110, 220, and 440 ng/μL (ultrapure water was used as the
Double- dilution solution). For samples with unknown concentrations, we Purification
Stranded DNA prepared the same DNA by ethanol precipitation. The TrayCell
using TrayCell can be changed to optical path lengths of 1.0 mm and 0.2 mm High Performance to Meet Diverse Needs
by using two different lids. In this test, we used a lid with an
optical path length of 1.0 mm, dropped 4 μL, and measured High Speed Scan Spectra can be acquired as fast as 29,000 nm/min. Ultra-fast scan is effective in tracking chemical
under the conditions shown in Table 1 (Fig. 1). reactions in a short time. In addition to the absorbance change at specified wavelengths, spectra can
Method for We prepared standard and unknown samples of ds DNA as well 4. After measurement, wipe the also be acquired in a short time with the UV-1900i.
measuring Double- as the above TrayCell. We also set the anallysis conditions as 3. Set the cell holder with Traycell sample with a Kim wipe. Therefore, more detailed behavior can be investigated by observing spectra with the UV-1900i.
Stranded DNA shown in Table 1. The optical path length of the Nano Stick was
using Nano Stick 0.5 mm, and the sample volume was measured at 3 μL (Fig. 2). Low Stray Light Stray light is at 0.5 % max. (198 nm), making accurate measurements are possible up to the vicinity
of 2 Abs even in the ultraviolet region. In addition, high-concentration samples can be quantified
Results Figures 3 and 4 show the calibration curve and UV spectrum
results when measured using TrayCell and Nano Stick. Both accurately.
results indicate a highly linear calibration curve. We repeatedly
measured 440 ng/μL 10 times. As a result, the correlation High Reproducibility and High repeatability of less than 0.0002 Abs reduces variation in measurement results. And it enables
function and CV value were calculated, and it was confirmed Repeatability Accuracy more accurate quantitative analysis and detection of lower concentrations of samples.
that the measurement was performed accurately.
Figure 2 What the Nano Stick Looks Like and How To Use It A Diversity UV-1900i features 6 measurement modes: Photometric, Spectral, Quantitative, Kinetics, Timecourse, Quality Control
of Measurement Modes and BioMethod.
Table 1 Analysis Conditions Characteristic analysis
Wavelength (calibration curve): 260 nm, 320 nm ー Unknown sample
concentration
Wavelength range: 220 nm ~ 330 nm ー 440ng/µL
ー Unknown concentration
Scan Speed : Low sample (3 times dilution) Full Support for Pharmacopeia, GLP/GMP, FDA 21 CFR Part 11 and Other Regulations
ー 220ng/µL
Sampling Pitch: 1 nm ー 110ng/µL Instrument Validation Functions This instrument can not only run checks for nine JIS items, but also those stipulated in the Japanese
ー 55ng/µL
ー 27.5ng/µL
Compliant with JP, USP, and EP Pharmacopoeia (JP), United States Pharmacopeia (USP), and the European Pharmacopoeia (EP).
Naturally, the hardware is also compliant with the specifications required by each Pharmacopeia. In
addition, the check conditions can be saved. As a result, once the conditions are saved, checks can
be performed easily just by calling them up as needed. Check results can also be saved. DDS
Figure 3 Results of measurements using TrayCell Resolution of 1 nm, In addition to achieving a resolution of 1 nm, the highest in its class, by using a monochromator
the Highest in its Class with a Czerny-Turner mounting, the UV-1900i also features a compact, bright optical system. The
instrument is more than capable of meeting the wavelength resolution required in the European Pharmacokinetics
ー Unknown sample
concentration Pharmacopoeia.
ー 440ng/µL
ー Unknown concentration Improved Security Functions An external control security function has been added to provide more support for compliance with
sample (3 times dilution)
ー 220ng/µL regulations. Three user authority levels, "Administrator", "Developer", and "Operator", can be set
ー 110ng/µL
ー 55ng/µL for instrument users.
ー 27.5ng/µL
Figure 1 TrayCell Appearance Support for Regulations and Ensuring the integrity of data (database management), including the user management, user
Guidelines authority management, and data audit trails required for compliance with FDA 21 CFR Part 11, PIC/S
GMP guidelines, and other ER/ES regulations is possible.
LabSolutions DB UV-Vis or UVProbe / LabSolutions DB System allows for data management and user
Figure 4 Results of measurements using the Nano Stick management with a database. Compliant with ER/ES regulations, the system is optimally configured
Conclusions for customers using a PC. Other
LabSolutions CS UV-Vis or UVProbe / LabSolutions CS System (Network System) is optimally
This application news has shown that the UV-1900i, TrayCell, and Nano Stick can be used to make accurate and simple measurements even for configured for customers who want to manage data on a server together with LC and GC data for
small samples of the order of several μL. ER/ES compliance.
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