You are on page 1of 3

NMR PROBES

  • I. GENERAL CONSIDERATIONS

>>>>probes.docx – 8/20/2013

  • A. PROBE TYPES: One of the most important details about a particular NMR spectrometer is the probe that is located inside the magnet where the sample sits. The probe contains the radio frequency (rf) circuitry that is used to pulse the appropriate frequency and detect the response. The frequency depends on the magnetic field strength and the gyromagnetic ratio of the nucleus. On any given spectrometer, the field strength doesn’t vary, but the nuclei to which the probe can be tuned is somewhat different from one spectrometer to the next.

    • 1. A “broadband” probe is one that has an rf coil that can be tuned to many different nuclei. These will always be nuclei other than 1 H or 19 F because the frequencies of 1 H and 19 F are much higher than all the others and require separate hardware from the lower frequency nuclei. These lower frequency nuclei are referred to as X-nuclei. The broadband coil is the inner coil (see below). Broadband probes also have an outer coil that is tuned to the 1 H frequency.

    • 2. A “dual” probe is one that is optimized for maximum sensitivity of a single nucleus. There is always some loss of sensitivity when a coil is tunable to more than one nucleus. One of the coils will be tuned to an X- nucleus and the other coil will be tuned to 1 H. Similarly, a “quadruple nucleus probe” (QNP) has a coil that can be tuned to three different nuclei and a coil that is tuned to 1 H.

    • 3. On virtually all probes, the 1 H coil is double-tuned to include 2 H, which is primarily for the lock channel but can also be used for direct observation of 2 H with the lock channel turned off.

    • 4. For historical reasons, probes that have 1 H on the inner coil and an X-nucleus on the outer coil are called “inverse” probes. For example, an “inverse broadband probe” is one with the inner coil tuned to 1 H and an outer coil that can be tuned to many X-nuclei.

    • 5. Cryoprobes have the rf coils and preamplifiers cooled to ~10 deg K. The sample area will still be at room temperature.

      • a. The advantage of a cryoprobe is a substantial increase in sensitivity because the signal-to-noise ratio (S/N) is increased by the reduction of electronic noise at very low temperature. Cryoprobes can be made in any of the above configurations.

      • b. A cryoprobe requires a large support system and is a very expensive probe to operate and complex to service. Thus special care must be taken to avoid damage to the probe. Make sure that you use only good quality NMR tubes that have not been abused. A broken sample in the probe could lead to extended down-time for the probe. Also, it is imperative that the outside of your NMR tube be absolutely clean.

  • B. INNER COILS VS. OUTER COILS

    • 1. The terms "inner coil" and "outer coil" refer to the fact that most probes have two different coils, one of which is wound on top of the other.

    • 2. All other things being equal, an inner coil will yield better sensitivity for observation of a given nucleus because the inner coil is more tightly coupled to the sample. This sensitivity difference is of the order of a factor of 2 which would require a factor of 4 in time to regain the original sensitivity. Whenever sensitivity

  • 1

    is an issue, you should always use a probe that has an inner coil tuned to the frequency of the nucleus to be observed. Examples:

    • a. To observe 13 C, use a probe with an inner coil tuned to 13 C and an outer coil tuned to 1 H for decoupling. If the sample is sufficiently concentrated to observe a 13 C spectrum, you will also be able to observe the 1 H spectrum on the outer coil. In this case, the sacrifice of 1 H sensitivity will be worth the convenience of not changing the probe to switch to 1 H observation.

    • b. If the amount of sample is small such that 1 H sensitivity is poor, use a probe with an inner coil tuned to 1 H. If there is also an outer coil that can be tuned to 13 C, you can obtain, for example, an HSQC-INEPT 2D spectrum, which is a 1 H observe experiment that yields 1 H - 13 C connectivity and multiplicity information with much better sensitivity compared to analogous experiments that utilize direct observation of 13 C.

    • c. The figure below shows the CH 2 peak from two spectra acquired on the AV600 with the 0.1% ethylbenzene standard sample that is used to measure 1 H sensitivity. All experimental conditions are the same except for the probe that was used. The spectrum on the left was acquired using the 5 mm triple broadband inverse (tbi5) probe and the one on the right was acquired using the 5 mm broadband (bb5) probe. The sensitivity of the tbi5 probe was measured to be 1.9 times higher than that of the bb5 probe.

    is an issue, you should always use a probe that has an inner coil tuned to
    • C. SAMPLE VOLUMES

      • 1. Whenever adequate sample is available, always use a relatively long column of liquid compared to the length of the coil used for observation. When the quantity of sample is limited, the length of sample should be somewhat longer than the observation coil. If this is a problem, you can use a Shigemi tube, which is a special tube that limits the sample region and avoids the problems associated with shimming short samples by using materials at the top and bottom of the tube that are magnetic susceptibility matched to the solvent in use.

      • 2. For 5 mm sample tubes, 50 l of liquid, corresponds to ~ 3.6 mm of length measured from the outside bottom of the tube to the bottom of the meniscus. A sample with a column of liquid that is 4 cm long in a standard 5 mm NMR tube requires 0.55 ml of sample.

    2

    II. PROBES AVAILABLE FOR THE UCLA MIC NMR SPECTROMETERS

    AV300

    The only probe is a 5 mm broadband probe with automatic tune and match.

    AV400

    The primary probe is a 5 mm broadband probe with automatic tune and match. There is also an inverse broadband probe, but it would only be used in the event of a significant problem with the broadband probe.

    DRX500 – The primary probe is a 5 mm broadband probe. There is also an inverse broadband probe. The 1 H sensitivity improvement is much greater for the AV600 which is a more recent spectrometer with improved sensitivity for 1 H. Thus this probe would probably only be used in the event of a significant problem with the broadband probe.

    AV500

    The primary probe is a 5 mm dual probe in which the inner coil is tuned to 13 C with automatic tune and match. This probe is a cryoprobe that is optimized for 13 C sensitivity. It is by far the most sensitive instrument we have for carbon observe, by almost a factor of 10. Proton sensitivity is also excellent, but the improvement is not as great as it is for carbon because proton is observed on the outer coil. There is a 5 mm broadband probe that was purchased with the previous console for this system. It will only be used if the cryoprobe is not available for an extended period.

    AV600

    There are two probes and we regularly change between the two. They are a 5 mm broadband (bb5) probe and a 5 mm inverse broadband (tbi5) probe that has an additional input for 13 C. This could be used for a triple resonance experiment and is thus called a “triple broadband inverse” probe. An inverse probe will give better water suppression than a broadband. The spectra of 2 mM sucrose in 10% D 2 O / 90% H 2 O below were acquired with presaturation of the water signal on the two AV600 probes using identical conditions. The inset of the anomeric proton of sucrose shows that the shimming is excellent in both cases. However, the larger volume of the 1 H coil of the bb5 probe picks up more signal from regions of the sample outside the coil where the magnetic field homogeneity is not as good. The very intense H 2 O signal overwhelms the spectrum in the region of the water resonance.

    II. PROBES AVAILABLE FOR THE UCLA MIC NMR SPECTROMETERS AV300 – The only probe is a

    3